A traditional Chinese medicine extract for anti-inflammatory and scar-reducing purposes, its preparation method, and its application.

Abstract

This invention discloses a traditional Chinese medicine extract for anti-inflammatory and scar-reducing purposes, its preparation method, and its application. The extract is obtained by mixing broken-cell wall Ganoderma lucidum spore powder and sea buckthorn seed powder in a weight ratio of 5-8:2-5, followed by dry granulation and supercritical CO2 extraction. This invention's traditional Chinese medicine extract for anti-inflammatory and scar-reducing purposes is prepared by mixing and granulating broken-cell wall Ganoderma lucidum spore powder and sea buckthorn seed oil, followed by supercritical CO2 extraction. The process is simple, energy-efficient, environmentally friendly, stable, and has high extraction efficiency. It exhibits significant anti-inflammatory and scar-reducing functions and has no toxic side effects.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, specifically relating to a traditional Chinese medicine extract for anti-inflammatory and scar-reducing purposes, its preparation method, and its application. Background Technology

[0002] Scars are fibrous tissue that replaces normal skin after injury and are an inevitable result of wound healing. Transforming growth factor-β1 (TGF-β1) is a pleiotropic cytokine involved in multiple biological processes of wound repair, including cell proliferation, cell migration, cell survival and apoptosis, immune regulation, epithelial-mesenchymal cell transition, and extracellular matrix formation. TGF-β1 can promote fibroblast expression, with a positive correlation to concentration, thus promoting fibroblast proliferation, increasing collagen secretion, and consequently leading to hypertrophic scars. Traditional Chinese medicine has a long history of anti-inflammatory and scar-reducing treatments, but its formulas are complex, and the products often have a strong taste, resulting in low public acceptance.

[0003] Currently, there are various scar-removing ointments on the market, but research on products formulated with natural plant oils is limited. There are no literature reports on the use of Ganoderma lucidum spore oil and sea buckthorn seed oil in the preparation of anti-inflammatory and scar-removing products. This invention involves mixing and granulating broken-cell wall Ganoderma lucidum spore powder and sea buckthorn seed powder, followed by supercritical CO2 extraction to obtain a traditional Chinese medicine extract containing both Ganoderma lucidum spore oil and sea buckthorn seed oil. The two ingredients work synergistically to prevent and treat local inflammation and abscesses caused by wound infections, inhibit fibroblast proliferation, and suppress the proliferative effect of TGF-β1 on fibroblasts, significantly enhancing the anti-inflammatory and scar-removing effects in animals.

[0004] Therefore, the present invention is of great significance in developing a product with anti-inflammatory and scar-reducing effects, which is formulated with Ganoderma lucidum spore oil and sea buckthorn seed oil. Summary of the Invention

[0005] The technical problem to be solved by this invention is to provide a traditional Chinese medicine extract containing Ganoderma lucidum spore oil and sea buckthorn seed oil with anti-inflammatory and scar-reducing properties. This extract, with natural plant oils as its components, is effective in reducing inflammation and scarring.

[0006] The objective of this invention is achieved through the following means:

[0007] A traditional Chinese medicine extract for anti-inflammatory and scar-reducing purposes is obtained by mixing broken-cell wall Ganoderma lucidum spore powder and sea buckthorn seed powder in a weight ratio of 5-8:2-5, followed by dry granulation and supercritical CO2 extraction.

[0008] Preferably, the broken-cell wall Ganoderma lucidum spore powder and sea buckthorn seed powder are mixed in a weight ratio of 5-7:3-5, and most preferably, the broken-cell wall Ganoderma lucidum spore powder and sea buckthorn seed powder are mixed in a weight ratio of 7:3.

[0009] Preferably, the drug extract is formulated into a topical preparation with pharmaceutically acceptable excipients.

[0010] Preferably, the topical preparation is a medicated oil or ointment.

[0011] A method for preparing a traditional Chinese medicine extract for anti-inflammatory and scar-reducing purposes, the method comprising the following steps:

[0012] (1) The cell wall of Ganoderma lucidum spore powder was broken, with a cell wall breaking rate of over 99%, to obtain broken cell wall Ganoderma lucidum spore powder;

[0013] (2) Grind the sea buckthorn seeds and pass them through a 40-50 mesh sieve to obtain sea buckthorn seed powder;

[0014] (3) Take broken-cell wall Ganoderma lucidum spore powder and sea buckthorn seed powder and mix them in a weight ratio of 5-8:2-5. Then granulate them by dry method to obtain mixed granules of broken-cell wall Ganoderma lucidum spore powder and sea buckthorn seed powder.

