CN115869349B - Traditional Chinese medicine composition for detoxifying and promoting tissue regeneration as well as preparation method and application thereof - Google Patents
Traditional Chinese medicine composition for detoxifying and promoting tissue regeneration as well as preparation method and application thereof Download PDFInfo
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Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Medicines Containing Plant Substances (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention relates to a traditional Chinese medicine composition for detoxification and granulation promotion, and a preparation method and application thereof. The traditional Chinese medicine composition for detoxification and promoting tissue regeneration is prepared from the following raw materials in parts by weight: 60-100 parts of lithospermum, 60-100 parts of angelica, 20-60 parts of angelica dahurica 20-60 parts of liquorice, 10-40 parts of frankincense and 10-50 parts of dried alum. According to the invention, the calcined alum is used for replacing calomel, so that 28 toxic traditional Chinese medicinal materials specified in the toxic traditional Chinese medicinal material catalogue are avoided, meanwhile, heavy metals are also avoided, and the modified formula is ensured to have a considerable curative effect.
Description
Technical Field
The invention relates to the field of traditional Chinese medicines, in particular to a traditional Chinese medicine composition for detoxifying and promoting granulation and a preparation method thereof.
Background
The ointment consists of lithospermum, angelica dahurica, liquorice, frankincense (processed with vinegar) and calomel, is a Chinese patent medicine approved by the Ministry of health, and the medicine standard is the eleventh book of Chinese patent medicine formulation. The ointment has effects of promoting blood circulation, removing blood stasis, relieving swelling and pain, removing toxic substance, promoting pus discharge, removing putrefaction, and promoting granulation, and can be used for treating various wound infection and second degree burn. The calomel in the ointment for detoxification and promoting granulation has the functions of killing insects and counteracting toxic substances for external use, the effect of healing sores, but as a heavy metal element, has liver and kidney toxicity, and can be accumulated in liver, kidney and blood tissue to damage organism. Secondly, the calomel as heavy metal brings inconvenience to the production and approval process. Therefore, the medicine material capable of replacing the calomel is found, the heavy metal elements are avoided, and the modified formula is ensured to still have good curative effect, so that the medicine material has important significance.
Disclosure of Invention
The invention aims to provide a traditional Chinese medicine composition for detoxification and granulation promotion, a preparation method and application thereof, and the dried alum is used for replacing calomel, so that the use of toxic traditional Chinese medicines specified in 28 toxic traditional Chinese medicine catalogues is avoided, meanwhile, heavy metals are also avoided, and the modified formula is ensured to still have a considerable curative effect.
The traditional Chinese medicine composition for detoxication and promoting tissue regeneration is prepared from the following raw materials in parts by weight:
60-100 parts of lithospermum, 60-100 parts of angelica, 20-60 parts of angelica dahurica, 20-60 parts of liquorice, 10-40 parts of frankincense and 10-50 parts of dried alum.
Preferably, the traditional Chinese medicine composition for detoxification and granulation promotion is prepared from the following raw materials in parts by weight:
70-90 parts of lithospermum, 70-90 parts of angelica, 30-50 parts of angelica dahurica, 30-50 parts of liquorice, 20-30 parts of vinegar processed frankincense and 30-40 parts of dried alum.
The preparation method of the active ingredients of the traditional Chinese medicine composition for detoxification and granulation promotion comprises the following steps:
pulverizing Olibanum and Alumen respectively into fine powder, and sieving;
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae and Glycyrrhrizae radix in 350-1000 weight parts of tea oil for 5-10 days, stirring for 1-3 times per day, and decocting at 105-110deg.C for 0.5-2 hr to obtain active component oil;
the olibanum, the dried alum and the active component oil are active components.
Preferably, the preparation method of the active ingredients of the traditional Chinese medicine composition for detoxification and granulation promotion comprises the following steps:
pulverizing Olibanum and Alumen respectively into fine powder, and sieving;
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae and Glycyrrhrizae radix in 400-700 weight parts of tea oil for 5-10 days, stirring for 1-3 times per day, and decocting at 105-110deg.C for 0.5-2 hr to obtain active component oil;
the olibanum, the dried alum and the active component oil are active components.
The preparation method of the ointment of the traditional Chinese medicine composition for detoxication and promoting tissue regeneration comprises the following steps:
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae, and Glycyrrhrizae radix in 350-1000 parts by weight of tea oil for 5-10 days, stirring for 1-3 times per day, decocting at 105-110deg.C for 0.5-2 hr, adding 60-200 parts by weight of matrix, dissolving, filtering, adding Olibanum and Alumen fine powder into the filtrate, and mixing well to obtain the final product;
the matrix is one or more of beeswax, lanolin, vaseline, polyethylene glycol and white wax.
Preferably, the preparation method of the ointment of the traditional Chinese medicine composition for detoxification and granulation promotion comprises the following steps:
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae, and Glycyrrhrizae radix in 400-700 weight parts of tea oil for 5-10 days, stirring for 1-3 times per day, decocting at 105-110deg.C for 0.5-2 hr, adding 80-120 weight parts of matrix, dissolving, filtering, adding Olibanum and Alumen fine powder into the filtrate, and mixing well to obtain the final product;
the matrix is one or more of beeswax, lanolin, vaseline, polyethylene glycol and white wax.
