CN105796630B - A kind of filial fan grass composition and its application - Google Patents

Abstract

The present invention relates to a composition of saffron chrysanthemum and an application thereof. The composition of chrysanthemum chrysanthemum consists of chrysanthemum and chrysanthemum with a mass ratio of 1 to 5:1. The beneficial effect of the application of the Filibulum chinensis composition of the present invention in the preparation of anti-rheumatoid arthritis is as follows: the composition of the present invention can effectively treat rheumatoid arthritis, and the effect is better than that of using Filiaria fructus alone, It has a good clinical application prospect.

Description

A kind of filial fan grass composition and its application

(1) Technical field

The present invention relates to a composition and application thereof.

(2) Background technology

Rheumatoid arthritis is a heterogeneous, systemic, chronic autoimmune dysfunction disease characterized by osteoarticular synovitis and symmetric polyarthritis as the main clinical manifestation. The incidence rate is 1% in the world, 0.3% in my country, and the disability rate is as high as 20% within one year of the onset, seriously affecting human health and quality of life. At present, the drugs used clinically to treat rheumatoid arthritis all have contradictions in toxicity and tolerance, and some new biological agents are limited in application due to their high price.

Filial fan grass is Calystegia soldanella (L.) R.Br. [Convolvulus soldanella L.]), a plant of the genus Calystegia in the Convolvulaceae family (Convolvulaceae). The stem is used as medicine. It is rich in resources and has a long history of medicinal use. It mainly has the functions of dispelling wind and dampness, resolving phlegm and relieving cough, clearing lung and stopping bleeding, diuresis and detoxification. For the treatment of cough, nephritis edema, rheumatic joint pain, arthralgia and so on.

Inula cappa (Buch.-Ham.) DC. is a plant of the genus Inula in the family Compositae. "Chinese Zhuang Pharmacy" and "Chinese Ethnic Medicine Processing Integration", have the functions of promoting qi and relieving pain, dispelling wind and reducing swelling, and mainly treating rheumatic bone pain, bruises, colds and colds, etc. In Guangxi, Yunnan, Guizhou and other provinces of my country, the people of the Dong, Dai, Jingpo, Lahu, Lisu, Miao, Yi, Wa, Zhuang and other ethnic groups have used it as the main drug in a variety of rheumatism and bone pain prescriptions, and have rich medicinal experience. In recent years, there have been few reports on the study of pharmacological activity of Fructus Fructus, and only anti-oxidation and antibacterial effects have been found so far.

(3) Contents of the invention

The object of the present invention is to provide a composition of fenugreek chrysanthemum and its application in the preparation of anti-rheumatoid arthritis drugs.

The technical scheme adopted in the present invention is:

A composition of chrysanthemum chrysanthemum and chrysanthemum chrysanthemum in mass ratio of 1-5:1.

The mass ratio of the filial leaf grass and the chrysanthemum chrysanthemum is preferably 2:1.

The present invention also relates to the application of the composition of fenugreek chrysanthemum in the preparation of medicine for treating rheumatoid arthritis.

Concretely, the application of the extract of Filiaria fragrans Fructus Chrysanthemi in the preparation of medicine for treating rheumatoid arthritis, the extract is water extract, alcohol extract or alcohol extraction ethyl acetate extraction part.

The preparation method of the water extract: take the dry coarse powder of the composition of the chrysanthemum chrysanthemum and the chrysanthemum chrysanthemum, the mass ratio of the chrysanthemum and the chrysanthemum powder is 1-5:1, add 8-10 times the amount of water of the medicinal material, and soak before extraction 1 to 2 hours, heated and refluxed for 2 to 3 times, each time for 1 to 2 hours, concentrated under reduced pressure and then obtained.

The preparation method of the alcohol extract: take the dry coarse powder of the composition of Fructus chinensis and Fructus chinensis, the mass ratio of Fenium persicae and Fructus chinensis coarse powder is 1-5:1, add 50% ethanol of 8-10 times the amount of medicinal materials, extract Pre-soak for 30min-2h, extract by heating and reflux for 2-3 times, each time for 1-2h, concentrate under reduced pressure and suction to obtain the product.

Through test comparison, it is found that the ethyl acetate extract of Fenugreek fruticosa has the best effect, therefore, the extract is preferably the ethyl acetate extract of Filium fructus Fructus Fructus, which is prepared by the following method: take its Dried grass and sheep ear chrysanthemum powder, the mass ratio of fennel grass and sheep ear chrysanthemum powder is 1 to 5:1, add 10 to 12 times the amount of 80 to 95% ethanol, soak for 1 to 2 hours, then heat and reflux to extract 2 to 3 once, 1-2 hours each time, combine the two filtrates, recover ethanol, concentrate until there is no alcohol smell, add appropriate amount of water to dissolve, first extract with petroleum ether until colorless, then extract the raffinate solution with ethyl acetate until colorless, take Ethyl acetate extraction part, the solvent is evaporated to get the ethyl acetate extract of Fenugreek chrysanthemum.

The medicine can be prepared into one of the following dosage forms by adding suitable pharmaceutical excipients: ① injection, ② tablet, ③ capsule, ④ granule, ⑤ decoction.

The present invention takes its activity as the forerunner to study the effective part of anti-inflammatory, analgesic, anti-rheumatism and anti-rheumatoid arthritis of the composition of Filiaria sativa, anti-rheumatism and anti-rheumatoid arthritis. The experiment starts with different extracts and extraction parts, and selects the classic Anti-inflammatory and analgesic models such as mouse hot plate method, writhing method, xylene-induced mouse ear swelling method and adjuvant arthritis rat model, anti-inflammatory and analgesic, anti-rheumatic and anti-rheumatoid at the overall level Pharmacodynamics study of acute arthritis, explored the effects of the composition of filial genus Fructus chrysanthemum on anti-inflammatory and analgesic model mice, and on the toe swelling degree, arthritis index and viscera of adjuvant arthritis rat model Influenced by the index, it is preliminarily determined that the best effective part of the composition of Fenugreek Fructus is the ethyl acetate extraction part of the composition of Fenugreek Fructus.

The beneficial effect of the application of the composition of Filiaria fragrans Fructus in the present invention in the preparation of anti-rheumatoid arthritis is as follows: the composition of the present invention can effectively treat rheumatoid arthritis, and the effect is better than that of Filiaria fragrans alone, and has Very good prospects for clinical application.

(4) Description of drawings

Fig. 1 is a rat synovial tissue diagram (10×10), A: blank; B: model; C: positive; D: administration;

Fig. 2 is a histological diagram of rat synovium (10×20), A: blank; B: model; C: positive; D: administration.

(5) Specific implementation methods

The present invention is further described below in conjunction with specific embodiment, but protection scope of the present invention is not limited thereto:

Example 1:

The preparation of the water extract of the chrysanthemum chrysanthemum: take the dry coarse powder of the chrysanthemum chrysanthemum composition, the mass ratio of the chrysanthemum chrysanthemum and the chrysanthemum powder is 2:1, add water with 8 times the mass of the medicinal material, and soak for 1 hour , heated and refluxed for 2 extractions, 1 hour each time, vacuum filtered and then concentrated to a relative density of 1.10.

Preparation of Alcohol Extract of Echinacea Echinacea: Take the dry coarse powder of Echinacea Echinacea Composition, the mass ratio of Echinacea Echinacea to coarse powder is 2:1, add 50% (v /v) ethanol, after soaking for 30 minutes, heating and refluxing to extract twice, each time for 1 hour, and concentrating to a relative density of 1.10 after vacuum filtration, to obtain the final product.

Preparation of the ethyl acetate extraction part (i.e., ethyl acetate extract) of P. chinensis: Take the dry coarse powder of the composition of P. chinensis, and the mass ratio of P. chinensis and P. chinensis coarse powder is 2:1, Add 10 times the mass of 95% ethanol, soak for 1 hour, heat and reflux for extraction twice, each time for 2 hours, combine the two filtrates, filter, recover ethanol, concentrate until no alcohol smell, add a small amount of water to dissolve, first extract with petroleum ether, The raffinate solution was then extracted with ethyl acetate until it was colorless, and the ethyl acetate extraction part was evaporated to dry the solvent to obtain the ethyl acetate extraction part of Saffron chrysanthemum.

