CN109846949B - A Chinese medicinal composition with anti-inflammatory and choleretic effects for treating cholecystitis and cholestasis - Google Patents
A Chinese medicinal composition with anti-inflammatory and choleretic effects for treating cholecystitis and cholestasis Download PDFInfo
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Abstract
The invention provides a traditional Chinese medicine composition with anti-inflammatory and cholagogic effects, which is mainly used for cholecystitis and cholestasis, and is prepared from the following raw material medicines in parts by weight: 3-300 parts of radix bupleuri, 3-300 parts of elecampane, 3-300 parts of fried immature bitter orange, 3-300 parts of green tangerine peel, 3-300 parts of raw hawthorn, 3-300 parts of raw rhubarb, 3-300 parts of mirabilite and 3-300 parts of radish seed. The invention clarifies and verifies the efficacy of the traditional Chinese medicine composition in the aspects of diminishing inflammation, benefiting gallbladder and treating intrahepatic cholestasis, and provides experimental data support for further clinical use and popularization.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicine preparations, and particularly relates to a traditional Chinese medicine composition for treating cholecystitis and cholestasis.
Background
Cholecystitis and cholestasis are common diseases of liver and gallbladder in clinic, and the incidence rate is high. Cholestasis (cholestasis) is a pathophysiological process caused by the secretion and excretion disorder of bile juice, which is manifested by excessive accumulation of bile components such as cholic acid, cholesterol and bilirubin in the liver and body circulation, causing damage to liver cells and body, and long-lasting cholestasis which will progress into hepatic fibrosis and even cirrhosis.
At present, chemical drugs for effectively treating cholecystitis and cholestasis are lacking clinically, the cholecystitis is mainly treated by surgical operation after the disease progresses to a certain stage, and for the treatment of the cholestasis, only one of relevant chemical treatment drugs approved by FDA is ursodeoxycholic acid, but the curative effect is unsatisfactory. The traditional Chinese medicine has obvious advantages in the aspect of treating cholecystitis and cholestasis, but the prescription composition of the related traditional Chinese medicine prescription preparation sold on the market at present is obviously different from the prescription composition, and the prescription has unique advantages in the aspect of treating cholecystitis and cholestasis.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a traditional Chinese medicine composition for treating cholestasis.
The invention provides a traditional Chinese medicine composition for treating cholestasis, which is prepared from the following raw material medicines in parts by weight: 3-300 parts of radix bupleuri, 3-300 parts of elecampane, 3-300 parts of fried immature bitter orange, 3-300 parts of green tangerine peel, 3-300 parts of raw hawthorn, 3-300 parts of raw rhubarb, 3-300 parts of mirabilite and 3-300 parts of radish seed.
Preferably, the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 100-200 parts of radix bupleuri, 100-200 parts of elecampane, 100-200 parts of fried immature bitter orange, 100-200 parts of green tangerine peel, 100-200 parts of raw hawthorn, 100-200 parts of raw rhubarb, 100-200 parts of mirabilite and 100-200 parts of radish seed.
Preferably, the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 150 parts of bupleurum, 100 parts of costustoot, 100 parts of fried immature bitter orange, 50 parts of green tangerine peel, 150 parts of raw hawthorn, 200 parts of raw rhubarb, 100 parts of mirabilite and 100 parts of radish seed.
The invention also provides a preparation method of the traditional Chinese medicine composition, which comprises the steps of mixing the traditional Chinese medicines, crushing, adding medically acceptable auxiliary materials, and preparing pills.
Preferably, the auxiliary material is refined honey.
The invention also provides application of the traditional Chinese medicine composition in preparing a medicine for treating cholestasis.
The prescription is as follows:
(1) radix bupleuri
[ PROPERTIES AND WEIGHT GUIDING CHANNEL ] is pungent, bitter and slightly cold. It enters liver, gallbladder and lung meridians.
[ FUNCTIONS AND INDICATIONS ] can dispel heat, sooth the liver, relieve depression, and elevate yang qi. Can be used for treating common cold, fever, abdominal distention, menoxenia, uterine prolapse, and proctoptosis.
