Traditional Chinese medicine composition and traditional Chinese medicine granules for treating acne as well as preparation method and application of traditional Chinese medicine granules

Abstract

The invention provides a traditional Chinese medicine composition and traditional Chinese medicine granules for treating acne as well as a preparation method and application thereof, and belongs to the technical field of traditional Chinese medicine compositions. The invention provides a traditional Chinese medicine composition for treating acne. In the traditional Chinese medicine composition, white mulberry root-bark, oldenlandia diffusa and the like are sweet and cold in nature, have the effect of clearing away heat and toxic materials and are monarch drugs; the scutellaria baicalensis, the honeysuckle, the selfheal and the like are cold and cool in nature, are medicines for clearing heat and removing dampness, are matched with the oldenlandia diffusa and the white mulberry root-bark to have a mutual promotion effect, and are ministerial medicines; the rhizoma alismatis is sweet and cold in nature and taste, has the effects of promoting diuresis, excreting dampness, discharging heat and enabling fire to flow out of the bladder, and has a positive and assistant effect; the radix rehmanniae recen, the radix polygonati officinalis, the charred hawthorn and the like have the effects of nourishing yin, moisturizing the lung, strengthening the body resistance and eliminating evil, have a counter-adjuvant effect and are used as assistants and guides; the traditional Chinese medicine composition has the effects of clearing away lung-heat, purging fire and removing dampness. The traditional Chinese medicine composition can be used for treating heat-toxic acne vulgaris and can improve and treat inflammation caused by propionibacterium acnes. And the traditional Chinese medicine composition is convenient to take orally and high in patient compliance, and has a remarkable acne treatment effect.

Description

Technical Field

[0001] The invention belongs to the technical field of Chinese medicine compositions, and in particular relates to a Chinese medicine composition for treating acne, Chinese medicine granules, and a preparation method and application thereof. Background Art

[0002] Acne is an inflammatory skin disease that affects the sebaceous glands and hair follicles. The main pathogenic mechanism is excessive keratinization of the sebaceous ducts, leading to obstruction of the sebaceous glands.

[0003] Traditional Chinese medicine believes that the causes of acne are wind-heat, blood-heat, stagnation of heat in the lung meridian, blood, yin deficiency, and Chong-Ren disorder, etc., and according to the different symptoms of the patients, acne is subdivided into different types such as damp-heat accumulation and wind-heat in the lung meridian. Therefore, traditional Chinese medicine treats acne by clearing the lungs, relieving heat, and removing dampness. However, different Chinese medicine formulas have different therapeutic effects. At present, the efficacy of Chinese medicine formulas for treating acne is mainly clearing heat and reducing inflammation, and the therapeutic effect needs to be further improved. Summary of the invention

[0004] The purpose of the present invention is to provide a Chinese medicine composition, Chinese medicine granules, preparation method and application for treating acne. The Chinese medicine composition of the present invention has a significant effect in treating acne.

[0005] The invention provides a traditional Chinese medicine composition for treating acne, comprising the following raw materials in parts by weight: 1-2 parts of mulberry bark, 4-6 parts of oldenlandia diffusa, 1-3 parts of scutellaria baicalensis, 1-3 parts of honeysuckle, 1-3 parts of qinpi, 1-2 parts of selfheal, 1-3 parts of oriental water plantain, 1-2 parts of rehmannia root, 1-2 parts of polygonatum odoratum and 1.5-3.5 parts of hawthorn charcoal.

[0006] Preferably, the following raw materials are included in parts by mass: 1 part of Morus alba bark, 5 parts of Hedyotis diffusa, 2 parts of Scutellaria baicalensis, 2 parts of Honeysuckle, 2 parts of Qinpi, 1.5 parts of Prunella Vulgaris, 2 parts of Alisma orientalis, 1.5 parts of Rehmannia root, 1.5 parts of Polygonatum odoratum and 2.5 parts of Hawthorn charcoal.

[0007] The present invention also provides a method for preparing the Chinese medicine composition described in the above scheme, comprising the following steps:

[0008] The Chinese medicine composition is mixed with water and boiled to obtain a decoction.

[0009] Preferably, after obtaining the decoction, the method further comprises filtering the decoction to obtain a filtrate; and concentrating the filtrate to obtain an extract.

[0010] The present invention also provides the use of the Chinese medicine composition described in the above scheme or the Chinese medicine composition prepared by the preparation method in preparing a drug for treating acne.

[0011] The present invention also provides the use of the Chinese medicine composition described in the above scheme or the Chinese medicine composition prepared by the preparation method in preparing a drug for inhibiting Propionibacterium acnes infection or treating or alleviating inflammation caused by Propionibacterium acnes.