Preparation of the petroleum ether extraction part of P. chinensis: Take the dry coarse powder of the composition of P. chinensis and S. chrysanthemum powder at a mass ratio of 2:1, add 10 times the mass of 95% ethanol, After soaking for 1 hour, heat and reflux to extract twice, each time for 2 hours, combine the two filtrates, filter, recover ethanol, concentrate until there is no alcohol smell, add a small amount of water to dissolve, then extract with petroleum ether until it is colorless, take the petroleum ether extraction part and volatilize Dry the solvent to obtain the petroleum ether extraction part of the filial sage chrysanthemum.

The preparation of the n-butanol extraction part and the remaining parts of the genus chrysanthemum chrysanthemum: take the dry coarse powder of the chrysanthemum chrysanthemum composition, the mass ratio of the chrysanthemum chrysanthemum and the chrysanthemum chrysanthemum powder is 2:1, and add 10 times the mass of 95% ethanol, after soaking for 1 hour, heat and reflux to extract twice, each time for 2 hours, combine the two filtrates, filter, recover ethanol, concentrate until there is no alcohol smell, add a small amount of water to dissolve, first extract with petroleum ether, and use the raffinate solution obtained Ethyl acetate extraction, and the obtained raffinate solution was extracted with n-butanol until it was colorless, and the n-butanol extraction part was evaporated to dry the solvent to obtain the n-butanol extraction part of Fenugreek chrysanthemum, and the remaining part after solvent extraction (i.e. n-butanol raffinate solution) for subsequent use.

Experimental method 1:

Mouse hot plate experiment: 50 male ICR mice, weighing (20±2) g (excluding hypersensitive and insensitive animals), were randomly divided into 5 groups with 10 mice in each group, which was the blank control group. The low, medium and high dose groups (respectively, crude drug amounts of 2.5g·kg ^-1 , 5.0g·kg ^-1 , 10.0g·kg ^-1 ) of the aqueous extract of the composition of Sheepearm chinensis, the positive drug (aspirin) group ( 0.25g·kg ^-1 ). All the drugs were adjusted to the corresponding concentration and administered by intragastric administration at 0.2ml·10g ^-1 once a day. The blank control group was given the same volume of distilled water for 7 consecutive days. One hour after the last administration, the mice were placed on a metal plate at 55°C to The latency of the mouse licking the hind paw or jumping response was used as the pain threshold index, and the time spent on the metal plate was recorded.

Experimental results:

Effects on the pain threshold of mice: the results showed that each dose group of the water extract of the filial genus Fructus chrysanthemum composition can improve the pain threshold of the mice, and there was a very significant difference (P<0.001) compared with the blank control group, suggesting that filial piety The water extract group of the composition of scalloped chrysanthemum has a very significant analgesic effect. See Table 1.

Table 1: Effects of the water extract of the filial scallop grass composition water extract on the pain threshold of mice ( n=10)

group Dose/g·kg^-1 Pain threshold 1h after administration blank group ---- 15.6±3.4 positive group 0.25 25.3±4.2^** high dose 10.0 35.9±4.7^** medium dose 5.0 29.6±5.9^** low dose 2.5 24.9±5.6^**

Note: Compared with blank control group, ^** P<0.001.

Experimental method 2:

Mouse acetic acid writhing experiment: 50 male ICR mice, weighing (20±2) g, grouped and administered as in Experiment 1, for 7 consecutive days, and 1 hour after the last administration, each mouse was intraperitoneally injected with 0.6% acetic acid 0.1ml·10g ^-1 , observe the number of writhing caused by acetic acid within 15min, and calculate the inhibition rate. Inhibition rate (%) = (writhing times in the normal saline group - writhing times in the test drug group)/writhing times in the normal saline group × 100%.

Experimental results:

Effects on the writhing response of mice induced by acetic acid: the results are shown in Table 2. The results showed that each dose group of the water extract of the chrysanthemum chrysanthemum composition can significantly reduce the number of writhing reactions in mice caused by acetic acid, and there is a very significant difference compared with the blank group (P<0.001). It shows that the water extract of the composition of Filiaria sativa has a very significant analgesic effect.

Table 2: Effects of the aqueous extract of the filial scalloped grass chrysanthemum composition on the writhing response of mice induced by acetic acid ( n=10)

group Dose/g·kg^-1 Number of twists/15min Inhibition rate/% blank group ---- 45.3±8.9 ---- positive group 0.25 18.8±5.7^** 58.5 high dose 10.0 25.6±8.4^** 43.49 medium dose 5.0 23.6±5.4^** 47.90 low dose 2.5 22.1±7.7^** 51.21

Note: Compared with blank control group, ^** P<0.001.

Experimental method 3:

Antagonism xylene-induced mouse ear swelling experiment: 50 male ICR mice, weighing (20±2) g, grouped and administered as in Experiment 1, 1 hour after the last administration, apply xylene on the front and back of the right ear of the mice 0.02ml/piece, the left ear is left untreated. After 4 hours, the animal was sacrificed by broken vertebrae, both ears were cut off, and round ears were punched in the same position with a 9mm diameter punch, and weighed with an electronic balance. The weight of the right ear of each mouse minus the weight of the left ear was the degree of swelling. Swelling inhibition rate=(average swelling degree of control group-average swelling degree of drug-administered group)/average swelling degree of control group×100%.

Experimental results:

Effect of xylene-induced ear swelling in mice: the results are shown in Table 3. The results showed that each dosage group of the water extract of the composition of Filiaria sativa has a very significant inhibitory effect on the swelling of the mouse auricle induced by xylene.

Table 3: The effect of the water extract of the composition of filial fan grass Fructus chrysanthemum on the swelling of the mouse auricle caused by xylene ( n=10)

group Dose/g·kg^-1 Swelling rate/% Ear swelling inhibition rate/% blank group ---- 35.7±12.5 ---- positive group 0.25 16.3±8.2^** 54.34 high dose 10.0 12.6±5.5^** 64.71 medium dose 5.0 8.1±6.8^** 77.31 low dose 2.5 15.7±3.9^** 56.02

Note: Compared with blank control group, ^** P<0.001.

Experimental method 4:

Capillary permeability experiment in mice: 50 male ICR mice, body weight (20±2) g, the grouping and administration method were the same as the experimental method 1, for 7 consecutive days, and 1 hour after the last administration, the mice were injected into the tail vein respectively 0.5% Evans blue normal saline solution 0.1ml·10g ^-1 , then intraperitoneally injected 0.6% acetic acid normal saline solution 0.1ml·10g ^-1 , put to death 20 minutes later, cut off the abdominal skin and muscles, washed the abdominal cavity with 6ml normal saline, sucked out and washed solution, add physiological saline to 10ml, centrifuge at 3000r·min ^-1 for 10min, take the supernatant and measure its absorbance at 576nm. The inhibition rate of the peritoneal exudation of the administered mice was calculated by taking the dye exudation amount of the mice in the blank control group as 100%. Permeability was calculated as inhibition rate value.

Inhibition rate=(blank control group exudation amount-medication group exudation amount)/blank control group exudation amount×100%.

Experimental results:

Effect on capillary permeability in mice: the results are shown in Table 4. The results show that, compared with the blank group, each dose group of the water extract of the chrysanthemum chrysanthemum composition can reduce the increase of the capillary permeability of mice caused by acetic acid, and the absorbance value of the peritoneal capillary permeate is significantly reduced. (P<0.001), that is, it has obvious inhibitory effect on acute inflammatory exudation.

Table 4: Effects of the water extract of the composition of filial fan grass Fructus chrysanthemum on the permeability of capillaries in mice ( n=10)

Note: Compared with blank control group, ^** P<0.001.

Experimental method 5:

Antagonism experiment of egg white-induced mouse paw swelling: 50 Kunming mice, weighing (20±2) g, grouped and administered in the same way as experimental method 1, for 7 consecutive days. 60 minutes after the last administration, the volume of the right hind toe of each mouse was measured, each mouse was subcutaneously injected with 0.1ml of egg white (room temperature 24°C), and the volume of the right hind toe of each mouse was measured 1h, 3h, and 6h after the injection. Toe volume, the swelling rate of the right hind toe of mice in each group at different times after drug treatment was compared. Calculate according to the following formula: swelling rate (%) = (volume value of toe after inflammation - volume value of toe before inflammation) / volume value of toe before inflammation × 100%, swelling inhibition rate (%) = (average swelling of the control group rate - the average swelling rate of the treatment group) / the average swelling rate of the control group × 100%.

