CN112773847A

CN112773847A - Preparation method and application of gel prepared from calyx seu fructus physalis extract

Info

Publication number: CN112773847A
Application number: CN202110283824.4A
Authority: CN
Inventors: 舒尊鹏
Original assignee: Guangdong Andao Medical Instrument Co ltd
Current assignee: Guangdong Andao Medical Instrument Co ltd
Priority date: 2021-03-17
Filing date: 2021-03-17
Publication date: 2021-05-11

Abstract

The invention discloses a preparation method and application of a gel prepared from calyx seu fructus physalis extract, wherein the preparation method comprises the following steps: preparing calyx seu fructus physalis extract from calyx seu fructus physalis; preparing a 2% carbomer solution; the calyx seu fructus physalis extract and the 2% carbomer solution are used as main raw materials to prepare the gel, the raw materials for preparing the gel are easy to obtain, the preparation method is simple, the cost is low, the invention also discloses the application of the gel prepared by the preparation method, the application comprises the application of the gel as an antiallergic drug and the application of the gel for treating acne, the gel prepared by the invention can inhibit anaphylaxis and has an antiallergic effect, and meanwhile, the gel prepared by the invention can also inhibit inflammatory reaction of the acne and has an anti-inflammatory effect.

Description

Preparation method and application of gel prepared from calyx seu fructus physalis extract

Technical Field

The invention discloses a preparation method and application of a gel prepared from calyx seu fructus physalis extract, and belongs to the technical field of medicines.

Background

Calyx seu fructus Physalis (Physicaceae) is dry calyx or calyx with fruit of Physalis of Solanaceae (Solanaceae), and is also named as calyx seu fructus Physalis, Physalis alkekengi, etc. It is cold in nature and bitter in taste, entering lung meridian. Native to China, mainly distributed in northeast, northwest, inner Mongolia and other places. It is recorded in Shen nong Ben Cao Jing (Shen nong's herbal), named sour sauce, listed as a Chinese medicine, and has the effects of clearing away heat and toxic materials, dispelling fire and relieving swelling. The herbs in the later generations are mostly collected and carried, and the Ming Dynasty Li Shizhen Ben Cao gang mu records that the calyx seu fructus physalis can remove dampness and heat, clear lung heat and treat cough, remove dampness and resolve phlegm, and treat deep-rooted carbuncle. Nowadays, the calyx seu fructus physalis is mainly used for treating pharyngalgia, hoarseness, phlegm-heat cough and dysuresia, and externally treating pemphigus, eczema and the like. The chemical components of the calyx seu fructus physalis mainly comprise steroids, sterols, alkaloids, flavonoids, amino acids, polysaccharides and the like. Modern pharmacological research shows that the crude extract and separated compounds of calyx seu fructus physalis have the effects of resisting inflammation, resisting cancer, killing bacteria, promoting urination, reducing blood sugar, reducing blood lipid, resisting oxidation, etc.

Allergy refers to the condition that people with allergic constitution contact with allergens including dust, pollen, chemicals, soap, detergent, ultraviolet rays, etc., so that the immune mechanism of the organism is activated and hypersensitive reaction is induced. According to the mechanism and characteristics of occurrence, the clinical classification of the cancer is four types, namely: type I hypersensitivity, type II hypersensitivity, type III hypersensitivity and type IV hypersensitivity. Among them, type I and type IV hypersensitivity reactions are the most common. Type I hypersensitivity, immediate hypersensitivity, is a series of immune responses resulting from the production of excessive atopic IgE antibodies by certain allergens after stimulation of the body. Clinically common skin type I hypersensitivity diseases include atopic dermatitis, urticaria, eczema and the like. Type I hypersensitivity has rapid onset, generally only causes functional disorder of the organism, has no organic damage, and although the life is rarely damaged, the life quality of patients is greatly influenced. Type IV hypersensitivity, i.e. delayed hypersensitivity, is mainly an immune dysfunction reaction mediated by T lymphocytes. Type IV hypersensitivity is mainly divided into 2 phases, the sensitization phase and the priming phase, which are regulated by different T cell populations and migrate to the skin, eventually producing various cytokines to trigger skin inflammation. The symptoms of the skin allergy can bring troubles to people in appearance, body and mind, and the anti-allergic drugs used clinically at present are hormones, so that the side effects of the drugs are obvious and the tolerance is easy to appear.