(2) Radix aucklandiae
[ PROPERTIES AND WEIGHT GUIDING GEN ] is pungent, bitter and warm. It enters spleen, stomach, large intestine, triple energizer and gallbladder meridians.
[ FUNCTIONS AND INDICATIONS ] can promote the circulation of qi, alleviate pain, invigorate the spleen and promote digestion. Can be used for treating distending pain in chest, hypochondrium and abdomen, diarrhea, dyspepsia, and anorexia.
(3) Parched fructus Aurantii Immaturus
[ PROPERTIES AND WEIGHT GUIJING ] is bitter, pungent, sour and slightly cold. It enters spleen and stomach meridians.
[ FUNCTIONS AND INDICATIONS ] can break qi, resolve food stagnation, resolve phlegm and disperse mass. Can be used for treating stagnation, abdominal fullness, hypochondriac pain, dysentery with tenesmus, constipation, phlegm stagnation, qi stagnation, thoracic obstruction, and viscera prolapse.
(4) Green tangerine peel
[ PROPERTIES AND WEIGHT GUIDING GEN ] is bitter, pungent and warm. It enters liver, gallbladder and stomach meridians.
[ FUNCTIONS AND INDICATIONS ] can soothe the liver, break qi, remove food stagnation and resolve stagnation. Can be used for treating distending pain in chest and hypochondrium, hernia pain, nodules of breast, mammary abscess, food stagnation, qi stagnation, and abdominal pain.
(5) Raw hawthorn
[ PROPERTIES AND WEIGHT GUIDING GEN ] is sour, sweet and slightly warm. It enters spleen, stomach and liver meridians.
[ FUNCTIONS AND INDICATIONS ] can promote digestion, invigorate stomach, promote the circulation of qi, remove blood stasis, reduce blood fat. Can be used for treating meat food stagnation, gastric cavity distention, dysentery, abdominal pain, blood stasis, amenorrhea, puerperal blood stasis, heart and abdomen stabbing pain, thoracic obstruction, cardialgia, hernia pain, and hyperlipidemia.
(6) Radix Et rhizoma Rhei
[ PROPERTIES, WESTERN AND GUIJING ] bitter and cold. It enters spleen, stomach, large intestine, liver and pericardium meridians.
[ FUNCTIONS AND INDICATIONS ] can purge pathogenic accumulation, clear heat and purge fire, remove heat from the blood and toxic material, remove blood stasis and dredge channels, remove dampness and alleviate jaundice. Can be used for treating constipation due to excessive heat accumulation, hematemesis, epistaxis, conjunctival congestion, pharyngeal swelling, carbuncle, furuncle, acute appendicitis, abdominal pain, amenorrhea due to blood stasis, puerperal blood stasis, traumatic injury, damp-heat dysentery, jaundice, dark urine, stranguria, and edema.
(7) Natrii sulfas
[ PROPERTIES, WESTERN AND GUIJING ] is salty, bitter and cold. It enters stomach and large intestine meridians.
[ FUNCTIONS AND INDICATIONS ] can relieve constipation by purgation, moisten dryness and soften hard mass, clear fire and relieve swelling. Can be used for treating retention of excess heat, abdominal distention, pain, constipation, and intestinal abscess with swelling and pain.
(8) Radish seed
[ PROPERTIES AND WEIGHT GUIDING CHANNEL ] is pungent, sweet and mild. It enters lung, spleen and stomach meridians.
[ FUNCTIONS AND INDICATIONS ] can promote digestion, relieve distention, direct qi downward and resolve phlegm. Can be used for treating food stagnation, abdominal distention and pain, constipation, stagnation, dysentery, phlegm stagnation, asthma, and cough.
In the prescription, the functions of each raw material medicine are respectively as follows: bupleuri radix, pericarpium Citri Reticulatae viride and parched fructus Aurantii Immaturus for soothing liver, relieving stagnant qi and removing food stagnation; the wood fragrance promotes qi stagnation of spleen and stomach, regulates middle energizer, relieves pain and promotes gallbladder; the raw hawthorn and the radish seed have the functions of promoting digestion and relieving distension; rhubarb, radix et rhizoma Rhei is bitter and cold in property and subsides, clears heat and facilitates gallbladder; mirabilite can moisten dryness, soften hardness and purge. The medicines are combined and mutually assisted, so that liver qi can reach, spleen and stomach qi can be regulated, and stagnated heat can be cleared, thereby being beneficial to smoothing bile flow.