[0012] Preferably, the dosage form of the drug includes an oral preparation.

[0013] The present invention also provides a Chinese medicine granule for treating acne, comprising a Chinese medicine composition for treating acne and pharmaceutically acceptable excipients; the Chinese medicine composition for treating acne is the Chinese medicine composition described in the above scheme or the Chinese medicine composition prepared by the preparation method.

[0014] Preferably, the auxiliary material includes one or more of dextrin, sucrose, lactose, soluble starch and mannitol.

[0015] Preferably, the auxiliary material is dextrin; when the traditional Chinese medicine composition is an extract with a relative density of 1.34, when the filtrate of the decoction is concentrated to a relative density of 1.34, the mass ratio of the traditional Chinese medicine composition to the auxiliary material is 1: (3-4).

[0016] The invention provides a Chinese medicine composition for treating acne. The Chinese medicine composition of the invention is based on the treatment principle of clearing heat and removing dampness, nourishing yin and moistening the lungs. Among them, Morus alba and Hedyotis diffusa are sweet and cold in nature, and have the effect of clearing heat and detoxicating, and are the main medicines; Scutellaria baicalensis, honeysuckle, Prunella vulgaris and the like are cold in nature, and are medicines for clearing heat and removing dampness, and have a complementary effect with Hedyotis diffusa and Morus alba, and are the ministerial medicines; and Alisma orientalis is sweet and cold in nature, and has the effect of diuresis, infiltration of dampness, and heat release, so that the fire comes out of the bladder, and has a positive adjuvant effect; Radix Rehmanniae, Polygonatum odoratum, Crataegus charcoal and the like have the function of nourishing yin and moistening the lungs, strengthening the body and eliminating evil, and have an anti-adjuvant effect, and are the adjuvants; the combination of the medicines has the effect of clearing the lungs and purging fire, and removing dampness, and can clear heat and purge fire, remove dampness and resolve phlegm, and treat acne caused by damp-heat in the lungs. The Chinese medicine composition of the invention can be used for common acne with heat toxicity, and can improve and treat inflammation caused by Propionibacterium acnes. Moreover, the invention is convenient to take orally, has strong patient compliance, and has a significant acne treatment effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0018] Figure 1 This is a statistical chart comparing the molding rates of Chinese medicine granules with different drug-excipient ratios;

[0019] Figure 2 It is a statistical chart comparing the IL-1β concentration in the cell supernatant;

[0020] Figure 3 It is a statistical graph comparing the concentration of IL-6 in the cell supernatant;

[0021] Figure 4 It is a statistical graph comparing the concentration of IL-8 in the cell supernatant;

[0022] Figure 5 It is a statistical graph comparing the concentration of TNF-α in the cell supernatant;

[0023] Figure 6 It is a statistical diagram for the comparison of CD80 expression in cells;

[0024] Figure 7 It is a statistical diagram for the comparison of CD86 expression in cells;

[0025] Figure 8 It is a statistical chart comparing the concentration of IL-1β in mouse serum;

[0026] Fig. 9 It is a statistical chart comparing the concentration of IL-6 in mouse serum;

[0027] Fig.10 It is a statistical chart comparing the concentration of IL-8 in mouse serum;

[0028] Fig.11 This is a statistical chart of the thickness of the stratum corneum on the back of the mouse;

[0029] Fig.12 Comparison of HE-stained sections of mouse back skin; A is the blank group, B is the model group, C is the positive drug group, D is the low-concentration group, E is the medium-concentration group, and F is the high-concentration group. The scale is 100 μm. DETAILED DESCRIPTION

[0030] The invention provides a traditional Chinese medicine composition for treating acne, comprising the following raw materials in parts by weight: 1-2 parts of mulberry bark, 4-6 parts of oldenlandia diffusa, 1-3 parts of scutellaria baicalensis, 1-3 parts of honeysuckle, 1-3 parts of qinpi, 1-2 parts of selfheal, 1-3 parts of oriental water plantain, 1-2 parts of rehmannia root, 1-2 parts of polygonatum odoratum and 1.5-3.5 parts of hawthorn charcoal.

[0031] In the specific implementation process of the present invention, the Chinese medicine composition includes the following raw materials in parts by weight: 1 part of mulberry bark, 5 parts of oldenlandia diffusa, 2 parts of scutellaria baicalensis, 2 parts of honeysuckle, 2 parts of qinpi, 1.5 parts of selfheal, 2 parts of oriental water plantain, 1.5 parts of rehmannia root, 1.5 parts of polygonatum odoratum and 2.5 parts of hawthorn charcoal.

