CN106038788B

CN106038788B - Compatible composition for reducing toxicity and preserving effects of gelsemium elegans and compatibility method thereof

Info

Publication number: CN106038788B
Application number: CN201610495381.4A
Authority: CN
Inventors: 王英豪; 吴水生
Original assignee: Fujian University of Traditional Chinese Medicine
Current assignee: Fujian University of Traditional Chinese Medicine
Priority date: 2016-06-29
Filing date: 2016-06-29
Publication date: 2019-12-13
Anticipated expiration: 2036-06-29
Also published as: CN106038788A

Abstract

The invention discloses a compatible composition for reducing toxicity and preserving effects of gelsemium elegans and a compatibility method thereof, wherein the composition is prepared by matching gelsemium elegans and components for reducing toxicity and preserving effects according to the weight part ratio of 1: 5-50, and the components for reducing toxicity and preserving effects are one of the following components: flos Daturae Metelis; sheep blood; or a mixture of houttuynia and peppermint. In the invention, the gelsemium elegans is combined with the datura flower, the herba houttuyniae and the mint to achieve the effect of attenuating the toxin of the gelsemium elegans to a certain extent, and particularly, the effect of the gelsemium elegans is combined with the datura flower to be better. Meanwhile, the gelsemium elegans compatible attenuation component can also play a role in easing pain for mice. Therefore, the compatible composition of gelsemium elegans of the invention has obvious effects of reducing toxicity and preserving effects.

Description

Compatible composition for reducing toxicity and preserving effects of gelsemium elegans and compatibility method thereof

Technical Field

The invention relates to compatibility research of gelsemium elegans attenuation and effect preservation, in particular to a compatible composition of gelsemium elegans attenuation and effect preservation and a compatibility method thereof.

Background

Gelsemium elegans benth, also known as Gelsemium elegans, gekko gecko and the like, is a whole plant of Gelsemium elegans benth, a plant of the genus calamus of the family marchancaceae, is bitter and pungent in taste, warm in nature, and toxic in nature, has the effects of dispelling wind and eliminating toxin, eliminating stagnation and reducing swelling and relieving pain, and is mainly used for treating eczema, carbuncle swelling, tinea corporis, traumatic injury, rheumatalgia, neuralgia and the like. Modern pharmacological experiments also show that the traditional Chinese medicine composition plays an important role in treating tumors, pains, inflammations, immunity, skin diseases and the like.

because the gelsemium elegans is extremely toxic, toxic components are mainly indole alkaloids, the treatment amount and the toxic amount are very close, the safety index is small, and the clinical report of gelsemium elegans poisoning is not lacked. However, indole alkaloids also have unique biological activity-anti-tumor, analgesic, and immune function influencing effects. The pharmaceutical effect component is also a toxic component, so how to reduce the toxicity of the gelsemium elegans and simultaneously keep the pharmacological activity of the gelsemium elegans is of great significance for improving the safety of gelsemium elegans in clinical application.

at present, the research on gelsemium elegans is mainly focused on the aspects of biology, chemistry, pharmacology, toxicology and the like, while the related research on the toxicity reducing and efficacy maintaining effects of gelsemium elegans is rarely reported, and the research on toxic components and toxic mechanisms of gelsemium elegans by compatibility is not seen. In view of the strong toxicity of gelsemium elegans, the development of compatible compositions with the effects of reducing and preserving the toxicity of gelsemium elegans is needed in the field, and the toxicity of gelsemium elegans is greatly reduced on the basis of ensuring the analgesic effect of gelsemium elegans.

Disclosure of Invention

In order to solve the problems of the traditional lubricating oil, a compatible composition with the effect of reducing toxicity and the existence of gelsemium elegans which can ensure the analgesic effect of gelsemium elegans and simultaneously reduce the toxicity of gelsemium elegans and a compatibility method thereof are needed.

Therefore, the inventor provides the following technical scheme:

The compatible composition for reducing and preserving effects of gelsemium elegans is prepared from gelsemium elegans and a component with the effect of reducing and preserving effects in a weight ratio of 1: 5-50, wherein the component with the effect of reducing and preserving effects is one of the following components: flos Daturae Metelis, sanguis Caprae Seu Ovis, or mixture of herba Houttuyniae and herba Menthae.

The invention combines the gelsemium elegans and the toxicity-reducing and efficacy-maintaining component, and can ensure the analgesic function of the gelsemium elegans under the condition of obviously reducing the toxicity of the gelsemium elegans.

Preferably, the toxicity-reducing and efficacy-maintaining component is Mussaenda pubescens. The golden camellia has the best toxicity reducing and effect storing effects when being matched with the gelsmium elegans.

Further, the kiss is pretreated before compatibility, and the pretreatment operation is as follows: firstly, crushing gelsemium elegans, and carrying out reflux extraction by using a solvent A to obtain a gelsemium elegans extract; the solvent A is water or 30-90% ethanol. Among them, the solvent a is preferably 90% ethanol. The gelsemium elegans extract extracted by 90% ethanol has the strongest toxicity.

