胃泌素释放肽受体拮抗剂RC―3095通过抑制TLR4途径减轻小鼠肝缺血再灌注损伤的研究

[摘要] 目的 本文^察胃泌素释放肽（GRP）受体（R）在肝缺血再灌注损伤模型中的表达情况，并探讨GRP及 GRPR拮抗剂RC-3095对小鼠肝缺血再灌注损伤是否具有保护作用。 方法 构建小鼠肝缺血再灌注损伤模型，将小鼠随机分为假手术组（sham组）、I/R再灌注后3 h组（3 h）、I/R 再灌注后6 h组（6 h）、I/R再灌注后 12 h组（12 h），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（I/R）、RC-3095干预组（RC-3095），HE染色观察小鼠肝组织坏死程度，检测各组小鼠外周血谷草转氨酶（AST）、谷丙转氨酶（ALT）水平，ELISA法检测小鼠外周血及肝脏组织中细胞因子TNF-α、IL-6、IL-1β的表达水平，RT-PCR法检测肝脏组织TLR4水平。 结果 小鼠肝脏缺血再灌注损伤GRP及GRPR表达增加，6 h达到高峰，随后表达降低；RC-3095能明显减轻肝缺血再灌注损伤模型小鼠肝脏坏死程度，抑制炎症因子（TNF-α、IL-6、IL-1β）的释放，抑制TLR4的表达。 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TLR4的表达。

[关键词] 胃泌素释放肽受体；RC-3095；肝缺血再灌注损伤；TLR4

[中图分类号] R657.3 [文献标识码] B [文章编号] 1673-9701（2017）04-0037-04

Study on the effect of gastrin-releasing peptide receptor antagonist RC-3095 on reducing hepatic ischemia reperfusion injury in mice by inhibiting TLR4 pathway

SUN Lu LU Yelan JIAN Chunyan FENG Hui CHEN Mingsheng CAO Lijun

Department of Anesthesiology， No. 113 Hospital of PLA， Ningbo 315040， China

[Abstract] Objective To observe the expression of gastrin-releasing peptide （GRP） receptor （R） in the model of hepatic ischemia reperfusion injury， and to explore whether GRP and GRPR antagonist RC-3095 has a protective effect on hepatic ischemia reperfusion injury in mice. Methods The model of hepatic ischemia reperfusion injury in mice was established， and the mice were randomly divided into sham group， 3h group after I/R reperfusion group （3 h）， 6 h group after I/R reperfusion （6 h） and 12 h group after I/R reperfusion （12 h）. The expression of GRP and GRPR in hepatic injury induced by ischemia reperfusion was observed. The mice were then randomly divided into sham group （sham group）， I/R group （I/R） and RC-3095 intervention group （RC-3095）， and HE staining was used to observe the degree of liver necrosis in mice. AST and ALT levels at the peripheral blood were measured in mice in each group， and the expression levels of cytokines of TNF-α， IL-6 and IL-1β in peripheral blood and liver tissues were tested by ELISA. The level of TLR4 in liver tissue was tested by RT-PCR. Results It was found that GRP and GRPR expression was increased in the mice with liver ischemia reperfusion injury， and the expression level reached a peak at 6 h， followed by a decreased expression； RC-3095 could significantly reduce the degree of liver necrosis in mice with the model of liver ischemia reperfusion injury， inhibit the release of inflammatory factors（TNF-α， IL-6 and IL-1β） and inhibit the expression level of TLR4. Conclusion RC-3095 can protect hepatic ischemia reperfusion injury by inhibiting the release of inflammatory factors， and its possible mechanism is to inhibit the expression of TLR4.

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（收稿日期：2016-11-9）