中文摘要：

目的探讨人miR-148a过表达对人脂肪间充质干细胞成脂分化过程中炎症及胰岛素敏感性相关蛋白IL-6、瘦素、抵抗素、肿瘤坏死因子a（TNF-a）分泌及表达的影响。方法采用人miR．148a过表达慢病毒及对照慢病毒分别感染人脂肪间充质干细胞，并进行诱导分化。收集第0天、第4天、第10天、第15天细胞培养基上清并提取各时间点的细胞RNA，采用酶联免疫吸附法检测各个时间点IL-6、瘦素、抵抗素、TNF-0I的分泌水平，采用实时定量PCR法检测IL-6mRNA、瘦素mRNA水平。结果人miR-148a过表达慢病毒感染人脂肪间充质干细胞并诱导其分化为成熟脂肪细胞，miR-148a过表达组脂肪细胞中IL-6[第10天：（50．92±14．93）ng／L比（136．86±35．53）ng／L，t=3．86，P=0．018；第15天：（123．64±18．76）ng／L比（178．59±7．54）ng／L．t=1．56，P=0．029]、瘦素[第4天：（12．28±0．56）．g／L比（14．38±0．84）ng／L，t=3．06，P=0．038；第15天：（34．45±4．30）ng／L比（115．85±28．81）ng／L，t=4，79，P=0．009]的表达量较对照组降低，抵抗素的分泌量较对照组升高，但量很低（〈0．20μg/L），在人脂肪细胞分化过程中未检测到TNF—a．分泌。与对照组比较，人miR-148a过表达组脂肪细胞中IL-6mRNA（第4天10．041±0．018比0．089±0．002，t=4．68，P=0．009；第15天：0．138±0．015比0．284±0．069，t=3．55，P=0．024）和瘦素mRNA（第10天：0．163±O．039比0．497±0．042，t=10．13，P=0．001；第15天：0．259±0．033比1．146-4-0．346，t=4．41，P=0．012）的表达水平降低。结论人miR-148a参与调节人脂肪间充质干细胞分化过程中炎症及胰岛素敏感性相关蛋白IL-6、瘦素、抵抗素、TNF—a的分泌和表达，提示人miR-148a可能参与肥胖相关炎症及胰岛素敏感性的调节。

英文摘要：

Objective To investigate the effect of over - expressed miR - 148a on secretion and expression of inflammatory and insulin sensitive protein, such as intedeukin 6 （ IL - 6 ） , leptin, resistin and tumor necrosis factor a （ TNF - a ） , during human adipose - derived mesenchymal stem cells （ hMSCs - Ad ） differentiation. Methods hMSCs - Ad was infected with the lentivirus of hsa - miR - 148a,then the secretion of inflammatory cytokines （ IL - 6, TNF - ct） and insulin sensitive protein （leptin, resistin） during differentiation process. The cells were collected at 0,4, 10 and 15 days for RNA extration. The expression levels of IL - 6, leptin, resistin and TNF - a were measm＇ed by using enzyme - linked immunosorbent assay, IL - 6 and leptin mRNA were examined by using real - time PCR. Results Compared with the control group, after hMSCs - Ad were infected with over - expressed miR - 148a lentivirus, both secretion of IL - 6 [ dayl0 ： （ 50.92 ±14.93 ） ng/L vs （ 136. 86 ± 35. 53 ） ng/L, t = 3. 86, P = 0. 018 ; day 15 ： （ 123.64 ± 18.76 ） ng/L vs （ 178.59 ± 7.54 ） ng/L, t = 1.56, P = 0. 029 ] and leptin day 4 ： （ 12.28 ±0.56 ） ng/L vs （14.38±0.84） ng/L,t =3.06,P=0.038;day 15：（34.45±4.30） ng/Lvs （115.85 ±28.81） ng/L,t =4,79,P= 0. 009 ] were reduced, while the concentration of resistin was higher than that of the control group and the concentration of resistin was less 0.20 μg/L in both groups, and TNF - a did not express, in hMSCs - Ad differentiation. Similarly, I L - 6 （day 4：0.041 ± 0. 018 vs 0. 089 ± 0. 002, t = 4.68, P = 0. 009 ; day 15 -0.138± 0.015 vs 0. 284 ± 0. 069, t = 3.55, P = 0. 024） and leptin （ day 10：0.163± 0.039 vs 0. 497 ± 0.042, t = 10.13, P = 0. 001 ; day 15：0. 259 ± 0.033 vs 1. 146 ± 0. 346, t = 4.41, P = 0. 012） mRNA expression levels were reduced in overexpressed miR - 148a group.Conclusions It suggests that miR - 148a can improve hMSCs - Ad inflammation status and improve insulin resistance through regulati