[0032] In one embodiment of the present invention, the Chinese medicine composition is composed of the following raw materials in parts by mass: 1 part of Morus alba bark, 5 parts of Hedyotis diffusa, 2 parts of Scutellaria baicalensis, 2 parts of Honeysuckle, 2 parts of Qinpi, 1.5 parts of Prunella vulgaris, 2 parts of Alisma orientalis, 1.5 parts of Rehmannia root, 1.5 parts of Polygonatum odoratum and 2.5 parts of Hawthorn charcoal.

[0033] In the specific implementation process of the present invention, among them, Fraxinus chinensis Roxb is the dried branch bark or dry bark of Fraxinus chinensis Roxb; Alisma plantago-aquatica Linn. is the dried tuber of Alisma plantago-aquatica Linn.; the origin of hawthorn charcoal is the dried mature fruit of Crataegus pinnatifida Bge.; the processing technology and quality standards of Hedyotis diffusa and hawthorn charcoal are implemented according to the 2018 edition of "Shanghai Traditional Chinese Medicine Piece Processing Specifications".

[0034] The Chinese medicine composition of the present invention is based on the treatment principle of clearing away heat and removing dampness, nourishing yin and moistening the lungs, wherein the mulberry bark and white flower oldenlandia diffusa are sweet and cold in nature, and have the effect of clearing away heat and detoxicating, and are the main medicines; the scutellaria baicalensis, honeysuckle, and selfheal are cold in nature, and are medicines for clearing away heat and removing dampness, and have a mutual effect with white flower oldenlandia diffusa and mulberry bark, and are the ministerial medicines; the oriental rhizome is sweet and cold in nature, and has the effect of diuresis, infiltration of dampness, and heat release, so that the fire comes out of the bladder, and has a positive auxiliary effect; the raw rehmannia root, polygonatum odoratum, and hawthorn charcoal have the merit of nourishing yin and moistening the lungs, strengthening the body and eliminating evil, and have an anti-adjuvant effect, and are the assistants; the combination of all the medicines has the effect of clearing the lungs and purging fire, and removing dampness, and can clear away heat and purge fire, remove dampness and resolve phlegm, and treat acne caused by damp-heat obstructing the lungs. The Chinese medicine composition of the present invention has a rigorous raw material combination, can be used for common acne with heat toxicity, can improve and treat inflammation caused by Propionibacterium acnes, and has a good therapeutic effect in clinical applications. In addition, the Chinese medicine composition of the present invention is convenient for oral administration, greatly reduces the cost of medication and the toxic and side effects of drugs compared to existing Western medicine treatments, and improves patient compliance.

[0035] The present invention also provides a method for preparing the Chinese medicine composition described in the above scheme, comprising the following steps: mixing the Chinese medicine composition with water, decocting the mixture, and obtaining a decoction.

[0036] In the specific implementation of the present invention, the number of decoctions is 1 to 3 times;

[0037] In one embodiment of the present invention, the decoction is as follows: a first decoction is performed, solid-liquid separation is performed, and a first extract and a solid component are obtained; the solid component is mixed with water, a second decoction is performed, solid-liquid separation is performed, and a second extract is collected; the first extract and the second extract are combined to obtain a decoction; the mass of water used in the first decoction is 8 to 11 times the mass of the traditional Chinese medicine composition, and further is 9 to 10 times; the time for the first decoction is 1 to 2.5 hours, and further is 1.5 to 2 hours; the mass of water used in the second decoction is 6 to 9 times the mass of the traditional Chinese medicine composition, and further is 7 to 8 times; the time for the second decoction is 0.5 to 1.5 hours, and further is 1 hour.

[0038] After obtaining the decoction, the present invention further comprises filtering the decoction to obtain a filtrate; concentrating the filtrate to obtain an extract; the concentration is to concentrate to a relative density of 1.2 to 1.4, further to 1.34; the concentration temperature is 80°C.

[0039] After obtaining the extract, the present invention freeze-dries the extract to obtain freeze-dried powder; or, the present invention mixes the extract with auxiliary materials, granulates, and obtains Chinese medicine granules; the vacuum degree of the freeze-drying is -0.01 to -0.06 MPa.

[0040] The present invention also provides the use of the Chinese medicine composition described in the above scheme or the Chinese medicine composition prepared by the preparation method in preparing a drug for treating acne.

[0041] In the present invention, the acne includes acne caused by damp-heat blocking the lungs or acne vulgaris caused by thermal toxicity; the acne vulgaris caused by thermal toxicity is acne vulgaris caused by the proliferation of Propionibacterium acnes.

[0042] The present invention also provides the use of the Chinese medicine composition described in the above scheme or the Chinese medicine composition prepared by the preparation method in preparing a drug for inhibiting Propionibacterium acnes infection or treating or alleviating inflammation caused by Propionibacterium acnes.