Further, before the compatibility, after the pretreatment operation, the gelsmium elegans is subjected to extraction operation, wherein the extraction operation comprises the following steps: extracting the gelsemium elegans extract by using a solvent B, and removing the solvent B to obtain gelsemium elegans extract; the solvent B is one of the following: petroleum ether, dichloromethane or ethyl acetate. The solvent B is preferably dichloromethane. The gelsemium elegans extract obtained by extracting with dichloromethane has the highest toxicity compared with the extract of other solvents.

Further, when the toxicity-reducing and effect-maintaining component is the mixture of the datura flower or the houttuynia cordata and the mint, the pretreatment is carried out before the compatibility, and the pretreatment operation is as follows: firstly, crushing the attenuation and efficacy-preserving component, and performing reflux extraction by using a solvent C to obtain an attenuation and efficacy-preserving component extracting solution; the solvent C is water or 30-90% ethanol. The solvent C is preferably 90% ethanol.

Further, the toxicity-reducing and effect-storing component is subjected to pretreatment operation before compatibility, and then is subjected to extraction operation, wherein the extraction operation comprises the following steps: extracting the attenuated effective component extracting solution by a solvent D to obtain attenuated effective component extracting solution or water phase solution, wherein the solvent D is one or more than two of the following components: petroleum ether, dichloromethane or ethyl acetate.

The invention also discloses a compatible method for reducing toxicity and preserving effects of gelsmium elegans, which comprises the following steps:

The gelsemium elegans and the toxicity-reducing and effect-preserving component are combined according to the weight ratio of 1: 5-50 to obtain a compatible composition of the gelsemium elegans with toxicity-reducing and effect-preserving effect; the toxicity-reducing and efficacy-maintaining component is one of the following components: flos Daturae Metelis, sanguis Caprae Seu Ovis, or mixture of herba Houttuyniae and herba Menthae.

Further, the kiss is subjected to pretreatment operation before compatibility, and the pretreatment operation is as follows: firstly, crushing gelsemium elegans, and carrying out reflux extraction by using a solvent A to obtain a gelsemium elegans extract; the solvent A is water or 30-90% ethanol; when the toxicity-reducing and effect-maintaining component is the mussaenda pubescens or the mixture of the houttuynia cordata and the mint, the components are pretreated before being compatible, and the pretreatment operation is as follows: firstly, crushing the attenuation and efficacy-preserving component, and performing reflux extraction by using a solvent C to obtain an attenuation and efficacy-preserving component extracting solution; the solvent C is water or 30-90% ethanol. The solvent C is preferably 90% ethanol. When the component with toxicity-reducing and effect-preserving effects is sheep blood, the component can be directly mixed with the gelsemium elegans extract.

Further, before the compatibility, after the pretreatment operation, the gelsmium elegans is subjected to extraction operation, wherein the extraction operation comprises the following steps: extracting the gelsemium elegans extract by using a solvent B, and removing the solvent B to obtain gelsemium elegans extract; the solvent B is one of the following: petroleum ether, dichloromethane or ethyl acetate; the toxicity-reducing and effect-maintaining components are subjected to pretreatment operation before compatibility, and then are subjected to extraction operation, wherein the extraction operation comprises the following steps: extracting the attenuated effective component extracting solution by a solvent D to obtain attenuated effective component extracting solution or water phase solution, wherein the solvent D is one or more than two of the following components: petroleum ether, dichloromethane or ethyl acetate. The solvent B is preferably dichloromethane.

Preferably, the compatible method for reducing toxicity and preserving effects of gelsmium elegans comprises the following steps:

1) and performing kiss pretreatment: firstly, crushing gelsemium elegans, and carrying out reflux extraction by using a solvent A to obtain a gelsemium elegans extract; the solvent A is 90% ethanol;

2) And performing gelsemium elegans extraction: extracting the gelsemium elegans extract by using a solvent B, and removing the solvent B to obtain gelsemium elegans extract; the solvent B is dichloromethane;

3) pretreating the mussaenda hemsl: firstly, crushing the mussaenda pubescens, and carrying out reflux extraction by using a solvent C to obtain a mussaenda pubescens extract; the solvent C is 90% ethanol;

4) Extracting the mussaenda hemsleyana: extracting the mussaenda pubescens extract by using a solvent D, and removing an extract liquid to obtain a mussaenda pubescens aqueous phase solution; the solvent D is one or more than two of the following solvents: petroleum ether, dichloromethane or ethyl acetate;

5) And the gelsemium elegans extract and the datura flower aqueous phase solution are mixed according to the weight part of 1:40 to obtain the compatible composition with the effect of reducing the toxicity and preserving the effects of the gelsemium elegans.

