流体剪切应力对肝星状细胞的活化和相关基因表达的影响

Effect of fluid shear stress on activation of hepatic stellate cells and expression of α-SMA,collogen-Ⅰ,collogen-Ⅲ,MMP-2,and MMP-9

目的:探讨不同大小的流体剪切应力对大鼠肝星状细胞株HSC-T6的活化和基因表达的影响.方法:对种植于鼠尾胶原上的HSC-T6细胞施以剪切应力处理,剪切应力的大小分别为6、12、20dyn/cm2,干预时间为3h.TRIzol法提取mRNA,采用SYBR Green荧光定量RT-PCR方法分别检测各组-SMA、Collogen-Ⅰ、Collogen-Ⅲ、MMP-2及MMP-9基因的表达情况,流式细胞术检测细胞内蛋白-SMA的表达.结果:剪切应力处理3h后,与对照组相比,6dyn/cm2剪切应力组-SMA、Collogen-Ⅲ的表达降低,但随着剪切应力的增加(12dyn/cm2、20dyn/cm2),-SMAmRNA较对照组表达升高;20dyn/cm2剪切应力组Collogen-Ⅲ基因表达较对照组升高;3种剪切应力组之间Collogen-ⅠmRNA的表达无统计学差异;3种剪切应力均上调MMP-2mRNA的表达.结论:低剪切应力对HSC-T6的活化有一定的抑制作用,相反高剪切应力可以促进HSC-T6的活化,促进Collogen-Ⅲ、MMP-2的表达.

AIM：To investigate the effects of different types of fluid shear stress on the activation of hepatic stellate cells-T6（HSC-T6） and expression of-SMA,collogen-Ⅰ,collogen-Ⅲ,MMP-2,and MMP-9.METHODS：HSC-T6 cells were seeded on slides precoated with rat tail collagen and exposedto different types of fluid shear stress（6,12,20 dyn/cm2） for 3 h,and static cells served as controls.The gene expression of-SMA,collogen-Ⅰ,collogen-Ⅲ,MMP-2 and MMP-9 was assayed by real-time RT-PCR.Intracellular-SMA protein was analyzed by FACS.RESULTS：After treatment by fluid shear stress,the expression of-SMA and collogen-Ⅲ was lower in the low shear stress（6 dyn/cm2） group than that in the control group.However,the expression of-SMA was higher in the moderate（12 dyn/cm2） and high（20 dyn/cm2） shear stress groups than that in the control group,and the expression of collogen-Ⅲ was higher in the high shear stress group than that in the control group.There was no significant difference in the expression of collogen-Ⅰ among different fluid shear stress groups.The expression of MMP-2 was higher that in three fluid shear stress groups than in the control group.CONCLUSION：High fluid shear stress can lead to the activation of HSC-T6 cells and promote the mRNA expression of-SMA,collogen-Ⅲ and MMP-2.