[0043] Propionibacterium acnes (scientific name Cutibacterium acnes) is a species of the genus Propionibacterium. Its proliferation in an anaerobic environment can cause an inflammatory response in acne and lead to the onset of the disease.

[0044] In the specific implementation of the present invention, the Chinese medicine composition is the active ingredient of the drug, and further, the Chinese medicine composition is the only active ingredient of the drug; the dosage form of the drug includes oral preparations; the oral preparations include granules or powders.

[0045] In the specific implementation process of the present invention, the drug is a unit dose preparation; based on mice, the unit dose preparation is prepared to facilitate administration of 0.17 to 13 g of the Chinese medicine composition / kg; based on humans, the content of the Chinese medicine composition in the unit dose preparation is 30 g.

[0046] The present invention also provides a Chinese medicine granule for treating acne, comprising a Chinese medicine composition for treating acne and pharmaceutically acceptable excipients; the Chinese medicine composition for treating acne is the Chinese medicine composition described in the above scheme or the Chinese medicine composition prepared by the preparation method.

[0047] In the specific implementation of the present invention, the auxiliary material includes one or more of dextrin, sucrose, lactose, soluble starch and mannitol.

[0048] In one embodiment of the present invention, the auxiliary material is dextrin; when the Chinese medicine composition is an extract with a relative density of 1.34, when the filtrate of the decoction is concentrated to a relative density of 1.34, the mass ratio of the Chinese medicine composition extract and the auxiliary material is 1: (3-4), further 1: (3.3-3.7), and further 1: 3.5, and this ratio has a high molding rate.

[0049] The Chinese medicine granules of the present invention can improve the medication compliance and safety of patients.

[0050] To further illustrate the present invention, a Chinese medicine composition for treating acne, Chinese medicine granules, preparation method and application provided by the present invention are described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0051] Example 1

[0052] A method for preparing Chinese medicine granules for treating acne, accurately weighing the following raw materials:

[0053] 150g of mulberry bark, 500g of oldenlandia diffusa, 200g of scutellaria baicalensis, 200g of honeysuckle, 200g of qinpi, 150g of selfheal, 200g of oriental water plantain, 150g of rehmannia root, 150g of polygonatum odoratum and 250g of hawthorn charcoal.

[0054] Preparation process:

[0055] Take the above mass of raw materials, add 10 times the amount of water, i.e. 21500 mL of water, decoct for 2 hours and filter for later use, add 8 times the amount of water for the second decoction, decoct for 1 hour and filter, combine the two filtrates, and concentrate at 80°C to a relative density of 1.34 to obtain a Chinese medicine composition extract; add the auxiliary material dextrin to the Chinese medicine composition extract, mix and granulate at a mass ratio of the Chinese medicine composition extract to the auxiliary material of 1:3, dry and granulate to obtain granules 1.

[0056] Example 2

[0057] A method for preparing Chinese medicine granules for treating acne, accurately weighing the following raw materials:

[0058] 150g of mulberry bark, 500g of oldenlandia diffusa, 200g of scutellaria baicalensis, 200g of honeysuckle, 200g of qinpi, 150g of selfheal, 200g of oriental water plantain, 150g of rehmannia root, 150g of polygonatum odoratum and 250g of hawthorn charcoal.

[0059] Preparation process:

[0060] Except that the mass ratio of the Chinese medicine composition extract to the auxiliary material is 1:3.3, the rest is the same as Example 1 to obtain granules 2.

[0061] Example 3

[0062] A method for preparing Chinese medicine granules for treating acne, accurately weighing the following raw materials:

[0063] 150g of mulberry bark, 500g of oldenlandia diffusa, 200g of scutellaria baicalensis, 200g of honeysuckle, 200g of qinpi, 150g of selfheal, 200g of oriental water plantain, 150g of rehmannia glutinosa, 150g of polygonatum odoratum and 250g of hawthorn charcoal.

[0064] Preparation process:

[0065] Except that the mass ratio of the Chinese medicine composition extract to the auxiliary materials is 1:3.5, the rest is the same as Example 1 to obtain granules 3.

[0066] Example 4

[0067] A method for preparing Chinese medicine granules for treating acne, accurately weighing the following raw materials:

[0068] 150g of mulberry bark, 500g of oldenlandia diffusa, 200g of scutellaria baicalensis, 200g of honeysuckle, 200g of qinpi, 150g of selfheal, 200g of oriental water plantain, 150g of rehmannia glutinosa, 150g of polygonatum odoratum and 250g of hawthorn charcoal.

[0069] Preparation process:

[0070] Except that the mass ratio of the Chinese medicine composition extract to the auxiliary material is 1:3.7, the rest is the same as Example 1 to obtain granules 4.