The invention also discloses application of the traditional Chinese medicine composition for detoxification and granulation promotion in preparation of anti-infective and antibacterial medicines.
The invention also discloses application of the traditional Chinese medicine composition for detoxifying and promoting tissue regeneration in preparing medicines for treating gynecological inflammation caused by bacteria.
The invention also discloses application of the traditional Chinese medicine composition for detoxifying and promoting tissue regeneration in preparing a medicine for treating skin itch caused by bacteria.
The invention also discloses a traditional Chinese medicine preparation for detoxification and granulation promotion, which is prepared by matching the traditional Chinese medicine composition for detoxification and granulation promotion with a pharmaceutically acceptable carrier, and various common preparations which can be prepared include, but are not limited to: decoction, granule, capsule, tablet, pill, oral liquid, tincture, syrup injection, suppository, spray, ointment, patch, cataplasma, gel, etc.
According to the invention, a large number of experimental screening is carried out on a plurality of medicinal materials, and finally, the selection of the dried alum to replace the calomel is determined, so that the defect that the heavy metal element calomel is used in the existing prescription of the instant detoxification and granulation promoting ointment is overcome, and pharmacodynamics and in-vitro bacteriostasis experiments of examples 9 and 10 prove that the traditional Chinese medicine composition has good curative effect on rats with secondary burn and infection, has more obvious bacteriostasis compared with the original prescription, and can resist more bacteria.
Drawings
FIG. 1 is a graph showing the comparison of the effect of the treatment on the scalded epidermis of rats in each experimental group;
FIG. 2 is a graph of wound healing rates for each group on day 25;
FIG. 3 is a graph of rat skin HE staining, 200 x;
FIG. 4 is a map of Masson staining of rat skin, 100 x;
FIG. 5 is a graph showing the results of detection of serum VEGF, IL-1. Beta., TNF-alpha, IL-6 levels and the contents of Hyp and TGF-beta by ELISA.
Detailed Description
The invention is further illustrated by the following examples. It should be understood that the examples of the present invention are intended to illustrate the present invention and not to limit the present invention. Simple modifications of the invention in accordance with the essence of the invention are all within the scope of the invention as claimed. Unless otherwise indicated, the percentage of ethanol in the present invention is a volume percentage, and v/v represents the volume ratio of the solution.
Example 1
The traditional Chinese medicine composition for detoxification and promoting tissue regeneration is prepared from the following raw materials:
6kg of lithospermum, 6kg of angelica, 2kg of angelica dahurica, 2kg of liquorice, 1kg of frankincense and 1kg of dried alum.
The preparation method of the active ingredients comprises the following steps:
pulverizing Olibanum and Alumen respectively into fine powder, and sieving;
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae, and Glycyrrhrizae radix in 35kg tea oil for 5 days, stirring for 1 time every day, and decocting at 105deg.C for 0.5 hr to obtain active ingredient oil;
the olibanum, the dried alum and the active component oil are active components.
Example 2
The traditional Chinese medicine composition for detoxification and promoting tissue regeneration is prepared from the following raw materials:
10kg of lithospermum, 10kg of angelica, 6kg of angelica dahurica, 6kg of liquorice, 4kg of frankincense and 5kg of dried alum.
The preparation method of the active ingredients of the traditional Chinese medicine composition for detoxification and granulation promotion comprises the following steps:
pulverizing Olibanum and Alumen respectively into fine powder, and sieving;
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae, and Glycyrrhrizae radix in 100kg tea oil for 10 days, stirring for 3 times per day, and decocting at 110deg.C for 2 hr to obtain active component oil;
the olibanum, the dried alum and the active component oil are active components.
Example 3
The traditional Chinese medicine composition for detoxification and promoting tissue regeneration is prepared from the following raw materials:
8kg of lithospermum, 8kg of angelica, 4kg of angelica dahurica, 4kg of liquorice, 3kg of frankincense and 2kg of dried alum.
The preparation method of the active ingredients of the traditional Chinese medicine composition for detoxification and granulation promotion comprises the following steps:
pulverizing Olibanum and Alumen respectively into fine powder, and sieving;
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae, and Glycyrrhrizae radix in 50kg tea oil for 7 days, stirring for 2 times per day, and decocting at 108deg.C for 1 hr to obtain active component oil;
the olibanum, the dried alum and the active component oil are active components.
Example 4
The traditional Chinese medicine composition for detoxification and granulation promotion is prepared from the following raw materials:
7kg of lithospermum, 7kg of angelica, 3kg of angelica dahurica, 3kg of liquorice, 2kg of vinegar processed frankincense and 3kg of dried alum.
The preparation method of the ointment comprises the following steps:
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae, and Glycyrrhrizae radix in 40kg tea oil for 6 days, stirring for 1 time every day, decocting at 106 deg.C for 0.6 hr, adding 7kg Cera flava for dissolving, filtering, adding Olibanum and Alumen fine powder into the filtrate, and mixing.
Example 5
The traditional Chinese medicine composition for detoxification and granulation promotion is prepared from the following raw materials:
9kg of lithospermum, 9kg of angelica, 5kg of angelica dahurica, 5kg of liquorice, 3kg of vinegar processed frankincense and 4kg of dried alum.