[0015] (4) The mixed particles of broken Ganoderma lucidum spore powder and sea buckthorn seed powder were subjected to supercritical CO2 extraction. The extraction conditions were: extraction vessel temperature 40-55℃, extraction vessel pressure 25-30MPa, and extraction time 2-4 hours to obtain a mixed extract of Ganoderma lucidum spore oil and sea buckthorn seed oil.

[0016] Preferably, the extraction conditions are: extraction vessel temperature 42-48℃, extraction vessel pressure 25-30MPa, and extraction time 2.5-4 hours.

[0017] The above-mentioned Chinese herbal extracts have shown significant effects in the preparation of anti-inflammatory and scar-reducing drugs.

[0018] Both Ganoderma lucidum spore oil and sea buckthorn seed oil are rich in nutrients. The combination of Ganoderma lucidum spore oil and sea buckthorn seed oil in extraction produces an extract that is a natural plant ingredient. This extract can reduce the risk of skin allergies and adverse reactions. It can work synergistically to reduce inflammation, promote healing and repair, help improve skin condition and reduce scar formation.

[0019] Compared with the prior art, the beneficial effects of the present invention are as follows: The present invention is a traditional Chinese medicine extract prepared by mixing and granulating broken Ganoderma lucidum spore powder and sea buckthorn seed oil and then using CO2 supercritical extraction. The process is simple, energy consumption is low, it is green and environmentally friendly, has good stability and high extraction efficiency, has significant anti-inflammatory and scar-reducing functions, and has no toxic side effects. Detailed Implementation

[0020] The present invention will be further explained and illustrated below through specific embodiments, but the scope of protection of the present invention is not limited to the following embodiments.

[0021] Example 1

[0022] (1) The cell wall of Ganoderma lucidum spore powder was broken, with a cell wall breaking rate of 99.5%, to obtain broken cell wall Ganoderma lucidum spore powder;

[0023] (2) Grind the sea buckthorn seeds and pass them through a 40-mesh sieve to obtain sea buckthorn seed powder;

[0024] (3) Take broken-cell wall Ganoderma lucidum spore powder and sea buckthorn seed powder and mix them in a weight ratio of 7:3. Then granulate them by dry method to obtain mixed granules of broken-cell wall Ganoderma lucidum spore powder and sea buckthorn seed powder.

[0025] (4) The mixed particles of broken Ganoderma lucidum spore powder and sea buckthorn seed powder were subjected to supercritical CO2 extraction. The extraction conditions were: extraction vessel temperature 45℃, extraction vessel pressure 25MPa, and extraction time 3 hours. The mixed extract of Ganoderma lucidum spore oil and sea buckthorn seed oil was obtained with an extraction rate of 24.2%.

[0026] Example 2

[0027] (1) The cell wall of Ganoderma lucidum spore powder was broken, with a cell wall breaking rate of 99.7%, to obtain broken cell wall Ganoderma lucidum spore powder;

[0028] (2) Grind the sea buckthorn seeds through a 50-mesh sieve to obtain sea buckthorn seed powder;

[0029] (3) Take broken-cell wall Ganoderma lucidum spore powder and sea buckthorn seed powder and mix them in a weight ratio of 6:4. Then granulate them by dry method to obtain mixed granules of broken-cell wall Ganoderma lucidum spore powder and sea buckthorn seed powder.

[0030] (4) The mixed particles of broken Ganoderma lucidum spore powder and sea buckthorn seed powder were subjected to supercritical CO2 extraction. The extraction conditions were: extraction vessel temperature 42℃, extraction vessel pressure 30MPa, and extraction time 4 hours. The mixed extract of Ganoderma lucidum spore oil and sea buckthorn seed oil was obtained with an extraction rate of 25.1%.

[0031] Example 3

[0032] (1) The cell wall of Ganoderma lucidum spore powder was broken, with a cell wall breaking rate of 99.3%, to obtain broken cell wall Ganoderma lucidum spore powder;

[0033] (2) Grind the sea buckthorn seeds and pass them through a 40-mesh sieve to obtain sea buckthorn seed powder;

[0034] (3) Take broken-cell wall Ganoderma lucidum spore powder and sea buckthorn seed powder and mix them in a weight ratio of 5:5. Then granulate them by dry method to obtain mixed granules of broken-cell wall Ganoderma lucidum spore powder and sea buckthorn seed powder.

