Kallistatin对大鼠肝缺血再灌注损伤的保护及机制研究

Effects and mechanisms of Kallistatin on liver ischemia reperfusion injury in rats

目的:探讨Kallistatin对大鼠肝缺血再灌注后肝损伤的保护作用及机制。方法:建立大鼠肝脏缺血再灌注模型,随机将Wistar大鼠分成假手术组(S组)、缺血再灌注组(I/R组)、空壳腺病毒组(O组)和Kallistatin组(K组)。测定血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平评价肝功能。HE染色观察肝组织病理学改变。TUNEL法观察肝细胞凋亡情况。Westen-blot法检测肝组织中Caspase-3、Bcl-2、Bax的表达。结果:与S组比较,I/R组AST、ALT水平明显升高(P<0.05),病理损伤严重,Cas-pase-3和Bax表达也相应增加(P<0.05),而Bcl-2表达减少(P<0.05)。与I/R组比较,K组各项指标均明显改善(均为P<0.05)。O组与I/R组间无明显差别(P>0.05)。结论:Kallistatin对大鼠肝脏缺血再灌注后具有明显的保护作用,其机制可能通过促进Bcl-2表达、抑制Bax表达实现。

Objective： To investigate the protective effects and the underlying mechanisms of Kallistatin on hepatic ischemia-reperfusion injury in rats. Methods： The models of hepatic ischemia-reperfusion injury were randomly divided into four groups which were established in male wistar rats. The Sham（S） group, ischemia reperfusion（t/R） group, empty adenovirus（O） group and Kallistatin （K） group. Liver function and histology change in rats with hepatic ischemia reperfusion injury after 6h were determined by detecting the ALT, AST in serum and HE staining respectively. Hepatocellular apoptosis was determined by terminal deoxynucleotidyl transferees mediated dUTP nick end labeling （TUNEL） method and the expression of caspase-3, BCL-2 and Bax were detected by Western blotting. Results： Compared with Sham group, the levels of ALT and AST were significant higher, the expression of caspase-3 and Bax were increasing dramatically while the expression of BCL-2 was decreasing significantly in ischemia-reperfusion group （P〈 0.05）.The path morphological changes were also more clearly in ischemia-reperfusion group than sham group by HE staining. Compared with ischemia-reperfusion group, all results were obviously reduced in Kallistatin group （P〈 0.05）, but there were no significant differences in empty adenovirus group （P 〉 0.05）. Conclusion; Kallistatin has an obvious protective effect on hepatic ischemia reperfusion injury in rats, and the underlying mechanisms maybe inhibit hepatocellular apoptosis by up-regulating Bcl-2 and down-regulating Bax and Caspase-3.