Acne is a chronic inflammatory skin disease of hair follicles and sebaceous glands caused by various factors including psychological factors, environmental factors, endocrine, improper diet and the like, which is particularly expressed as papules, comedones, pustules, nodules, cysts and scars and is better developed on parts rich in sebaceous glands, such as the face, the chest, the back and the like. Teenagers often become high-incidence people of acne due to excessive sebum secretion, and the disease is a common disease and frequently encountered disease in outpatient clinics of dermatology and has high recurrence rate. Although the clinical treatment scheme for acne is more, the effect is poor and the acne is easy to relapse, and most commonly, antibiotics and anti-inflammatory drugs are used and have a certain treatment effect, but the long-term administration has more adverse reactions and the compliance of patients is poor.

In conclusion, no safe and effective therapeutic drug is clinically available for skin allergy or acne. Therefore, it is an urgent need to solve the problems of the art to provide a natural extract for treating skin allergy and acne with natural ingredients, good therapeutic effect, and no toxic or side effects. However, the detailed research on the medical application of the calyx seu fructus physalis extract, particularly the treatment of skin allergy and acne is not reported at present, so that the calyx seu fructus physalis extract is taken as a research object to research the effect of the calyx seu fructus physalis extract on the treatment of skin allergy and acne.

Disclosure of Invention

The first purpose of the invention is to provide a preparation method for preparing gel by using calyx seu fructus physalis extract.

The second purpose of the invention is to provide the application of the gel prepared by the preparation method as an antiallergic medicine.

The third purpose of the invention is to provide the application of the gel prepared by the preparation method for treating acne.

In order to realize the first purpose of the invention, the following technical scheme is adopted:

a method for preparing gel by using calyx seu fructus physalis extract is characterized in that: the preparation method comprises the following steps:

the method comprises the following steps: preparation method of calyx seu fructus physalis extract

Cutting calyx seu fructus physalis, and placing into flask; adding 70% ethanol into the flask, performing first reflux extraction operation on the cut calyx seu fructus physalis medicinal material in the flask, starting boiling 70% ethanol in the flask to keep the 70% ethanol in the flask in a boiling state for 2h, then extracting a first filtrate in a first part of reflux liquid by using a suction filtration method, and keeping filter residue in the first part of reflux liquid; adding the filter residue into the flask, performing second reflux extraction operation on the filter residue in the flask after the filter residue is added by using the residual 70% ethanol in the flask after the first reflux extraction operation, starting boiling the residual 70% ethanol in the flask to keep the residual 70% ethanol in the flask in a boiling state for 2 hours, then extracting a second filtrate in a second part of reflux liquid by using a suction filtration method, and combining the first filtrate and the second filtrate to obtain a third filtrate; and (3) carrying out vacuum concentration on the volume of the third filtrate, and adding 95% ethanol into the vacuum concentrated third filtrate for mixing to obtain the calyx seu fructus physalis extract.

Step two: preparation of 2% carbomer solution

And (3) putting carbomer 940 into pure water, stirring until the carbomer 940 is completely dissolved, and swelling for 12h to obtain a 2% carbomer solution.

Step three: preparation of gel

Putting the 2% carbomer solution into a beaker, and putting the beaker on a magnetic stirrer to stir the 2% carbomer solution in the beaker at a constant speed; taking glycerol and propylene glycol, fully dissolving the glycerol and the propylene glycol mutually, and adding the glycerol and the propylene glycol into the first mixed solution which is mixed and stirred with the 2% carbomer solution in the beaker; taking the calyx seu fructus physalis extract, adding ethylparaben and menthol into the calyx seu fructus physalis extract, stirring until the ethylparaben and the menthol are completely dissolved to obtain a second mixed solution, adding the second mixed solution into the beaker, mixing with the first mixed solution, stirring to obtain a third mixed solution, stirring until the third mixed solution in the beaker is uniform in color, adding a saturated sodium bicarbonate solution into the beaker to adjust the pH of the third mixed solution to obtain a gel, adding water into the beaker, and stirring the gel in the beaker until the gel is completely dissolved to obtain a gel.

Preferably, the 70% ethanol is prepared by 95% ethanol or recovered alcohol.