[0071] Example 5

[0072] A method for preparing Chinese medicine granules for treating acne, accurately weighing the following raw materials:

[0073] 150g of mulberry bark, 500g of oldenlandia diffusa, 200g of scutellaria baicalensis, 200g of honeysuckle, 200g of qinpi, 150g of selfheal, 200g of oriental water plantain, 150g of rehmannia glutinosa, 150g of polygonatum odoratum and 250g of hawthorn charcoal.

[0074] Preparation process:

[0075] Except that the mass ratio of the Chinese medicine composition extract to the auxiliary material is 1:4, the rest is the same as Example 1 to obtain granules 5.

[0076] Test Example 1

[0077] The forming rate of granules 1, 2, 3, 4 and 5 obtained by the preparation method described in Examples 1 to 5 was measured. The determination method refers to the Chinese Pharmacopoeia General Rules for Preparations 0982 Particle Size and Particle Size Distribution Determination Method Second Method Double Sieving Method, the sum of the particles that cannot pass through the No. 1 sieve and the particles that can pass through the No. 5 sieve shall not exceed 15%. Take the particles that can pass through the No. 1 sieve and the particles that cannot pass through the No. 5 sieve, weigh them, and calculate the forming rate. The calculation formula is as follows:

[0078] Forming rate = particle mass after sieving / particle mass before sieving × 100%.

[0079] See Table 1 for the results.

[0080] Table 1 Forming rate of Chinese medicine granules

[0081]

[0082]

[0083] Note: The mass ratio of extract and excipients of each Chinese medicine composition in the table was tested repeatedly 6 times, and each data is expressed as mean ± standard deviation (SD).

[0084] In traditional Chinese medicine preparations, granules not only improve the stability of decoctions and retain the effective ingredients in traditional decoctions, but also overcome the trouble of decoction, and are convenient to carry and transport, and also improve patient compliance. In addition, the uniformity and formability of the granule particle size mainly affect the water absorption and solubility of the granule. In order to enable the extract of the Chinese medicine composition of the present invention to be evenly mixed with the excipients and avoid wasting the Chinese medicine composition of the present invention, the formability of the granule is particularly important. According to the above content, it can be seen that the particle size and formability of the granule play an important role in the Chinese medicine preparation of the present invention. The forming rate of the Chinese medicine granules of the present invention prepared with different drug-excipient ratios is analyzed by Table 1. The results show that the best preparation method of the Chinese medicine granules of the present invention is granule 3, with a drug-excipient ratio of 1:3.5, and its forming rate is higher than that of granules 1, granules 2, granules 4 and granules 5.

[0085] The mass ratio of extracts and excipients of different Chinese medicine compositions and the particle size comparison of Chinese medicine granules are shown in the following table. Figure 1 ,Depend on Figure 1 It can be seen that the molding rate is optimal when the mass ratio of the Chinese medicine composition extract to the excipients is 1:3.5.

[0086] Example 6

[0087] A Chinese medicine composition (A) for treating acne, accurately weighing the following raw materials:

[0088] 150g of mulberry bark, 500g of oldenlandia diffusa, 200g of scutellaria baicalensis, 200g of honeysuckle, 200g of qinpi, 150g of selfheal, 200g of oriental water plantain, 150g of rehmannia glutinosa, 150g of polygonatum odoratum and 250g of hawthorn charcoal.

[0089] Prepare according to the above-mentioned preparation method of Chinese medicine granules:

[0090] Take the above mass of raw materials, add 10 times the amount of water, i.e. 21500mL of water, decoct for 2h and filter for later use. Add 8 times the amount of water for the second decoction, decoct for 1h and filter, combine the two filtrates, and concentrate at 80℃ to a relative density of 1.34 for later use.

[0091] Comparative Example 1 is relative to Example 6 in which Hedyotis diffusa is omitted.

[0092] A Chinese medicine composition (B) for treating acne, accurately weigh the following raw materials:

[0093] 150g of mulberry bark, 200g of scutellaria baicalensis, 200g of honeysuckle, 200g of qinpi, 150g of selfheal, 200g of oriental water plantain, 150g of rehmannia glutinosa, 150g of polygonatum odoratum, and 250g of hawthorn charcoal.

[0094] The preparation was carried out according to the preparation method of the Chinese medicine composition in Example 6.

[0095] In Comparative Example 2, relative to Example 6, Cortex Fraxini and Herba Prunellae were omitted.

[0096] A Chinese medicine composition (C) for treating acne, accurately weigh the following raw materials:

[0097] 150g of mulberry bark, 500g of oldenlandia diffusa, 200g of scutellaria baicalensis, 200g of honeysuckle, 200g of oriental water plantain, 150g of rehmannia glutinosa, 150g of polygonatum odoratum and 250g of hawthorn charcoal.

