Effect Of Qingxiao Method On Sebum Metabolism And SREBP-1 Signal Pathway In Golden Hamster

Part one:Regulation of qingxiao method on the function of sebaceous gland and the state of sebaceous metabolism in golden hamsterObjective and significance:To investigate the effect of qingxiao method on the metabolism of cortex lipid in golden hamster by observing the changes of serum hormone level,the morphology of sebaceous gland spot and the level of lipid secretion.Methods:The male golden hamster was used as the animal model.Because of the hyperplasia of sebaceous gland spots on the back and the high dependence on androgen,the control group,the low dose group,the middle dose group,the high dose group and the Western Medicine（spironolactone）group were set up.The control group was given 10 m L/（kg?d）distilled water,the low dose group,the middle dose group and the high dose group were given5.63,11.25 and 22.5g/（kg·d）loquat Qingfei Decoction respectively,and the western group was given 6.25 mg/（kg·d）spironolactone for 4 weeks.The area and weight of sebaceous gland spots of golden hamster were measured,the levels of T and E₂ in serum were detected by radioimmunoassay,the content of NEFA and TG in sebaceous gland spots were detected by colorimetry,the testicular coefficient of each group was calculated,and the sebaceous gland spots group was observed by HE staining and oil red O staining Histopathological changes.Results:1.Weight and testicular coefficientThere was no significant difference in the body weight of golden hamster in each group;compared with the control group,there was no significant difference in the testicular coefficient between the low-dose group,the middle dose group and the high-dose group（all P>0.05）.The testicular coefficient of the western medicine group was significantly reduced,and the difference was statistically significant（P<0.01）.2.Appearance and area measurement of sebaceous gland spotsIn the control group,the color and area of the sebaceous gland spots were dark and large.In the middle dose group,the high dose group and the western medicine group,the color and area of the sebaceous gland spots became lighter and smaller in varying degrees.In the middle dose group and the western medicine group,the color and area of the sebaceous gland spots changed most significantly.There was no significant difference in the area of sebaceous gland spots between the groups before and after administration（all P>0.05）;after administration,compared with the control group,the area of sebaceous gland spots in the middle dose group,the high dose group and the western medicine group decreased significantly（all P<0.01）;compared with the western medicine group,there was no significant difference in the area of sebaceous gland spots in the middle dose group（P>0.05）,and the high dose group increased significantly（P<0.01）;compared with the middle dose group,the area of sebaceous gland in the high dose group increased significantly（P<0.05）.3.Changes of serum hormone levelCompared with the control group,the middle dose group,the high dose group and the western medicine group showed a significant decrease in serum T-water（all P<0.01）;compared with the western medicine group,the middle dose group showed no significant difference in serum T-level（P>0.05）,the high dose group showed a significant increase in serum T-level（P<0.01）;compared with the middle dose group,the serum T level in the high dose group increased significantly（P<0.05）.ompared with the control group,the level of serum E₂ in the low dose group,the middle dose group,the high dose group and the western medicine group increased slightly,there was no significant difference（all P>0.05）.4.HE staining of sebaceous plaquesIn the control group,the sebaceous glands were divided into two or three layers.Compared with the control group,there was no significant difference in the thickness of the sebaceous glands in the low dose group of traditional Chinese medicine,and the sebaceous glands in the middle dose group and the western medicine group were arranged closely;the thickness of the sebaceous glands in the middle dose group and the western medicine group was obviously thinner,arranged loosely,and the number of sebaceous glands was reduced,mostly one layer.In the high dose group,the thickness of sebaceous glands was thinner than that in the control group,and the number of sebaceous glands was reduced,mostly 1-2 layers.5.Oil red O staining of sebaceous gland spotIn the control group,because of the hyperplasia of sebaceous glands,the secretion of sebaceous glands increased,and there were red lipid blobs in the cytoplasm of sebaceous gland cells,which even fused into pieces to form