Preferably, the weight ratio of the ethylparaben to the menthol is 1: 1.

Preferably, the amount of the saturated sodium bicarbonate solution added is 10% by weight of the second mixed solution.

Preferably, the pH is 6.5.

The inventor experiments show that the gel prepared by the invention has the function of an anti-allergic medicament and can treat skin allergy, so the invention provides the following components according to the invention: (1) a gel prepared from calyx seu fructus physalis extract is used as antiallergic agent, and is applied to classical animal models for research on mouse auricle and IV type hypersensitivity.

In addition, the inventor also finds that the gel prepared by the invention has the function of treating acne, so the invention also provides: (2) a gel prepared from calyx Seu fructus Physalis extract for treating acne is applied in model of rat acne caused by Propionibacterium acnes.

The invention is further illustrated below:

in the invention, 95% ethanol is used for preparing 70% ethanol, so that the purity of the 70% ethanol can be improved, and in the invention, the recovered ethanol can be used for preparing 70% ethanol, thereby being beneficial to the recovery and reutilization of resources.

In the method of preparing the gel of the present invention, in order to achieve better gel effect and increase the consistency of the gel, a 2% carbomer solution is prepared with carbomer 940.

In the invention, in order to enhance the bacteriostatic performance of the prepared gel, a proper bacteriostatic agent is added, and in the invention, ethylparaben is used as the bacteriostatic agent, so that the gel has a good bacteriostatic effect.

In the process of preparing the gel, menthol is added, so that the gel has mint fragrance, and meanwhile, the menthol also has a cooling effect, so that the medication experience can be improved.

In the invention, the weight ratio of the ethyl paraben to the added menthol is 1: 1.

In the present invention, in order to obtain a better gel, the PH of the second mixed solution needs to be adjusted, and the present invention adopts a method of adding a saturated sodium bicarbonate solution into the second mixed solution, and the addition amount of the saturated sodium bicarbonate solution is related to the weight of the second mixed solution, and the inventor finds that the effect is best when the addition amount of the saturated sodium bicarbonate solution is 10% of the weight of the second mixed solution in the experimental process, and also finds that the gel is thick when the PH of the second mixed solution is adjusted to 6.5 by using the saturated sodium bicarbonate solution, and the effect of the gel is best at this time, so the preferred PH of the present invention is 6.5.

The invention has the following beneficial effects:

1) the calyx seu fructus physalis extract prepared from calyx seu fructus physalis medicinal materials has the advantages of simple preparation method and easily-obtained raw materials, and the prepared calyx seu fructus physalis extract has natural components, is nontoxic and has no side effect.

2) The raw material for preparing the 70 percent ethanol can be recovered alcohol, thereby being beneficial to resource recovery and reutilization.

3) In the process of preparing the gel, the required raw materials are simple and easy to obtain, and the cost is low.

4) The prepared gel can be used as antiallergic medicine for inhibiting anaphylaxis, and has antiallergic effect.

5) The prepared gel can be used for treating acne, inhibiting inflammatory reaction of acne, and resisting inflammation.

Drawings

FIG. 1: the effect of the gel of example seven on the levels of IL-1 α, IL-6, IL-2 and TNF- α in serum is shown in the experimental results.

Detailed Description

The present invention will be further understood from the specific examples given below, which are not intended to limit the present invention.

Examples one to two are examples of a preparation method of a gel using calyx seu fructus physalis extract.

Examples three to five and comparative examples one to three are applications of the gel prepared by the present invention as an antiallergic agent for passive skin allergy (skin type I hypersensitivity).

In a specific embodiment, the calculation of the inhibition rate of the blue spot is related to the following formula: the blue spot inhibition (%) is (control absorbance-drug absorbance)/control absorbance × 100%.

The sixth embodiment is the application of the gel prepared by the invention as an antiallergic drug in an Allergic Contact Dermatitis (ACD) model (a classic animal model for type IV hypersensitivity study).

The seventh example is the application of the gel prepared by the invention in treating a model of rat acne caused by propionibacterium acnes.