The preparation method of the active ingredients of the traditional Chinese medicine composition for detoxification and granulation promotion comprises the following steps:
the preparation method of the ointment comprises the following steps:
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae, and Glycyrrhrizae radix in 80kg tea oil for 9 days, stirring for 3 times per day, decocting at 109 deg.C for 1.6 hr, adding 10kg white wax for dissolving, filtering, adding Olibanum and Alumen fine powder into the filtrate, and mixing.
Implementation of the embodiments example 6
The traditional Chinese medicine composition for detoxification and granulation promotion is prepared from the following raw materials:
8kg of lithospermum, 8kg of angelica, 4kg of angelica dahurica, 4kg of liquorice, 2.66kg of vinegar processed frankincense and 3.05kg of dried alum.
The preparation method of the ointment comprises the following steps:
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae, and Glycyrrhrizae radix in 50kg tea oil for 7 days, stirring for 2 times per day, decocting at 106 deg.C for 1 hr, adding 10kg white wax for dissolving, filtering, adding Olibanum and Alumen fine powder into the filtrate, and mixing.
Example 7
The traditional Chinese medicine composition for detoxification and granulation promotion is prepared from the following raw materials:
7.5kg of lithospermum, 8.5kg of angelica, 3kg of angelica dahurica, 3kg of liquorice, 3.2kg of vinegar-processed frankincense and 2.5kg of dried alum.
The preparation method of the ointment comprises the following steps:
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae, and Glycyrrhrizae radix in 70kg tea oil for 7 days, stirring for 2 times per day, decocting at 106 deg.C for 1 hr, adding 8kg vaseline for dissolving, filtering, adding Olibanum and Alumen fine powder into the filtrate, and mixing.
Example 8
The traditional Chinese medicine composition for detoxification and granulation promotion is prepared from the following raw materials:
9kg of lithospermum, 9kg of angelica, 4kg of angelica dahurica, 5kg of liquorice, 3.5kg of frankincense and 2.5kg of dried alum.
The preparation method of the ointment comprises the following steps:
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae, and Glycyrrhrizae radix in 80kg tea oil for 7 days, stirring for 2 times per day, decocting at 106 deg.C for 0.8 hr, adding lanolin and polyethylene glycol, dissolving, filtering, adding Olibanum and Alumen fine powder into the filtrate, and mixing.
Example 9
The traditional Chinese medicine composition for detoxification and granulation promotion is prepared from the following raw materials:
6.5kg of lithospermum, 5.5kg of angelica, 3.5kg of angelica dahurica, 5.5kg of liquorice, 3.5kg of frankincense and 1.5kg of dried alum.
The preparation method of the ointment comprises the following steps:
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae, and Glycyrrhrizae radix in 60kg tea oil for 7 days, stirring for 2 times per day, decocting at 106 deg.C for 1.2 hr, adding 12kg Cera flava, lanolin, and vaseline, dissolving, filtering, adding Olibanum and Alumen fine powder into the filtrate, and mixing.
Example 10
The traditional Chinese medicine composition for detoxification and granulation promotion is prepared from the following raw materials:
7kg of lithospermum, 7kg of angelica, 5kg of angelica dahurica, 5kg of liquorice, 1.8kg of vinegar processed frankincense and 2kg of dried alum.
The preparation method of the ointment comprises the following steps:
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae, and Glycyrrhrizae radix in 50kg tea oil for 7 days, stirring for 2 times per day, decocting at 106 deg.C for 1.5 hr, adding 11kg Cera flava, lanolin, and vaseline, dissolving, filtering, adding Olibanum and Alumen fine powder into the filtrate, and mixing.
Example 11
The traditional Chinese medicine composition for detoxification and granulation promotion is prepared from the following raw materials:
8.5kg of lithospermum, 6.5kg of angelica, 3.5kg of angelica dahurica 2.5kg of liquorice, 1.5kg of frankincense and 2.5kg of dried alum.
Preparation of its active ingredient the method comprises the following steps:
pulverizing Olibanum and Alumen respectively into fine powder, and sieving;
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae, and Glycyrrhrizae radix in 60kg tea oil for 8 days, stirring for 2 times per day, and decocting at 110deg.C for 2 hr to obtain active component oil;
the olibanum, the dried alum and the active component oil are active components.
The active ingredients and the conventional film forming materials and plasticizers are taken together to prepare the film coating agent.
Example 12
The traditional Chinese medicine composition for detoxification and granulation promotion is prepared from the following raw materials:
7kg of lithospermum, 9kg of angelica, 3kg of angelica dahurica, 5kg of liquorice, 2kg of vinegar processed frankincense and 4.5kg of dried alum.
The preparation method of the active ingredients comprises the following steps:
pulverizing Olibanum and Alumen respectively into fine powder, and sieving;
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae, and Glycyrrhrizae radix in 70kg tea oil for 7 days, stirring for 1 time every day, and decocting at 103 deg.C for 0.8 hr to obtain active ingredient oil;
the olibanum, the dried alum and the active component oil are active components.
Mixing the active ingredients with conventional penetrant, binder, humectant and filler, and making into gel paste.
