氢溴酸樟柳碱对慢性脑缺血损伤大鼠氧化应激及细胞凋亡的影响

Oxidant stress and opoptotic effects of anisodine hydromide on rats with chronic cerebral ischemic injury

目的研究氢溴酸樟柳碱对慢性脑缺血损伤大鼠氧化应激及细胞凋亡的影响。方法体内实验中,双侧颈总动脉结扎建立大鼠慢性脑缺血模型,大鼠分为假手术组,模型组,阳性对照组,氢溴酸樟柳碱高、中、低剂量组(1.2、0.6、0.3 mg/kg),测定脑组织与血清中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、乳酸脱氢酶(LDH)、一氧化氮合酶(iNOS)活性、微量还原型谷胱甘肽(GSH)、一氧化氮(NO)含量。体外实验中,CoCl_2处理PC12细胞以建立化学性缺氧模型,大鼠分为对照组,模型组,阳性组,氢溴酸樟柳碱高、中、低剂量组(100、50、25μmol/L),采用Hochest33342荧光染色,荧光显微镜观察细胞凋亡情况,免疫荧光染色、Western blotting法检测P53蛋白表达。结果体内实验显示,氢溴酸樟柳碱干预后大鼠血清SOD活性增加,iNOS活性降低,脑组织GSH含量升高。体外实验显示,氢溴酸樟柳碱干预后能降低LDH、NO释放,减少凋亡细胞数目及P53表达。结论氢溴酸樟柳碱能提高慢性脑缺血大鼠抗氧化作用,减少神经细胞凋亡,其作用机制与减少P53蛋白表达有关。

AIM To observe the oxidant stress and opoptotic effects of anisodine hydromide（ AH） on chronic cerebral hypoperfusion（ CCH） rats. METHODS In vivo CCH models were established in adult male SpragueDawley rats by permanent ligation of bilateral common carotid arteries [two-vessel occlusion（ 2-VO） ] surgery.Rats were randomly divided into six groups,sham group,model group,positive group of n-butylphthalide and sodium chloride injection,and AH groups（ 1. 2 mg/kg high-dose group,0. 6 mg/kg medium-dose group,and0. 3 mg/kg low-dose group）. Antioxidant indices including the activity of SOD,CAT,LDH and i NOS and the content of GSH and NO were measured. In the in vitro trial,PC12 cells were divided into control group,model group,positive group of n-butylphthalide,and AH groups（ 100 μmol/L high-dose group,50 μmol/L mediumdose group,and 25 μmol/L low-dose group）,and the hypoxic models were established by treating PC12 cells with CoCl2.The cells had their release of NO and LDH detected,their cellular apoptosis determined by Hochest 33342 fluorescence staining,and the expression of P53 protein identified by IF（ immunofluorescence） and Western blotting method. RESULTS The in vivo trial revealed AH＇ s enhancement in serum SOD activity and inhibition in serum i NOS activityof the CCH rats,and its power in the cerebral GSH and LDH release reduction. The in vitro trial showed the resultant lower LDH and NO release,decreased number of neuro-apoptosis,and inhibited P53 protein expression after AH intervention. CONCLUSION The antioxidant and antiapoptotic effects of AH on CCH rats may be associated with down regulation of P53 protein.