Effects Of Safflower Decoction On Skin And Lung Fibrosis And The Expression Of TGF-?1 And IL-10 In Scleroderma Mice

ObjectivesTo observe the effect of safflower decoction on the expression of TGF-?1 and IL-10 in skin and lung tissue of scleroderma mice,and to explore the mechanism of safflower decoction on anti-SSc tissue fibrosis.Methods128 BALB/C mice were randomly divided into 4 groups:blank control group,model control group,prednisone acetate group,safflower decoction group,32 mice in each group.To establish scleroderma mouse model,meanwhile,the control group and the model control group were given normal saline,the prednisone acetate group and the safflower decoction group were respectively treated with prednisone acetate suspension and safflower decoction for drug intervention.Eight mice in each group were sacrificed at week 1,week 2,week 3 and week 4.The pathological changes of skin and lung tissues were observed by HE staining.Masson staining,Sirius Red staining and Van Gieson staining were used to observe the distribution of collagen fibers.The changes of collagen fibers in skin and lung tissues were measured.The localization and expression of TGF-?1 and IL-10 in skin and lung tissue were detected by IHC.The expression of TGF-?1 and IL-10 in serum was detected by ELISA.Results1.HE staining of skin and lung tissueThere was no significant change in the skin and lung tissue of the blank control group at all observation time points,and the structures of the skin and lung tissue were clear and complete,without obvious inflammatory cell infiltration.In the model control group,there was a slight infiltration of inflammatory cells in the skin and lung tissue at week 1,and the dermal thickening was not obvious.In the second week,the dermis was thickened and a lot of inflammatory cells were infiltrated.The third week of dermal thickening was further,the inflammatory cells decreased and collagen fiber hyperplasia occured,also fibroblasts increased;The dermal thickening went a step further with inflammatory cell infiltration decreased,collagen fibers and the number of fibroblastsincreased much more at 4^thh week.The levels of inflammatory cells and collagen fibers in the prednisone acetate group and the safflower decoction group were less than those in the model control group at each time point.2.Masson staining,Sirius Red staining,Van Gieson staining of skin and lung tissueThere was no significant change in the skin and lung tissue of the blank control group at each observation time point,without or with little collagen fibrillation.In the 1^stt week,the collagen fiber became disordered with less collagen hyperplasia in the skin and lung tissues of the model control group,and the number of fibroblasts increased gradually at the 2nd,3rd and 4th week.Compared with the model control group,the collagen fibers and fibroblasts were decreased in the skin and lung tissues of the prednisone acetate group and the safflower decoction group.3.The inflammation score of skin and lung tissueThere was no significant change in tissue inflammation score at each time point in the blank control group?P>0.05?.The inflammation of the model control group gradually increased in the first and second week,reached the peak in the second week;and gradually decreased in 3^rd,4^thh weeks,but still higher than the blank control group mice.The inflammation of prednisone acetate group and safflower intervention group were lower than that of model control group?P<0.05?.4.Collagen fiber content changes of skin and lung tissueThere was no significant change in collagen fibers in the blank control group?P>0.05?.Compared with the blank control group,the collagen fibers of all the observation mice in the model control group increased?P<0.05?,and gradually increased with the modeling time.The content of collagen fibers in prednisone acetate group and safflower decoction group decreased compared with the model control group?P<0.05?.5.Skin and lung tissue TGF-?1,IL-10 IHC detectionThere was no significant change in the expression of TGF-?1 and IL-10protein in blank control group?P>0.05?.Compared with the blank control group,the expression of TGF-?1 and IL-10 in the mice were increased at each time point?P<0.05?.Compared with model control group,the expression of TGF-?1 and IL-10 in prednisone acetate group and safflower decoction group decreased at each time point,the difference was statistically significant?P<0.05?.6.The ELISAdetection of TGF-?1 and IL-10 in serumThe levels of TGF-?1 and IL-10 in the blank control group were lower at each time point?P>0.05?.Compared with the blank control group,the levels of TGF-?1and IL-10 in the model control group increased at each time point?P<0.05?.Compared with model control group,the expression of TGF-?1 and IL-10 in prednisone acetate group and safflower decoction group decreased at each time point,the difference was statistically significant?P<0.05?.Conclusions1.The expression of TGF-?1 and IL-10 increased in the skin and lung tissues of bleomycin-induced scleroderma mice.2.Safflower decoction may reduce the expression levels of TGF-?1,IL-10 and immune inflammation,also relieve scleroderma tissue fibrosis process.