骨成形蛋白7对大鼠肝星状细胞增殖的抑制作用  

The inhibitory effect of bone morphogenetic protein 7 on proliferation of rat hepatic stellate cells

作　　者：王春莹[1] 陈达之 刘俊[1] 李春艳[1] 林镯[1] 阳韬[1] 陆小蒟[1] 陈永平[1] 

机构地区：[1]温州医学院附属第一医院感染科,325000 [2]温州医学院仁济学院临床一系

出　　处：《中华传染病杂志》2012年第4期195-199,共5页Chinese Journal of Infectious Diseases

基　　金：基金项目：浙江省自然科学基金资助项目（Y207464）;王宝恩肝纤维化研究基金资助项目（20080017）

摘　　要：目的研究骨成形蛋白7（BMP7）对大鼠肝星状细胞（HSC）增殖的抑制作用。方法体外培养HSC—T6细胞，分为对照组、TGFβ1组、TGFβ1＋BMP7组和TGFβ1＋BMP7＋Noggin组。RT—PCR法检测果蝇蛋白质1（smadl）、α-平滑肌肌动蛋白（mSMA）、gremlin和纤维连接蛋白（FN）mRNA的表达，Western印迹法检测磷酸化smadl／5／8（P-smadl／5／8）、α-SMA、gremlin蛋白的表达。采用单因素方差分析、LSD检验、Dunnett T3检验和Pearson直线相关分析进行统计学处理。结果对照组HSC—T6细胞中可见P—smadl／5／8、α-SMA、gremlin及FN的表达，TGFβ1刺激后表达量明显增加，差异有统计学意义（均P〈0．05）。与TGFβ1组比较，TGFβ1＋BMP7组α—SMA、gremlin及FN表达下调，但P—smadl／5／8蛋白表达增加（1．613±0．031比2．503±0．014±34．89，P〈0．05）。TGFβ1＋BMP7＋Noggin组α-SMA、gremlin及FN表达较TGFβ1＋BMP7组增加，而P-smadl／5／8蛋白表达下调（t=34．05，P〈0．05）。smad1mRNA在各组表达均无显著变化。结论BMP7对α—SMA、gremlin和FN均有较强的抑制作用。Objective To investigate the inhibitory effect of bone morphogenetic protein 7 （BMP7） on proliferation of rat hepatic stellate cells （HSC）. Methods HSC-T6 cells were cultured in vitro, and divided into four groups： control group, transforming growth factor （TGF）β1 group, TGFβ1 ＋ BMP7 group, and TGFβ1 ＋ BMP7 ＋ Noggin group. The mRNA expressions of smadl, α-smooth muscle actin （α-SMA）, gremlin and fibronectin （FN） were determined by semi-quantitative reverse transcriptase polymerase chain reaction （ RT-PCR ）. The protein expressions of phosphorylated-smadl /5 /8 （P-smadl/5/8）, α-SMA and gremlin were detected by Western blot. The statistical analysis were done using one-factor analysis of variance, LSD-t test, Dunnett＇s T3 test and Pearson linear correlation analysis. Results Phosphorylated-smadl/5/8, α-SMA, gremlin and FN were expressed on HSC--T6 cells in control group, which were significantly up-regulated after TGFβ1 stimulation （P〈0.05）. The expressions of α-SMA, gremlin and FN were significantly down-regulated in TGFβ1 ＋ BMP7 group compared with TGFβ1 group, while P-smadl/5/8 expression was upregulated （1.613±0.031 vs. 2. 503±0.014; t=34.89, P〈0.05）. The expressions of α-SMA,gremlin and FN in TGFβ1 ＋ BMP7 ＋ Noggin group were up-regulated than those in TGFβ1 ＋ BMP7 group, while P-smadl/5/8 was down-regulated （t= 34.05, P〈0.05）. There was no difference of total smadl mRNA expression among each group. Conclusion These results collectively suggest that BMP7 strongly inhibits the expressions of α-SMA, gremlin and FN.

关 键 词：骨形态发生蛋白7 果蝇蛋白质类 磷酰化 肌 平滑肌动蛋白 纤连蛋白类 

分 类 号：R575.2[医药卫生—消化系统]