Example 13
Medicine effect study of dried alum prescription granulation promoting ointment and calomel prescription granulation promoting ointment group on rats with secondary burn complicated with infection
1. Material
1.1. Experimental reagent and equipment
LB liquid culture medium, an ultra clean bench, a biochemical incubator, a constant temperature water bath, a centrifuge, weights, 10% chloral hydrate and positive medicines (recombinant bovine basic fibroblast growth factor gel).
The dried alum prescription granulation promoting ointment, the calomel prescription granulation promoting ointment and the blank matrix are provided by the company of Guangxi Zhuang Huahong pharmaceutical industry group.
The Elisa kit was purchased from Jiangsu enzyme technology Co., ltd.
1.2. Experimental animal
SPF-grade SD male rats (200-170 g) were purchased from the university of south medical science animal experiment center.
1.3. Bacteria and method for producing same
Staphylococcus aureus (ATCC 25923), pseudomonas aeruginosa (ATCC 27853) was purchased from the microorganism technologies limited of the ciclesonide.
2. Method of
2.1. Bacterial culture
Activating frozen bacteria solution of Staphylococcus aureus and Pseudomonas aeruginosa at-80deg.C, adding 100 μL of the activated frozen bacteria solution into fresh LB culture solution at a culture concentration of 1:100, and culturing in biochemical incubator at 37deg.C for 24 hr according to culture conditions and method on specification to obtain Staphylococcus aureus with a quantity of 9.47 x 10 8 CFU, the number of Pseudomonas aeruginosa was 4.35X 10 11 CFU。
2.2. Animal model
Rats were anesthetized with 10% chloral hydrate and dehaired, exposing back skin. Except for the blank group, the rest rats are used for forming a scald model, the rats are put into boiling water with the temperature of 100 ℃ for 10 minutes by using weights with the diameter of 2.5cm, and the rats are taken out to be contacted with the exposed skin part for 15 seconds for 3 times, so that the scald is formed. After the wound is cooled, 200 mu L of bacterial liquid cultured under the item "1.3" is dripped on the surface of the wound, and the bacterial liquid is uniformly smeared.
2.3. Animal administration
SD rats 25 groups randomly classifying into normal groups; model control group; blank matrix groups; a positive drug group; high and low dose groups of the dried alum prescription myogenic paste prepared in example 6; the Calomelas prescription has high granulation promoting ointment and low dosage. Animals in the model group and normal group were not treated. The blank matrix group was given 0.4g of blank matrix paste and uniformly smeared on the scald epidermis. The positive medicine group is smeared with positive medicine with the thickness of 0.2 cm. The high and low dosage groups of the dried alum prescription granulation promoting ointment are respectively given with 0.4g and 0.2g of the dried alum prescription granulation promoting ointment and uniformly coated, and the high and low dosage groups of the calomel prescription granulation promoting ointment are given with 0.4g and 0.2g of the calomel prescription granulation promoting ointment and uniformly coated. The administration was continued for 25 days.
Mice were sacrificed on day 25 and blood, skin, liver, kidney tissues were harvested and stored at-80 ℃ for use.
2.4. Wound size assessment
SD rats were photographed and wound healing recorded on day 0,5,10,15,20,25, and the wound healing rate was calculated by Image J software analysis, the calculation formula:
wound healing Rate= (S0-SN)/S0
Wherein S0 represents a 0d wound surface area, and SN represents an Nd-th wound surface area in units of: cm 2 Wherein N is = 0,5,10,15,20,25.
HE staining and Masson staining
Taking a rat wound skin specimen, fixing 4% paraformaldehyde, dehydrating and transparentizing, embedding paraffin, slicing, staining HE and Masson, carrying out microscopic histopathological observation, and observing the healing degree of the rat skin.
Elisa assay
SD rat abdominal aorta blood collection, standing at room temperature for 30 min, and centrifuging at 4deg.C (3500 r/min,15 min) to obtain serum solution.
The Elisa kit detects the contents of endothelial growth factor (VEGF), interleukin 1 beta (IL-1 beta), tumor necrosis factor alpha (TNF-alpha) and interleukin 1 (IL-6) in the serum of the rat.
The wound surface and the surrounding skin of the rat are taken for measuring the content of hydroxyproline (Hyp) and transforming growth factor beta (TGF-beta), and the measurement is carried out by referring to the instruction book of the kit.
3. Results
3.1. Wound size assessment
The extent of wound healing for each group is shown in figure 1. Model group wound healing was slower 25 days after dosing. The blank matrix had no statistical difference from the model group in wound healing extent. Compared with the model group, the positive medicine has the best wound healing effect (P is less than 0.01), and the wounds of the dried alum prescription granulation promoting ointment group (P is less than 0.01) and the calomel prescription granulation promoting ointment group (P is less than 0.05) are obviously healed. The wound healing rate for each group on day 25 is shown in figure 2.
HE staining and Masson staining
The results of rat skin HE staining are shown in FIG. 3, and it is clear from the results that the epidermis of the rat in the model group is obviously thickened, the dermis and subcutaneous tissues are thickened, inflammatory factor infiltration is increased, and necrotic tissues are increased. After the external administration of the skin for 25 days, the skin surface edema of rats in the dried alum prescription granulation promoting ointment group and the calomel prescription granulation promoting ointment group is reduced, the inflammatory factors of the dermis and subcutaneous tissues are reduced, and the necrotic tissues are reduced.