According to the invention, 17 alpha-ethinylestradiol is given to a rat through neck subcutaneous injection to induce an intrahepatic cholestasis model, and after high, medium and low doses of the traditional Chinese medicine composition suspension are given through intragastric perfusion during the period, the returning capacity of the traditional Chinese medicine composition on serum transaminase, bilirubin and total bile salt level of the model rat and pathological changes of the liver are examined, so that the pharmacodynamic effect of the anti-inflammatory choleretic pill on the aspect of treating cholestasis is clarified. The invention clarifies and verifies the efficacy of the traditional Chinese medicine composition in treating intrahepatic cholestasis, and provides experimental data support for further clinical use and popularization.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention and not to limit the invention. In the drawings:
FIG. 1 is the level of TNF α in rat serum.
Figure 2 shows rat serum transaminase, total bilirubin, and total bile acid salt levels (x ± s, n ═ 6).
Fig. 3 is liver histopathology.
Fig. 4 is a liver histopathological score compared to normal group.
Detailed Description
The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples are commercially available unless otherwise specified.
The traditional Chinese medicine composition for treating cholestasis is prepared from the following raw material medicines in parts by weight:
3-300 parts of radix bupleuri, 3-300 parts of elecampane, 3-300 parts of fried immature bitter orange, 3-300 parts of green tangerine peel, 3-300 parts of raw hawthorn, 3-300 parts of raw rhubarb, 3-300 parts of mirabilite and 3-300 parts of radish seed.
The preparation method of the traditional Chinese medicine composition for treating cholestasis comprises the following steps:
mixing the above traditional Chinese medicines, pulverizing, sieving with 100 mesh sieve, and mixing fine powder according to a ratio of 1: 1.2-1: 2, adding refined honey according to the weight ratio, mixing and stirring uniformly until a moist mass block is obtained, and preparing the Chinese medicinal big honeyed pill by related equipment.
Example 1
The traditional Chinese medicine composition for treating cholestasis is prepared from the following raw material medicines in parts by weight:
120g of radix bupleuri, 100g of costustoot, 100g of fried immature bitter orange, 100g of green tangerine peel, 120g of raw hawthorn, 100g of raw rhubarb, 100g of mirabilite and 100g of radish seed.
Mixing the above traditional Chinese medicines, pulverizing, sieving with 100 mesh sieve, and mixing fine powder according to a ratio of 1: adding refined honey at a ratio of 1.6, mixing and stirring uniformly to obtain moist mass, and preparing into Chinese medicinal big honeyed pill with related equipment.
Example 2
The traditional Chinese medicine composition for treating cholestasis is prepared from the following raw material medicines in parts by weight:
250g of bupleurum, 150g of costustoot, 150g of fried immature bitter orange, 120g of green tangerine orange peel, 120g of raw hawthorn, 120g of raw rhubarb, 100g of mirabilite and 100g of radish seed.
Mixing the above traditional Chinese medicines, pulverizing, sieving with 100 mesh sieve, and mixing fine powder according to a ratio of 1: 1.2, adding refined honey, mixing and stirring uniformly until a moist block mass is obtained, and preparing the Chinese medicinal big honeyed pill by related equipment.
Example 3
The traditional Chinese medicine composition for treating cholestasis is prepared from the following raw material medicines in parts by weight:
300g of radix bupleuri, 200g of costustoot, 120g of fried immature bitter orange, 120g of green tangerine peel, 100g of raw hawthorn, 100g of raw rhubarb, 50g of mirabilite and 50g of radish seed.
Mixing the above traditional Chinese medicines, pulverizing, sieving with 100 mesh sieve, and mixing fine powder according to a ratio of 1: adding refined honey at a ratio of 1.5, mixing and stirring uniformly to obtain moist mass, and preparing into Chinese medicinal big honeyed pill with related equipment.