In the invention, the gelsemium elegans is combined with the datura flower, the herba houttuyniae and the mint to achieve the effect of attenuating the toxin of the gelsemium elegans to a certain extent, and particularly, the effect of the gelsemium elegans is combined with the datura flower to be better. Meanwhile, the gelsemium elegans compatible attenuation component can also play a role in easing pain for mice. Therefore, the compatible composition of gelsemium elegans of the invention has obvious effects of reducing toxicity and preserving effects.

Description of the drawingsthe present invention will be described in further detail below with reference to the following figures and detailed description:

FIG. 1 is a graph showing the effect of the herba Gelsemii Elegantis dichloromethane extract on the body twisting frequency of mice before and after the herba Gelsemii Elegantis extract is combined with the aqueous phase solution of radix Mussaendae;

FIG. 2 is a graph showing the effect of the herba Gelsemii Elegantis dichloromethane extract on the analgesic rate of mice before and after being combined with the aqueous phase solution of Mussaenda Furetii.

Detailed Description

In order to explain technical contents, structural features, and objects and effects of the present invention in detail, the following description is given in detail with reference to the embodiments.

The invention discloses a compatible composition of gelsemium elegans for reducing toxicity and preserving effects, which is prepared by matching gelsemium elegans and an attenuation and preserving effect component in a weight part ratio of 1: 5-50, wherein the attenuation and preserving effect component is one of the following components: flos Daturae Metelis, sanguis Caprae Seu Ovis, or mixture of herba Houttuyniae and herba Menthae.

Before the compatibility of medicines, the gelsmium elegans is subjected to pretreatment operation, wherein the pretreatment operation comprises the following steps: firstly, crushing gelsemium elegans, and carrying out reflux extraction by using a solvent A to obtain a gelsemium elegans extract; the solvent A is water or 30-90% ethanol; the pretreatment method of the attenuated effective component comprises the following steps: firstly, crushing the attenuation and efficacy-preserving component, and performing reflux extraction by using a solvent C to obtain an attenuation and efficacy-preserving component extracting solution; the solvent C is water or 30-90% ethanol.

In the invention, when the toxicity-reducing and effect-preserving component is the mussaenda virginiana, the gelsmium elegans and the mussaenda virginiana can be mixed and then extracted together, or the gelsmium elegans and the mussaenda virginiana can be respectively extracted and then the extracting solutions are mixed. When the toxicity-reducing and efficacy-maintaining component is a mixture of houttuynia cordata and mint, the operation is similar to that of Mussaenda pubescens. Wherein, the heartleaf houttuynia herb and the mint are mixed according to the weight ratio of 1: 1. When the component with toxicity-reducing and effect-maintaining effects is sheep blood, the gelsemium elegans needs to be extracted first, and then gelsemium elegans extract is mixed with the sheep blood.

The method for pretreating gelsmium elegans by using different solvents A is shown in examples 1-4.

Example 1

Placing 10.0g of herba Gelsemii Elegantis powder in 150mL round bottom flask, soaking in 50mL water for 0.5 hr, reflux extracting for 1 hr, extracting for 2 times, mixing supernatant, recovering under reduced pressure, concentrating, dissolving with distilled water, transferring, and diluting to constant volume to 10mL to obtain herba Gelsemii Elegantis extract 1 g/mL^-1(in terms of gelsemium dose).

Example 2

Placing 10.0g of herba Gelsemii Elegantis powder in 150mL round bottom flask, soaking in 50mL of 30% ethanol for 0.5 hr, reflux extracting for 1 hr, extracting for 2 times, mixing the supernatant, recovering under reduced pressure, concentrating, dissolving in distilled water, transferring, and metering to volume of 10mL to obtain herba Gelsemii Elegantis extractive solution 1 g/mL^-1(in terms of gelsemium dose).

example 3 and example 4

Example 3 and example 4 were performed in the same manner and in the same amount as in example 2, wherein the extraction solvent was replaced, and in example 3, the extraction solvent was 60% ethanol; in example 4, the extraction solvent was 90% ethanol.

The toxicity test of the gelsemium elegans extract was performed according to the gelsemium elegans extract prepared in examples 1 to 4, and the following specific procedures were performed:

Dividing 50 KM mice into 5 groups (blank control group, water extract group, 30% alcohol extract group, 60% alcohol extract group and 90% alcohol extract group)10 per group, marked by picric acid, fasting before the test for 12h without water supply, and then performing disposable gastric lavage according to 0.6 g.kg^-1The corresponding drugs were administered (in terms of gelsemium amount), and the same volume of physiological saline was administered to the blank control group. Signs of toxicity and mortality within 24h of mice were recorded immediately following dosing. The toxicity test results are shown in Table 1.

after the gelsemium elegans extract extracted in the examples 1 to 4 is administered to the mice, silence, trembling, limb weakness and dyskinesia appear in sequence; further turning to an excited state, frequent tapping and accompanied spasm; the spasm intensity is larger at the later stage, then intermittent convulsion and jump occur, then systemic severe tonic convulsion appears immediately, and finally the breathing is exhausted and died. The death peak period is 0.5-2 h after the administration, and the death phenomenon is within 24 h. According to table 1, the results of acute toxicity tests show that: the gelsemium elegans aqueous extract in example 1 has no death and has the minimum toxicity; however, the toxicity of the gelsemium elegans alcohol extract gradually increases with the increase of the ethanol concentration, and the death rate of the mice with 90% gelsemium elegans alcohol extract in example 4 is 100%, and the toxicity is the highest. Therefore, 90% ethanol is preferably used as the extraction solvent of gelsemium elegans.