In FIG. 3, NC represents a normal group, MC represents a model group, KB represents a blank matrix, YX represents a positive medicine group, QG-L represents a low-dose group of the dried alum prescription granulation promoting ointment, QG-H represents the high-dose group of the calcined alum prescription myogenic paste, YG-L represents the low-dose group of the calomel prescription myogenic paste, and YG-H represents the high-dose group of the calomel prescription myogenic paste
The high-dose and the high-dose of the calcined alum prescription granulation promoting ointment have no difference in wound healing degree, so that one of the two dose groups is selected for the subsequent experiment.
As shown in figure 4, the skin collagen fibers (blue color under Masson dyeing) of the model group are obviously less than those of the normal group, and the skin collagen fibers of the rats in the dried alum prescription granulation promoting ointment group and the dried calomel prescription granulation promoting ointment group are obviously increased 25 days after administration.
In FIG. 4, NC represents a normal group, MC represents a model group, KB represents a blank matrix, YX represents a positive drug group, QG represents a dried alum prescription granulation promoting ointment group, and YG represents a calomel prescription granulation promoting ointment group
Elisa assay
Serum VEGF, IL-1. Beta., TNF-alpha, IL-6 levels were measured by ELISA and the results are shown in FIG. 5.
From FIG. 5, it can be seen that the serum levels of VEGF, hyp and TGF-beta were significantly reduced in the rats of the model group, and the levels of the pro-inflammatory factors IL-1 beta, TNF-alpha, IL-6 were significantly increased, as compared to the normal group. 25 days after skin administration, the dried alum prescription granulation promoting ointment and the calomel prescription granulation promoting ointment increase VEGF in rat serum, hyp and TGF-beta levels to reduce the levels of the pro-inflammatory factors IL-1 beta, TNF-alpha, IL-6. And the dried alum prescription granulation promoting ointment and the calomel prescription granulation promoting ointment have no obvious difference.
FIG. 5. Rat Elisa test (NC represents normal group, MC represents model group, KB represents blank matrix, YX represents positive drug group, QG represents dried alum prescription granulation promoting ointment, YG represents calomel prescription granulation promoting ointment group).
** P < 0.01, ns represents no statistical difference, P > 0.05
4. Discussion of the invention
In the study, the selection of an animal skin injury model is a key link, the skin anatomical structure of an SD rat is similar to that of a human, and the skin injury model has less subcutaneous fat on the back and is smoother and easy to process. SD rats were thus selected as experimental animals. Moulding method for early moulding stage refers to literature second degree burn and scald and wound infection model [1,2] The two methods are combined, and after the modeling damage mode, the modeling wound size, the infection source inoculation mode and the inoculation amount are examined through a pre-experiment, the improved modeling method of the second-degree burn and scald infection model is obtained, and the method is simple and safe to operate. After molding, the back wound of the rat is red and swollen, the yellowish white pus and tissue fluid appear in the early stage, the scab is formed in the later stage, the pus is arranged under the scab, and the healing is slow. The pathological section microscope shows that the model group rats have increased epidermal inflammatory factor infiltration, necrotic tissue is increased, and the cortex is thickened to indicate that the wound has edema. Subsequent Elisa analysis results show that compared with a normal group, the inflammatory factors of a model group are obviously increased, the level of the epidermal growth factor is obviously lower than that of other groups, and the results together indicate that the modeling is successful.
The administration components are dried alum prescription granulation promoting ointment and calomel prescription granulation promoting ointment. The results show that in the evaluation of the wound size, no significant difference exists between the high and low doses of the dried alum prescription granulation promoting ointment and the high and low doses of the calomel prescription granulation promoting ointment. The wound healing rate of the Calomelas prescription granulation promoting ointment group is not statistically different from that of the dried alum prescription granulation promoting ointment group, and the section dyeing result also shows that the two administration groups reduce the inflammation of skin tissues, the structure is repaired, and the skin collagen fibers are increased.
Elisa analysis results show that the dried alum prescription granulation promoting ointment and the calomel prescription granulation promoting ointment have the functions of resisting inflammation and promoting growth factors at the wound of the epidermis, and the reduction of the pro-inflammatory factors leads to improvement of wound redness and swelling and is beneficial to wound healing. In addition, the result of the inflammatory factor level shows that the anti-inflammatory effect of the dried alum prescription granulation promoting ointment is slightly better than that of the calomel prescription granulation promoting ointment, but the two groups of serum inflammatory factor levels have no significant difference. VEGF is involved in the overall process of angiogenesis. VEGF promotes basal lamina decomposition during the process of angiogenesis, and can accelerate angiogenesis. Angiogenesis can provide nutrition and oxygen for granulation tissue and cells of wound surface, and plays an important role in the wound surface repair process [3] . TGF-beta mediated signaling pathways are currently widely recognized as the most closely related mechanism to promote wound healing, but overactivation can progress to pathological scarring [4] . One main appearance of wound healing is deposition and repair of collagen, the main component of collagen is hydroxyproline, the content of which is constant in collagen, and the measurement of the content of the hydroxyproline of the wound can reflect the content of the collagen [5] . The experimental result shows that compared with the three indexes of the dried alum prescription granulation promoting ointment group and the calomel prescription granulation promoting ointment group, the model group is obviously improved, and the dried alum prescription granulation promoting ointment and the calomel prescription granulation promoting ointment group play a role in promoting wound healing by increasing the expression level of skin conversion growth factors and hydroxyproline.