Experimental results:

Effect on mouse paw swelling caused by egg white: see Table 5 for the results. The results showed that in the acute non-infectious inflammation model of mouse foot edema induced by egg white, the swelling peak was 2 hours after the inflammation. It can be seen from Table 5 that each dosage group of the water extract of the composition of F. miltiorrhiza chrysanthemum has a very significant inhibitory effect on mouse paw swelling induced by egg white (P<0.001).

Table 5: The effect of the water extract of the composition of filial fan grass chrysanthemum chrysanthemum on egg white-induced mouse paw swelling ( n=10)

Note: Compared with blank control group, ^** P<0.001.

Example 2:

Experimental method 1: randomly divided into 5 groups, 10 in each group, namely the blank control group, low, medium and high dose groups (respectively crude drug amount 2.5g kg ^-1 , 5.0g·kg ^-1 , 10.0g·kg ^-1 ), positive drug (aspirin) group (0.25g·kg ^-1 ). All the other methods are the same as in Example 1.

Experimental results:

Effects on the pain threshold of mice: the results showed that each dose group of the alcohol extract of the composition of Filiaria sativa can significantly improve the pain threshold of heat-induced pain mice, which is very significantly different from that of the blank group (P<0.001), Its pain threshold increase rate is slightly higher than that of aspirin. It is suggested that the alcohol extract group of the composition of Fenugreek chrysanthemum has a very significant analgesic effect. See Table 6.

Table 6: Effects of the alcoholic extract of the composition of filial leaf grass Fructus chrysanthemum on the pain threshold of mice ( n=10)

group Dose/g·kg^-1 Pain threshold 1h after administration blank group ---- 16.64±7.23 positive group 0.25 30.11±9.01^** high dose 10.0 35.97±8.45^** medium dose 5.0 39.54±9.11^** low dose 2.5 34.58±8.01^*

Note: Compared with blank control group, ^** P<0.001.

Experimental method 2: The method is the same as that in Example 1, except that the administration is replaced by the alcohol extract of the composition of Fenugreek chrysanthemum.

Experimental results:

Effect on the writhing response of mice induced by acetic acid: the results are shown in Table 7. The results showed that each dose group of the alcohol extract of the composition of Filiaria sylvestris chrysanthemum can significantly reduce the number of writhing reactions in mice caused by acetic acid, and there is a very significant difference compared with the blank group (P<0.01, P<0.001) , suggesting that each dosage group of Alcohol extract of Fenugreek Fructus has a very obvious inhibitory effect on the writhing response of mice induced by acetic acid, indicating that the alcohol extract of Fructus Fructus Fructus composition has a very significant analgesic effect.

Table 7: Effects of the alcoholic extract of the composition of filial leaf grass Fructus chrysanthemum on the writhing reaction of mice induced by acetic acid ( n=10)

group Dose/g·kg^-1 Number of twists/15min Inhibition rate/% blank group ---- 46.3±8.3 ---- positive group 0.25 16.4±5.4^* 64.58 high dose 10.0 2.0±7.1^** 95.68 medium dose 5.0 1.9±6.4^** 95.90 low dose 2.5 7.9±8.1^** 82.94

Note: Compared with blank control group, ^* P<0.01, ^** P<0.001.

Experimental method 3: The method is the same as that in Example 1, except that the administration is replaced by the alcohol extract of the composition of Fenugreek chrysanthemum.

Experimental results:

Effect of xylene-induced ear swelling in mice: the results are shown in Table 8. The results showed that each dose group of the ethanol extract of the composition of Fenugreek chrysanthemum had a very significant inhibitory effect on the swelling of the mouse auricle induced by xylene.

Table 8: The effect of the alcohol extract of the composition of Filiaria sativa on the ear swelling of mice caused by xylene ( n=10)

group Dose/g·kg^-1 Swelling rate/% Ear swelling inhibition rate/% blank group ---- 37.98±11.0 ---- positive group 0.25 13.33±8.10^** 64.90 high dose 10.0 14.11±2.99^** 62.85 medium dose 5.0 13.01±3.7^** 65.75 low dose 2.5 16.29±3.91^** 57.11

Note: Compared with blank control group, ^** P<0.001.

Experimental method 4:

The method is the same as that in Example 1, except that the administration is replaced by the alcohol extract of the composition of Fenugreek chrysanthemum.

Experimental results:

Effects on the permeability of mouse capillaries: the results are shown in Table 9. The results show that, compared with the blank group, each dose group of the alcohol extract of the composition of Fenugreek chrysanthemum can reduce the increase of the capillary permeability of mice caused by acetic acid, and the absorbance value of the peritoneal capillary permeate fluid is significantly reduced. (P<0.01, P<0.001), that is, it has obvious inhibitory effect on acute inflammatory exudation.

Table 9: Effects of the alcoholic extract of the composition of filial piety fennel chrysanthemum on the permeability of capillaries in mice ( n=10)

group Dose/g·kg^-1 Absorbance/A Inhibition rate/% blank group ---- 0.112±0.014 ---- positive group 0.25 0.045±0.010^** 59.82 high dose 10.0 0.061±0.017^** 45.54 medium dose 5.0 0.068±0.011^** 39.29 low dose 2.5 0.089±0.023^* 20.54

Note: Compared with blank control group, ^* P<0.01, ^** P<0.001.

Example 3:

Experimental method 1: The method is the same as that in Example 1, except that the administration is replaced by alcohol extraction of the ethyl acetate extraction part of the Fructus fruticosa composition.

Experimental results:

The effect on the pain threshold of mice: the results show that the group of ethyl acetate extraction parts (group Ⅱ) and the remaining parts after solvent extraction (group Ⅳ) can significantly improve the pain threshold of mice , compared with the blank group (P<0.05, P<0.01). The results are shown in Table 10.

Table 10 Effects of Alcohol Extraction of Each Extraction Parts of Filiaria sativa Composition on the Pain Threshold of Mice (x±s, n=10)

group Dose/g·kg^-1 Pain threshold 1h after administration/s blank group / 19.16±7.2 Group I 10 20.24±6.98 Group II 10 30.87±11.19^** Group III 10 21.08±9.54 Group IV 10 25.99±7.98^* positive group 0.25 30.19±7.55^**

Note: Compared with the blank group, ^* P<0.05, ^** P<0.01.

Experimental method 2: The method is the same as that in Example 1, except that the administration is replaced by alcohol extraction of the ethyl acetate extraction part of the Fructus fruticosa composition.

Experimental results:

Effects on the writhing response of mice induced by acetic acid: the results are shown in Table 11. The results showed that the alcohol extraction of each extraction part of the composition of Fenugreek chrysanthemum can significantly reduce the number of writhing reactions in mice induced by acetic acid, and there is a significant difference compared with the blank group (P<0.05, P<0.01). It shows that the alcohol extraction of each extraction part of the composition of F. miltiorrhiza has a significant analgesic effect.

Table 11: Effects of alcohol extraction of each extraction part of the composition of Fructus chinensis chrysanthemum on the writhing response of mice induced by acetic acid (x±s, n=12)

group Dose/g·kg^-1 Twisting times/time Inhibition rate/% blank group / 38.74±6.66 / Group I 10 16.39±7.79^* 57.69 Group II 10 7.02±5.59^** 81.88 Group III 10 7.19±6.22^** 81.44 Group IV 10 10.17±7.44^* 73.74 positive group 0.25 14.07±3.99^* 63.68

Note: Compared with the blank group, ^* P<0.05, ^** P<0.01.

Experimental method 3: The method is the same as that in Example 1, except that the administration is replaced by alcohol extraction of the ethyl acetate extraction part of the composition of Fenugreek chrysanthemum.

Experimental results:

Effect of p-xylene-induced auricle swelling in mice: the results are shown in Table 12. The results show that the alcohol-extracted ethyl acetate extraction part group (Group Ⅱ) of the Xylene-induced auricle swelling of the mice has a significant effect. Significant inhibitory effect, compared with the blank group, there was a significant difference (P<0.01).

Table 12: The effect of the alcohol extraction of each extraction part of the composition of Filiaria sativa on the ear swelling of mice induced by xylene ( n=10)

group Dose/g·kg^-1 swelling rate% Ear swelling inhibition rate/% blank group / 37.20±10.25 / Group I 10 25.47±6.54 31.53 Group II 10 12.45±8.01^** 66.53 Group III 10 24.59±7.77 33.90 Group IV 10 28.46±5.98 23.49 positive group 0.25 15.77±8.54^** 57.61

Note: Compared with the blank group, ^* P<0.01.