Table 1 toxicity test of gelsemium elegans extract of examples 1 to 4

Note a: the dosage is based on the weight of gelsemium elegans, and the same is given below.

Taking the components with toxicity reducing and effect preserving functions as the example, the pretreatment method of the mussaenda pubescens can be seen in examples 5 to 8 according to different solvents C. When the toxicity-reducing and efficacy-maintaining component is a mixture of houttuynia cordata and mint, the extraction operation is similar to that of Mussaenda pubescens, and the houttuynia cordata and the mint are mixed according to the weight ratio of 1: 1.

Example 5

Placing 50g crude drug of Mussaenda Nitrilensis in 500ml round bottom flask, soaking in 150ml water for 0.5 hr, reflux extracting for 1 hr, extracting for 2 times, mixing supernatant, recovering under reduced pressure, concentrating, dissolving with distilled water, transferring, and metering to volume of 50ml to obtain Mussaenda Nitrilensis extractive solution 1 g/ml^-1(with jade)Crude drug amount of datura flower).

Example 6

Placing 50g crude drug of Mussaenda Nitrilensis into 500ml round bottom flask, soaking in 150ml 30% ethanol for 0.5 hr, reflux extracting for 1 hr, extracting for 2 times, mixing supernatants, recovering under reduced pressure, concentrating, dissolving with distilled water, transferring, and metering to volume of 50ml to obtain Mussaenda Nitrilensis extractive solution 1 g/ml^-1(calculated by crude drug amount of Mussaenda pubescens).

Example 7 and example 8

Example 7 and example 8 were performed in the same manner and in the same amount as in example 5 or 6, wherein the extraction solvent was replaced, and in example 7, the extraction solvent was 60% ethanol; in example 8, the extraction solvent was 90% ethanol.

In order to examine the attenuation effect of the gelsmium elegans extract and the datura flower extract in the best ratio, the following examples of different ratios of the gelsmium elegans extract and the datura flower extract were performed.

Example 9

the gelsemium elegans extract prepared in the example 4 is mixed with the mussaenda pubescens extract prepared in the example 8 according to the weight part ratio of 1: 5.

Example 10

The gelsemium elegans extract prepared in the example 4 is mixed with the mussaenda pubescens extract prepared in the example 8 according to the weight part ratio of 1: 10.

Example 11

The gelsemium elegans extract prepared in the example 4 is mixed with the mussaenda pubescens extract prepared in the example 8 according to the weight part ratio of 1: 20.

example 12

The gelsemium elegans extract prepared in the example 4 is mixed with the mussaenda pubescens extract prepared in the example 8 according to the weight part ratio of 1: 30.

Example 13

The gelsemium elegans extract prepared in example 4 is mixed with the mussaenda pubescens extract prepared in example 8 according to the weight ratio of 1: 35.

Example 14

The gelsemium elegans extract prepared in the example 4 is mixed with the mussaenda pubescens extract prepared in the example 8 according to the weight part ratio of 1: 40.

Example 15

The gelsemium elegans extract prepared in the example 4 is mixed with the mussaenda pubescens extract prepared in the example 8 according to the weight part ratio of 1: 50.

In actual operation, according to requirements, the crude drug of gelsemium elegans and the crude drug of mussaenda pubescens can be completely mixed according to the required parts by weight, then the reflux extraction is carried out by using a solvent A or a solvent C, then the decompression recovery and the concentration are carried out, then the distilled water is used for dissolving and transferring and fixing the volume, and the extract of the gelsemium elegans and the mussaenda pubescens is obtained.

The composition prepared from the gelsemium elegans extract and the datura flower extract in the embodiments 9-15 in different parts by weight is subjected to acute toxicity test, and the specific operation is as follows:

90 KM mice were weighed and randomly divided into 9 groups: blank control group, gelsemium elegans extract group, examples 9-15, the weight parts of the gelsemium elegans extract and the datura flower extract are different (1:5, 1:10, 1:20, 1:30, 1:35, 1:40 and 1:50), each group contains 10 drugs, the drugs are marked by picric acid, fasting is not prohibited for 12 hours before test, and the drug is 0.8 g.kg.^-1Gavage (all in terms of gelsemium dose) and administration of the same volume of physiological saline was given to the blank control group. Signs of intoxication and death were recorded in each group of mice 24h immediately after dosing and the results are shown in table 2.

as shown in Table 2 and the test results, the amount of the gelsemium elegans extract alone was 0.8 g/kg^-1The death rate of the mice is 100%, and in the different weight part proportioning groups of the gelsemium elegans extract and the mussaenda pubescens extract, the death rate of the mice is reduced along with the increase of the weight part of the mussaenda pubescens extract. When the weight part ratio of the gelsemium elegans extract to the mussaenda pubescens extract is more than 1:40, the death rate of mice is not reduced. Therefore, in example 14, the weight ratio of the gelsmium elegans extract to the mussaenda pubescens extract is 1:40, which is the optimal ratio.