[0098] The preparation was carried out according to the preparation method of the Chinese medicine composition in Example 6.

[0099] Comparative Example 3 omitted Alisma orientalis relative to Example 6.

[0100] A Chinese medicine composition for treating acne (D), accurately weigh the following raw materials:

[0101] 150g of mulberry bark, 500g of oldenlandia diffusa, 200g of scutellaria baicalensis, 200g of honeysuckle, 200g of qinpi, 150g of selfheal, 150g of rehmannia glutinosa, 150g of polygonatum odoratum and 250g of hawthorn charcoal.

[0102] The preparation was carried out according to the preparation method of the Chinese medicine composition in Example 6.

[0103] Comparative Example 4 omits Rehmannia root, Polygonatum odoratum and Crataegus pinnatifida charcoal relative to Example 6.

[0104] A Chinese medicine composition (E) for treating acne, accurately weigh the following raw materials:

[0105] 150g of mulberry bark, 500g of oldenlandia diffusa, 200g of scutellaria baicalensis, 200g of honeysuckle, 200g of qinpi, 150g of selfheal and 200g of oriental water plantain.

[0106] The preparation was carried out according to the preparation method of the Chinese medicine composition in Example 6.

[0107] Test Example 2 The therapeutic effect of the freeze-dried powder of the Chinese medicine composition of the present invention on inflammation caused by Propionibacterium acnes.

[0108] Test cells: mouse mononuclear macrophage leukemia cells (RAW264.7).

[0109] Test drugs: The Chinese medicine compositions A, B, C, D, and E of Example 6 and Comparative Examples 1 to 4 were placed in freeze dryers to prepare freeze-dried powders for later use.

[0110] Preparation groups: blank group, model group, Chinese medicine composition group A (Group A), Chinese medicine composition group B (Group B), Chinese medicine composition group C (Group C), Chinese medicine composition group D (Group D), and Chinese medicine composition group E (Group E).

[0111] Test process:

[0112] 1. Take the lyophilized powder of Chinese medicine composition A, Chinese medicine composition B, Chinese medicine composition C, Chinese medicine composition D and Chinese medicine composition E respectively, add DMSO to prepare 50 μg / mL drug solution.

[0113] 1. Preparation of inactivated Propionibacterium acnes (C. acne): 10 μL of C. acne strain (purchased from Shuji Biotechnology Co., Ltd., catalog number 336649) was added to 50 mL of brain heart infusion medium and cultured at 37°C under anaerobic conditions for 24 h. The bacterial solution was taken out and the concentration was adjusted to 1×10 8 CFU / mL, discard the supernatant after centrifugation, add PBS to resuspend, and heat at 80℃ for 30min to inactivate for later use.

[0114] 2. RAW.264.7 cell plating and administration: RAW.264.7 cells were gently scraped from the culture flask to form a cell suspension, which was then centrifuged. After the centrifuge was discarded, DMEM medium was added to resuspend the cells for cell counting. The cells were plated at 1×10 6The number of cells was seeded in a 6-well plate and cultured for 24 hours. After 24 hours of culture, the blank group was not treated, and 10 μL of C. acne inactivation material was added to the other groups except the blank group, 1 μL of Chinese medicine composition A solution was added to the Chinese medicine composition A group, 1 μL of Chinese medicine composition B solution was added to the Chinese medicine composition B group, 1 μL of Chinese medicine composition C solution was added to the Chinese medicine composition C group, 1 μl of Chinese medicine composition D solution was added to the Chinese medicine composition D group, and 1 μL of Chinese medicine composition E solution was added to the Chinese medicine composition E group.

[0115] 1) Enzyme-linked immunosorbent assay (ELISA)

[0116] Collect the supernatant of each group of cells, centrifuge for 20 minutes, collect the supernatant, and detect the secretion changes of IL-1β, IL-6, IL-8, and TNF-α in the cells. Add 50μL of the cell supernatant sample after administration to the pre-coated ELISA plate, stick the sealing tape on it, mix well, and incubate at 37℃ for 50min. After washing, add the biotinylated antibody working solution, mix well, incubate at 37℃ for 50min, and then wash again. After adding the SABC complex, incubate at 37℃ for 30min. After washing three times, add the TMB colorimetric solution and incubate at 37℃ for 20min. After adding the stop solution, detect the absorbance at a wavelength of 450nm. The results of the secretion changes of IL-1β, IL-6, IL-8, and TNF-α in cells are respectively shown in Figure 2 to Figure 5 As shown in Table 2, all the Chinese medicine compositions showed a significant effect of reducing the release of inflammatory factors compared with the model group, and the ability of Chinese medicine composition A to reduce the release of inflammatory factors was significantly stronger than that of other Chinese medicine compositions.

