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目的观察参附注射液对大鼠肢体缺血再灌注损伤后肝功能、血红素加氧酶-1(HO-1)表达的影响,并对其保护机制做一初步探讨。方法 64只清洁级SD雄性大鼠,用随机数字表法随机分为4组,每组16只,分别为假手术组、缺血再灌注组、参附干预组、参附+锌原卟啉Ⅸ(Znpp)干预组。假手术组:大鼠麻醉后仅分离不夹闭股动脉,分离血管前10 min以7.5 mL/kg腹腔注射生理盐水;缺血再灌注组:夹闭股动脉前10 min以7.5 mL/kg腹腔注射生理盐水,夹闭股动脉缺血3 h,再灌注4 h;参附干预组:夹闭股动脉前10 min以7.5 mL/kg腹腔注射参附注射液,夹闭股动脉缺血3 h,再灌注4 h;参附+Znpp干预组:术前30 min腹腔注射Znpp 5 mg/kg,余同参附干预组。再灌注完毕后取材,取外周静脉血测血清谷丙转氨酶(GPT)、谷草转氨酶(GOT)含量;取肝组织测定肝组织中丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性;采用免疫组织化学法测定肝脏组织中HO-1蛋白表达;光镜下观察肝脏病理学改变。结果 1与假手术组比较,各肢体缺血再灌注造模组MDA含量均明显升高(P<0.05),SOD活性明显降低(除参附干预组外,P<0.05),GPT、GOT含量均明显升高(P<0.05),HO-1蛋白表达明显升高(P<0.05)。2与缺血再灌注组比较,参附干预组MDA含量明显降低(P<0.05),SOD活性明显升高(P<0.05),血清GPT、GOT含量明显降低(P<0.05),肝组织中HO-1蛋白表达升高(P<0.05)。3与参附干预组比较,参附+Znpp干预组MDA含量明显升高(P<0.05),SOD活性明显降低(P<0.05),血清GPT、GOT含量明显升高(P<0.05),而肝组织HO-1蛋白表达差异无统计学意义(P>0.05)。结论肢体缺血再灌注可造成肝脏功能损伤,给予参附注射液预处理可以减轻肝脏损害程度,这种保护作用可能与参附注射液预处理上调HO-1蛋白在肝组织中的表达、抑制氧自由基生成有关。
Objective To observe the effect of Shenfu injection on liver function and expression of heme oxygenase-1 (HO-1) after limb ischemia-reperfusion injury in rats and its protective mechanism. Methods Sixty-four SD male rats were randomly divided into 4 groups (n = 16): sham operation group, ischemia / reperfusion group, Shenfu intervention group, Shenfu + Zinc Protoporphyrin Ⅸ (Znpp) intervention group. Sham-operated group: rats were anesthetized only isolated without femoral artery separation, 10 min before the separation of blood vessels to 7.5 mL / kg intraperitoneal injection of saline; ischemia reperfusion group: 10 minutes before clamping the femoral artery to 7.5 mL / kg intraperitoneal Injected with saline, occluded femoral artery ischemia 3 h, reperfusion 4 h; Shenfu intervention group: 10 minutes before clamping the femoral artery with 7.5 mL / kg intraperitoneal injection of Shenfu injection, femoral artery ischemia 3 h , And reperfusion for 4 h. Participation + Znpp intervention group: Znpp 5 mg / kg was injected intraperitoneally 30 minutes before operation, and more than the same reference group. After reperfusion, the contents of GPT and GOT were measured by peripheral venous blood. The contents of malondialdehyde (MDA), superoxide dismutase (SOD) The expression of HO-1 protein in liver tissue was detected by immunohistochemistry. The pathological changes of liver were observed under light microscope. Results Compared with the sham operation group, the content of MDA in each limb ischemia-reperfusion group was significantly increased (P <0.05) and SOD activity was significantly decreased (P <0.05 except the intervention group) (P <0.05), and HO-1 protein expression was significantly increased (P <0.05). Compared with ischemia-reperfusion group, the content of MDA in Shen-Fu intervention group was significantly decreased (P <0.05), the activity of SOD was significantly increased (P <0.05), the content of GPT and GOT in serum was significantly decreased (P <0.05) HO-1 protein expression increased (P <0.05). Compared with Shenfu group, the content of MDA and the activity of SOD in Ginseng + Znpp intervention group were significantly increased (P <0.05), while the activity of SOD was significantly lower (P <0.05), and the levels of GPT and GOT in serum were significantly increased There was no significant difference in HO-1 protein expression in liver tissue (P> 0.05). Conclusion Limb ischemia-reperfusion can cause hepatic injury. Pretreatment with Shenfu Injection can reduce the degree of hepatic injury. This protective effect may be related to the increase of HO-1 protein expression and inhibition Oxygen free radical generation related.