large lipid blobs.Compared with the control group,the accumulation of sebum in the sebaceous gland cells of golden hamster in the middle dose group and western medicine group was significantly reduced,while that in the low dose group and high dose group was slightly reduced.6.The content of NEFA and TG in local sebaceous plaquesCompared with the control group,the content of NEFA and TG in the middle dose group,the high dose group and the western medicine group decreased significantly（all P<0.01）;compared with the western medicine group,the content of NEFA and TG in the middle dose group had no significant difference（all P>0.05）,the content of NEFA and TG in the high dose group of traditional Chinese medicine increased significantly（all P<0.01）;compared with the middle dose group,the content of NEFA and TG in the high dose group of traditional Chinese medicine increased significantly（P<0.05,P<0.01）;compared with the middle dose group,the expression levels of AR,SREBP-1c and ACC1m RNA in the low dose group and the high dose group were significantly higher（P<0.01）.Conclusion:1.Qingxiao method can effectively reduce the serum T level of golden hamster,inhibit the hyperplasia of sebaceous gland,and reduce the secretion of sebum.2.The mechanism of qingxiao method in the treatment of acne may be related to the reduction of T level.Part two Based on the SREBP-1 signal pathway,the mechanism of Qingxiao method regulating sebum metabolism was discussedObjective and significance:By detecting the m RNA and protein expression of AR,SREBP-1,ACC1 in the sebaceous plaques of golden hamster,to explore the mechanism of Qingxiao method in regulating sebaceous metabolism.Methods:RT-q PCR was used to detect the relative expression of AR,SREBP-1c and ACC1 m RNA in the sebaceous plaques of golden hamsters;immunohistochemistry was used to detect the distribution of AR,SREBP-1and ACC1 protein in the sebaceous plaques of golden hamsters;Western blot was used to detect the relative expression of AR,SREBP-1 and ACC1 protein in the sebaceous plaques of golden hamsters.Results:1.Immunohistochemical detection of AR,SREBP-1,ACC1 protein expression in sebaceous plaquesCompared with the control group,the relative expression of AR,SREBP-1 and ACC1 protein in the sebaceous plaques of the middle dose group,the high dose group and the western medicine group decreased significantly（all P<0.01）;compared with the western medicine group,the relative expression of AR,SREBP-1 and ACC1 protein in the sebaceous plaques of the middle dose group had no significant change（all P>0.05）;the relative expression of AR,SREBP-1 and ACC1 protein in sebaceous plaques of high dose group was significantly increased（P<0.01）;compared with the middle dose group,the relative expression of AR,SREBP-1 and ACC1 protein increased significantly（P<0.05,P<0.01）.2.Western blot analysis of AR,SREBP-1 and ACC1 protein expression in the sebaceous plaques of golden hamsterCompared with the control group,the relative expression of AR and ACC1 protein in the low dose group was significantly reduced（all P<0.05）,the relative expression of AR,SREBP-1 and ACC1 protein in sebaceous plaques of middle dose group,high dose group and Western medicine group decreased significantly（P<0.05,P<0.01）;compared with the western medicine group,the relative expression of AR,ACC1 protein increased significantly（all P<0.01）in the low dose group,and the relative expression of AR,SREBP-1,ACC1 protein increased significantly（all P<0.01）in the middle dose group and the high dose group;compared with the middle dose group,the relative expression of AR and ACC1 protein in the low dose group was significantly increased（P<0.01）,and the relative expression of AR,SREBP-1 and ACC1protein in the high dose group was significantly increased（P<0.01）.3.Expression level of AR,SREBP-1c and ACC1m RNA in sebaceous plaques of golden hamsterCompared with the control group,the expression levels of AR,SREBP-1c and ACC1m RNA in the low dose group,the middle dose group,the high dose group and the western medicine group were significantly lower（all P<0.01）;compared with western medicine group,the expression of AR,SREBP-1c and ACC1m RNA in low dose group,middle dose group and high dose group were significantly higher（P<0.05,P<0.01）;compared with the middle dose group,the expression levels of AR,SREBP-1c and ACC1m RNA in the low dose group and the high dose group were significantly higher（P<0.01）.Conclusion:Qingxiao method can reduce the T level and inhibit the over activation of SREBP-1 signal pathway,so as to improve the abnormal metabolism of sebum in the sebaceous gland spots of golden hamster,so as to achieve the purpose of acne treatment.