Example one

Taking 50g of calyx seu fructus physalis, cutting into pieces, and placing into a flask; adding 1.5L of 70% ethanol prepared from 95% ethanol into a flask, performing first reflux extraction on the cut calyx seu fructus physalis medicinal material in the flask, starting boiling 70% ethanol in the flask to keep 70% ethanol in the flask at boiling state for 2h, pumping out the first filtrate in the first part of reflux liquid by suction filtration, and retaining the filter residue in the first part of reflux liquid; adding filter residues into a flask, performing secondary reflux extraction on the filter residues in the flask after the filter residues are added by using the residual 70% ethanol in the flask after the primary reflux extraction operation, starting boiling the residual 70% ethanol in the flask to keep the residual 70% ethanol in the flask in a boiling state for 2 hours, then extracting a second filtrate in a second part of reflux liquid by using a suction filtration method, and combining the first filtrate and the second filtrate to obtain a third filtrate, wherein the volume of the third filtrate is 500-600 ml; and concentrating the volume of the third filtrate under reduced pressure to 300-400 ml, and adding 95% ethanol into the third filtrate after the third filtrate is concentrated under reduced pressure to mix and dissolve the third filtrate until the volume of the mixed and dissolved solution is 650ml, so as to obtain the calyx seu fructus physalis extract.

Step two: preparation of 2% carbomer solution

2g of carbomer 940 is put into 100ml of pure water, stirred until the carbomer 940 is completely dissolved, and then swelled for 12 hours to obtain 2% carbomer solution.

Step three: preparation of gel

Putting 40g of 2% carbomer solution into a beaker, and putting the beaker on a magnetic stirrer to stir the 2% carbomer solution in the beaker at a constant speed; taking 10g of glycerol and 5g of propylene glycol, completely dissolving the glycerol and the propylene glycol mutually, adding the glycerol and the propylene glycol into a beaker, and mixing and stirring the glycerol and the propylene glycol with 2% of carbomer solution to obtain a first mixed solution; taking 30ml of calyx seu fructus physalis extract, adding 0.1g of ethylparaben and 0.1g of menthol into the calyx seu fructus physalis extract, stirring until the ethylparaben and the menthol are completely dissolved to obtain a second mixed solution, adding the first mixed solution into a beaker, mixing and stirring with the first mixed solution to obtain a third mixed solution, stirring until the third mixed solution in the beaker is uniform in color, adding 10g of saturated sodium bicarbonate solution into the beaker to adjust the pH of the third mixed solution to 6.5 to obtain a gel, adding water into the beaker until the total weight of the water and the gel in the beaker is 100g, and stirring the gel in the beaker until the gel is completely dissolved to obtain 100g of gel.

Example two

Taking 40g of calyx seu fructus physalis, cutting into pieces, and placing into a flask; adding 1.5L of 70% ethanol prepared from recycled ethanol into a flask, performing first reflux extraction on the cut calyx seu fructus physalis medicinal material in the flask, starting boiling 70% ethanol in the flask to keep 70% ethanol in the flask at boiling state for 2h, extracting a first filtrate from a first part of reflux liquid by suction filtration, and keeping filter residue in the first part of reflux liquid; adding filter residues into a flask, performing secondary reflux extraction on the filter residues in the flask after the filter residues are added by using the residual 70% ethanol in the flask after the primary reflux extraction operation, starting boiling the residual 70% ethanol in the flask to keep the residual 70% ethanol in the flask in a boiling state for 2 hours, then extracting a second filtrate in a second part of reflux liquid by using a suction filtration method, and combining the first filtrate and the second filtrate to obtain a third filtrate, wherein the volume of the third filtrate is 450-500 ml; and concentrating the volume of the third filtrate under reduced pressure to 300-400 ml, and adding 95% ethanol into the third filtrate after the third filtrate is concentrated under reduced pressure to mix and dissolve the third filtrate until the volume of the mixed and dissolved solution is 650ml, so as to obtain the calyx seu fructus physalis extract.

Step two: preparation of 2% carbomer solution

4g of carbomer 940 is put into 200ml of pure water, stirred until the carbomer 940 is completely dissolved, and then swelled for 12 hours to obtain a 2% carbomer solution.