Example 4
The traditional Chinese medicine composition for treating cholestasis is prepared from the following raw material medicines in parts by weight:
50g of radix bupleuri, 150g of costustoot, 300g of fried immature bitter orange, 200g of green tangerine peel, 200g of raw hawthorn, 50g of raw rhubarb, 200g of mirabilite and 200g of radish seed.
Mixing the above traditional Chinese medicines, pulverizing, sieving with 100 mesh sieve, and mixing fine powder according to a ratio of 1: adding refined honey at a ratio of 1.6, mixing and stirring uniformly to obtain moist mass, and preparing into Chinese medicinal big honeyed pill with related equipment.
Example 5
The traditional Chinese medicine composition for treating cholestasis is prepared from the following raw material medicines in parts by weight:
80g of radix bupleuri, 300g of costustoot, 200g of fried immature bitter orange, 300g of green tangerine peel, 300g of raw hawthorn, 50g of raw rhubarb, 150g of mirabilite and 100g of radish seed.
Mixing the above traditional Chinese medicines, pulverizing, sieving with 100 mesh sieve, and mixing fine powder according to a ratio of 1: 2, adding refined honey according to the proportion, mixing and stirring uniformly until a moist mass block is obtained, and preparing the Chinese medicinal big honeyed pill by related equipment.
Example 6
The traditional Chinese medicine composition for treating cholestasis is prepared from the following raw material medicines in parts by weight:
250g of radix bupleuri, 30g of costustoot, 30g of fried immature bitter orange, 30g of green tangerine peel, 50g of raw hawthorn, 300g of raw rhubarb, 300g of mirabilite and 30g of radish seed.
Mixing the above traditional Chinese medicines, pulverizing, sieving with 100 mesh sieve, and mixing fine powder according to a ratio of 1: 1.3, adding refined honey, mixing and stirring uniformly until a moist block mass is obtained, and preparing the Chinese medicinal big honeyed pill by related equipment.
Example 7
The traditional Chinese medicine composition for treating cholestasis is prepared from the following raw material medicines in parts by weight:
160g of bupleurum, 80g of costustoot, 30g of fried immature bitter orange, 30g of green tangerine peel, 250g of raw hawthorn, 250g of raw rhubarb, 240g of mirabilite and 300g of radish seed.
Mixing the above traditional Chinese medicines, pulverizing, sieving with 100 mesh sieve, and mixing fine powder according to a ratio of 1: adding refined honey at a ratio of 1.7, mixing and stirring uniformly to obtain moist mass, and preparing into Chinese medicinal big honeyed pill with related equipment.
Example 8 Effect of the Chinese medicinal composition of the present invention on intrahepatic cholestasis rat serum inflammatory factor TNF α 1.1 pharmaceutical Agents
The traditional Chinese medicine composition is prepared according to the method of example 1; 17 α -ethinylestradiol was purchased from Sigma, usa; the rat tumor necrosis factor alpha (TNF alpha) assay kit was purchased from Shanghai enzyme-linked Biotech, Inc. (Cat. No.: ml 002859). XIAOYANLIDAN tablet is obtained from Guangzhou Baiyunshan and Megaoku Chinese medicine GmbH (batch number: L17A 010); propylene glycol was purchased from Jiangxi Yi general crude drug industry Co., Ltd (Jiangxi food Standard 2009006).
1.2 Experimental animals
Healthy male Wistar rats, SPF grade, weight 280-300 g.
1.3 instrumentation
The Varioskan Flash multifunctional enzyme-labeling instrument is manufactured by Thermo company of America; the 3K15 high speed centrifuge was purchased from Sgima centrifuge, germany; the LT-224S analytical balance is manufactured by Sidolis, Beijing.
1.4 Experimental methods
1.4.1 establishment of intrahepatic cholestasis rat model
A rat intrahepatic cholestasis model was induced by subcutaneous injection of a solution of 17 α -ethinylestradiol in propylene glycol at a dose of 5mg/kg/d (q.d.) into the neck of the rat for 6 consecutive days.