[0035] (4) The mixed particles of broken Ganoderma lucidum spore powder and sea buckthorn seed powder were subjected to supercritical CO2 extraction. The extraction conditions were: extraction vessel temperature 48℃, extraction vessel pressure 28MPa, and extraction time 2.5 hours. The mixed extract of Ganoderma lucidum spore oil and sea buckthorn seed oil was obtained with an extraction rate of 22.6%.

[0036] Comparative Example 1

[0037] (1) The cell wall of Ganoderma lucidum spore powder was broken, with a cell wall breaking rate of 99.5%, to obtain broken cell wall Ganoderma lucidum spore powder;

[0038] (2) Dry granulation of broken-cell wall Ganoderma lucidum spore powder to obtain broken-cell wall Ganoderma lucidum spore powder granules;

[0039] (3) The broken Ganoderma lucidum spore powder particles were subjected to supercritical CO2 extraction. The extraction conditions were: extraction vessel temperature 40℃, extraction vessel pressure 30MPa, and extraction time 2.5 hours to obtain Ganoderma lucidum spore oil with an extraction rate of 30%.

[0040] Comparative Example 2

[0041] (1) Grind the sea buckthorn seeds and pass them through a 40-mesh sieve to obtain sea buckthorn seed powder;

[0042] (2) Dry granulation of sea buckthorn seed powder to obtain sea buckthorn seed powder granules;

[0043] (3) The sea buckthorn seed powder particles were subjected to supercritical CO2 extraction. The extraction conditions were: extraction vessel temperature 45℃, extraction vessel pressure 25MPa, and extraction time 3 hours to obtain sea buckthorn seed oil with an extraction rate of 20%.

[0044] The invention will be further illustrated by the following experiments:

[0045] Experimental Example 1: The anti-inflammatory and hemostatic effects of the present invention are further illustrated by the following experiment:

[0046] 1. Experimental Materials

[0047] 1.1 Test drugs: Product of Example 1 (hereinafter referred to as No. 1), Product of Example 2 (hereinafter referred to as No. 2), Product of Example 3 (hereinafter referred to as No. 3), Product of Comparative Example 1 (hereinafter referred to as No. 4); Product of Comparative Example 2 (hereinafter referred to as No. 5), Jiuhua Ointment (commercially available), xylene, 10% egg white solution.

[0048] 1.2 Animals: SPF mice, weighing 18-22g, all healthy males; SD rats, half male and half female, weighing 180-220g; provided by the laboratory of China Pharmaceutical University, with housing conditions of 18-24℃ and 55-70% humidity.

[0049] 2 Dosage Design

[0050] Eight dosage groups were set up, including a blank control group, a low-dose group (No. 1): 0.5 mL / kg; a high-dose group (No. 1): 2 mL / kg; a high-dose group (No. 2): 2 mL / kg; a high-dose group (No. 3): 2 mL / kg; a high-dose group (No. 4): 2 mL / kg; a high-dose group (No. 5): 2 mL / kg; and Jiuhua ointment: normal dosage.

[0051] 3. Test methods

[0052] 3.1 Mouse ear swelling test

[0053] Eighty mice were randomly divided into eight groups of ten each. The corresponding drug was applied to the reaction surfaces of the left and right ears of each mouse, once daily for three consecutive days. Thirty minutes after the last administration, 0.1 mL of xylene was applied to both sides of the right auricle of each mouse, with the left ear serving as a control. Two hours later, the mice were sacrificed, and both auricles were cut off, overlapped, and ear pieces were harvested using an 8 mm diameter punch. The difference between the two ears was calculated as the degree of inhibition.

[0054] 3.2 Rat paw swelling test

[0055] Eighty SD rats, half male and half female, were randomly divided into eight groups of ten each. The corresponding drug was applied to the right hind paw of each rat. Before each drug administration, a clear horizontal line was drawn at the ankle joint of the left hind paw of each rat, and the normal volume of the right hind paw of each rat was accurately measured. Forty minutes after drug administration, 0.1 mL of 10% egg white solution was subcutaneously injected into the right hind paw of each rat. The volume of the right hind paw of each rat was measured at 1 h, 2 h, and 4 h after injection of the 10% egg white solution, and the difference before and after inflammation was calculated as the degree of swelling.