Step three: preparation of gel

Putting 30g of 2% carbomer solution into a beaker, and putting the beaker on a magnetic stirrer to stir the 2% carbomer solution in the beaker at a constant speed; taking 8g of glycerol and 4g of propylene glycol, completely dissolving the glycerol and the propylene glycol mutually, adding the glycerol and the propylene glycol into a beaker, and mixing and stirring the glycerol and the propylene glycol with 2% of carbomer solution to obtain a first mixed solution; taking 30ml of calyx seu fructus physalis extract, adding 0.08g of ethylparaben and 0.08g of menthol into the calyx seu fructus physalis extract, stirring until the ethylparaben and the menthol are completely dissolved to obtain a second mixed solution, adding the first mixed solution into a beaker, mixing and stirring with the first mixed solution to obtain a third mixed solution, stirring until the third mixed solution in the beaker is uniform in color, adding 8g of saturated sodium bicarbonate solution into the beaker to adjust the pH of the third mixed solution to 6.5 to obtain a gel, adding water into the beaker until the total weight of the water and the gel in the beaker is 100g, and stirring the gel in the beaker until the gel is completely dissolved to obtain 100g of gel.

EXAMPLE III

Gel low dose group

Preparation of anti-ovalbumin serum

Taking 3 male SD rats, injecting egg albumin solution with concentration of 10mg/mL, 0.5 mL/rat, 2 days 1 time, 5 times, 14 days after last sensitization, collecting blood from abdominal aorta of SD rat, centrifuging at 3000r/min for 10min, separating serum to obtain anti-egg albumin serum, and placing at-20 deg.C for use.

Passive cutaneous anaphylaxis (RCA reaction) assay

Taking 8 healthy KM mice, each half of the mice are male and female, (1) on the first day of experiment, smearing the gel prepared in the first example or the second example (the same below) on the inner and outer surfaces of auricles of all the mice according to the low dose of 2.5mg/kg, wherein the smearing method is 2 times per day, and each time is 0.5g per mouse, and the smearing is continuously carried out for 7 days; (2) on the 5 th day of the experiment, each mouse was anesthetized with sodium pentobarbital 30mg/kg, and the mice were given passive cutaneous hypersensitivity by intradermal injection of 20 μ L of anti-ovalbumin serum into the right ear of all mice; (3) injecting 0.25ml of 0.5% Ewensa blue egg protein mixed solution into tail vein of mice 48h after sensitization, removing neck after 30min, killing all mice, and cutting auricles of the mice; (4) the excised mouse auricles were placed in 0.75ml of a 1mol/L KOH solution and digested overnight at 37 ℃; (5) 3.5ml of 0.2mol/LH was added to the KOH solution₃PO₄Mixing the solution with 9.1ml acetone solution to obtain mixed solution, shaking with a vortex device, centrifuging the mixed solution at 2500r/min for 15min, and extracting supernatant; (6) the absorbance of the supernatant was measured at a wavelength of 610nm, and the rate of blue spot inhibition was calculated. The results of this example III are shown in Table 1.

Example four

Gel medium dosage group

(1) The gel prepared in example one or example two (same below) was applied to the inner and outer surfaces of the auricle of all mice at a medium dose of 5mg/kg for 2 times per day (0.5 g/mouse per time) for 7 consecutive days. The rest of the steps are the same as those in the embodiment, and are not described herein. The results of this example four are shown in Table 1.

EXAMPLE five

Gel high dose group

(1) The gel prepared in example one or example two (same below) was applied to the inner and outer surfaces of the auricle of all mice at a high dose of 10mg/kg for 7 days at a dose of 0.5 g/mouse per application for 2 times per day. The rest of the steps are the same as those in the embodiment, and are not described herein. The results of this example five are shown in Table 1.

Comparative example 1

Blank group

Preparation of anti-ovalbumin serum according to preparation method of example III

Passive cutaneous anaphylaxis (RCA reaction) assay

Taking 8 healthy KM mice, half each mouse, and (1) injecting 10 mu L of physiological saline into each mouse every day for 7 days continuously; (2) on the 5 th day of the experiment, each mouse was anesthetized with sodium pentobarbital 30mg/kg, and 20. mu.L of physiological saline was intradermally injected into the right ear of all mice; (3) after 48h, 0.25ml of 0.5% Ewensa blue egg protein mixed solution is injected into the tail vein of each mouse, all the mice are killed by removing necks after 30min, and the auricles of the mice are cut off; (4) the excised mouse auricles were placed in 0.75ml of a 1mol/L KOH solution and digested overnight at 37 ℃; (5) 3.5ml of 0.2mol/LH was added to the KOH solution₃PO₄Mixing the solution with 9.1ml acetone solution to obtain mixed solution, shaking with a vortex device, centrifuging at 2500r/min for 15min, and collecting supernatant; (6) the absorbance of the supernatant was measured at a wavelength of 610nm, and the rate of blue spot inhibition was calculated. The results of this comparative example one are shown in table 1.

