siRNA腺病毒载体对兔骨髓基质细胞成脂分化的干扰效应

目的 观察靶向过氧化物酶体增殖子活化受体-γ(PPARγ)基因siRNA腺病毒载体阻断乙醇诱导兔骨髓基质细胞(MSCs)的成脂分化.方法 培养兔MSCs随机分为7组:N:对照组,M:模型组,CO:感染空载体组,C1:感染无关siRNA组,S1、S2、S3:干扰1、2、3组.将重组腺病毒载体转染细胞,检测MSCs内PPARγ mRNA、osteocalcin mRNA、PPARγ蛋白表达、甘油三酯、ALP活性和培养液骨钙素含量,并计数脂肪细胞.结果 N、M、CO、C1组PPARγ mRNA和osteocalcinmR-NA表达值分别为(0.39±0.02)、(0.75±0.03)、(0.74±0.03)、(0.73±0.02)和(1.09±0.19)、(0.50±0.10)、(0.46±0.12)、(0.49±0.13),M、CO、C1组脂肪细胞多,甘油三酯高,ALP活性与骨钙素量均低.S1、S2、S3组PPARγ mRNA和osteocalcin mRNA表达值分别为(0.28±0.03)、(0.30±0.03)、(0.31±0.01)和(0.92±0.09)、(0.87±0.32)、(0.93±0.25),脂肪细胞数、甘油三酯量、ALP活性值与骨钙素含量均接近正常,与M、CO、C1组间差异有统计学意义(P<0.05),与N组间差异无统计学意义(P>0.05).结论 该腺病毒载体能阻断乙醇诱导的MSCs成脂分化.

Interfering effect of siRNA adenovirus vector on adipogenic differentiation in marrow stromal cells of rabbits

Objective To observe the inhibitory effects of siRNA adenovirus vector targeting per-oxisome proliferator-activated receptor-γ (PPARγ) gene on adipogenic differentiation induced by alcohol in marrow stromal cells (MSCs) of rabbits. Methods MSCs were cultured, and randomly divided into 7 groups:normal (N) group,model (M) group,empty infection (CO) group, irrelative array to the siRNA (CI) group, RNA interference 1,2 and 3 (S1, S2 and S3 ) groups. Recombinant adenovirus vector was transfected into MSCs. The expression levels of PPARγ mRNA, osteocalcin mRNA and PPARγ protein, content of intracellular triglyceride, ALP activity in cells, and osteoealein in culture media were deter-mined. The number of adipocytes was counted under light microscope. Results The expression levels of PPARγ mRNA in groups N, M, CO and C1 were (0.39±0.02), (0.75±0.03), (0.74±0.03), and (0.73±0.02),and those of osteocalcin mRNA (1.09±0.19),(0.50±0.10), (0.46±0.12),and (0.49±0.13),respectively. The number of adipocytes, content of triglyceride in groups M, CO and C1 were increased markedly, but the ALP activity and ostcoealcin in culture media decreased significantly. The expression levels of PPARγ mRNA in groups S1,S2 and S3 were (0.28±0.03), (0.30±0.03), and (0.31±0.01), whereas those of osteocalcin mRNA (0.92±0.09), (0.87±0.32), and (0.93± 0.25), respectively. The number of adipoeytes, content of triglyceride, ALP activity, and ostcocalcin in cul-ture media in groups S1, S2 and S3 approximated to the results in group N,which had statistically signifi-cant differences with the results in groups M, CO and C1 (P<0.05), but had no statistically significant differences with those in group N (P>0.05). Conclusion siRNA adenovims vector could suppress the differentiation of MSCs into adipocytes induced by alcohol.