Therapeutic Effect Of Adipose Tissue-derived Stem Cells On Bleomycin-induced Mice Of Scleroderma

Objective To investigate the effects and mechanisms of adipose-derived stem cells(ADSCs)on bleomycin-induced mice of scleroderma.Method 24 C57BL/6 female mice were randomly divided into control group,bleomycin group,ADSCs(hypodermic injection)group and ADSCs(intravenous injection)group.bleomycin [2mg/(kg·day)] was injected into mice to establishment model of scleroderma.There are 6 mice in each group.Control group mice which were injected with normal saline 2 m L/(kg.d)by subcutaneous.The rest of the three groups mice which were injected with BLM.Three weeks later,Control group and bleomycin group mice which were injected with normal saline 2 m L/(kg.d)by subcutaneous.ADSCs groups mice which were injected with ADSCs(2×105)by subcutaneous and intravenous injection respectively.Once every four days,repeat this method three times.T-helper 17(Th17)and regulatory T cell(Treg cell)of spleen cells were detected by flow cytometry.The levels of cytokines in lung tissue and in serum were detected by real-time fluorescence quantification.Real-time polymerase chain reaction(PCR)and enzyme-linked immuno sorbent assay(ELISA).The pathology change of skin and lung tissue was observed by hematoxylin eosin(HE)staining.Results The proportion of Th17 and Treg increased in BLM group than control group(P<0.05),the expression of Th17 significantly decreased(P<0.05)and the expression of Treg increase after the ADSCs therapy(P<0.05).The expression levels of IL-17,IL-6,tumor necrosis factor-?(TNF-?)m RNA in the lung tissue and IL-6 in the serum increased in BLM group than control group(P<0.05).The expression of these cytokines significant decreased after the ADSCs therapy(P<0.05).But there were no obvious difference between ADSCs(hypodermic injection)group and ADSCs(intravenous injection)group.The expression of TGF-? in the serum increased in BLM group than control group(P<0.05),however,the expression of TGF-? m RNA has no significant differences in BLM group than control group.The expression of TGF-? m RNA and protein have no significant differences in ADSCs(hypodermic injection)group and ADSCs(intravenous injection)group than BLM group.HE staining showed inflammation of lung tissue was relieved and the dermal thickness and collagen deposition were decreased after the ADSCs therapy.Conclusions ADSCs could effectively alleviate inflammation of the lungs and fibrosis of skin;the effects of anti-inflammatory and anti-fibrosis were associated with immune regulating function.