1.4.2 grouping and administration
36 healthy male Wistar rats (280-300 g) are randomly divided into six groups of a normal group, a model group, a low-dosage group, a medium-dosage group and a high-dosage group of the traditional Chinese medicine composition pill and a negative control group (a positive control group) (the negative control group is a commercially available medicine and is used as a positive control) according to the weight, and each group comprises 6 rats, and the specific intervention scheme is as follows:
preparation of a traditional Chinese medicine composition pill suspension: weighing Chinese medicinal composition, namely honey pills, adding a certain amount of purified water, and shaking to obtain a pill suspension water solution; preparing an anti-inflammatory and cholagogic tablet suspension: the inflammation diminishing and gallbladder benefiting tablet sold in the market is ground into fine powder, weighed, added with a certain amount of purified water and shaken to obtain the suspension water solution of the inflammation diminishing and gallbladder benefiting tablet.
(1) Normal group: injecting propylene glycol solution subcutaneously at neck of rat for 6 days, and feeding purified water twice per day for 7 days;
(2) model group: purified water was administered twice daily by gavage for 7 days during induction model under item "1.4.1";
(3) pill low dose group: performing intragastric administration on the Chinese medicinal composition pill suspension twice a day for 7 days at a dose of 0.67g/kg/d during induction model period under item 1.4.1;
(4) pill medium dose groups: during the induction model period under the item of '1.4.1', the traditional Chinese medicine composition pill suspension is administrated twice a day by intragastric administration for 7 days according to the dosage of 1 g/kg/d;
(5) bolus high dose group: performing intragastric administration on the Chinese medicinal composition pill suspension twice a day for 7 days at a dose of 1.5g/kg/d during induction model period under the item of 1.4.1;
(6) inflammation diminishing and cholagogic tablet group (positive traditional Chinese medicine control group): the suspension of the anti-inflammatory and cholagogic tablet is administrated twice a day by gavage for 7 days at a dose of 0.7g/kg/d during an induction model period under the item of '1.4.1'.
1.4.3 serum sample Collection and index determination
After the intervention of each group of rats according to the scheme under the item '1.4.2', after fasting for 12h before the experiment, ether anesthesia is carried out, 5mL of blood sample is collected in an EDTA anticoagulation tube through an abdominal aorta, 12000r/min and is centrifuged at a high speed for 5min, and after the serum sample is separated, the kit determines the level of inflammatory factor TNF alpha in the serum.
1.4.4 statistical analysis
The data of the measured data are expressed by mean +/-standard deviation, statistical analysis and comparison between two groups are carried out by student t test, and P is less than 0.05 and P is less than 0.01, which is statistically significant.
1.5 results of the experiment
The experimental results are shown in fig. 1: the level of the proinflammatory factor TNF alpha in rat serum is obviously increased compared with the normal group in the model group (P < 0.01); compared with the model group, the level model of TNF alpha in the serum of rats is reduced (P is less than 0.05) after the treatment of low, medium and high doses of pills and positive drug anti-inflammatory cholagogic tablets.
FIG. 1 is the level of TNF α in rat serum.
1.6 conclusion
TNF α is a pro-inflammatory cytokine produced primarily by macrophages and monocytes and is involved in inflammation and immune responses in the body. Normally, TNF α activity in vivo is extremely low, and in some pathological conditions, TNF α is released continuously or in large amounts into local tissue and body fluids and mediates the damage of pathogenic factors to the body. Kupffer cells in the liver are both the primary site for TNF α synthesis and the primary site for TNF α clearance. Factors such as bacteria, virus infection, immune complex, chemical drugs and the like can cause Kupffer cells to release a large amount of TNF alpha, and participate in the process of liver injury. The research shows that the content of serum TNF alpha is obviously increased during the cholestasis induced by ethinylestradiol, and the experimental result also finds that the level of TNF alpha in the serum of rats with intrahepatic cholestasis induced by estrogen is obviously increased. After the traditional Chinese medicine composition and the positive medicine anti-inflammatory and cholagogic tablet with different doses are applied for intervening for 7 days, the content of serum TNF alpha can be obviously reduced, which indicates that the traditional Chinese medicine composition has obvious anti-inflammatory effect, and the anti-inflammatory effect is equivalent to that of the anti-inflammatory and cholagogic tablet.