Reference to the literature
[1] Liu Wenya, qian Wenhui, su Hua, etc. the effect of compound wound healing lotion on repair of wound of bacterial infection is studied [ J ]. Chinese pharmacist, 2019,22 (07): 1246-1249.
[2] Zhang Dawei establishment of animal model for pharmacodynamics evaluation of burn wound treatment drug [ D ]. Third army university of medical science, 2011.
[3] Kong Lingzhen, xing Jie. Influence of BAZHEN decoction on expression of human skin fibroblast epidermal growth factor, transforming growth factor beta 1 and vascular endothelial growth factor A [ J ]. J.J.of Chinese-Western medicine-incorporated surgery, 2022,28 (01): 17-21.
[4] Zhang Liangping, wang Yang, lei Rui, et al nuclear factor I-C reduces the sensitivity of dermal fibroblasts to TGF-beta [ J ]. University of southern medical science, 2015,35 (09): 1245-1250.
[5] Xie Meina, li Jing, yang Lixia, etc. the effect of decellularized biological dressing on the healing of scalded wound [ J ]. Experimental animal science 2020,37 (02): 19-25.
[6] Wang Xiao, lin Ruichao, dong Shifen, et al, toxicity studies of mercury-containing mineral medicaments, J. Chinese J.traditional Chinese medicine, 2017,42 (07): 1258-1264.
[7] He Rong and Peng Bo, beautiful road, etc. the effects of the powder and its prescription on the histomorphology of damaged skin of rats are observed [ J ]. J. Chinese J.Chinese medicine, 2012,37 (06): 715-718.
Example 14
Detoxicating and tissue regeneration promoting ointment external bacteriostasis experiment
Sample name: detoxicating and tissue regeneration promoting ointment
The preparation method of the samples 1-7 comprises the following steps:
80kg of lithospermum, 80kg of angelica, 40kg of angelica dahurica, 40kg of liquorice, 26.6kg of vinegar-processed frankincense and the dosage of dried alum are changed.
The preparation method comprises the following steps: soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae, and Glycyrrhrizae radix in 500kg tea oil for 7 days, stirring for 2 times per day, decocting at 106 deg.C for 1 hr, adding 100kg white wax for dissolving, filtering, adding Olibanum and Alumen fine powder into the filtrate, and mixing.
The weight percentage concentrations of the dried alum in samples 1-7 were: 0.8%, 1.2%, 1.6%, 2.0%, 5.0%, 10.0%.
Sample 8 was a sample prepared according to the ministry standards and did not contain dried alum.
1. Detection results and conclusions
1 detection results
The test adopts escherichia coli, pseudomonas aeruginosa, bacillus vulgaris, staphylococcus epidermidis and staphylococcus aureus to carry out bacteriostasis test, and as a result, a negative control group has no bacteriostasis ring, and the diameter of the bacteriostasis ring of a sample 1, a sample 2, a sample 3, a sample 4 and a sample 8 to test bacteria is smaller than or equal to 7mm; the diameters of the antibacterial rings of the sample 6 and the sample 7 are larger than 7mm, which shows that the sample has antibacterial effect on the test bacteria 5. Sample dried alum amount screening test: when the concentration of the dried alum is 4.0%, the diameter of a bacteria inhibition ring of the sample to the test bacteria is larger than 7mm, which shows that the concentration has a bacteria inhibition effect on the test bacteria.
2 conclusion of detection
Under the test condition, the in-vitro antibacterial test is carried out by adopting test sample liquid, and both the sample 6 and the sample 7 have antibacterial effects on escherichia coli, pseudomonas aeruginosa, bacillus vulgaris, staphylococcus epidermidis and staphylococcus aureus, and have stronger antibacterial effects.
When the concentration of the dried alum is 4.0%, the sample has an antibacterial effect on escherichia coli, pseudomonas aeruginosa, bacillus vulgaris, staphylococcus epidermidis and staphylococcus aureus.
2. Detection process
1. Purpose of test
Determining antibacterial effect of the sample of the detoxication granulation promoting ointment on Escherichia coli, pseudomonas aeruginosa, proteus vulgaris, staphylococcus epidermidis and Staphylococcus aureus, the effective bacteriostatic concentration of dried alum in the sample was determined.
2. Test article
2.1 name (or code): detoxicating and promoting tissue regeneration ointment;
sample number: TC22096 to TC22103 (samples 1 to 8);
lot number: ys-20211202, ys-20211203, ys-20211204, ys-20211205, ys-20211206, ys-20211207, ys-20211208, ys-20211201;
specification of: 10 g-a branch;
content/concentration: 10 g/min;
### Supporting;
traits: a brownish red paste;
preservation conditions: sealing, avoiding light, and placing in a shade place;
production date: 20211222, 20211223, 20211224, 20211227, 20211228, 20211229, 20211230, 20211221;
production unit: guangxi Zhuang autonomous region Huahong pharmaceutical Congress company;
3. main instrument and reagent
3.1 major instrumentation
3.2 major reagents
3.2.1 Main Medium and reagent basic information
4. Experimental system
4.1 Strain: escherichia coli [ CMCC (B) 44 102 ], pseudomonas aeruginosa [ CMCC (B) 10 104 ], proteus vulgaris [ ATCC 6896 ], staphylococcus epidermidis [ ATCC 12228 ], staphylococcus aureus [ CMCC (B) 26003 ] were purchased from the Guangdong province microorganism strain collection.
