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目的探索库普弗细胞Toll样受体2(TLR2)在肝脏缺血再灌注损伤中的表达变化,分析库普弗细胞TLR2信号通路参与肝脏缺血再灌注损伤的机制。方法动物分假手术对照(SH)组、缺血再灌注(I/R)组及氯化钆处理(Gd)组,复制小鼠肝脏部分缺血1 h再灌注4 h损伤模型,结束再灌注后,取缺血叶肝脏组织及其库普弗细胞,提取总RNA及膜蛋白质分析TLR 2 mRNA及蛋白的表达,同时提取缺血肝叶组织核蛋白分析核因子(NF—κB),并检测门静脉血中肿瘤坏死因子(TNF α)、丙氨酸氨基转移酶(ALT) 及内毒素水平。结果再灌注4 h,(1)I/R组缺血肝叶TLR2 mRNA及蛋白表达水平较SH组高,△Ct值分别为1.06±0.91和5.08±1.32,t=7.80,P<0.01;A值分别为433.91±25.53和102.86±13.58,t=28.04, P<0.01。Gd组缺血肝叶TLR2 mRNA及蛋白表达水平较I/R组下降,△Ct值分别为4.22±0.84和1.06±0.91,t=7.56,P<0.01;A值分别为125.89±15.49和433.91±25.53,t=25.27,P<0.01。(2)I/R组缺血肝叶库普弗细胞中TLR2 mRNA及蛋白表达水平较SH组高,△Ct值分别为0.52±0.23和2.61±0.1 8, t=17.47,P<0.01;A值分别为379.70±34.16和114.98±21.90,t=15.98,P<0.01。Gd组缺血肝叶库普弗细胞中TLR2 mRNA及蛋白表达水平则较I/R组下降,△Ct值分别为1.90±0.14和0.52±0.23,t= 12.
Objective To explore the changes of TLR2 in hepatic ischemia-reperfusion injury in Kupffer cells and to explore the mechanism of TLR2 signaling pathway in Kupffer cells involved in hepatic ischemia-reperfusion injury. Methods Animals were divided into sham operation group (SH), ischemia / reperfusion (I / R) group and Gd group (Gd group) Then, the liver tissue and its Kupffer cells were extracted from the ischemic lobule. The total RNA and membrane proteins were extracted to analyze the expression of TLR2 mRNA and protein. At the same time, nuclear factor-kappaB (NF-κB) Tumor necrosis factor (TNFα), alanine aminotransferase (ALT) and endotoxin levels in portal vein blood. Results At 4 h after reperfusion, (1) TLR2 mRNA and protein expression in the ischemic liver lobe of I / R group were higher than those in SH group, with △ Ct values of 1.06 ± 0.91 and 5.08 ± 1.32, respectively, t = 7.80, P <0.01; A The values were 433.91 ± 25.53 and 102.86 ± 13.58, respectively, t = 28.04, P <0.01. Compared with I / R group, the expression of TLR2 mRNA and protein in hepatic lobule of Gd group was lower than that of I / R group (△ 4.12 ± 0.84 and 1.06 ± 0.91 respectively, t = 7.56, P <0.01); A values were 125.89 ± 15.49 and 433.91 ± 25.53, t = 25.27, P <0.01. (2) TLR2 mRNA and protein expression in Kupffer cells of I / R group were higher than those in SH group, with △ Ct values of 0.52 ± 0.23 and 2.61 ± 0.1 8, respectively, t = 17.47, P <0.01; A Values were 379.70 ± 34.16 and 114.98 ± 21.90, respectively, t = 15.98, P <0.01. The expression of TLR2 mRNA and protein in Kupffer cells of ischemic liver lobule in Gd group was lower than that in I / R group, with △ Ct values of 1.90 ± 0.14 and 0.52 ± 0.23, respectively, t = 12.