| Objective:Objective To observe the changes of skin lesions and pathomorphology of Sanhuang gel on auricular compound acne model in rats,so as to provide experimental basis for clinical treatment and prevention of external mechanisms of acne scars.Methods:Sixty two SPF Wistar rats,female,were randomly divided into two groups: blank group and model group.There were 10 in the blank group and 52 in the model group.After successful replication of the model,the model group was divided into model group,positive control group(0.025% dimensional A yogurt cream)and Sanhuang gel treatment group(high,medium and low)by random number table method,with 10 rats in each group.In addition to the blank group,on the basis of Kligman method,100% oleic acid solution was applied once a day at the opening of the inner ear duct on the right side of the rat,0.5-1ml each time,and 0.1ml Propionibacterium acnes solution was injected into the auricle skin of the rat every other day for 4 weeks to establish the compound acne model of the rat auricle.End of modeling: if the skin lesions show inflammatory reactions such as redness,swelling and desquamation,and the pores of hair follicles can be seen to expand,with white content blocked at the mouth of hair follicles,two of them are randomly taken out of the model group,and the auricle tissue at the right ear Modeling Site is cut off for HE staining.If excessive keratosis of cuticle is seen,the mouth of hair follicles and the horn plug of funnel are blocked,and the expansion is in the shape of a pot,it indicates that the animal model is well established.After successful modeling,they were treated with 0.025% dimensional A yogurt,three yellow gel(0.08g/kg)and medium(0.04g/kg)low dose(0.02g/kg)respectively,and 1 times /d for 28 consecutive days.After administration of 4W,the rats were anesthetized with 10% chloral hydrate,and the heart was sampled for 5ml.After centrifugation,the supernatant was stored at-20 ?,and the levels of IL-6,IL-10,CTGF and VEGF in serum were detected by ELISA;rats were killed by cervical dislocation,and auricle tissues were taken,washed with PBS(phosphate buffer)for several times,some of them were fixed in 10% paraformaldehyde for he staining,the rest were frozen in liquid nitrogen and stored in-80 ? refrigerator,and then western staining was used The expression of IL-6,IL-10,CTGF,VEGF and their genes were detected by RT-PCR and blot respectively.Results:1 Macroscopic Observation:(1)blank group: the auricle tissue of rats is soft,without swelling and desquamation,with light red appearance and capillaries;the hair follicle has no keratosis,without acne,pustule,nodule,cyst,which is the same as the normal tissue.(2)model group: the epidermis is obviously thickened,rough,a lot of desquamation,skin temperature is high,swelling is obvious,there is papule like protuberance,accompanied by cyst or pustule.(3)after 4 weeks of treatment,auricle of each group: compared with model group,there is obvious difference in auricle tissue of each treatment group,among which the auricle tissue of positive control group(0.025% retinoic acid)is soft,light red,with capillaries,without acne,pustule The appearance of nodules and cysts was close to the blank group.The auricular tissue of Sanhuang gel(high,medium and low)groups were significantly improved,especially in the high and middle dose groups.The auricle tissue of the rats was soft and light red,showing capillaries,no pimples,pustules,nodules,cysts,and the appearance was close to the blank group.2 Pathomorphological Changes:(1)Compared with the blank group,the epidermis of the model group,the Sanhuang gel high and low dose group were thickened obviously,the glands in the subcutaneous interstitium were proliferated,fibrous tissue hyperplasia,hyperkeratosis or incomplete,intercellular edema and lymphocytic infiltration were still obvious.The 0.025% dimensional A cream group and the three yellow gel medium dose group were close to normal.(2)compared with the pathological morphology of the model group,there were differences between the 0.025% dimensional A yogurt group and the Sanhuang gel group in high,medium and low dose groups.(3)compared with the 0.025% dimensional A yogurt group,the tissue morphology of the Sanhuang gel medium dose group and blank group were similar.3 ELISA Test Results: The content of IL-6,IL-10,CTGF and VEGF in the ear tissue of the model group was significantly higher than that of the blank group(P The content of IL-6,IL-10 and VEGF in the ear tissue of rats decreased significantly(P < 0.05),but there was no significant difference(P> 0.05).(2)Compared with the blank group,the CTGF content in the ear tissue of the model group was significantly increased,with a statistically significant difference(P<0.05);compared with the model group,the CTGF content in the ear tissue of the rats in the administration group was decreased to varying degrees,especially in the positive group and the high-dose group,with a statistically significant difference(P< 0.05),and the CTGF content in the middle and low-dose groups of the traditional Chinese medicine was significantly reduced Although the CTGF content in the ear tissue of the dose group decreased,there was no significant difference(P > 0.05).4 Western-blotting Test Results:the expression of IL-6,IL-10,CTGF and VEGF in the ear tissue of the model group was significantly higher than that of the blank group(P< 0.05),The expression of IL-6 in the tissue decreased significantly(P< 0.05),but there was no significant difference(P> 0.05).(2)Compared with the model group,the expression of IL-10,CTGF and VEGF in the ear tissue of the rats in the administration group decreased in varying degrees,especially in the high and medium dose groups of traditional Chinese medicine and the positive group,the expression of IL-10,CTGF and VEGF in the ear tissue of the rats in the low dose group of traditional Chinese medicine decreased significantly(P<0.05)There was no statistical significance(P> 0.05).5 RT-PCR Test Results: changes of IL-6,IL-10,CTGF and VEGF gene expression in the ear tissue:(1)compared with the blank group,the expression of IL-6,IL-10,CTGF and VEGF gene in the ear tissue of the model group was significantly higher,with statistical significance(P< 0.05);compared with the model group,the expression of IL-6 gene in the ear tissue of the treated group was decreased in different degrees,especially in the high dose group and the positive group of traditional Chinese medicine The expression of IL-6 gene in rat ear decreased significantly(P< 0.05),but there was no significant difference(P > 0.05).(2)Compared with the blank group,the expression of IL-10,CTGF and VEGF gene in the ear tissue of the model group increased significantly(P< 0.05);compared with the model group,the expression of IL-10,CTGF and VEGF gene in the ear tissue of the rats in the treatment group decreased to varying degrees,especially in the high,medium dose and positive groups of Chinese medicine,The difference was statistically significant(P< 0.05).Although the expression of IL-10,CTGF and VEGF gene in the ear tissue of the low dose group of traditional Chinese medicine decreased,the difference was not statistically significant(P> 0.05).Conclusion:1 The expression of IL-6,IL-10,CTGF and VEGF in the model group was higher than that in the blank group,suggesting that they may be involved in the formation of acne scars.2 Sanhuang gel can effectively reduce edema,reduce inflammatory infiltration among tissues,prevent desquamation and correct the pathological changes of epidermal keratinization.It can reduce the expression of IL-6,IL-10,CTGF and VEGF and the expression of mRNA,and the dose is equivalent to that of 0.025% dimensional A acid.It has a certain dose effect relationship,which is an effective mechanism for the treatment and prevention of auricular Acne Complex Model in rats. |
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