5. Test method
5.1 preparation of strains and bacterial liquids
The reserve strains of the 5 bacteria qualified by resuscitation are prepared into working strains and bacterial suspensions according to the following operation steps: inoculating fresh cultures of Escherichia coli, pseudomonas aeruginosa, proteus vulgaris, staphylococcus epidermidis and Staphylococcus aureus into brain heart leaching liquid medium, culturing at 37deg.C for 24h, preparing the above culture into bacterial suspension containing about 1.5X108 cfu/mL (colony forming unit) per l mL with 0.9% sterile sodium chloride solution, and continuously diluting the bacterial suspension to 5×106cfu/mL with sterile physiological saline 10 times before inoculating.
5.2 in vitro bacteriostasis test
5.2.1 doses design and grouping
Taking samples 1-8, and respectively preparing sample extracting solutions as groups 1-8; sample 6 and sample 7 were prepared as groups 9 and 10 according to 80% of the stock solution, respectively, as a result of the pre-test; distilled water was used instead of the sample for the test as a negative control group.
5.2.2 sample preparation of the extractive solution
Selecting samples 1-8, and respectively preparing 1-8 groups of sample solutions: taking 1g of a sample, adding 2mL of isopropyl myristate, shaking to form a flowing liquid, adding 1mL of sterile distilled water, placing in a water bath at 80 ℃ for 20 minutes, shaking to form an emulsified suspension, standing, and taking a lower water phase as a sample extracting solution for later use.
5.2.3 screening test for amount of dried alum in sample:
taking a sample 6 and a sample 7, and respectively preparing 9-10 groups of sample liquids: 0.8g of sample is added with 2mL of isopropyl myristate and shaken to form flowing liquid, 1mL of sterile distilled water is added, the mixture is placed in a water bath at 80 ℃ for 20 minutes, the mixture is shaken to form emulsified suspension, and the suspension is placed still, and the lower water phase is taken as sample extracting solution for standby.
5.2.4 preparation of antibacterial test sample (preparation on the same day)
Test group sample preparation: taking sterile dry filter paper sheets with the diameter of 6mm, respectively putting the sterile dry filter paper sheets into sample extracting solutions with different concentrations, soaking, respectively preparing 15 sheets with each concentration, respectively putting the 15 sheets in a clean sterile empty plate, and naturally drying for later use.
And taking an aseptic drying filter paper sheet, soaking the aseptic drying filter paper sheet in aseptic distilled water, and taking the aseptic drying filter paper sheet as a negative control sample sheet for standby.
5.2.5 bacteriostasis test
(1) Inoculation of test bacteria: 0.1mL of test bacteria suspension with the concentration of 5 multiplied by 106cfu/mL is uniformly coated on the surface of a nutrient agar culture medium flat plate, and 3 plates are inoculated for each test bacteria. The plates were covered and naturally dried in a biosafety cabinet.
(2) Placing a bacteriostasis test sample piece: 1 bacteria-staining plate was attached to each test, 4 test pieces, 1 negative control piece were attached to each plate, and each bacteria was repeated 3 times. The sample piece was attached to the surface of the plate using sterile forceps. The distance between the centers of the sample pieces is more than 25mm, and the distance between the centers of the sample pieces and the periphery of the flat plate is more than 15 mm. After the sticking, the sample wafer is lightly pressed by sterile forceps to be tightly attached to the surface of the flat plate.
(3) Culturing: plates were covered and incubated in a 37℃incubator for 24h for observation. The diameter of the inhibition ring (including the patch) was measured with a vernier caliper and recorded.
6. Result determination method
6.1, the diameter of the antibacterial ring is larger than 7mm, and the antibacterial ring is judged to have antibacterial effect.
And 6.2, judging that the antibacterial effect is absent when the diameter of the antibacterial ring is smaller than or equal to 7 mm.
And 6.3 repeated tests are judged to be qualified if the result of the bacteriostasis effect is obtained.
6.4 negative control should have no inhibition zone generated, otherwise the test is ineffective.
7. Results and analysis
Sample pair test strains is effective in inhibiting bacteria
7.1 sample 1, sample 2, sample 3, sample 4, sample 8 have a diameter of less than 7mm on the antibacterial ring of Escherichia coli, pseudomonas aeruginosa, proteus vulgaris, staphylococcus epidermidis, staphylococcus aureus, and show that the above samples have no antibacterial effect on the above 5 bacteria. The negative control group has no inhibition ring, and the test is effective.
7.2 sample 6 and sample 7 have a diameter of the inhibition ring of Escherichia coli, pseudomonas aeruginosa, proteus vulgaris, staphylococcus epidermidis and Staphylococcus aureus of more than 7mm, the sample has antibacterial effect on the above 5 bacteria.