TABLE 2 acute toxicity test of different weight ratio of herba Gelsemii Elegantis extractive solution and radix Mussaendae extract

Note: hooking: the jade is' the weight parts of gelsemium elegans extract: shorthand of the weight parts of the mussaenda pubescens extract.

The optimal proportion of the gelsemium elegans extract and the mussaenda pubescens extract is subjected to an acute toxicity verification test, and the method specifically comprises the following operation:

50 KM mice were weighed and randomly divided into 5 groups: blank control group, single gelsemium elegans extract group, example 14 optimal weight part ratio of gelsemium elegans extract and datura flower extract 1:40 groups (repeat 3 groups), each group 10, bitter acid mark, fasting before test for 12h without water prohibition, and disposable after test for 0.8g.kg^-1gavage (all in terms of gelsemium dose) and administration of the same volume of physiological saline was given to the blank control group. Signs of toxicity and mortality within 24h of each group of mice were recorded immediately after dosing. The specific results are shown in Table 3.

TABLE 3 test for the verification of acute toxicity of the extract of gelsmium elegans and the extract of mussaenda

Note: hooking: the jade is the weight parts of the gelsmium elegans extract: the weight parts of the datura flower extract are abbreviated.

As can be seen from Table 3, the optimal ratio of the gelsemium elegans extract to the datura flower extract in example 14 of 1:40 was used for acute toxicity tests, and the average mortality of the mice was confirmed to be 23.33 + -5.78%. Under the condition of the same concentration, the gelsemium elegans extract and the datura flower extract are combined according to the weight portion of 1:40, and the effects of attenuation and efficacy preservation are optimal.

before the compatibility, after the pretreatment operation, the gelsemium elegans is subjected to extraction operation, wherein the extraction operation comprises the following steps: extracting the gelsemium elegans extract by using a solvent B, and removing the solvent B to obtain gelsemium elegans extract; the solvent B is one of the following: petroleum ether, dichloromethane or ethyl acetate. In the invention, the gelsemium elegans extract is extracted by a plurality of solvents respectively, so as to find out the best toxic component of gelsemium elegans.

Example 16

Extracting the gelsemium elegans extract obtained in the embodiment 4 by petroleum ether for 3 times, combining the petroleum ether extract, and carrying out reduced pressure recovery and concentration to obtain the gelsemium elegans petroleum ether extract.

Example 17

And (3) extracting the gelsemium elegans extract obtained in the embodiment 4 by dichloromethane, extracting for 3 times, combining dichloromethane extract, and carrying out reduced pressure recovery and concentration to obtain the gelsemium elegans dichloromethane extract.

Example 18

And (3) extracting the gelsemium elegans extract obtained in the embodiment 4 by ethyl acetate for 3 times, combining ethyl acetate extract, and carrying out reduced pressure recovery and concentration to obtain the gelsemium elegans ethyl acetate extract.

Example 19

And (3) sequentially extracting the gelsemium elegans extract obtained in the embodiment 4 by using petroleum ether, dichloromethane and ethyl acetate, extracting each solvent for 3 times, discarding the extract liquor, collecting the water phase, and performing reduced pressure recovery and concentration to obtain the gelsemium elegans water phase solution. The specific operation is as follows: extracting the gelsmium elegans extract by petroleum ether, removing petroleum ether extract, and collecting water phase; then extracting the aqueous phase solution after petroleum ether extraction by dichloromethane, removing dichloromethane extraction liquid, and collecting the aqueous phase; and extracting the water phase solution after the extraction of the dichloromethane with ethyl acetate, discarding ethyl acetate extract, collecting the water phase, and performing reduced pressure recovery and concentration to obtain the gelsemium elegans water phase solution.

Each gelsmium elegans extract of examples 16-19 was subjected to an acute toxicity test, and 60 KM mice were weighed and randomly divided into 6 groups: respectively setting blank control group, herba Gelsemii Elegantis extract control group, herba Gelsemii Elegantis petroleum ether extract group, herba Gelsemii Elegantis dichloromethane extract group, herba Gelsemii Elegantis ethyl acetate extract group, and herba Gelsemii Elegantis aqueous solution group, marking with picric acid, fasting for 12 hr before test, and then once administering at a dose of 0.01 ml/g^-1Gavage was performed, and the same volume of physiological saline was administered to the placebo group. Signs of intoxication and death were recorded in each group of mice 24h immediately after dosing and the results are shown in table 4.