Example 4:

experimental method:

Animal grouping: 60 SD male rats, body weight (160±10) g, after a week of adaptation to feeding, they were randomly divided into 6 groups of 10 rats in each group: blank group, model group, and water extraction of chrysanthemum chrysanthemum composition Low, medium and high dose groups, positive control group.

Modeling and administration: Positive group, model group, low-, medium-, and high-dose groups of the extracts of the composition of Fennelia sativa, medium, and high doses were injected intradermally with 0.1ml/rat's complete Freund's adjuvant into the right hind foot pad of the rats to establish the model , the blank group was intradermally injected with 0.1ml/rat normal saline on the right hind foot pad; while the model was being established, the positive group was given tripterygium glycosides tablets and fenugreek chrysanthemum composition by intragastric administration respectively at 15 mg·kg ^-1 The low-, medium-, and high-dose groups were administered intragastrically with 2.5 g·kg ^-1 , 5.0 g·kg ^-1 , and 10.0 g·kg ^-1 of the water extracts of the composition of the chrysanthemum chrysanthemum. To the corresponding concentration, 1ml·100g ^-1 was given by intragastric administration, and the model group and the blank group were given equal volume of distilled water, once a day, for 28 days in total.

Detection Indicator:

① Observation of animal state: including coat color, gloss and activity.

② Effects on rat body weight: Weighed at the first ld of the experiment, and weighed once every 7 days thereafter. The body weight of the last time minus the body weight of the previous time is the weight gain of rats in each group.

③Measurement of swelling degree of paws: before modeling, the rats’ hind limbs were straightened, marked with picric acid at 0.5 cm above the ankle joint, and then put into the toe volume measuring instrument, so that the paw marks were level with the water surface, and measured Before the modeling, the left and right paw volumes (mL) of the rats in each group were recorded 5 times, and the average value was taken as the swelling degree of the forefoot of the modeling. And use the same method to measure the volume of the right hind toe at 6h, 12h, 24h, 36h, 48h, 60h, 72h, 7d, and 11d after modeling, and measure the volume of the left and right hind toes at 14d, 18d, 21d, 24d, and 27d and record. The degree of foot swelling was calculated according to the volume of the left and right hind toes and the volume before modeling.

Paw swelling degree (mL) = toe volume after modeling - toe volume before modeling.

④Measurement of main organ index: Animals were sacrificed 4 hours after the last administration (administration on the 28th day), thymus and spleen were taken, weighed, and organ index was calculated, organ index (mg·g ^-1 )=(organ weight×1000 )/weight.

⑤Pathological section of synovial tissue: After the last administration, the animals were sacrificed, and the synovial membrane of the knee joint was taken, fixed in formalin, and routine HE pathological section examination was performed. Synovial pathological scoring criteria: inflammatory cells: 0 points, none; 1 point, sparse, scattered; 2 points, relatively dense; 3 points, a large number. Fibrous tissue hyperplasia: 0 points, no; 1 point, a small amount of hyperplasia; 2 points, moderate; 3 points, a large amount. Synovial hyperplasia: 0, no; 1 point, single layer; 2 points, 2 layers; 3 points, 3 layers.

Experimental results:

①Observation of general situation

After modeling, the acute inflammatory reaction of the rats was obvious, and the side injected with complete Freund's adjuvant was obviously swollen. Compared with the blank group, the activity of the model rats was reduced, and when obvious inflammatory reactions occurred, the food intake was significantly reduced, the coat color was not smooth, and the ankle joints were swollen and festered; the rats in the drug treatment group had no ulceration; difference.

② Changes in body weight of rats in each group

Within 28 days of modeling, the weight gain of the rats in the model group was significantly lower than that in the normal group, and the difference was significant (P<0.05, P<0.01). There was a significant difference (P<0.05, P<0.01) in the body weight gain of AA rats in each dose group of the water extract of the composition of F. syphilis and the model group (P<0.05, P<0.01), indicating that the water extract of the composition of F. All dose groups can improve the effect of inflammation on the body weight of AA rats to varying degrees. The results are shown in Table 13.

Table 13: Changes in body weight of rats in each group (unit: g; n=10)

Note: Compared with the blank group, ^# P<0.05, ^## P<0.01; compared with the model group, ^★ P<0.05, ^★★ P<0.01.

③Measurement of rat joint swelling

Before the experiment, there was no significant difference in the toe volume of all the rats between the groups (P>0.05). During the experiment period, the rats in the blank group had no redness and swelling in the joints of the limbs, and they moved freely. After modeling, the acute primary inflammatory reaction of the rats was obvious, and the toes were severely red and swollen. Secondary inflammatory reactions also occurred. On the 28th day of modeling, most of the ankle joints of the rats in the model group were ulcerated to varying degrees, and there was no ulceration in the positive control group and the treatment group treated with the water extract of the chrysanthemum chrysanthemum composition. See Tables 14 and 15 for the effects of the composition of Filibulum chinensis on the primary inflammation and secondary inflammation of the toe swelling of rats.

Table 14: The effect of the water extract of the composition of filial scalloped chrysanthemum chrysanthemum on the swelling degree of the right foot of AA rats ( n=10)

(Continued) Table 14: Effects of the water extract of the composition of Filiaria syphilis chrysanthemum on the swelling degree of the right toe of AA rats ( n=10)

Note: Compared with the blank group, ^# P<0.05, ^## P<0.01; compared with the model group, ^★ P<0.05, ^★★ P<0.01.

Table 15: The effect of the water extract of the composition of filial scalloped chrysanthemum chrysanthemum on the degree of swelling of the left toe of AA rats ( n=10)

Note: Compared with the blank group, ^# P<0.05, ^## P<0.01; compared with the model group, ^★ P<0.05, ^★★ P<0.01.

④ Determination of major organ indexes

The results are shown in Table 16. The results showed that each dosage group of the water extract of the composition of F. filalis sylvestris chrysanthemum had obvious inhibitory effect on thymus, spleen and adrenal gland of AA rats.

Table 16: Effects of the water extract of the filial sage chrysanthemum composition water extract on the weight of the thymus, spleen and adrenal glands of AA rats ( n=10)

Note: Compared with the blank group, ^# P<0.05, ^## P<0.01; compared with the model group, ^★ P<0.05, ^★★ P<0.01.

⑤ Synovial pathology

No obvious neovascularization and inflammatory cell infiltration were found in the synovial tissue of normal rats, and the synovial cells were arranged in 1-2 layers. In the model group, there were obvious angiogenesis, synovial cell volume increase, synovial thickening, and inflammatory cell infiltration. In the synovial tissue of the treatment group, a small amount of new blood vessels and inflammatory cell infiltration were seen, and the synovial membrane was slightly thickened. The pathological scores of the model group were significantly higher than those of the normal group (P<0.01); the pathological scores of the inflammatory cells and synovial tissue hyperplasia in the low, medium and high dose groups of the composition group were lower than those of the model group (P<0.01). The results are shown in Table 17, accompanying drawings 1 and 2.

Table 17: Effects of the aqueous extract of the filial genus Fructus chrysanthemum composition on the pathological score of the synovial membrane of AA rats ( n=10)

group Dose/g·kg^-1 Inflammatory cell infiltration synovial tissue hyperplasia Fibroplasia blank group ----- 0.53±0.12^★★ 0.56±0.33^★★ 0.49±0.24^★★ model group ----- 1.29±0.11^## 1.53±0.45^## 1.28±0.24^## positive group 0.015 0.77±0.14^##★★ 0.68±0.33^#★★ 0.78±0.31^##★★ high dose 10.0 0.79±0.22^##★★ 0.75±0.34^##★★ 0.84±0.36^##★★ medium dose 5.0 0.80±0.44^##★★ 0.70±0.22^##★★ 0.85±0.23^##★★ low dose 2.5 0.83±0.22^##★★ 0.72±0.40^##★★ 0.86±0.20^##★★

Note: Compared with the blank group, ^# P<0.05, ^## P<0.01; compared with the model group, ^★ P<0.05, ^★★ P<0.01.

Example 5:

experimental method:

Animal grouping: 60 SD male rats, body weight (160±10) g, after one week of adaptation to feeding, were randomly divided into 6 groups of 10 rats in each group: blank group, model group, ethanol extraction of the composition of saffron chrysanthemum chrysanthemum Low, medium and high dose groups, positive control group.

Modeling and administration: the same as in Example 4.

Detection Indicator:

①Observation of the state of the animal: the same as in Example 4.

② Effect of rat body weight: same as in Example 4.