Comparative example No. two

Model set

Passive cutaneous anaphylaxis (RCA reaction) assay

Taking 8 healthy KM mice, half each mouse, and (1) injecting 10 mu L of physiological saline into each mouse every day for 7 days continuously; (2) on the 5 th day of the experiment, each mouse was anesthetized with sodium pentobarbital 30mg/kg, and the mice were given passive cutaneous hypersensitivity by intradermal injection of 20 μ L of anti-ovalbumin serum into the right ear of all mice; (3) injecting 0.25ml of 0.5% Evans blue egg protein mixed solution into tail vein of each mouse after the mice are sensitized for 48h, removing neck after 30min, killing all mice, and cutting auricles of the mice; (4) the excised mouse auricles were placed in 0.75ml of a 1mol/L KOH solution and digested overnight at 37 ℃; (5) 3.5ml of 0.2mol/LH was added to the KOH solution₃PO₄Solution and 9.1ml acetone solution setShaking the resultant mixed solution with a vortex device, centrifuging at 2500r/min for 15min, and extracting supernatant; (6) the absorbance of the supernatant was measured at a wavelength of 610nm, and the rate of blue spot inhibition was calculated. The results of this comparative example two are shown in table 1.

Comparative example No. three

Positive control group

Passive cutaneous anaphylaxis (RCA reaction) assay

Taking 8 healthy KM mice, half each mouse, and (1) injecting mometasone furoate gel into each mouse according to the dose of 5mg/kg every day for 7 days continuously; (2) on the 5 th day of the experiment, each mouse was anesthetized with sodium pentobarbital 30mg/kg, and the mice were given passive cutaneous hypersensitivity by intradermal injection of 20 μ L of anti-ovalbumin serum into the right ear of all mice; (3) injecting 0.25ml of 0.5% Evans blue egg protein mixed solution into tail vein of each mouse after the mice are sensitized for 48h, removing neck after 30min, killing all mice, and cutting auricles of the mice; (4) the excised mouse auricles were placed in 0.75ml of a 1mol/L KOH solution and digested overnight at 37 ℃; (5) 3.5ml of 0.2mol/LH was added to the KOH solution₃PO₄Mixing the solution with 9.1ml acetone solution to obtain mixed solution, shaking with a vortex device, centrifuging at 2500r/min for 15min, and collecting supernatant; (6) the absorbance of the supernatant was measured at a wavelength of 610nm, and the rate of blue spot inhibition was calculated. The results of this comparative example three are shown in table 1.

Is metered by

Shown, statistically processed using the t-test. When P is less than 0.05, the difference between the two groups has statistical significance.

TABLE 1 Effect of gelling Agents on RCA reaction (

n＝8)

Note: compared with the blank control group, the composition of the composition,^**p is less than 0.01; compared with the model control group,^##P＜0.01。

the anti-ovalbumin serum is prepared by taking ovalbumin as an antigen. And then injecting the gel into the skin of the same animal, smearing corresponding medicines, injecting a mixed solution of evans blue dye and antigen intravenously after 24 hours to dye the antiserum injection part in the skin, judging the intensity of the allergic reaction of each group of passive skin according to the optical density value of the dyed skin, wherein the result is shown in the table 1, and after the gel is smeared for 7 days, compared with the comparative example II, the blue spot area of the skin of the auricular corridor of the mouse in the example III, the example IV and the example V is reduced, the color is obviously weakened, the absorbance value is reduced, and the difference is obvious (P is less than 0.01), so that the gel with low, medium and high doses can obviously inhibit the allergic reaction of the heterogeneous passive skin of the mouse ear, and the gel prepared by the invention has the antiallergic effect.