[0056] 4. Experimental Results

[0057] 4.1 Mouse ear swelling test

[0058] No. 1, No. 2, and No. 3 all inhibited mouse ear swelling to varying degrees, showing significant differences compared to the blank control group (P < 0.05). No. 1, at a high dose, showed a significant effect (P < 0.01), with a stronger inhibitory effect than comparative examples 1 and 2 and the positive control Jiuhua ointment. The results are shown in Table 1.

[0059] Table 1. Results of the effect of the test drugs on ear swelling in mice (n=10)

[0060]

[0061] Note: Compared with the control group, **P < 0.01, *P < 0.05

[0062] 4.2 Rat paw swelling test

[0063] The high-dose group (Group 1) effectively inhibited egg white-induced toe swelling, showing a significant difference compared with the blank control group (P < 0.05). The results are shown in Table 2.

[0064] Table 2 Results of toe swelling test (X±SD) (n=10)

[0065]

[0066] Note: Compared with the blank control group, *P<0.05

[0067] Experimental Example 2 further illustrates the scar-removing effect of the present invention through a rabbit ear hypertrophic scar experiment:

[0068] 1. Experimental Materials

[0069] 1.1 Test drugs: Model group: 0.9% sodium chloride solution; Treatment group: Product of Example 1 of this invention (hereinafter referred to as No. 1), Product of Example 2 (hereinafter referred to as No. 2), Product of Example 3 (hereinafter referred to as No. 3), Product of Comparative Example 1 (hereinafter referred to as No. 4); Product of Comparative Example 2 (hereinafter referred to as No. 5).

[0070] 1.2 Animals: Adult SD white rabbits, clean grade, weighing 2.3-2.5kg, male or female, provided by the laboratory of China Pharmaceutical University. Husbandry conditions: separate cages, normal diet, acclimatization feeding for 24 hours before the experiment.

[0071] 2. Experimental Methods

[0072] 2.1 Rabbit ear scar modeling

[0073] Six large-eared white rabbits were removed from their cages and secured. After disinfecting one side of the ear margin with an alcohol swab, 3% sodium pentobarbital was slowly injected via the rabbit's ear vein at a dose of 1 mL / kg. The ventral side of the rabbit's ear was disinfected with an alcohol swab. Four 1cm square incisions were made on both sides of the ventral ear, spaced 2cm apart, removing the full thickness of skin down to the perichondrium surface, and completely removing the perichondrium. The wounds were pressure-sealed with dry cotton balls to stop bleeding, and then aureomycin eye ointment was applied. The wounds were left exposed without bandaging. Wound healing and scar formation were observed starting on the first postoperative day. Simultaneously, two pieces of excised skin tissue from each rabbit were preserved and fixed in 10% formalin. After 24 hours of fixation, they were routinely preserved in paraffin for HE staining and immunohistochemical staining to observe TGF-β1 expression. A total of 12 samples were used as the normal control group.

[0074] 2.2 Grouping and Treatment

[0075] One month later, four rabbits were randomly selected, and one proliferative mass from each was taken for pathological section observation, which showed characteristics consistent with granulation tissue. Two months later, the same method was used to excise the proliferative mass for histopathological observation, which showed characteristics consistent with hypertrophic scar tissue. Seventy-two scars with obvious hypertrophic growth were selected and divided into a model group and a treatment group. The model group received topical application of 0.9% sodium chloride solution or the test drug of this invention twice daily for one month. Each group had 12 samples. Scar tissue samples were fixed in 10% neutral formalin, embedded in paraffin, and sectioned continuously at a thickness of 4 μm for HE staining. The sections were then dewaxed, antigen-blocked, subjected to antigen heat retrieval, and immunohistochemically stained. The scar hyperplasia index (HI), fibroblast density (NA), and cumulative optical density (AOI) were calculated.

[0076] 3. Experimental Results

[0077] 3.1 Scar hyperplasia index and fibroblast density

[0078] The results showed a significant difference in the hyperplastic scar hyperplasia index between the treatment group and the model group (P < 0.05). The number of fibroblasts per unit area in the treatment group was significantly reduced compared with that in the model group (P < 0.05), especially in group 1, which was superior to other groups. The results are shown in Table 3.