Example 9 Effect of the Chinese medicinal composition of the present invention on Biochemical indicators of serum associated with liver of intrahepatic cholestatic rat
1.1 pharmaceutical Agents
The traditional Chinese medicine composition is prepared according to the method of example 1; 17 α -ethinylestradiol was purchased from Sigma, usa; glutamic-oxaloacetic transaminase (AST), glutamic-pyruvic transaminase (ALT), alkaline phosphatase (ALP), glutamyl transpeptidase (GGT), Total Bilirubin (TBIL) and total cholate (TBAs) determination kits were purchased from Nanjing institute of bioengineering. Propylene glycol was purchased from Jiangxi Yi crude drug industry Co., Ltd (Jiangxi food Standard 2009006).
1.2 Experimental animals
Healthy male Wistar rats, SPF grade, weight 280-300 g.
1.3 instrumentation
An Olympus full-automatic biochemical analyzer of AU400 type is manufactured by Olympus of Japan; the 3K15 high speed centrifuge was purchased from Sgima centrifuge, germany; the LT-224S analytical balance is manufactured by Sidolis, Beijing.
1.4 Experimental methods
The establishment of intrahepatic cholestasis rat models and the grouping and administration of the same are described in example 8 under "1.4".
1.4.3 serum sample Collection and index determination
After the intervention of each group of rats according to the experimental scheme, after fasting for 12h before the experiment, ether anesthesia is carried out, 5mL of blood sample is collected in an EDTA anticoagulation tube through abdominal aorta, 12000r/min and is centrifuged at high speed for 5min, after the serum sample is separated, the levels of AST, ALT, ALP, GGT, TBIL and TBAs in the serum are analyzed and determined by an Olympus full-automatic biochemical analyzer.
1.4.4 statistical analysis
The data of the measured data are expressed by mean +/-standard deviation, statistical analysis and comparison between two groups are carried out by student t test, and P is less than 0.05 and P is less than 0.01, which is statistically significant.
1.5 results of the experiment
1.5.117 alpha-ethinylestradiol induced intrahepatic cholestasis rat model
Compared with the normal group, the water levels of AST, ALT, ALP, GGT, TBIL and TBAs in the serum of the model group rat are obviously increased (P is less than 0.05/0.01, see table 1), which indicates that the intrahepatic cholestasis model of the rat can be successfully induced by adopting a mode of injecting 17 alpha-ethinylestradiol with 5mg/kg/d subcutaneously into the neck of the rat for 6 days.
1.5.2 pharmacodynamic evaluation of the Chinese medicinal composition of the present invention
The low-dose group of the traditional Chinese medicine composition pills is compared with the model group (see table 1-1, table 1-2 and figure 2): the levels of AST, TBIL and TBAs in rat serum are respectively reduced by about 37%, 16% and 28% (P <0.05), which indicates that the Chinese medicinal composition pill with low dose can relieve the symptoms of intrahepatic cholestasis induced by 17 alpha-ethinylestradiol.
The traditional Chinese medicine composition pill medium dosage group is compared with the model group (see table 1-1, table 1-2 and figure 2): except that AST, TBIL and TBAs in the serum of rats are respectively reduced by 32 percent, 15 percent and 38 percent (P <0.05), the level of ALP is also reduced by 17 percent (P <0.05), which indicates that the traditional Chinese medicine composition pill with the medium dosage can also effectively relieve the symptom of intrahepatic cholestasis induced by EE, and the curative effect of the traditional Chinese medicine composition pill is better than that of the low-dosage group.
The traditional Chinese medicine composition pill high dose group is compared with the model group (see table 1-1, table 1-2 and figure 2): ALP and TBIL in rat serum are respectively reduced by 18 percent and 28 percent (P <0.05), TBAs have no obvious change (P >0.05), and GGT is conversely increased by 40 percent (P <0.05), which shows that the high-dose traditional Chinese medicine composition pill has certain curative effect on EE-induced cholestasis, but has no obvious reduction effect on cholate in vivo, and the increase of the medicine dose generates certain damage to liver.