③Measurement of paw swelling: same as in Example 4.

④ Arthritis index score: From the 14th day after the rat model was established, the clinical inflammation score was regularly recorded according to the degree of redness, swelling and hardening of the joint inflammation of the rat limbs, so as to evaluate the degree of secondary lesions of the AA rat model. The highest score for a single limb of a rat is 4 points, and the highest score for all four limbs is 16 points. The following table 18 scoring criteria.

Table 18: Arthritis Index Score Form

⑤ Determination of main organ index: same as Example 4.

Experimental results:

①Observation of general situation

After modeling, the acute inflammatory reaction of the rats was obvious, and the side injected with Freund's complete adjuvant was obviously swollen. The rats in the blank group were in a good mental state during the 28-day test period, with shiny hair, quick movements, and normal diet and drinking water. Compared with the blank group, the model rats were listless, and when obvious inflammatory reactions occurred, the food intake was significantly reduced, the coat color was dull, the weight gain was slow, and the ankle joints were swollen and ulcerated; the rats in the drug treatment group basically had no ulceration; the rats in the blank group There is basically no difference from before modeling.

② Changes in body weight of rats in each group

The body weight of rats in the blank group increased progressively during the 28-day test period. Within 28 days of modeling, the weight gain of rats in the model group was significantly different from that in the blank group (P<0.01), and the weight gain of AA rats in each dose group of Alcohol Extract of Fructus Fructus Fructus was significantly different from that in the model group. The differences (P<0.01, P<0.05) indicated that each dosage group of Alcohol Extract of Fenugreek Fructus can improve the effect of inflammation on the body weight of AA rats to varying degrees. The results are shown in Table 19.

Table 19: Changes in body weight of rats in each group (unit: g; n=10)

group Before modeling W/g 7d(ΔW)/g 14d(ΔW)/g 21d(ΔW)/g 27d(ΔW)/g blank group 175.14±7.21 47.11±11.91^★★ 50.29±12.99^★★ 48.67±7.26^★★ 43.29±7.71^★★ model group 178.91±6.44 26.12±9.8^## 27.15±16.01^## 29.00±7.89^## 32.08±11.98^## positive group 176.11±9.47 29.81±7.9^# 32.11±18.74^##★ 30.01±6.09^## 39.77±13.31^##★ high dose 179.29±6.89 38.77±11.57^★★ 40.02±11.08^#★★ 38.74±11.54^★★ 33.99±10.08^## medium dose 180.33±4.07 32.64±15.71^#★ 39.94±11.34^#★★ 38.90±11.29^★★ 33.77±10.48^## low dose 175.47±9.87 32.01±11.00^#★ 38.88±8.02^#★★ 34.33±10.74^#★ 29.08±7.15^##

Note: Compared with the blank group, ^# P<0.05, ^## P<0.01; compared with the model group, ^★ P<0.05, ^★★ P<0.01.

③Measurement of rat joint swelling

Before the experiment, there was no significant difference in the toe volume of all the rats between the groups (P>0.05). During the experiment period, the rats in the blank group had no redness and swelling in the joints of the limbs, and they moved freely. After modeling, the rats had obvious inflammation, and the toes were severely red and swollen. Swelling began to appear 6 hours after modeling, and secondary inflammatory reactions also occurred. Each dose group of the alcohol extract of the composition of Filiaria sativa has different degrees of inhibitory effect on the primary acute swelling of the toes of AA rats. Table 20 and Table 21 show the effects of the composition of Filibulum chinensis on the primary inflammation and secondary inflammation of the toe swelling of rats.

Table 20: The effect of the alcohol extract of the composition of filial piety fennel chrysanthemum chrysanthemum on the swelling degree of the right toe of AA rats ( n=10)

(Continued) Table 20: Effects of the alcohol extract of the composition of Fenugreek chrysanthemum on the swelling degree of the right toe of AA rats ( n=10)

(Continued) Table 20: Effects of the alcohol extract of the composition of Fenugreek chrysanthemum on the swelling degree of the right toe of AA rats ( n=10)

Note: Compared with the blank group, ^# P<0.05, ^## P<0.01; compared with the model group, ^★ P<0.05, ^★★ P<0.01.

Table 21: The effect of the alcohol extract of the composition of filial leaf grass Fructus chrysanthemum on the swelling degree of the left toe of AA rats ( n=10)

(Continued) Table 21: The effect of the alcohol extract of the composition of Fenugreek chrysanthemum on the swelling degree of the left toe of AA rats ( n=10)

Note: Compared with the blank group, ^# P<0.05, ^## P<0.01; compared with the model group, ^★ P<0.05, ^★★ P<0.01.

④ Rat Arthritis Index Score

The experimental results are shown in Table 22. The results showed that the toes, foot pads, and ankle joints of all the rats in the model group had different degrees of redness and swelling. Due to the aggravation of secondary swelling in the later stage, the number of animals with joint deformities and dysfunction increased. The joint scores of rats in the model group The index keeps rising. Compared with the model group, there were significant differences (P<0.01, P<0.05).

Table 22: Effect of Alcoholic Extract of Sheafflower on the Arthritis Score Index of AA Rats ( n=10)

Note: Compared with the blank group, ^## P<0.01; compared with the model group, ^★ P<0.05, ^★★ P<0.01.

⑤ Determination of main organ index

The results are shown in Table 23. The results showed that each dose group of the alcohol extract of the composition of F. filariae Fructus chrysanthemum had obvious inhibitory effect on the thymus, spleen and adrenal gland of AA rats.

Table 23: Effects of the alcoholic extract of the composition of filial leaf grass Fructus chrysanthemum on the weight of thymus, spleen and adrenal gland in AA rats ( n=10)

group Dose/g·kg^-1 Thymus index/mg·g^-1 Spleen index/mg g^-1 Adrenal index/mg·g^-1 blank group ----- 1.64±0.19^★★ 1.98±0.25^★★ 0.137±0.05^★★ model group ----- 1.30±0.18^## 2.41±0.27^## 0.177±0.03^## positive group 0.015 1.60±0.23^★★ 2.00±0.24^★★ 0.140±0.02^★★ high dose 10.0 1.48±0.25^#★★ 2.00±0.15^★★ 0.148±0.03^★ medium dose 5.0 1.50±0.24^#★★ 1.99±0.14^★★ 0.140±0.07^★★ low dose 2.50 1.49±0.22^#★★ 2.01±0.19^★ 0.144±0.04^★

Note: Compared with the blank group, ^# P<0.05, ^## P<0.01; compared with the model group, ^★ P<0.05, ^★★ P<0.01.

Embodiment 6:

experimental method:

Animal grouping: 60 SD male rats, body weight (160±10) g, adapted to feeding for a week, were randomly divided into 6 groups according to 10 rats in each group: blank group (group I), model group (group II), positive group (Group III), Alcohol Extraction of Ethyl Acetate Extraction Parts Group (Group IV), Alcohol Extraction of Butanol Extraction Parts Group (Group V) and Each Solvent Extraction After the rest of the site group (VI group).

Modeling and administration: blank group (group I), model group (group II), positive group (group III), alcohol extraction of ethyl acetate extraction part group (group IV), The composition of sheep ear chrysanthemum was extracted with alcohol and n-butanol extraction group (group V) and the remaining parts group after each solvent extraction (group VI) were intradermally injected with 0.1ml/piece of complete Freund's adjuvant in the right hind foot pad of rats. The rats in the blank group were intradermally injected with 0.1ml of normal saline on the right hind footpad; at the same time, the rats in the positive group were given tripterygium glycosides tablets and fenugreek chrysanthemum combination by intragastric administration at 15 mg·kg ^-1 Ethyl acetate extraction group (group IV), alcohol extraction group of n-butanol extraction group (group V) and the remaining parts group after each solvent extraction (group VI) were given by intragastric administration, respectively. 10g·kg ^-1 liquid medicine, all the drugs were adjusted to the corresponding concentration and administered by intragastric administration at 1ml·100g ^-1 , and the model group and blank group were given equal volume of distilled water once a day for a total of 28 days.

Detection Indicator:

① Observation of animal state: including coat color, gloss and activity.

② Effects on rat body weight: Weighed at the first ld of the experiment, and weighed once every 7 days thereafter. The body weight of the last time minus the body weight of the previous time is the weight gain of rats in each group.