EXAMPLE six

48 SD rats were divided into 6 groups of 8 rats each. After adaptive feeding for 3 days, the gel is randomly divided into a blank group and a model group, the gel prepared in the first embodiment or the second embodiment is divided into a high-dose 10mg/kg gel group according to the dosage, the group with the medium dose of 5mg/kg gel, the group with the low dose of 2.5mg/kg gel and a positive control drug group, 6 groups all have hair removal on two parts of the back one day before the experiment, the area of a first part is about 2cm multiplied by 3cm, the area of a second part is about 4cm multiplied by 4cm, a blank group is coated with distilled water, an ACD model is established by the model group, after 24 hours of hair removal, 7% of 2, 4-Dinitrofluorobenzene (DNFB) solution is coated on the first part of a hair removal area for sensitization, after 2 weeks, 0.1% of DNFB solution is coated on the second part of the hair removal area for excitation, a rat is irritated and has harassing behavior, and the skin on the second part of the back has obvious erythema, swelling and pimple, and the ACD model of the rat is obtained. Each administration group is coated with 0.5g of gel agent 12h after molding, and is gently massaged for 10 circles, 2 times per day with 1 week of treatment course, and positive control group is coated with mometasone furoate gel (5mg/kg, purchased from Calif.) outside. Exciting the second place with 0.1% DNFB solution after 1 week, externally coating gel for 12h after excitation, externally coating mometasone furoate gel for positive control group, collecting blood of experimental animal under anesthesia state in EP tube after 48h, centrifuging, processing, separating serum, and detecting IL-4 and TNF-alpha in serum by ELISA; after the blood sampling experiment is completed, rats are immediately dislocated and killed, sensitized skin with the same area (2cm multiplied by 2cm) as that of the second part is cut off, and the histamine concentration of the tissue homogenate is determined by an enzyme-linked immunosorbent assay (ELISA) kit in the tissue homogenate determination.

The results of this example six are shown in Table 2.

Is metered by

Shown, statistically processed using the t-test. The difference between the two groups is significant when P is less than 0.05.

TABLE 2 Effect of gels on ACD rats: (

n＝8)

Note: compared with the blank control group, the composition of the composition,^**p is less than 0.01; in comparison with the set of models,^#P＜0.05，^##P＜0.01。

TNF-alpha is a mediator involved in mediating the development of allergic reactions, and IL-4 inhibits the development of allergic reactions. Histamine is one of the most characteristic blood vessel strong active substances, has strong correlation with anaphylactic reaction, can cause skin itch, increase vascular permeability and other anaphylactic reactions, and the change of the histamine level can be used as an important index for detecting the anaphylactic reaction, so the anti-allergic effect of the brocade gel preparation is reflected by detecting the indexes. The ACD model group rats have obvious erythema, swelling and papules after skin sensitization on the back A, obvious diffuse erythema, unclear border and slight swelling are observed on the skin on the back B of the rats 48 hours after excitation, and then the concentration of IL-4, TNF-alpha and histamine in serum is detected, and the result is shown in table 2, compared with the blank group, the IL-4 content of the model group rats is obviously reduced (P is less than 0.01), and the content of TNF-alpha and the concentration of histamine are obviously increased (P is less than 0.01); compared with the model group, the low, medium and high dose groups of the gel can obviously reduce the content of TNF-alpha and the concentration of histamine (P is less than 0.01) and increase the content of IL-4 (P is less than 0.05). The result shows that the gel can inhibit the release of histamine by reducing the release of TNF-alpha and increasing the secretion of IL-4, and finally inhibit the occurrence of anaphylactic reaction and inflammatory reaction, thereby playing the role of antianaphylaxis.

EXAMPLE seven

Taking 40 healthy Wistar rats, half male and half female, randomly dividing into 4 groups, namely a blank group, a model group, a positive drug group (smearing tretinoin cream for 0.1g times per day, purchased from Hubei Kangzheng pharmaceutical Co., Ltd.) and a gel group consisting of the gel prepared in the first embodiment or the second embodiment; except for the blank group, a rat otic acne model was prepared using classical rat modeling, by injecting propionibacterium acnes intradermally into auricles (purchased from institute of microbiology, department of Chinese academy of sciences, accession number: 112236) to simulate the in vitro manifestation of acne and the onset of inflammatory reactions. Acetobacter acidi strain liquid (1X 10) is injected into auricle of right ear of each rat⁶cfu/mL) 50. mu.L, 1 time per day, for 3 consecutive days. Day 3, each rat was administered another intraperitoneal injection of propionibacterium solution (1X 10)⁶cfu/mL)1mL, 1 time per day for 7 consecutive days. Each group was 10 rats and administered for 14 days.

