缺血后预处理对缺血再灌注大鼠肝内胆管细胞损伤的影响及机制

发布时间：2019-06-12 13:09

【摘要】：目的观察缺血后预处理对缺血再灌注大鼠肝内胆管细胞损伤的影响并探讨其机制。方法取健康雄性SD大鼠24只,随机分为假手术组、缺血再灌注组、缺血后处理组,每组8只。假手术组:大鼠麻醉后于上腹部正中取一长3.5 cm纵切口,将腹壁各层依次切开,肝脏显露后进一步游离出肝十二指肠韧带,关腹,无其他干预。缺血再灌注组:在假手术组基础上,游离肝十二指肠韧带,用无损伤动脉夹将肝门区域夹闭,缺血30 min后取下血管夹,使血管开放,对肝脏持续再灌注,最后关腹。缺血后处理组:在缺血再灌注组基础上,血供完全恢复之前再灌注10 s,夹闭10 s,如此反复6个循环(共2 min)后完全恢复血供。比色法测定血清γ-GT、SOD活力,免疫组化法测定胆管组织中Bcl-2和Bax蛋白表达,采用原位缺口末端标记法测算胆管细胞凋亡指数(AI),HE染色后光镜下观察胆管上皮细胞基本形态学变化。结果 IR组、IPO组、SO组血清γ-GT值分别为(34.64±3.28)、(26.30±3.32)、(17.33±2.08)U/L,三组间两两比较,P均0.05。SO组、IR组、IPO组大鼠血清SOD活力值分别为(58.26±3.00)、(60.28±3.73)、(61.53±7.99)U/m L,各组间比较,P均0.05。IR组、IPO组、SO组胆管组织中Bcl-2蛋白阳性表达率分别为(42.67±3.73)%、(77.94±5.25)%、(18.21±4.16)%,三组间两两比较,P均0.05。IR组、IPO组、SO组胆管组织中Bax蛋白阳性表达率分别为(90.17±2.12)%、(70.10±4.27)%、(30.63±3.00)%,三组间两两比较,P均0.05。IR组、SO组、IPO组AI分别为79.31±1.12、26.06±2.27、58.90±2.75,三组间两两比较,P均0.05。IPO组病理改变较IR组明显减轻,肝小叶结构较完整,肝细胞变性程度减轻,汇管区胆管上皮连续完整,炎细胞浸润少。结论缺血后预处理通过促进肝内胆管组织中Bcl-2蛋白表达、抑制Bax蛋白表达,抑制SD大鼠肝脏缺血再灌注过程中肝内胆管细胞凋亡,从而减轻肝脏缺血再灌注过程中肝内胆管细胞的损伤。
[Abstract]:Objective To observe the effect of pre-ischemic preconditioning on the hepatic bile duct cell injury in rats with ischemia-reperfusion and to explore its mechanism. Methods 24 healthy male SD rats were randomly divided into the sham operation group, the ischemia-reperfusion group and the ischemic post-treatment group. Sham operation group: after the rats were anesthetized, a long 3.5 cm longitudinal incision was taken in the middle of the upper abdomen, the layers of the abdominal wall were cut in sequence, and the hepatic and duodenal ligament was further free after the liver was exposed, and the abdomen was closed without any other intervention. Ischemia-reperfusion group: on the basis of the sham-operation group, the free-hepatic duodenal ligament was closed, the area of the liver-door was clipped by the non-invasive valve clamp, and the lower blood vessel clamp was taken after the ischemia for 30 min, so that the blood vessel was opened, the liver was continuously reperfused, and the abdomen was finally closed. Post-ischemic post-treatment group: on the basis of the ischemia-reperfusion group, the blood supply was reperfused for 10 s before the blood supply was fully recovered, and the blood supply was completely recovered after repeated 6 cycles (2 min). The expression of Bcl-2 and Bax in the bile duct was determined by colorimetric method, and the expression of Bcl-2 and Bax in the bile duct tissue was determined by immunohistochemistry. The apoptosis index (AI) of the bile duct was measured by the in-situ nick end labeling method. The basic morphological changes of the bile duct epithelial cells were observed under the light microscope after HE staining. Results The serum SOD-GT values of the IR group, the IPO group and the SO group were (34.64-3.28), (26.30-3.32), (17.33-2.08) U/ L, and the serum SOD activity values of the group were (58.26-3.00), (60.28-3.73), (61.53-7.99) U/ m-L, respectively. The positive rate of Bcl-2 protein in the group was 42.67 (3.73)%, (77.94-5.25)%, (18.21-4.16)%, and P was 0.05. IR group and IPO group, respectively. The positive expression rate of Bax in the bile duct of the SO group was (90.17-2.12)%, (70.10-4.27)%, (30.63-3.00)%, and between the three groups, P was 0.05. IR group, SO group and IPO group AI were 79.31-1.12, 26.06-2.27, 58.90-2.75 respectively. P was 0.05. The pathological changes of the IPO group were significantly reduced in the IR group, the structure of the hepatic lobule was relatively complete, the degree of degeneration of the liver cells was reduced, the epithelium of the bile duct of the foreign exchange tube was continuous and intact, and the infiltration of the inflammatory cells was less. Conclusion The post-ischemic preconditioning can inhibit the expression of Bcl-2 protein in the hepatic bile duct tissue, inhibit the expression of Bax protein, and inhibit the apoptosis of the intrahepatic bile duct cells in the hepatic ischemia-reperfusion process of the SD rats, thereby reducing the damage of the hepatic bile duct cells in the hepatic ischemia-reperfusion process.
【作者单位】：河北省人民医院;
【分类号】：R575

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