摘要
目的探讨B7H3分子在肝脏缺血再灌注损伤中的作用。方法采用H7H3野生型(WT)、基因敲除(KO)小鼠以及利用特异抗体阻断B7H3功能小鼠制作肝脏70%缺血再灌注损伤模型。再灌注6h收集各组小鼠血清和肝组织标本。通过检测血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平、肝组织HE染色、TUNEL染色和蛋白免疫印迹法检测凋亡蛋白判断肝脏损伤程度,用免疫组织化学染色检测肝内中性粒细胞浸润情况,采用q PCR检测肝脏内趋化因子及炎症因子基因表达。结果缺血90min,再灌注6h后,KO小鼠及抗体阻断组小鼠ALT及AST水平显著高于WT组,且肝细胞嗜酸性变数目、细胞肿胀程度、凝固性坏死范围均更为严重。TUNEL染色及Cleaved caspase-3检测均提示KO组及抗体阻断组小鼠肝内凋亡明显高于WT组。而且,肝组织内中性粒细胞浸润数目、多种趋化因子CXCL1、CXCL5、CXCL10和CCL2以及炎症因子IL6的mRNA表达量KO组及抗体阻断组均显著高于WT组,具有统计学意义。结论 B7H3可通过抑制趋化因子的表达减少肝内中性粒细胞浸润而发挥保护作用。
Objective To study the role of B7H3 in mouse model of hepatic ischemia reperfusion injury. Methods B7H3 wild type ( WT), knock out ( KO) and B7H3 antibody blocked mice were used to build a mouse model of 70% hepatic ischemia reperfusion injury. Samples were collected 6h post reperfusion. Serum level of ALT and AST, liver tissue HE stain, TUNEL stain and apoptosis-regulated proteins were used to estimate hepatic injury. Neutrophils infiltration was measured by immuniohistochemical staining. The inflammatory cytokines and chemotactic factors were detected by qPCR analyze. Results The serum level of ALT and AST were significant higher in KO and antibody blocked group compared to WT group. The histopathologic injury of liver including portal inflammation ,hepatocyte swelling and coagulative necrosis were also more severe in KO and antibody blocked group, as well as liver apoptosis. The WT mice had lower expression of CXCLI , CXCL5 , CXCL10, CCL2 and IL6. Conclusion B7H3 may alleviate mouse hepatic ischemia reperfusion injury by inhibiting neutrophils recruitment via reduction of chemotactic factors.
作者
李势辉
郑旭
潘国政
LI Shi-hui;ZHENG Xu;PAN Guo-zheng(The Provical Hospital, Hefei 230001, China)
出处
《肝胆外科杂志》
2019年第1期70-73,79,共5页
Journal of Hepatobiliary Surgery
基金
"科大新医学"联合基金(WK9110000052)