胃泌素释放肽受体拮抗剂通过NF-κB途径减轻小鼠肝缺血再灌注损伤实验研究

Gastrin-releasing peptide receptor antagonist RC-3095 attenuates hepatic ischemia-reperfusion injury in mice via NF-kB pathway

目的观察胃泌素释放肽(GRP)受体(R)在肝缺血再灌注损伤(I/R)模型中表达情况,并探讨GRP及GRPR拮抗剂RC-3095对小鼠I/R的保护作用。方法构建小鼠I/R模型,将小鼠随机分为假手术组(sham组)、I/R再灌注后3 h组(I/R 3h组)、I/R再灌注后6h组(I/R 6h组)、I/R再灌注后12 h组(I/R 12h组),各6只小鼠。观察GRP及GRPR在缺血再灌注引起的肝损伤中的表达情况;然后将小鼠随机分为假手术组(sham组)、I/R组及RC-3095干预组,HE染色观察小鼠肝组织坏死程度,检测各组小鼠外周血天门冬氨酸氨基转移酶(AST)及丙氨酸氨基转移酶(ALT)水平,ELISA法检测小鼠外周血及肝脏组织中细胞因子肿瘤坏死因子(TNF-α)、白介素-6(IL-6)及IL-1β的表达水平,Western-blot法检测各组小鼠肝脏组织NF-κB磷酸化水平。结果小鼠肝脏缺血再灌注损伤GRP表达增加,6h达到高峰;RC-3095能明显减轻小鼠肝脏坏死程度,抑制炎症因子的释放,抑制NF-κB的磷酸化。结论 RC-3095可以通过抑制炎症因子的释放起到保护肝缺血再灌注损伤,其可能的机制是抑制了NF-κB的磷酸化。

Objective To observe the expression of gastrin-releasing peptide （GRP） receptor （R） in hepatic ischemia- reperfusion injury model, and to explore the protective effect of GRPR antagonist RC-3095 on liver ischemia-reperfusion injury in mice. Methods Twenty-four mice were randomly divided into four groups sham group, I/R 3h group, I/R 6h group, and I/R 12h group, with six mice in each group. The expression of GRP and GRPR was detected in liver injury induced by ischemia-reperfusion. Then, these mice were randomly divided into three groups： sham group, I/R group and RC-3095 group. The changes of liver damage were observed by pathological section and blood of aspartate aminotrans-ferase （AST） and alanine aminotransferase （ALT） content were detected. ELASA method was used to detect the levels of tumor necrosis factor-α （TNF-α）, interleukin-6 （IL-6 ）and interleukin-lp （IL-1β） in the blood. RT-PCR was used to detect content of TNF-α, IL-6 and IL-1β in the liver tissue. Western-blot was used to detect the phosphorylation of NF- kB in liver tissue. Results The results showed that the expressions of GRP and GRPR in model of hepatic ischemia- reperfusion injury were increased and the peak time was at the sixth hour. RC-3095 could significantly reduce the degree of liver necrosis by inhibition of inflammatory factor release and inhibition of NF-κB activation. Conclusions The pro-tective effect of RC-3095 on hepatic ischemia-reperfusion injrny by inhibiting NF-κB phosphorylation.