③Measurement of swelling degree of paws: before modeling, the rats’ hind limbs were straightened, marked with picric acid at 0.5 cm above the ankle joint, and then put into the toe volume measuring instrument, so that the paw marks were level with the water surface, and measured Before the modeling, the left and right paw volumes (mL) of the rats in each group were recorded 5 times, and the average value was taken as the swelling degree of the forefoot of the modeling. And use the same method to measure the volume of the right hind foot at 6h, 18h, 24h, 48h, 4d, 8d, 12d, 16d, 20d, 24d, and 28d after modeling, and measure the volume of the left and right hind feet at 12d, 16d, 20d, 24d, and 28d. Full toe volume and record. The degree of foot swelling was calculated according to the volume of the left and right hind toes and the volume before modeling.

Paw swelling degree (mL) = toe volume after modeling - toe volume before modeling.

④ Arthritis index score: From the 14th day after the rat model was established, the clinical inflammation score was regularly recorded according to the degree of redness, swelling and hardening of the joint inflammation of the rat limbs, so as to evaluate the degree of secondary lesions of the AA rat model. The highest score for a single limb of a rat is 4 points, and the highest score for all four limbs is 16 points. Scoring criteria as in Table 18 above.

⑤ Determination of main organ index: Animals were sacrificed 4 hours after the last administration, thymus and spleen were taken out, weighed, and organ index was calculated, organ index (mg·g ^-1 )=(organ weight×1000)/body weight.

Experimental results:

①Observation of general situation

After modeling, the acute inflammatory reaction of the rats was obvious, and the side injected with complete Freund's adjuvant was obviously swollen. The blank group was in a good mental state during the 28-day test period, with shiny hair, quick movements, and normal diet and drinking water. Compared with the blank group, the model rats were less active, and when obvious inflammatory reactions occurred, the food intake was significantly reduced, the hair was dull, the weight gain was slow, and the ankle joints were swollen and ulcerated; the rats in the drug treatment group basically had no ulceration; There was basically no difference between the mice and those before modeling.

② Changes in body weight of rats in each group

Within 28 days of modeling, the rats in the model group (Group II) and the rats in the blank group (Group I) were in good condition, with shiny hair, weight gain, free movement and sensitive response. After 28 days of modeling, the weight gain of rats in the modeling group (Group II) was significantly different from that in the blank group (Group I) (P<0.01, P<0.05). Decrease, slow weight gain. Modeling 14d, 21d, 27d The weight gain of AA rats in the Alcohol-extracted ethyl acetate extraction part group (Group Ⅳ) was significantly different from that in the model group (Group Ⅱ) (P<0.01) . There was a very significant difference (P<0.01) in the weight gain of the rats in the group (Group IV) compared with the model group (Group II) in the alcohol extraction of the ethyl acetate extraction part of the composition of the filial leaf grass and chrysanthemum chrysanthemum on the 21d and 27d of modeling. See Table 25.

Table 25: Changes in body weight of rats in each group ( n=10)

group Before modeling W/g 7d(△W)/g 14d(△W)/g 20d(△W)/g 28d(△W)/g Group I 170.44±5.88 40.90±5.66^▲▲ 45.1±5.11^▲▲ 46.92±7.97^▲▲ 45.97±6.94^▲▲ Group II 169.98±8.01 24.11±6.77^** 32.19±6.11^** 28.60±7.11^** 38.11±6.89^* Group III 171.55±9.01 21.83±7.77^** 32.88±5.93^** 39.61±12.45^**▲▲ 42.47±12.01^▲▲ Group IV 169.88±5.69 28.60±9.70^* 49.89±9.01^▲▲ 51.27±9.11^*▲▲ 64.02±11.01^*▲▲ Group V 168.41±6.99 26.79±5.17^* 39.99±4.81^**▲ 27.73±5.87^** 36.20±6.51^** Group VI 168.47±8.22 22.55±6.19^** 33.87±4.17^** 37.46±5.11^**▲ 39.05±11.37^**

Note: Compared with the blank group ^* P<0.05, ^** P<0.01, compared with the model group ^▲ P<0.05, ^▲▲ P<0.01.

③Measurement of rat joint swelling

Before the experiment, there was no significant difference in the toe volume of all the rats between the groups (P>0.05). During the experiment period, the rats in the blank group had no redness and swelling in the joints of the limbs, and they moved freely. After modeling, the acute primary inflammation of rats was obvious, and the toes were severely red and swollen, and swelling began to appear 6 hours after modeling. The rats in the model group had swelling, ulceration, edema, joint deformity on both sides of the toes, red, swollen and inflamed ears, and nodular lumps on the tail and other secondary lesions. After 28 days of modeling, the ankle joints of the modeling rats all appeared to varying degrees. fester. The experimental results of the effect of the alcohol extraction of each extraction part of the composition of Fenugreek chrysanthemum on the swelling degree of the right toe of AA rats are shown in Table 17. See Table 26 for the experimental results of degree influence.

Table 26: The effect of the alcohol extraction of each extraction part of the composition of filial leaf grass Fructus chrysanthemum on the swelling degree of the right toe of AA rats ( n=10)

group 6 hours 18h 24h 48h 4d Group I 0.02±0.05^▲▲ 0.03±0.01^▲▲ 0.03±0.03^▲▲ 0.04±0.05^▲▲ 0.04±0.22^▲▲ Group II 0.43±0.09^** 0.76±0.29^** 0.78±0.11^** 0.88±0.22^** 0.90±0.22^** Group III 0.28±0.09^**▲ 0.74±0.11^** 0.74±0.18^** 0.74±0.11^**▲ 0.80±0.11^**▲ Group IV 0.20±0.05^**▲ 0.35±0.27^**▲▲ 0.55±0.17^**▲▲ 0.67±0.19^**▲▲ 0.77±0.22^**▲ Group V 0.23±0.05^**▲ 0.64±0.33^**▲ 0.62±0.27^**▲▲ 0.70±0.33^**▲ 0.84±0.19^** Group VI 0.30±0.09^** 0.50±0.19^**▲▲ 0.48±0.08^**▲▲ 0.81±0.47^** 0.94±0.27^**

(Continued) Table 26: The effect of the alcohol extraction of the composition of Fenugreek chrysanthemum on the swelling degree of the right foot toes of AA rats ( n=10)

Note: Compared with the blank group ^* P<0.05, ^** P<0.01, compared with the model group ^▲ P<0.05, ^▲▲ P<0.01.

Table 27: The effect of the alcohol extraction of each extraction part of the composition of filial piety chrysanthemum chrysanthemum on the swelling degree of the left foot of AA rats ( n=10)

group d12 d16 d20 d24 d28 Group I 0.11±0.03 0.18±0.06^▲▲ 0.19±0.05^▲▲ 0.23±0.05^▲▲ 0.23±0.07^▲▲ Group II 0.16±0.05 0.20±0.41^** 0.25±0.39^** 0.37±0.51^** 0.49±0.29^** Group III 0.14±0.02^* 0.18±0.22 0.19±0.18^▲▲ 0.23±0.49^▲▲ 0.25±0.21^*▲▲ Group IV 0.10±0.04^**▲ 0.14±0.21^*▲▲ 0.24±0.22^** 0.22±0.38^▲▲ 0.17±0.15^**▲▲ Group V 0.15±0.05^** 0.29±0.22^* 0.31±0.11^**▲ 0.69±0.51^**▲▲ 0.87±0.451^**▲▲ Group VI 0.18±0.07^** 0.77±0.29^**▲▲ 0.86±0.27^**▲▲ 1.73±0.41^**▲▲ 1.71±0.47^**▲▲

Note: Compared with the blank group ^* P<0.05, ^** P<0.01, compared with the model group ^▲ P<0.05, ^▲▲ P<0.01.

④ Rat Arthritis Index Score

The experimental results are shown in Table 28. The results showed that all the joints of the modeled rats had varying degrees of inflammation, and there were extremely significant differences (P<0.01) from those in the blank group (Group I). The positive group (Ⅲ group) and the alcohol-extracted ethyl acetate extraction part group (group Ⅳ) of Fenugreek chrysanthemum composition can significantly improve joint inflammation, reduce the arthritis index of AA rats, and inhibit secondary inflammation, compared with the model group ( Group Ⅱ) were significantly different (P<0.01).