Compared with the model group, the traditional Chinese medicine composition tablet group (shown in the table 1-1, the table 1-2 and the figure 2): the AST, TBIL and TBAs in the rat serum are respectively reduced by 32 percent, 27 percent and 46 percent (P is less than 0.05), which shows that the traditional Chinese medicine composition tablet has obvious curative effect on 17 alpha-ethinylestradiol-induced liver stasis of rats as a positive traditional Chinese medicine.
Figure 2 shows rat serum transaminase, total bilirubin, and total bile acid salt levels (x ± s, n ═ 6). P <0.05, P <0.01 indicates statistical differences compared to the model group.
TABLE 1-1 Biochemical index of Chinese medicinal composition pill for treating intrahepatic cholestasis in rat serum
Influence (mean ± SD, n ═ 6)
Table 1-2 effect of the Chinese medicinal composition pill on serum biochemical index of intrahepatic cholestasis rat (mean ± SD, n ═ 6)
*P<0.05,**P<0.01 indicates a statistical difference compared to the model group.
1.6 conclusion
The research result shows that the traditional Chinese medicine composition pill has an obvious treatment effect on 17 alpha-ethinylestradiol-induced intrahepatic cholestasis of rats, wherein the traditional Chinese medicine composition pill has the most obvious curative effect and no obvious toxicity when the traditional Chinese medicine composition pill is used in a medium dose (1 g/kg/d).
Example 10 Effect of the Chinese medicinal composition of the present invention on liver pathology in intrahepatic cholestatic rats
1.1 Experimental reagents and apparatus
Hydroextractor (model TP1020, Lycra, Germany) Paraffin embedding machine (model HBM-200 LF, Shanghai atlantoaxi medical instruments Co., Ltd.), Formaldehyde, Anhydrous ethanol and Paraffin were purchased from Bailingwei science and technology, China.
1.2 Experimental methods
1.2.1 section preparation
Taking out liver tissue from 10% formaldehyde solution, cutting into small pieces of 0.5cm, soaking in water for a period of time, removing part of formaldehyde, sequentially passing through 70%, 80%, 90%, 95% and 100% alcohol, and gradient dehydrating for 2.5 hr; after dehydration, the tissue was immersed in xylene and subjected to a clearing treatment for about 50 min. And (3) putting the treated liver tissues into dissolved paraffin wax No. 1, 2 and 3, keeping the temperature constant at 60-65 ℃, and soaking for about 2 hours. After paraffin immersion, paraffin embedding was carried out, and the embedded wax block was sliced with a thickness of 5 μm using a microtome.
1.2.2 HE staining
Firstly, placing the tissue slices into a thermostat (the temperature is 90 ℃) for 25min, then sequentially soaking the slices into dimethylbenzene I, II and III for 10min respectively and absolute ethyl alcohol I and absolute ethyl alcohol II for 15min respectively, and carrying out dewaxing treatment. Then the slices are washed with distilled water for three times, then hematoxylin staining is carried out, the soaking time is about 5min, then differentiation and bluing are carried out, namely, the slices are firstly put into hydrochloric acid alcohol differentiation liquid for 30s and then into tap water at 40 ℃ for 5min, and then are put into an acid red pulp-coating dye eosin staining liquid tank for 1min, and then the slices are washed with distilled water for three times. Finally, after dehydration by gradient alcohol and xylene and transparency, the piece is sealed by neutral gum. Pathological observations were then made under an optical microscope and pictures were taken.
1.2.3 pathological Scoring method
Reference is made to the Knodell System (Table 2)
The "hepatic Histology Activity Index (HAI)" was scored and the degree of bile duct proliferation was evaluated, and the score results of each group were expressed as mean ± standard error (x ± s, n ═ 6), and t-test was performed between groups.