7.3 dried alum usage screening test in samples: when the concentration of the dried alum is 4.0%, the diameter of the antibacterial ring of the sample on escherichia coli, pseudomonas aeruginosa, bacillus vulgaris, staphylococcus epidermidis and staphylococcus aureus is larger than 7mm, which shows that the concentration of the sample has antibacterial effect on the above 5 bacteria.
Conclusion 8
Under the test condition, the in-vitro antibacterial test is carried out by adopting test sample liquid, and both the sample 6 and the sample 7 have antibacterial effects on escherichia coli, pseudomonas aeruginosa, bacillus vulgaris, staphylococcus epidermidis and staphylococcus aureus, and have stronger antibacterial effects.
When the concentration of the dried alum is 4.0%, the sample has an antibacterial effect on escherichia coli, pseudomonas aeruginosa, bacillus vulgaris, staphylococcus epidermidis and staphylococcus aureus.
While the invention has been described in detail in the foregoing general description, embodiments and experiments, it will be apparent to those skilled in the art that modifications and improvements can be made thereto. Accordingly, such modifications or improvements may be made without departing from the spirit of the invention and are intended to be within the scope of the invention as claimed.
Claims (8)
1. A traditional Chinese medicine composition for detoxification, granulation promotion and bacteriostasis is characterized by being prepared from the following raw materials in parts by weight:
60-100 parts of lithospermum, 60-100 parts of angelica, 20-60 parts of angelica dahurica, 20-60 parts of liquorice, 10-40 parts of frankincense and 10-50 parts of dried alum; wherein the weight percentage concentration of the dried alum is more than or equal to 4.0 percent.
2. The traditional Chinese medicine composition for detoxification, granulation promotion and bacteriostasis according to claim 1, which is characterized by being prepared from the following raw materials in parts by weight:
70-90 parts of lithospermum, 70-90 parts of angelica, 30-50 parts of angelica dahurica, 30-50 parts of liquorice, 20-30 parts of vinegar processed frankincense and 30-40 parts of dried alum.
3. The traditional Chinese medicine composition for detoxification, granulation promotion and bacteriostasis according to claim 1 or 2, characterized in that the preparation method of the active ingredients comprises the following steps:
pulverizing Olibanum and Alumen respectively into fine powder, and sieving;
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae and Glycyrrhrizae radix in 350-1000 weight parts of tea oil for 5-10 days, stirring for 1-3 times per day, and decocting at 105-110deg.C for 0.5-2 hr to obtain active component oil;
the olibanum, the dried alum and the active component oil are active components.
4. The traditional Chinese medicine composition for detoxification, granulation promotion and bacteriostasis according to claim 3, wherein the preparation method of the active ingredients comprises the following steps:
pulverizing Olibanum and Alumen respectively into fine powder, and sieving;
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae and Glycyrrhrizae radix in 400-700 weight parts of tea oil for 5-10 days, stirring for 1-3 times per day, and decocting at 105-110deg.C for 0.5-2 hr to obtain active component oil;
the olibanum, the dried alum and the active component oil are active components.
5. The traditional Chinese medicine composition for detoxification, granulation promotion and bacteriostasis according to claim 1 or 2, wherein the preparation method of the ointment comprises the following steps:
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae, and Glycyrrhrizae radix in 350-1000 parts by weight of tea oil for 5-10 days, stirring for 1-3 times per day, decocting at 105-110deg.C for 0.5-2 hr, adding 60-200 parts by weight of matrix, dissolving, filtering, adding Olibanum and Alumen fine powder into the filtrate, and mixing well to obtain the final product;
the matrix is one or more of beeswax, lanolin, vaseline, polyethylene glycol and white wax.
6. The traditional Chinese medicine composition for detoxification, granulation promotion and bacteriostasis according to claim 5, wherein the preparation method of the ointment comprises the following steps:
soaking radix Arnebiae, radix Angelicae sinensis, radix Angelicae Dahuricae, and Glycyrrhrizae radix in 400-700 weight parts of tea oil for 5-10 days, stirring for 1-3 times per day, decocting at 105-110deg.C for 0.5-2 hr, adding 80-120 weight parts of matrix, dissolving, filtering, adding Olibanum and Alumen fine powder into the filtrate, and mixing well to obtain the final product;
the matrix is one or more of beeswax, lanolin, vaseline, polyethylene glycol and white wax.
7. Use of a traditional Chinese medicine composition for detoxification, tissue regeneration and bacteriostasis according to any one of claims 1-6 in the preparation of anti-infective and bacteriostasis medicines.
8. A traditional Chinese medicine preparation for detoxification and granulation promotion, which is prepared by matching the traditional Chinese medicine composition for detoxification, granulation promotion and bacteriostasis according to any one of claims 1-4 with a pharmaceutically acceptable carrier.
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Denomination of invention: A traditional Chinese medicine composition for detoxification and muscle growth, its preparation method and use Granted publication date: 20230912 Pledgee: Guilin Bank Co.,Ltd. Liuzhou Branch Pledgor: Huahong Pharmaceutical Group Co.,Ltd. of Guangxi Zhuang Autonomous Region Registration number: Y2024450000033 |