Table 28: The effect of alcohol extraction of each extraction part of the composition of filial leaf grass Fructus chrysanthemum on the arthritis index of AA rats ( n=10)

group 14d 21d 27d Group I 0 0 0 Group II 5.50±1.22^** 5.69±0.89^** 5.70±0.77^** Group III 3.00±1.11^**▲▲ 4.11±1.08^**▲▲ 4.08±0.69^**▲▲ Group IV 3.14±1.11^**▲▲ 3.67±1.02^**▲▲ 3.87±0.74^**▲▲ Group V 5.21±1.27^** 5.37±0.99^** 5.62±0.66^** Group VI 5.39±0.99^** 5.54±1.12^** 5.60±0.69^**

Note: Compared with the blank group ^* P<0.05, ^** P<0.01; compared with the model group ^▲ P<0.05, ^▲▲ P<0.01.

⑤ Determination of main organ index

The experimental results are shown in Table 29. The results showed that the thymus of the AA rats in the model group (Group II) atrophied, the index of the thymus decreased, and the index of the spleen increased. Compared with the blank control group (Group I), there were extremely significant differences (P<0.01, P<0.01), the positive group (group III), the ethyl acetate extraction part group (group IV) and the n-butanol extraction part group (group V) of the fenugreek chrysanthemum composition can significantly inhibit the atrophy of the thymus, Increased thymus index in rats, compared with the model group (Group II), there was a very significant difference (P<0.01, P<0.01, P<0.01). Both the positive group (Ⅲ group) and the alcohol-extracted ethyl acetate extraction part group (group Ⅳ) can significantly increase the spleen index, and there is a significant difference compared with the model group (group Ⅱ) (P<0.01 , P<0.05).

Table 29 Effects of Alcoholic Extraction of Alcohol Extraction Parts on AA Rat Visceral Index of AA Rats (x±s, n=10)

group Thymus index/g kg^-1 Spleen index/g·kg^-1 Group I 1.74±0.06^▲▲ 2.06±0.20^▲▲ Group II 1.38±0.13^** 2.93±0.44^** Group III 1.61±0.11^*▲▲ 2.54±0.40^**▲▲ Group IV 1.68±0.19^*▲▲ 2.60±0.40^**▲ Group V 1.63±0.33^*▲▲ 2.79±0.22^** Group VI 1.57±0.18^** 2.90±0.41^**

Note: Compared with the blank group ^* P<0.05, ^** P<0.01, compared with the model group ^▲ P<0.05, ^▲▲ P<0.01.

Implementation case 7:

experimental method:

Animal grouping: 60 SD male rats with a body weight of (160±10) g, after being fed for one week, they were randomly divided into 6 groups of 10 rats in each group: blank group, model group, and each extraction part group of Alcohol Extraction ( Respectively, petroleum ether extraction parts (group Ⅰ), ethyl acetate extraction parts (group Ⅱ), n-butanol extraction parts (group Ⅲ) and the remaining parts after each solvent extraction (group Ⅳ), the positive control group, and the The preparation method of the extract is the same as that in Example 1, except that the fescue component in the composition is removed.

Modeling and drug administration: positive group, model group, each extraction part group of Alcohol Extraction (respectively, petroleum ether extraction part (Group Ⅰ), ethyl acetate extraction part (Group Ⅱ), n-butanol extraction part (Group Ⅲ) group) and the remaining parts after each solvent extraction (Group IV)), the positive control group was intradermally injected with 0.1ml/piece Freund's complete adjuvant into the right hind foot pad of rats to establish a model, and the blank group was injected into the skin of the right hind foot pad of rats. 0.1ml/saline was injected internally; at the same time as the model was established, Tripterygium wilfordii Polyglycoside Tablets were given intragastrically at 15 mg·kg ^-1 in the positive group, and each extraction site group (respectively, petroleum ether extraction sites (group Ⅰ) , ethyl acetate extraction site (group Ⅱ), n-butanol extraction site (group Ⅲ) and the remaining site after each solvent extraction (group Ⅳ)) were given 10.0 g·kg ^-1 crude drug by intragastric administration, and the drug was adjusted to The corresponding concentration was given by intragastric administration of 1ml·100g ^-1 , and the model group and the blank group were given equal volumes of distilled water once a day for 28 days in total.

Detection Indicator:

① Observation of animal state: including coat color, gloss and activity.

② Effects on rat body weight: Weighed at the first ld of the experiment, and weighed once every 7 days thereafter. The body weight of the last time minus the body weight of the previous time is the weight gain of rats in each group.

③Measurement of swelling degree of paws: before modeling, the rats’ hind limbs were straightened, marked with picric acid at 0.5 cm above the ankle joint, and then put into the toe volume measuring instrument, so that the paw marks were level with the water surface, and measured Before the modeling, the left and right paw volumes (mL) of the rats in each group were recorded 5 times, and the average value was taken as the swelling degree of the forefoot of the modeling. And use the same method to measure the volume of the right hind toe at 6h, 12h, 24h, 36h, 48h, 60h, 72h, 7d, and 11d after modeling, and measure the volume of the left and right hind toes at 14d, 18d, 21d, 24d, and 27d and record. The degree of foot swelling was calculated according to the volume of the left and right hind toes and the volume before modeling.

Paw swelling degree (mL) = toe volume after modeling - toe volume before modeling.

④ Arthritis index score: From the 14th day after the rat model was established, the clinical inflammation score was regularly recorded according to the degree of redness, swelling and hardening of the joint inflammation of the rat limbs, so as to evaluate the degree of secondary lesions of the AA rat model. The highest score for a single limb of a rat is 4 points, and the highest score for all four limbs is 16 points. Scoring criteria as in Table 18 above.

⑤ Determination of main organ index: Animals were sacrificed 4 hours after the last administration, thymus and spleen were taken out, weighed, and organ index was calculated, organ index (mg·g ^-1 )=(organ weight×1000)/body weight.

Experimental results:

①Observation of general situation

After modeling, the acute inflammatory reaction of the rats was obvious, and the side injected with complete Freund's adjuvant was obviously swollen. The rats in the blank group were in a good mental state during the 28-day test period, with shiny hair, quick movements, normal diet and drinking water, and a progressive increase in body weight. Compared with the blank group, the model rats were depressed, and when obvious inflammatory reactions occurred, that is, on the 18th to 24th day after modeling, the food intake was significantly reduced, the hair color was dull, the weight growth was slow, and the ankle joints were swollen and festered; the rats in the drug treatment group There was basically no ulceration; the rats in the blank group were basically the same as those before modeling.

② Changes in body weight of rats in each group

Within 28 days of modeling, the weight gain of rats in the model group was significantly different from that in the blank group (P<0.01, P<0.05). There were significant differences (P<0.01, P<0.05), which indicated that the alcohol extraction of Filiaria miltiorrhiza could improve the effect of inflammation on the body weight of AA rats to varying degrees. The results are shown in Table 30.

During the whole experiment, the rats in the blank group (group I) were in good condition, with shiny hair, weight gain, free movement and sensitive response. After 28 days of modeling, the weight gain of rats in all modeling groups (group II) was significantly different from that in the blank group (group I) (P<0.01). The body weight increased slowly, and the body weight increased slightly on the 14th day of modeling. At the same time, the results showed that the weight gain of AA rats in the group (Group IV) was significantly higher than that in the model group (Group II). Sex difference (P<0.05). There was a very significant difference (P<0.01, P<0.01) in the weight gain of the rats in the 20d and 28d model-making days compared with the model group (Group Ⅱ) in the rats of the ethyl acetate extraction part group (Group Ⅳ). Compared with the group of extract parts extracted by alcohol extraction of F. miltiorrhiza, AA rats whose extract parts were extracted by alcohol alone had a slower weight gain, a smaller increase, a lower state of depression, and a lower desire to eat. Slightly sluggish in response. The results are shown in Table 30.

Table 30 The effect of each extraction part of Alcohol Extraction on AA Rat Body Weight Gain ( n=10)

group Before modeling W/g 7d(ΔW)/g 14d(△W)/g 21d(△W)/g 27d(△W)/g Group I 174.84±7.87 24.84±5.63 48.7±5.02^▲▲ 60.91±8.30^▲▲ 84.81±7.44^▲▲ Group II 169.52±7.91 24.71±6.86 39.45±6.05^** 29.60±7.01^** 40.79±7.66^** Group III 171.69±9.01 20.82±8.23 31.5±5.92^** 36.61±11.46^**▲ 50.48±11.05^**▲ Group IV 170.82±6.94 27.55±3.98 44.67±3.01^*▲ 41.43±9.21^**▲▲ 56.19±12.23^**▲▲ Group V 165.75±8.25 25.66±5.27 37.25±4.88^** 25.73±4.87^** 36.91±6.58^** Group VI 170.51±11.66 20.52±6.39 31.78±4.16^** 23.23±4.96^** 33.48±10.37^**

Note: Compared with the blank group ^* P<0.05, ^** P<0.01, compared with the model group ^▲ P<0.05, ^▲▲ P<0.01.

