SOCS2基因转染对人肾小球系膜细胞TGF-β、Ⅳ型胶原蛋白、纤维连接蛋白表达的影响

Effect of SOCS2 gene transfection on expression of TGF- β,Collagen Ⅳ and FN in human glomerular mesangial cells

目的观察过表达细胞因子信号转导抑制因子(SOCS2)对人肾小球系膜细胞(HMCs)中转化生长因子β(TGF-β)、Ⅳ型胶原蛋白(CollagenⅣ)、纤维连接蛋白(FN)表达的影响,并探讨其可能机制。方法体外培养HMCs,分为A、B、C、D、E、F组,A、B组分别加入Ad-SOCS2病毒液(含SOCS2的腺病毒病毒液)、Ad-null病毒液(只含腺病毒的病毒液),24 h后将培养液更换成高糖培养液;C组加入30 mmol/L的D-葡萄糖;D、E组分别加入Ad-SOCS2病毒液、Ad-null病毒液,24 h后将培养液更换成常规培养液;F组加入5.5 mmol/L的D-葡萄糖+24.5mmol的D-甘露醇。Western blotting法检测各组TGF-β、CollagenⅣ、FN及贾纳斯激酶2(JAK2)、信号传导及转录激活因子3(STAT3)、磷酸化贾纳斯激酶2(p-JAK2)、磷酸化信号传导及转录激活因子3(p-STAT3)。结果与F组比较,A、B、C组TGF-β、CollagenⅣ、FN、p-JAK2、p-STAT3的相对表达量升高;与C组比较,A组TGF-β、CollagenⅣ、FN、p-JAK2、p-STAT3的相对表达量降低;P均<0.01。结论 SOCS2通过抑制JAK/STAT信号通路来下调HMCs中TGF-β、CollagenⅣ、FN表达。

Objective To observe the effect of overexpressing suppressor of cytokine signaling 2 （SOCS2） on the ex- pression of transforming growth factor β（TGF-β）, Collagen IV and fibronectin （FN） in human glomerular mesangial cells （HMCs） and to investigate its possible mechanisms. Methods HMCs euhured in vitro were divided into groups A, B, C, D, E and F： Groups A and B were respectively added with Ad-SOCS2 venom （containing SOCS2 adenovirus） and Ad-null venom （ containing adenovirus）, after 24 h, we changed the nutrient solution into high glucose solution; group C was added with 30 mmol/L D-glucose; groups D and E were respectively added with Ad-SOCS2 venom and Ad-null venom, after 24 h, we changed the venom into conventional solution; group F was added with 5.5 mmol/L D-glucose ＋ 24.5 mmol D- mannitol. Western blotting was used to detect the TGF-β, Collagen IV, FN and Janus kinase 2 （JAK2）, signal transducer and activator of transcription 3 （STAT3）, phosphorylated Janus kinase 2 （p-JAK2） and phosphorylated signal transducer and activator of transcription 3 （p-STAT3）. Results Compared with group F, the expression of TGF-β, Collagen IV, FN, p-JAK2, and p-STAT3 was significantly increased in the groups A, B and C; compared with group C, the expression of TGF-β, Collagen IV, FN, p-JAK2 and p-STAT3 in the group A was significantly decreased （all P 〈 0.01 ）. Conclusion SOCS2 can down-regulate the expression of TGF-β, Collagen IV and FN in HMCs by inhibiting JAK/STAT sigaling pathway.