[0117] Table 2 Concentration of inflammatory factors in cell supernatant

[0118]

[0119] 2) Real-time quantitative polymerase chain reaction (qRT-PCR)

[0120] Pretreatment solution was added to the collected cells to extract total RNA, and the total RNA was quantified. The total RNA was reverse transcribed using the HiScriptIII RT SuperMix for qPCR (+gDNAwiper) kit to obtain template DNA (cDNA), and qRT-PCR was performed on the LightCycler 480 System (Roche) using the ChamQ Universal SYBR qPCRMasterMix kit. The reaction conditions included pre-denaturation at 95.0°C for 30 seconds, followed by denaturation at 95.0°C for 6 seconds for 40 cycles, and annealing at 60.0°C for 30 seconds. GAPDH was used as an internal reference, and the relative quantification of the target gene was performed using 2-ΔΔCT The primers used are shown in Table 3.

[0121] Table 3 Sequences of PCR primers

[0122]

[0123]

[0124] All data are expressed as mean ± standard deviation (SD). Data were analyzed using one-way analysis of variance (ANOVA). Statistical analysis was performed using GraphPad Prism (9.0.0). P values ​​less than 0.05 were considered statistically significant (***P<0.001, ****P<0.0001).

[0125] The comparative statistical diagrams of CD80 and CD86 expression in cells are shown in Figure 6 and Figure 7 It can be seen that all Chinese medicine compositions can significantly reduce the expression of CD80 and CD86, and the ability of Chinese medicine composition A to reduce the expression of these two proteins is significantly stronger than that of the other groups.

[0126] Test Example 3 The therapeutic effect of the Chinese medicine granules of the present invention on acne.

[0127] Experimental animals: BABL / C female mice, 8 weeks old, weighing 17-19 g, and raised under specific pathogen-free conditions (SPF grade).

[0128] Test drug: According to the preparation method of the Chinese medicine granules in Example 3, dextrin is added to the Chinese medicine composition of the present invention and mixed to prepare Chinese medicine granules for use.

[0129] The experimental groups were: blank group, model group, positive drug (Compound Pearl Acne Tablets) group, high concentration group, medium concentration group and low concentration group.

[0130] Test process:

[0131] 1. Dosage design: The human dosage of the Chinese medicine granules of the present invention is 30g / time. According to the dosage conversion formula between humans and mice, the dosage of mice is 30g×0.0026 / 0.18=4.33g / kg. In this test, the human-mouse equivalent dose is the middle dose, three times the middle dose is the high dose, i.e. 13g / kg, and 1 / 3 times the middle dose is the low dose, i.e. 1.44g / kg. The positive drug used in this test is the compound pearl acne tablets (purchased from Shenyang Dongxin Pharmaceutical Co., Ltd., each tablet weighs 0.3g), and the dosage is 0.17g / kg after calculation according to the above formula.

[0132] 2. Preparation of Propionibacterium acnes: Take 10 μL of C. acne strains and add them to 50 mL of brain heart infusion medium. Incubate at 37°C under anaerobic conditions for 24 h. Take out the bacterial solution and adjust the concentration to 1×10 8 CFU / mL, discard the supernatant after centrifugation, add PBS to resuspend, and heat at 80℃ for 30min to inactivate for later use.

[0133] 3. The mice were depilated on their backs with a range of 2×2 cm and randomly divided into 6 groups, with 6 mice in each group: high concentration group, medium concentration group, low concentration group, model group, positive drug group and blank group. Except for the blank group, all other groups were subcutaneously injected with 100 μL of 1×10 8 CFU / mL of Propionibacterium acnes liquid was administered after 21 consecutive days of modeling. The concentration of the positive drug group was 0.006g / mL (0.17g / kg), the concentration of the low concentration group was 0.052g / mL (1.44g / kg), the medium concentration group was 0.154g / mL (4.3g / kg), and the high concentration group was 0.468g / mL (13g / kg). The model group and the blank group were given an equal amount of pure water by gavage. Each group of mice was gavaged once a day, 0.5mL each time, and subsequent experiments were carried out after 21 consecutive days of treatment.

