CAJ | 학술논문

目的探讨低氧预适应诱导热休克蛋白70(HSP70)在大鼠肝癌肝切除术后对残肝细胞凋亡的影响及可能的作用机制.方法采用SD大鼠移植性肝癌模型,根据随机表将60只模型鼠分为A组(假手术组),B组(缺血状态下肝叶切除组),C组(低氧预适应+缺血状态下肝叶切除组),每组20只.C组的处理方式:术前给予10％氮氧混合气体处理90 min,Pringle法完全阻断入肝血流15 min,同时切除肿瘤所在的肝左外叶.各组分别于术后1,8,12,24 h取残余肝脏标本,用Western blotting免疫印迹法检测HSP70、Bax蛋白的表达,应用流式细胞仪检测肝细胞凋亡率,下腔静脉取血检测ALT、AST,并在光镜下观察各组组织形态学改变,对相关数据进行统计学处理.结果 (1)与A、B组比较,C组检测的各同时相点HSP70的相对量表达明显增加,组间比较差异有统计学意义(均P＜0.05),A、B组肝组织中仅有微量HSP70表达,两组比较差异无统计学意义(P＞0.05);(2)与A组比较,B组各同时相点Bax蛋白表达水平明显升高,肝细胞凋亡率明显增加,两组比较差异有统计学意义(均P＜0.01或0.05),而C组以上各项指标的变化均较B组明显减弱(均P ＜0.05);(3)C组肝功能较B组改善明显,病理示A组肝组织形态基本正常,C组呈轻微病理学改变,而B组肝组织损伤明显加重.结论低氧预适应对缺血条件下肝癌肝切除术产生的缺血再灌注损伤具有保护作用,其可能的机制之一是通过上调HSP70蛋白的表达,而HSP70通过降低Bax的表达,抑制细胞凋亡,从而发挥肝脏保护作用.
목적 탐토저양예괄응유도열휴극단백70(HSP70)재대서간암간절제술후대잔간세포조망적영향급가능적작용궤제.방법 채용SD대서이식성간암모형,근거수궤표장60지모형서분위A조(가수술조),B조(결혈상태하간협절제조),C조(저양예괄응+결혈상태하간협절제조),매조20지.C조적처리방식:술전급여10％담양혼합기체처리90 min,Pringle법완전조단입간혈류15 min,동시절제종류소재적간좌외협.각조분별우술후1,8,12,24 h취잔여간장표본,용Western blotting면역인적법검측HSP70、Bax단백적표체,응용류식세포의검측간세포조망솔,하강정맥취혈검측ALT、AST,병재광경하관찰각조조직형태학개변,대상관수거진행통계학처리.결과 (1)여A、B조비교,C조검측적각동시상점HSP70적상대량표체명현증가,조간비교차이유통계학의의(균P＜0.05),A、B조간조직중부유미량HSP70표체,량조비교차이무통계학의의(P＞0.05);(2)여A조비교,B조각동시상점Bax단백표체수평명현승고,간세포조망솔명현증가,량조비교차이유통계학의의(균P＜0.01혹0.05),이C조이상각항지표적변화균교B조명현감약(균P ＜0.05);(3)C조간공능교B조개선명현,병리시A조간조직형태기본정상,C조정경미병이학개변,이B조간조직손상명현가중.결론 저양예괄응대결혈조건하간암간절제술산생적결혈재관주손상구유보호작용,기가능적궤제지일시통과상조HSP70단백적표체,이HSP70통과강저Bax적표체,억제세포조망,종이발휘간장보호작용.
Objective To investigate the effect of Heat shock protein 70 induced by hypoxic precondition on apoptosis of the remnant livers after the hepatectomy of transplanted hepatic cancer models in SD rants and it's possible mechanisms.Methods Sixty SD rats of transplanted hepatic cancer models were divided into group A (sham operation group),group B (IR + hepatectomy group),and group C (hypoxia precondition + IR + hepatectomy group),with 20 rats in each group.C group was given 10％ oxygen atmosphere for 90 minutes before operation,In 15 minutes of hepatic vascular occlusion with pringle method,Left lateral of liver which tumor was located was resected.At 1,8,12,24 hours after operation,Five rats of each group were killed respectively,then the remnant liver tissues were sampled to measured the hepatocytes apoptosis rate with flow cytometry,expression of heat shock protein 70 (HSP70) and Bax were detected by Western blotting.The serum AST and ALT activities were measured by an automatic analyzer.The histological changes of the hepatocytes were also observed by optical microscopy.All the data were statistically analyzed.Results (1) Compared with A and B group,the HSP70 expression increased obviously in C group at each time point (all P ＜ 0.05).The protein showed a weak expression in group A and B group,while the index had no significant differences between two groups (P ＞ 0.05).(2) In B group compared with A group,the expression of Bax and the hepatocytes apoptosis rate were significantly increased (all P ＜ 0.01 or P ＜0.05).However,amplitudes of the above changes in C group were significantly lower than those of B group (all P ＜ 0.05).(3)The liver function was improved in group HP.The pathological examination showed that the liver tissues in A group were general normal and in C group were showed mild changes,but in B group presented evident pathological changes.Conclusions HP can protect against ischemia-reperfusion injury after the hepatectomy of transplanted hepatic cancer models in SD rants,one of possible mechanisms is that HP can induce HSP70 expression at great intensity,which may decrease expression of Bax protein and the apoptosis of hepatocytes.