【摘要】 目的探讨自噬信号与NLRP3炎症小体在硬化肝组织肝部分切除术后肝再生进程中的表达情况及可能的作用,期望能为促进肝硬化术后肝再生提供新思路。方法 C57BL/6小鼠110只随机分为对照组和实验组,每组55只,通过建立正常肝与硬化肝-50%及70%肝切除-肝再生模型,观察两组小鼠肝部分切除术后生存率、残肝重量变化、肝组织形态学变化及增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)表达情况,测定小鼠血清谷丙转氨酶(glutamic-pyruvic transaminase,GPT)、谷草转氨酶(glutamic-oxaloacetic transaminase,GOT)及白介素18(interleukin-18,IL-18)、白介素1β(interleukin-1β,IL-1β)水平,并检测小鼠自噬相关蛋白及炎症小体NLRP3、胱天蛋白酶1(Caspase1)蛋白的表达情况。结果对照组小鼠术后残肝重量明显高于实验组小鼠,差异有统计学意义(P=0.02)。对照组术后48h细胞核分裂象最多,PCNA表达最高。实验组在术前可见较多细胞核分裂象及PCNA阳性细胞核,术后24～48h之间细胞核分裂象较少,PCNA少量表达,72～120h缓慢逐渐增加。两组小鼠肝部分切除后血清GOT、GPT、IL-18、IL-1β水平均较术前升高(P<0.05),实验组术后各时点显著高于对照组(P<0.05)。实验组自噬相关蛋白Beclin1、LC3B术前表达高于对照组,术后24h、48h升高不明显,低于同时点的对照组自噬水平,而Caspase1、NLRP3表达水平术前、术后均高于对照组。结论实验组肝部分切除术前及术后早期阶段NLRP3炎症小体均活化,术后活化水平显著高于对照组,但自噬水平下调,肝再生能力下降,肝再生启动延缓,这种下降可能与NLRP3炎症小体活化、自噬下调有关,但其具体作用机制有待进一步研究。更多还原

【Abstract】 Objective To explore the expression of autophagy and NLRP3 inflammasome activation in the liver regeneration,hoping to provide new ideas for promoting liver regeneration of liver cirrhosis.Methods Models undergoing 50% and 70% hepatectomy was respectively established to calculate the survival rate and liver weight/body weight ratio,to observe the liver histomorphology and the expression of proliferating cell nuclear antigen (PCNA) after hepatectomy,to measure the serum levels of glutamic-pyruvic transaminase (GPT),glutamic-oxaloacetic transaminase (GOT),interleukin-18(IL-18) and interleukin-1β (IL-1β),and to detect the expressions of autophagy,NLR family pyrin domain containing 3 (NLRP3) and Caspase-1.Results The postoperative liver weight of mice in the control group was significantly higher than that in the liver cirrhosis group.The expression of PCNA was the highest in the control group at 48h.In the cirrhosis group,more mitotic figures and PCNA positive nuclei were seen before surgery,and fewer mitotic figures were seen between 24h and 48h after surgery with PCNA gradually increasing from 72h to 120h.Serum GOT,GPT,IL-18 and IL-1β levels in both groups were significantly higher than those before partial hepatectomy (P﹤ 0.05).Furthermore,these biomarkers at each postoperative timepoints in the cirrhosis group were significantly higher than those in the normal group (P ﹤ 0.05).The expression of autophagy-related proteins in the cirrhosis group were lower than that in the control group after partial hepatectomy,but the expression levels of caspase-1 and NLRP3 were higher in the cirrhosis group than those in the control group preoperatively and postoperatively (P ﹤ 0.05).Conclusion The activation of NLRP3 inflammasome was significantly enhanced in the cirrhosis group,while the level of autophagy was down regulated.They may be linked to activation of NLRP3 inflammasome and the decrease of liver regeneration ability,but the specific mechanism needs further research.更多还原