The thickness of the auricle was measured with a vernier caliper before each group of mice was modeled. After modeling, observing the skin state of the right ear of each group of rats, and determining whether the modeling is successful. Continuously applying for 14 days, weighing, measuring auricle thickness, taking blood from abdominal aorta, standing for 30min, centrifuging at 4 deg.C in a low temperature centrifuge at 12000 rpm for 10 min. Collecting the upper layer serum, and storing in a refrigerator at-20 deg.C. The contents of IL-1 alpha, IL-1 beta, IL-6 and TNF-alpha in rat serum are measured by enzyme-linked immunosorbent assay (ELISA).

The results of this example seven are shown in Table 3.

Table 3 ear swelling rate measurements for each group of rats (n-10,

)

note: in comparison with the blank set, the results,^*p is less than 0.01; in comparison with the set of models,^#P＜0.01；

mean ± standard deviation.

Example seven results analysis

(1) The gel has therapeutic and relieving effects on auricle swelling

Compared with the blank group, the auricle of the model group rat has obvious swelling phenomenon. Compared with the blank group, the auricle swelling rate of the model group rat is obviously increased, and the difference has statistical significance (P is less than 0.01); after administration, auricle swelling rates of rats of the positive drug group and the gel group are obviously lower than that of a model group (P is less than 0.01), which shows that the positive drug and the gel have obvious inhibition effect on inflammation of the rats of the acne model.

(2) The gel can inhibit inflammatory factor level in serum of rat with acne model

Note: the brocade lantern group shown in figure 1 is the gel group of the seventh embodiment

As shown in FIG. 1, compared with the blank group, the model group has a significant increase in inflammatory factors and a significant decrease in anti-inflammatory factors in the rat serum, and it can be seen that IL-1 alpha, TNF-alpha and IL-6 in the rat serum are all significantly increased and IL-2 is significantly decreased (P < 0.01). Compared with the model group, IL-1 alpha, TNF-alpha and IL-6 in the serum of the rat in the gel group are all obviously reduced, and IL-2 is obviously increased (P is less than 0.01); IL-1 alpha, TNF-alpha and IL-6 in the serum of rats of the PC group are all obviously reduced, IL-2 is obviously increased, and the difference has statistical significance (P is less than 0.05).

The seven results of the embodiment prove that the gel can reduce the contents of inflammatory factors IL-1 alpha, IL-6 and TNF-alpha in serum of rats of an acne model, can inhibit the content of the inflammatory factor IL-2, and shows that the gel has obvious anti-inflammatory effect in the treatment of acne.

The above description is only an embodiment of the present invention, and the protection scope of the present invention is not limited to the above embodiment, and all technical solutions belonging to the idea of the present invention belong to the protection scope of the present invention. It should be noted that modifications and embellishments within the scope of the present invention may be made by those skilled in the art without departing from the principle of the present invention, and such modifications and embellishments should also be considered as within the scope of the present invention.

Claims

1. A method for preparing gel by using calyx seu fructus physalis extract is characterized in that: the preparation method comprises the following steps:

Step two: preparation of 2% carbomer solution

Step three: preparation of gel

2. The method for preparing the gel from the calyx seu fructus physalis extract as claimed in claim 1, which is characterized in that: the 70% ethanol is prepared by 95% ethanol or recovered alcohol.

3. The method for preparing the gel from the calyx seu fructus physalis extract as claimed in claim 1, which is characterized in that: the weight ratio of the ethylparaben to the menthol is 1: 1.

4. The method for preparing the gel from the calyx seu fructus physalis extract as claimed in claim 1, which is characterized in that: the amount of the saturated sodium bicarbonate solution added was 10% by weight of the second mixed solution.

5. The method for preparing the gel from the calyx seu fructus physalis extract as claimed in claim 1, which is characterized in that: the pH was 6.5.

6. Use of the gel prepared by the preparation method of any one of claims 1 to 5 as an antiallergic agent.

7. Use of the gel prepared by the preparation method of any one of claims 1 to 5 for treating acne.