[0134] 1)ELISA

[0135] Collect peripheral blood from mice, let it stand at room temperature for 24 hours, centrifuge it for 20 minutes, collect the supernatant (serum) and detect the secretion of IL-1β, IL-6, and IL-8 in mouse blood. Add 50μL of standard protein / sample diluent to the blank wells, and add 50μL of standard protein or test sample to each of the remaining wells, mix well and incubate at 37°C for 50 minutes. After washing, add biotinylated antibody working solution, mix well, incubate at 37°C for 50 minutes, wash again, add SABC complex and incubate at 37°C for 30 minutes, wash three times, add TMB colorimetric solution and incubate at 37°C for 20 minutes, add stop solution and detect absorbance at 450nm wavelength. The secretion of IL-1β, IL-6, and IL-8 in mouse blood can be found in the table below. Figure 8 to Figure 10 As shown in Table 4, the Chinese herbal granules of the present invention can exert the same anti-inflammatory effect as the positive drugs, and the anti-inflammatory effect of the Chinese herbal granules of the present invention shows obvious concentration dependence. This result shows that the Chinese herbal granules of the present invention can effectively alleviate the inflammation caused by acne vulgaris.

[0136] Table 4 Concentration of inflammatory factors in mouse serum

[0137]

[0138] 2) Hematoxylin-eosin staining (HE)

[0139] 2×2cm skin tissue was cut from the back of the mouse, wrapped in tin foil to prevent tissue curling from affecting the slices, and fixed in 4% paraformaldehyde solution for later use. After paraffin embedding and sectioning, the skin samples were stained with hematoxylin-eosin. The changes in the thickness of the stratum corneum and the inflammatory infiltration of the mouse skin tissue were observed under a 40× microscope.

[0140] All data are expressed as mean ± standard deviation (SD). Data were analyzed using one-way analysis of variance (ANOVA). Statistical analysis was performed using GraphPad Prism (9.0.0). P values ​​less than 0.05 were considered statistically significant (***P<0.001, ****P<0.0001).

[0141] Results Fig.11 , Fig.12 And Table 5. It can be seen that the Chinese medicine granules of the present invention and the positive drugs can reduce the thickness of the stratum corneum in the lesion skin of mice, which indicates that the Chinese medicine granules of the present invention have an alleviating effect on the thickening of the stratum corneum caused by acne vulgaris.

[0142] Table 5 Comparison of stratum corneum thickness

[0143]

[0144]

[0145] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A Chinese medicine composition for treating acne, characterized in that: The invention comprises the following raw materials in parts by weight: 1-2 parts of mulberry bark, 4-6 parts of oldenlandia diffusa, 1-3 parts of scutellaria baicalensis, 1-3 parts of honeysuckle, 1-3 parts of qinpi, 1-2 parts of selfheal, 1-3 parts of oriental water plantain, 1-2 parts of rehmannia root, 1-2 parts of polygonatum odoratum and 1.5-3.5 parts of hawthorn charcoal.

2. The Chinese medicine composition according to claim 1, characterized in that: The invention comprises the following raw materials in parts by weight: 1 part of mulberry bark, 5 parts of oldenlandia diffusa, 2 parts of scutellaria baicalensis, 2 parts of honeysuckle, 2 parts of qinpi, 1.5 parts of selfheal, 2 parts of oriental water plantain, 1.5 parts of rehmannia root, 1.5 parts of polygonatum odoratum and 2.5 parts of hawthorn charcoal.

3. The method for preparing the Chinese medicine composition according to claim 1 or 2, characterized in that: The following steps are involved: The Chinese medicine composition is mixed with water and boiled to obtain a decoction.

4. The preparation method according to claim 3, characterized in that: After obtaining the decoction, the method further includes filtering the decoction to obtain a filtrate; and concentrating the filtrate to obtain an extract.

5. Use of the Chinese medicine composition according to claim 1 or 2 or the Chinese medicine composition prepared by the preparation method according to claim 3 or 4 in preparing a drug for treating acne.

6. Use of the Chinese medicine composition according to claim 1 or 2 or the Chinese medicine composition prepared by the preparation method according to claim 3 or 4 in preparing a drug for inhibiting Propionibacterium acnes infection or treating or alleviating inflammation caused by Propionibacterium acnes.

7. The use according to claim 5 or 6, characterized in that: The dosage form of the drug includes oral preparations.

8. A Chinese medicine granule for treating acne, characterized in that: It includes a Chinese medicine composition for treating acne and pharmaceutically acceptable excipients; The Chinese medicine composition for treating acne is the Chinese medicine composition according to claim 1 or 2 or the Chinese medicine composition prepared by the preparation method according to claim 3 or 4.

9. The Chinese medicine granules according to claim 8, characterized in that: The auxiliary materials include one or more of dextrin, sucrose, lactose, soluble starch and mannitol.

10. The Chinese medicine granules according to claim 9, characterized in that: The auxiliary material is dextrin; when the Chinese medicine composition is an extract with a relative density of 1.34, when the filtrate of the decoction is concentrated to a relative density of 1.34, the mass ratio of the Chinese medicine composition to the auxiliary material is 1: (3-4).

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