TABLE 4 determination of optimal extraction solvent for gelsemium elegans

As can be seen from table 4, the results of the mouse acute toxicity tests of the extract and aqueous phase solutions (petroleum ether extract, dichloromethane extract, ethyl acetate extract, aqueous phase solution) obtained by extracting the gelsemium elegans 90% ethanol extract with three solvents show that: example 17 gelsemium elegans dichloromethane extract mice showed 100% mortality, but none of the gelsemium elegans petroleum ether extract, the gelsemium acetate extract, and the gelsemium aqueous solution died (see table 4). The experimental result shows that the gelsemium elegans dichloromethane extract in example 17 is the main toxic component of gelsemium elegans, and the gelsemium elegans toxic component may be total indole alkaloid.

in the invention, when the toxicity-reducing and effect-maintaining component is the mussaenda or the mixture of the houttuynia cordata and the mint, the toxicity-reducing and effect-maintaining component is subjected to extraction operation after pretreatment operation before compatibility, and the extraction operation is as follows: extracting the attenuated effective component extracting solution by a solvent D to obtain attenuated effective component extracting solution or water phase solution, wherein the solvent D is one or more than two of the following components: petroleum ether, dichloromethane or ethyl acetate. Wherein, taking the components with toxicity reducing and effect preserving as the example to carry out the extraction operation, the specific operation is shown in the examples 20 to 23.

Example 20

Extracting the mussaenda pubescens extract obtained in the example 9 by using petroleum ether, extracting for 3 times, combining the petroleum ether extract, and performing reduced pressure recovery and concentration to obtain the petroleum ether extract of the mussaenda pubescens.

Example 21

Extracting the mussaenda pubescens extract obtained in example 9 with dichloromethane, extracting for 3 times, combining dichloromethane extract, and recovering and concentrating under reduced pressure to obtain the dichloromethane extract of the mussaenda pubescens.

Example 22

Extracting the extractive solution of Mussaenda pubescens from example 9 with ethyl acetate for 3 times, mixing the extractive solutions, and recovering under reduced pressure to obtain the extractive solution.

Example 23

extracting the mussaenda pubescens extract obtained in the example 9 by using petroleum ether, dichloromethane and ethyl acetate in sequence, extracting for 3 times, discarding the extract, collecting the water phase, and recovering and concentrating under reduced pressure to obtain the mussaenda pubescens water phase solution. The specific operation is as follows: extracting the mussaenda pubescens extract by petroleum ether, removing the petroleum ether extract, and collecting a water phase; then extracting the aqueous phase solution extracted by the petroleum ether by dichloromethane, discarding dichloromethane extraction liquid, and collecting the aqueous phase; and extracting the water phase solution after the extraction of the dichloromethane with ethyl acetate, discarding ethyl acetate extract, collecting the water phase, and performing reduced pressure recovery and concentration to obtain the datura flower water phase solution.

In other embodiments, the aqueous solution of datura flower can also be an aqueous solution extracted with a single solvent (petroleum ether, dichloromethane, or ethyl acetate).

The gelsemium elegans methylene dichloride extract obtained in example 17 is respectively compatible with the various groups of the datura flower extract and the aqueous phase solution obtained in examples 20 to 23 according to the weight part of 1:40, and the following examples are specifically carried out:

Example 24

The gelsemium elegans dichloromethane extract obtained in example 17 and the mussaenda pubescens petroleum ether extract obtained in example 20 are combined according to the weight part ratio of 1:40 (the concentration of the gelsemium elegans dichloromethane extract and the concentration of the mussaenda pubescens petroleum ether extract are the same).

Example 25

The gelsemium elegans dichloromethane extract obtained in example 17 and the mussaenda pubescens dichloromethane extract obtained in example 21 are combined according to the weight part of 1: 40.

Example 26

The gelsemium elegans dichloromethane extraction liquid obtained in the embodiment 17 and the mussaenda pubescens ethyl acetate extraction liquid obtained in the embodiment 22 are combined according to the weight part ratio of 1: 40.

Example 27

the gelsemium elegans dichloromethane extract obtained in the example 17 and the aqueous phase solution of the mussaenda fragrans obtained in the example 23 are combined according to the weight part of 1: 40.

The combinations obtained by combining the above examples 24 to 27 were subjected to acute toxicity test. 60 KM mice were weighed and randomly divided into 6 groups: the test was conducted in a blank control group, a gelsemium chloride extract group, example 24, example 25, example 26, and example 27, respectively, under picric acid labeling, fasting was not performed for 12 hours before the test, and the administration volume of the test was 0.01ml g^-1Gavage was performed, and the same volume of physiological saline was administered to the placebo group. Signs of intoxication and death were recorded in each group of mice 24h immediately after dosing and the results of the specific tests are shown in table 5.

Table 5 and the test results show that the mortality rate of acute toxicity of mice in the gelsemium dichloromethane extract of example 17 is 100%, but the mortality rate is reduced after the gelsemium dichloromethane extract and the 90% alcohol extract of the mussaenda dissimalis of examples 24 to 27 are combined with aqueous phase solutions (petroleum ether extract, dichloromethane extract, ethyl acetate extract and aqueous phase solution). In example 27, the mortality of the mice is the lowest after the aqueous phase solution of the datura flower and the gelsemium elegans extract are combined, which indicates that the mutual compatibility of the gelsemium elegans extract and the aqueous phase solution of the datura flower has the best effect.

