小鼠肝脏部分缺血再灌注过程中Toll样受体2的激活及意义(英文)

Activation of TLR2 in mouse partial hepatic ischemia/reperfusion injury model and it′s relation with liver function impairment

目的探索Toll样受体2(Toll-likereceptor2,TLR2)在小鼠肝脏部分缺血再灌注过程中在缺血肝叶中的激活,分析TLR2的激活与肝功能损伤之间的关系。方法BALB/c小鼠复制肝脏部分缺血再灌注(缺血1h再灌注4h)损伤动物模型(I/R组),假手术对照组(SH组)同样行开腹手术但不夹闭血管。采用实时荧光定量PCR(Real-time-PCR)方法定量检测缺血肝叶中TLR2mRNA的表达变化,同时采用Westernblot检测缺血肝脏组织中TLR2蛋白的表达变化,检测门静脉血浆丙氨酸氨基转移酶(pALT)水平。结果肝脏部分缺血1h再灌注4h后,I/R组与SH组小鼠缺血肝叶TLR2mRNA的表达(ΔCt值)分别为(1.06±0.91)vs(5.08±1.32)(P<0.01),由于ΔCt值越小则基因表达水平越高,说明缺血再灌注过程中缺血肝叶TLR2mRNA的表达水平增高。且I/R组小鼠缺血肝叶TLR2蛋白的表达(OD值)水平较SH组高[(433.91±25.53)vs(102.86±13.58),P<0.01],I/R组中门静脉血清ALT水平升高[(848.33±271.37)μ/Lvs(42.39±14.75)μ/L,P<0.01]。结论TLR2在肝脏缺血再灌注过程中缺血肝叶的表达增强,且这种变化与肝功能的损伤相关。

To explore TLR2 (Toll-like receptor 2) expression under the condition of partial liver ischemia/reperfusion injury in BALB/c mice and discuss its relationship with liver function impairment. The hepatic partial ischemia/reperfusion injury model was established as the following, the portal vein and hepatic artery supplying to the media and left lobes of the liver were obstructed by atraumatic artery microclip, the obstruction lasted for about 60 min, then reperfusion was fulfilled by removal of the clip. The liver samples were collected at the 4th hour after the restoration of blood inflow. TLR2 mRNA and membrane protein were extracted from the liver samples and analyzed quantitatively by method of real-time PCR(indicated as the value of ΔCt: the greater the value, the less the TLR2 mRNA expressed) and western blot, respectively. At the same time point portal vein serum was taken for further detection of the level of ALT. TLR2 mRNA and protein were up regulated markedly in the mice partial liver ischemia/reperfusion injury model compared to the sham operation group (value of ΔCt: (1.06±0.91) vs (5.08±1.32) (P <0.01); value of OD: (433.91±25.53) vs (102.86±13.58), P <0.01),as well as the level of portal vein ALT (848.33±271.37) μ/L vs (42.39±14.75) μ/L, P <0.01). [Conclusion] TLR2 expression was up regulated in the model of mice partial ischemia/reperfusion injury, which was correlated with the variation of liver function impairment. It suggests the involvement of TLR2 in ischemia/reperfusion injury pathological process.