| Purpose:Based on the efficacy of Taohongsiwutang,the effects of Taohongsiwutang on the changes of blood flow of mouse skin tissue,the changes of i NOS,CD206,VEGF protein content and the polarization of macrophages were observed,and the mechanism of Taohongsiwutang in regulating the photoaging process of mouse skin was discussed,which provided theoretical basis and basis for the clinical treatment of photoaging with Chinese medicine,and also provided new ideas for developing new anti-aging products of Chinese medicine in the field of medicine and beauty.Material and method:1.Experimental animal grouping,modeling and evaluationSixty 2~3-month-old female SD mice with SPF were randomly divided into four groups:blank group,model group,high-dose group of rosé soup,and vitamin E group.Except for the blank group,the mice in the other groups were irradiated with ultraviolet lamps,and the cumulative irradiation intensity of UVA was 261.79J/cm2 and the cumulative irradiation intensity of UVB was 6.54J/cm2,and a photoaging mouse model was constructed.Starting from the day after the successful photoaging modeling,each medication group was given the corresponding drug continuous intervention for 4W.The mouse state was observed,evaluated and recorded throughout the experiment.2.Test indicatorsThe camera photographs the skin state of the mouse;The laser speckle contrast imaging system was used to detect skin microcirculation and blood flow changes;The content of collagen fibers in the skin dermis of photoaged mice was observed by HE staining method and Masson three-color staining method;The distribution of vascular endothelial growth factor(VEGF)protein expression in the skin tissues of each group of mice was observed by immunohistochemical staining(IHC)and semi-quantitatively analyzed;The levels of induced nitric oxide synthase(i NOS)and mannose receptor(CD206)in each group were detected by enzyme-linked immunosorbent assay(ELISA);Western blotting(WB)was used to detect the expression of i NOS and CD206 proteins in the skin tissues of each group of mice.3.Statistical methodsSPSS 25,Image j,and Graph Pad Prism8 software are used for data analysis,image analysis,and mapping.P<0.05 is statistically significant,and P<0.01 is statistically significant.Results:1.General situation: After successful modeling,it was observed that the mice had symptoms of photoaging: body sleepiness,curling,uneven skin color,wilting and dryness,coarse texture,rough and thick,desquamation,dark red color or ecchymosis.After 4W of drug intervention,it was observed that compared with the model group,the photoaging performance of mice in each drug group was improved to different degrees.2.Skin pathology: HE staining found that rosé soup can effectively improve the skin epidermis and the thickening of the stratum corneum,and skin keratosis is inhibited;Masson staining found that the content of collagen fibers in the dermis of Taohongsiwutang group increased significantly,the density was also significantly increased,and the arrangement was neatly arranged compared with the model group.3.The results of laser speckle scattering showed that compared with the model group,the skin blood flow in the Taohongsiwutang treatment group was significantly increased.4.Enzyme-linked immunosorbent assay(ELISA): 4 weeks after administration,compared with the blank group,i NOS and CD206 in the model group increased significantly(p<0.01);compared with the model group,the i NOS and CD206 in the Taohongsiwutang group were significantly reduced(p<0.01).5.Immunohistochemical staining(IHC): Compared with the blank group,the average optical density value of VEGF protein in the model group was significantly reduced(P<0.01);compared with the model group,the average optical density value of VEGF protein in each drug group was increased to different degrees.6.Western blotting(WB): Compared with the pre-treatment model group and the post-treatment model group,the expression of M1 macrophage-associated protein i NOS was significantly reduced(p<0.01),and the protein expression of M2 macrophage marker CD206 was significantly increased(p<0.01);after 4 weeks of medication,it was found that Taohongsiwutang decoction could significantly inhibit the expression of i NOS protein(p<0.01)and significantly increase the expression of CD206 protein(p<0.01).Conclusion:1.Taohongsiwutang can effectively improve skin blood flow,accelerate blood flow rate,enhance body metabolism,and achieve the effect of improving skin photoaging.2.Taohongsiwutang can regulate the polarization of M1 macrophages into M2 macrophages,so as to repair the skin and achieve the effect of anti-skin photoaging. |
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