Effects of RNAi for target on PPARγ to suppress adipogenic differentiation of MSCs of rabbit induced by ethanol
WANG Shaohua1),WANG Yisheng1),LI Yuebai2),ZHAO Guoqiang3)1)Departmemt of Orthopaedic Surgery,the First Affiliated Hospital,Zhengzhou University;Open Laboratory of Unode Science of Clinical Medicine of Henan Province,Zhengzhou 450052 2)Department of Biochemistry,College of Basic Medical Sciences,Zhengzhou University,Zhengzhou 450001 3)Department of Microbiology and Immunology,College of Basic Medical Sciences,Zhengzhou University,Zhengzhou 450001
Aim:To observe effects of RNAi for the target on peroxisome proliferator-activated receptor-γ(PPARγ) suppress the adipogenic differentiation of bone marrow stromal cells (MSCs) induced by ethanol.Methods:2nd dynasty MSCs were selected and randomly separated into 7 groups.Group normal (N):the cells were treated without the adenovirous and alcohol as control. Group model (M):the cells were treated with 0.09 mol/L ethanol. Group empty infection (C0):the cells were treated with adenovirous 1×105 U and 0.09 mol/L ethanol. Group irrelative array to the siRNA (C1):the cells were treated with irrelative array to the siRNA adenovirous 1×105 U and 0.09 mol/L ethanol. Group RNA interference 1,2,3 (S1,S2,S3):the cells were correspondingly treated with relative siRNA adenovirous 1,2,3,1×105 U,and with 0.09 mol/L ethanol respectively. After the cells being treated 7 days,the expression of PPARγ mRNA was measured with RT-PCR for 7 groups. After 14 days,the content of triglyceride in cells was measured. After 21 days,the cells were stained with Sudan Ⅲ,and then the number of adipocytes was counted on a light microscope. Results:There was high expression of PPAR mRNA in the cells,the number of adipocytes in culture increased,and the content of triglyceride in cells increased in group M,C0,and C1,and there were significant differences compared with group N (P0.05). There was low expression of PPAR mRNA in the cells,the few adipocytes in culture,and the content of triglyceride in cells slightly increased in group S1,S2,and S3,and there were significant differences compared with group M,C0,and C1 (P0.05),as well as no significant differences compared with group N (P0.05).Conclusion:RNAi for the target on PPARγ could efficaciously suppress the adipogenic differentiation of the MSCs induced by alcohol,which establishes a fine experimental fundament for the prevention to the ethanol-induced osteonecrosis of the femoral head.