[0079] Table 3 Comparison of the effects of the test drugs on HI and NA in rabbit ear scars (X±SD) (n=12)

[0080] Group Scar hyperplasia index <![CDATA[Fibroblast density (cells / mm 2 )]]> Model group 3.21±0.47 4366.52±278.50 Treatment Group 1 2.34±0.23* 3418.65±311.18* Treatment Group 2 2.55±0.17* 3625.44±481.11* Treatment Group No. 3 2.62±0.32* 3689.51±298.32* Treatment Group No. 4 2.93±0.24 3921.57±119.43 Treatment Group No. 5 3.03±0.11 4073.56±524.18

[0081] Note: Compared with the model group, *P < 0.05

[0082] 3.2 Immunohistochemical staining results of TGF-β1 expression

[0083] Compared with the normal group, the expression of TGF-β1 in fibroblasts of both the model group and the treatment group was significantly increased (P<0.05, P<0.01). However, the expression of TGF-β1 in fibroblasts No. 1, No. 2, and No. 3 of the treatment group was significantly decreased compared with that of the model group (P<0.05). The results are shown in Table 4.

[0084] Table 4. Immunohistochemical results of TGF-β1 in rabbit ear tissues of each group (X±SD) (n=12)

[0085] Group Cumulative optical density value normal group 627.56±123.77 Model group 1783.44±328.19** Treatment Group 1 <![CDATA[1054.27±264.73* Δ ]]> Treatment Group 2 <![CDATA[1172.59±341.89* Δ <!-- 5 -->]]> Treatment Group No. 3 <![CDATA[1255.43±415.09* Δ ]]> Treatment Group No. 4 1477.56±299.13* Treatment Group No. 5 1596.94±501.38*

[0086] Note: Compared with the normal group, *P<0.05, **P<0.01; compared with the model group, Δ P < 0.05.

Claims

1. A traditional Chinese medicine extract for anti-inflammatory and scar removal, characterized in that Ganoderma lucidum spore powder and sea buckthorn seed powder were mixed in a weight ratio of 7:3, then granulated by dry method and extracted by supercritical CO2. The extraction conditions were: extraction vessel temperature 40-55℃, extraction vessel pressure 25-30MPa, and extraction time 2-4 hours, to obtain a mixed extract of Ganoderma lucidum spore oil and sea buckthorn seed oil.

2. The traditional Chinese medicine extract for anti-inflammatory and scar removal according to claim 1, characterized in that The drug extract is formulated into a topical preparation with pharmaceutically acceptable excipients.

3. The traditional Chinese medicine extract for anti-inflammatory and scar-removing purposes according to claim 2, characterized in that the topical preparation is a medicinal oil or ointment.

4. A method for preparing the traditional Chinese medicine extract for anti-inflammatory and scar removal according to claim 1, characterized in that The method includes the following steps: (1) Break the cell wall of Ganoderma lucidum spore powder to obtain broken cell wall Ganoderma lucidum spore powder with a cell wall breaking rate of over 99%; (2) Grind the sea buckthorn seeds and pass them through a 40-50 mesh sieve to obtain sea buckthorn seed powder; (3) Take broken-cell wall Ganoderma lucidum spore powder and sea buckthorn seed powder and mix them in a weight ratio of 7:

3. Then granulate them by dry method to obtain mixed granules of broken-cell wall Ganoderma lucidum spore powder and sea buckthorn seed powder. (4) The mixed particles of broken Ganoderma lucidum spore powder and sea buckthorn seed powder were subjected to supercritical CO2 extraction. The extraction conditions were: extraction vessel temperature 40-55℃, extraction vessel pressure 25-30MPa, and extraction time 2-4 hours to obtain a mixed extract of Ganoderma lucidum spore oil and sea buckthorn seed oil.

5. The method for preparing the traditional Chinese medicine extract for anti-inflammatory and scar-reducing purposes according to claim 4, characterized in that... The extraction conditions are as follows: extraction vessel temperature 42-48℃, extraction vessel pressure 25-30MPa, and extraction time 2.5-4 hours.

6. The use of the traditional Chinese medicine extract according to any one of claims 1-3 in the preparation of anti-inflammatory and scar-reducing drugs.

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