TABLE 2 liver Histology Activity Index (HAI) scoring System
1.3 results of the experiment
1.3.1 Effect of the Chinese medicinal composition of the invention on histopathology of model rat liver
As shown in FIG. 3, the normal group had normal liver tissue structure and cell morphology, clear hepatic lobule structure, and well-arranged liver cell cords. The model group showed more severe liver cell eosinophilia, inflammatory infiltration, biliary sinus dilation and bile duct hyperplasia. Compared with a model group, the Chinese medicinal composition pill obviously reduces the inflammatory infiltration, the biliary sinus dilation and the bile duct hyperplasia degree in a low-dose group, a medium-dose group and a high-dose group, wherein the effect is most obvious in the medium-dose group; the positive group also reduces the degree of inflammatory infiltration of liver tissues, expansion of biliary sinuses and proliferation of bile ducts; compared with the positive group, the low-dose group, the medium-dose group and the high-dose group of the traditional Chinese medicine composition pill have no obvious difference in liver tissue pathological changes.
Fig. 3 is liver histopathology. Wherein, A: blank group; b: a model group; c: a low dose group of pills of a traditional Chinese medicine composition; d: traditional Chinese medicine composition pill medium dosage group; e: a high-dose group of pills of a traditional Chinese medicine composition; f: and (4) a positive group. In the figure, the liver cell is changed into acidophilic liver cell and condensed nucleus; ② liver sinus dilation. The microscope magnification was 200 times.
1.3.2 Effect of the pill of Chinese medicinal composition of the present invention on the evaluation of pathological changes of liver tissue of model rat
As shown in fig. 4, the histopathological score of liver and the number of hyperplasia of bile duct were significantly increased in the model group compared to the normal group (P < 0.01); compared with the model group, the liver histopathology score and the bile duct hyperplasia number of the traditional Chinese medicine composition pill low-dose group, the medium-dose group, the high-dose group and the traditional Chinese medicine composition tablet are significantly reduced (P < 0.05); compared with the positive group, the traditional Chinese medicine composition pill has no significant difference in liver histopathology scores and bile duct hyperplasia numbers of the low-dose group, the medium-dose group and the high-dose group.
Fig. 4 is a liver histopathological score compared to normal group. Wherein, P<0.05；**P<0.01; in comparison to the set of models,#P<0.05；##P<0.01。
1.4 conclusion
Experimental results show that the low-dose group, the medium-dose group and the high-dose group of the traditional Chinese medicine composition pill can obviously reduce the pathological change degree of liver histopathology of rat liver injury induced by estrogen, and is beneficial to the regression of liver function.
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments described in the foregoing embodiments, or that equivalents may be substituted for elements thereof. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (4)
1. The application of a traditional Chinese medicine composition in preparing a medicine for treating intrahepatic cholestasis induced by estrogen is disclosed, wherein the traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 100-200 parts of radix bupleuri, 100-200 parts of elecampane, 100-200 parts of fried immature bitter orange, 100-200 parts of green tangerine peel, 100-200 parts of raw hawthorn, 100-200 parts of raw rhubarb, 100-200 parts of mirabilite and 100-200 parts of radish seed.
2. The application of a traditional Chinese medicine composition in preparing a medicine for treating intrahepatic cholestasis induced by estrogen is disclosed, wherein the traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 150 parts of bupleurum, 100 parts of costustoot, 100 parts of fried immature bitter orange, 50 parts of green tangerine peel, 150 parts of raw hawthorn, 200 parts of raw rhubarb, 100 parts of mirabilite and 100 parts of radish seed.
3. Use according to claim 1 or 2, characterized in that: the preparation method of the traditional Chinese medicine composition comprises the following steps: mixing the above Chinese medicinal materials, pulverizing, adding pharmaceutically acceptable adjuvants, and making into pill.
4. Use according to claim 3, characterized in that: the auxiliary material is refined honey.
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| 大黄木香泡服治疗胆绞痛45例;石坚;《中国中西医结合杂志》;19911231(第3期);第183页 * |
| 消炎利胆丸质量标准的初步研究;王文权;《兰州医学院学报》;20010930;第27卷(第3期);第26-27页，尤其是第26页左栏第1段 * |
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