TABLE 5 screening of herba Gelsemii Elegantis extractive solution, radix Mussaendae extractive solution and water phase solution

In order to further verify that the effect of the gecko reduction and the effect of the gecko are the best after the combination of the water-phase solution of the datura flower and the extract of the gelsemium chloride in example 27 is performed, the following verification test is performed. 70 KM mice were weighed and randomly divided into 7 groups: blank control group, gelsemium elegans dichloromethane extract group (repeat 3 groups) of example 17, and aqueous phase solution of Murraya koenigii and gelsemium elegans of example 27The dichloromethane extract liquid is 40 parts by weight: 1 compatibility group (repeating 3 groups), 10 per group, picric acid mark, fasting before test for 12h without water prohibition, and one-time intragastric administration of 0.5 g.kg^-1The corresponding drugs were administered, and the blank group was given the same volume of physiological saline. Signs of intoxication and death were recorded in mice 24h immediately after dosing and the results are shown in table 6.

TABLE 6 verification of sites of facial callosity

Note a: p is less than 0.05 compared with herba Gelsemii Elegantis dichloromethane extract group

As can be seen from table 6, the aqueous solution of datura flower of example 23 can effectively reduce the toxicity of the dichloromethane extract of gelsemium of example 17, and thus it is confirmed that the aqueous solution of datura flower of example 23 is an attenuated solution of gelsemium, and the average mortality rate is greatly reduced (P is less than 0.05) after the aqueous solution of datura flower of example 23 is combined with the dichloromethane extract of gelsemium of example 17, and it is further verified that the efficacy of the aqueous solution of datura flower of example 27 in reducing toxicity of the aqueous solution of datura flower and the dichloromethane extract of gelsemium is.

Example 28

The invention relates to a compatible method for reducing toxicity and preserving effects of gelsmium elegans, which specifically comprises the following steps:

1) And performing kiss pretreatment: weighing herba Gelsemii Elegantis 10g, pulverizing, adding 50ml 90% ethanol, soaking for 0.5 hr, reflux extracting for 1 hr, extracting for 2 times, mixing supernatant, recovering under reduced pressure, concentrating, and diluting to constant volume to obtain herba Gelsemii Elegantis extract with concentration of 0.05 g/ml^-1(based on the mass of the gelsemium elegans L.var.elegans L.var..

2) And performing gelsemium elegans extraction: extracting herba Gelsemii Elegantis extractive solution with dichloromethane 3 times of herba Gelsemii Elegantis crude drug by weight for 3 times, mixing dichloromethane extractive solutions, recovering under reduced pressure, concentrating to obtain herba Gelsemii Elegantis dichloromethane extractive solution, and storing in refrigerator at 4 deg.C for use.

3) Pretreating the mussaenda hemsl: weighing crude drug of flos Daturae Metelis 10g, pulverizing, adding 50ml 90% ethanol, soaking for 0.5 hr, reflux extracting for 1 hr, extracting for 2 times, mixing supernatant, recovering under reduced pressure, concentrating, and diluting to desired volumeThe concentration of the extractive solution is 2.0 g/ml^-1(calculated by crude drug amount of Mussaenda pubescens).

4) Extracting the mussaenda hemsleyana: sequentially extracting the extractive solution with petroleum ether, dichloromethane and ethyl acetate 3 times the mass of crude drug of radix Mussaendae, extracting for 3 times, discarding the extractive solution, collecting water phase, recovering water phase under reduced pressure, concentrating to obtain water phase solution of radix Mussaendae, and storing in refrigerator at 4 deg.C for use.

5) And matching the gelsemium elegans dichloromethane extract and the datura flower aqueous phase solution in equal volume to obtain the compatible composition with the gelsemium elegans toxicity-reducing effect.

Test for detecting writhing of mouse by using gelsemium elegans toxicity-reducing efficacy-maintaining compatible composition prepared in example 28

Taking 70 KM mice, each half of male and female, randomly dividing into 7 groups, and blank control group (giving 0.32 g.kg for blank control group)^-1Physiological saline) and indomethacin positive control group (0.01 g/kg)^-1) Gelsemium elegans methylene dichloride extract group (0.27g kg)^-1) Mussaenda pubescens aqueous phase solution group (7.12g kg)^-1) The herba Gelsemii Elegantis dichloromethane extractive solution is combined with flos Daturae Metelis aqueous phase solution at high dosage (0.41 g/kg based on herba Gelsemii Elegantis amount)^-1) The herba Gelsemii Elegantis dichloromethane extractive solution is combined with the dosage group (0.32 g.kg based on herba Gelsemii Elegantis crude drug) in the water phase solution of flos Daturae Metelis^-1) The herba Gelsemii Elegantis dichloromethane extractive solution is combined with low dose of flos Daturae Metelis aqueous phase solution (0.27 g/kg based on herba Gelsemii Elegantis crude drug amount)^-1) Each group had 10 picric acid labels. After each group is administrated by gastric lavage for 30min, 0.1ml/10g of 0.6% acetic acid solution is injected into abdominal cavity. The number of writhing reactions (hind limb extension, abdominal contraction and concavity, hip elevation) occurring within 15min after acetic acid solution injection was observed and recorded, and the analgesic rate was calculated according to the formula (1-1).