②Effects of various extraction parts of Alcohol Extraction of Filiaria miltiorrhiza on the degree of swelling of the toes of AA rats

After modeling, the acute primary inflammation of rats was obvious, and the toes were severely red and swollen, and swelling began to appear 6 hours after modeling. The primary lesion reached its peak on the 8th day after modeling. Secondary lesions began to appear on the 12th day of modeling, and the swelling degree of the left toe reached a peak on the 16th to 20th day. Swelling, ulceration, edema, and joint deformity appeared in the toes on both sides, red, swollen and inflamed ears, and nodular mass in the tail. On the 28th day of modeling, the ankle joints of the modeling rats all had varying degrees of ulceration. Compared with the different extractions of the Alcohol-extracted Alcohol-extracted Alcohol-extracted AA Rats, the swelling of the right toe of AA rats was more serious, the curative effect was slow, the effect was mild, and the secondary swelling degree is also more serious.

Table 31 shows the experimental results of the effect of the various extraction parts of the Alcohol Extraction on the swelling degree of the left toes of AA rats.

Table 31 The effect of each extraction part on the swelling degree of the right foot of the AA rat by the alcohol extraction of fennel grass ( n=10)

group 6 hours 18h 24h 48h Group I 0.14±0.06 0.08±0.05^▲▲ 0.08±0.02^▲▲ 0.08±0.05 Group II 0.25±0.10^* 0.88±0.29^** 0.65±0.12^** 0.78±0.27^** Group III 0.28±0.08^** 0.78±0.29^** 0.79±0.17^**▲ 0.76±0.15^** Group IV 0.30±0.07^** 0.83±0.31^** 0.70±0.16^** 0.69±0.18^** Group V 0.23±0.06^** 0.66±0.35^** 0.60±0.14^** 0.75±0.27^** Group VI 0.31±0.08^** 0.54±0.20^**▲▲ 0.51±0.09^**▲▲ 0.81±0.17^**

Table 31 The effect of each extraction part on the swelling degree of the right foot of the AA rat by the alcohol extraction of fennel grass ( n=10)

Note: Compared with the blank group ^* P<0.05, ^** P<0.01, compared with the model group ^▲ P<0.05, ^▲▲ P<0.01.

Table 31 The effect of each extraction part on the swelling degree of the left foot of AA rats by alcohol extraction n=10)

Note: Compared with the blank group ^* P<0.05, ^** P<0.01, compared with the model group ^▲ P<0.05, ^▲▲ P<0.01.

③Effects of different extraction parts of Alcohol Extraction of Filiaria miltiorrhiza on the arthritis index of AA rats

The experimental results are shown in Table 45. The results showed that all the joints of the modeled rats had varying degrees of inflammation, and there were extremely significant differences (P<0.01) from those in the blank group (Group I). The positive group (Ⅲ group) and the ethyl acetate extraction part group (group Ⅳ) can significantly improve the joint inflammation, reduce the arthritis index of AA rats, and inhibit the secondary inflammation, compared with the model group (group Ⅱ). There are extremely significant differences (P<0.01, P<0.01). Compared with the alcohol-extracted parts of S. chinensis chrysanthemum, AA rats with alcohol-extracted parts of S. syphilis alone generally have higher arthritis index and obvious joint inflammation.

Table 45 The influence of each extraction part of Alcohol Extraction on AA Rat Arthritis Index ( n=10)

group 12d 20d 28d Group I 0 0 0 Group II 5.20±1.32^** 5.60±0.70^** 5.60±0.84^** Group III 3.10±1.10^**▲▲ 4.30±1.06^**▲▲ 3.90±0.74^**▲▲ Group IV 3.70±1.06^**▲▲ 4.60±0.97^**▲▲ 3.90±0.74^**▲▲ Group V 5.30±1.25^** 5.70±0.82^** 5.20±0.63^** Group VI 5.70±0.95^** 6.20±0.92^** 5.80±0.79^**

Note: Compared with the blank group ^* P<0.05, ^** P<0.01; compared with the model group ^▲ P<0.05, ^▲▲ P<0.01.

④Effects of different extracting parts of Alcohol Extraction of Filiaria miltiorrhiza on the visceral index of AA rats

The experimental results are shown in Table 46. The results showed that the thymus of the AA rats in the model group (Group II) atrophied, the index of the thymus decreased, and the index of the spleen increased. Compared with the blank control group (Group I), there were extremely significant differences (P<0.01, P<0.01), the positive group (Ⅲ group) and the ethyl acetate extraction part group (group Ⅳ) can significantly inhibit the atrophy of the thymus and increase the thymus index of rats, compared with the model group (group Ⅱ) there is a significant Significant difference (P<0.01, P<0.01). Compared with the alcohol-extracted parts of AA rats with Alcohol-extracted extract parts of F. syphilis, the thymus atrophy of AA rats was more severe, the spleen index was higher, and the curative effect was mild.

Table 46 The effect of each extraction part on the viscera index of AA rats by alcohol extraction n=10)

group Thymus index/g kg^-1 Spleen index/g·kg^-1 Group I 1.73±0.16^▲▲ 2.07±0.18 Group II 1.40±0.12^** 2.88±0.38^** Group III 1.59±0.15^*▲▲ 2.60±0.43^** Group IV 1.62±0.22^▲▲ 2.73±0.45^** Group V 1.60±0.34 2.81±0.33^** Group VI 1.47±0.17^** 3.53±0.61^**▲▲

Note: Compared with the blank group ^* P<0.05, ^** P<0.01, compared with the model group ^▲ P<0.05, ^▲▲ P<0.01.

Embodiment 8:

experimental method:

To compare the effects of water extract, alcohol extract and ethyl acetate extraction part of the composition of Filiaria fragrans Fructus chrysanthemum on the swelling degree of right and left toes of AA rats. △Toe swelling degree (ml) = toe swelling degree on the 27th day - toe swelling degree on the 1st day.

The experimental results are shown in Table 47. The results show that the swelling degree of the right and left paws of the Ethyl Acetate Extraction Group with the alcohol extraction group is smaller than that of the Water Extraction Group and the Alcohol Extraction Group, and there is a significant difference. Difference (P<0.01, P<0.05). It shows that the alcohol-extracted ethyl acetate extract part of the composition of filial scalloped chrysanthemum has the best curative effect.

Table 47 The effect of different extracts of the filial piety scalloped chrysanthemum composition on the swelling degree of the right toe of AA rats ( n=10)

Note: ^** P<0.01 compared with the water extract group, ^▲ P<0.05 compared with the alcohol extract group.

Table 48 The effect of different extracts of the filial piety scalloped chrysanthemum composition on the swelling degree of the left foot of the AA rat ( n=10)

Note: ^* P<0.05 compared with water extract group, ^▲ P<0.05 compared with alcohol extract group.

Claims (5)

1. A filial piebal-inula flower composition for treating rheumatoid arthritis comprises filial piebal and inula flower in a mass ratio of 1 ~ 5: 1.

2. The composition of claim 1, wherein the ratio of the masses of the filial piebal and the inula cappa is 2: 1.

3. Application of the filial fantasy inula herb composition in preparing a medicament for treating rheumatoid arthritis.

4. The use as claimed in claim 3, wherein the application of the filial fantasy inula extract in preparing the medicament for treating rheumatoid arthritis is the water extract, the alcohol extract or the ethyl acetate extract.

5. The application of claim 4, wherein the extract is ethyl acetate extract of filial fanwort herb and inula cappa, and is prepared by the following steps of taking dry coarse powder of filial fanwort herb and inula cappa, adding 80 ~ 95% ethanol in an amount which is 10 ~ 12 times of that of the dry coarse powder, soaking for 1 ~ 2h, heating, refluxing and extracting for 2 times 2 ~ 3 times, 1 ~ 2h each time, combining two filtrates, recovering ethanol, concentrating until the ethanol smell is absent, adding a proper amount of water for dissolving, extracting with petroleum ether until the filtrate is colorless, extracting the residual solution with ethyl acetate until the filtrate is colorless, taking an ethyl acetate extraction part, and volatilizing the solvent to obtain the ethyl acetate extract of filial fanwort herb and inula cappa.