Pain rate (number of writhing in control group-number of writhing in administration group)/number of writhing in control group x 100% (1-1)

The test results are shown in FIGS. 1 and 2. As can be seen from fig. 1, the indomethacin group, the gelsemium dichloromethane extraction group and the gelsemium dichloromethane extraction liquid compatible datura flower aqueous phase solution in high, medium and low dosage groups can all significantly reduce the times of mouse body writhing caused by acetic acid within 15min, compared with the blank control normal saline group, the significant difference (P is less than 0.01) is very significant, the significant difference is not generated in the effect of the gelsemium dichloromethane extraction group and the gelsemium dichloromethane extraction liquid compatible datura flower aqueous phase solution in high, medium and low dosage groups, and the analgesic rate and the dosage of the gelsemium dichloromethane extraction liquid compatible with the datura flower aqueous phase solution are positively correlated. As can be seen from fig. 2, the analgesic rate of the gelsemium elegans and dichloromethane extract solution and the datura flower aqueous phase solution is significantly different from that of the normal saline solution (P is less than 0.05); the analgesic rate of the gelsemium elegans methylene dichloride extract and the golden pattern aqueous phase solution is very different from that of the indometacin group (delta P is less than 0.01). The result shows that the gelsemium elegans methylene dichloride extract is matched with the datura flower aqueous phase solution and still has remarkable inhibiting effect on the pain reaction of the mouse caused by the chemical stimulation method.

The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by the present specification, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.

Claims

1. The compatible composition of the gelsemium elegans for reducing and preserving effects is characterized in that the composition is prepared from gelsemium elegans and components for reducing and preserving effects in a crude drug weight ratio of 1:40, and the preparation of the gelsemium elegans comprises the following steps: performing reflux extraction on the crushed gelsemium elegans by 90% ethanol to obtain gelsemium elegans extract, performing extraction on the gelsemium elegans extract by dichloromethane, and removing the dichloromethane to obtain gelsemium elegans components; the toxicity-reducing and effect-maintaining component is Mussaenda pubescens, and the preparation steps comprise: and (2) performing reflux extraction on the crushed mussaenda leptinotarsa through 90% ethanol to obtain an attenuation efficacy-maintaining component extracting solution, sequentially extracting the attenuation efficacy-maintaining component extracting solution through petroleum ether, dichloromethane and ethyl acetate, discarding an extracting solution, collecting a water phase, and performing reduced pressure concentration to obtain the attenuation efficacy-maintaining component.

2. The gelsmium elegans attenuation and effect preservation compatible composition according to claim 1, wherein the attenuation and effect preservation component is obtained by sequentially extracting the attenuation and effect preservation component extracting solution with petroleum ether, dichloromethane and ethyl acetate for three times respectively, discarding the extracting solution, collecting the water phase, and concentrating under reduced pressure.

3. A preparation method of a compatible composition for reducing toxicity and preserving effects of gelsmium elegans is characterized by comprising the following steps:

The gelsemium elegans and the toxicity-reducing and efficacy-preserving component are combined according to the crude drug weight ratio of 1:40 to obtain a compatible composition of the gelsemium elegans with toxicity-reducing and efficacy-preserving effect; the preparation method of the gelsemium elegans comprises the following steps: performing reflux extraction on the crushed gelsemium elegans by 90% ethanol to obtain gelsemium elegans extract, performing extraction on the gelsemium elegans extract by dichloromethane, and removing the dichloromethane to obtain gelsemium elegans components; the toxicity-reducing and effect-maintaining component is Mussaenda pubescens, and the preparation steps comprise: and (2) performing reflux extraction on the crushed mussaenda leptinotarsa through 90% ethanol to obtain an attenuation efficacy-maintaining component extracting solution, sequentially extracting the attenuation efficacy-maintaining component extracting solution through petroleum ether, dichloromethane and ethyl acetate, discarding an extracting solution, collecting a water phase, and performing reduced pressure concentration to obtain the attenuation efficacy-maintaining component.

4. The preparation method of the gelsmium elegans toxicity-reducing effect-preserving compatible composition as claimed in claim 3, wherein the toxicity-reducing effect-preserving component extracting solution is sequentially extracted by petroleum ether, dichloromethane and ethyl acetate for three times respectively, the extracting solution is discarded, the water phase is collected, and the toxicity-reducing effect-preserving component is obtained by reduced pressure concentration.