三七总皂苷对大鼠肾缺血再灌注损伤的保护作用及其线粒体机制

Protective effects of total saponins of Panax notoginseng on renal ischemia/reperfusion injury in rats and the mitochondrial mechanism

目的:观察三七总皂苷(血栓通注射液的药物成分)对大鼠肾缺血再灌注损伤的保护作用,分析其可能的线粒体机制。方法:实验于2006-01/09在广州医学院病理生理学实验室完成。实验分组:取108只SD雄性大鼠,随机分为假手术组、缺血再灌注模型组、三七总皂苷组,每组36只。实验干预:①假手术组:实验前1h腹腔注射生理盐水,全麻下行右肾摘除,左肾暴露40min后取下。②缺血再灌注模型组:实验前1h腹腔注射生理盐水,全麻下行右肾摘除,左肾动脉夹闭缺血45min后再灌注1h。③三七总皂苷组:实验前1h腹腔注射血栓通注射液70mg/kg,其余方法同缺血再灌注模型组。实验评估:①再灌注后1h取肾组织匀浆检测肾组织超氧化物歧化酶活性、丙二醛含量。②测血浆肌酐、尿素氮、血清降钙素基因相关肽含量。③电镜观察肾组织线粒体超微结构的改变。④检测线粒体细胞色素氧化酶、琥珀酸脱氢酶活性,Ca2+含量。结果:108只大鼠均进入结果分析。①三七总皂苷组肾组织超氧化物歧化酶活性高于缺血再灌注模型组;丙二醛含量低于缺血再灌注模型组。②三七总皂苷组血浆肌酐、尿素氮含量低于缺血再灌注模型组;降钙素基因相关肽水平高于缺血再灌注模型组。③电镜观察三七总皂苷组线粒体结构有明显的改善,嵴数目减少不多,且排列较整齐,没有线粒体崩解的情况。④三七总皂苷组肾组织线粒体中细胞色素氧化酶、琥珀酸脱氢酶活性高于缺血再灌注模型组;Ca2+含量低于缺血再灌注模型组。结论:三七总皂苷对肾缺血再灌注损伤有明显保护作用,其保护作用机制与直接清除自由基,提高琥珀酸脱氢酶、细胞色素氧化酶活性、升高血清降钙素基因相关肽水平,降低钙超载,从而减轻线粒体损伤有关。

AIM： To observe the protective effects of total saponins of Panax notoginseng （PNS）, the drug component of Xueshuantong injection, on renal ischemia/reporfusion （I/R） injury in rats, and explore the possible mitochonddal mechanism. METHODS： The experiment was carried out in the Department of Pathophysiology, Guangzhou Medical College from January to September 2006. 108 male SD rats were randomly averaged into sham operation group, I/R group and PNS group. （1）Sham operation group was injected intraperitoneally normal saline one hour before experiment, and the right kidney was removed under general anesthesia, then the left kidney was removed after exposed for 40 minutes. （2）In I/R group, the rats were injected intraperitoneally normal saline one hour before experiment, and their right kidneys were removed under general anesthesia, then the left renal artery was clamped for 45 minutes immediately followed by 1 hour blood reperfusion. （3）The rats of PNS group were intraperitoneally injected PNS one hour before the experiment, following by the same procedure in the I/R group. After one-hour reperfusion, the renal tissue homogenate was harvested to examine the superoxide dismutase （SOD） activity, and malondialdehyde （MDA）, serum creatinine （SCr）, blood urea nitrogen （BUN）, and serum calcitonin gene-related peptide （CGRP）. The mitochondria ultrastructural alterations of renal tissue were observed by electron microscope. Meanwhile, the renal mitochondrial activities of cytochrome oxidase （COX） and succinic dehydrogenase （SDH） and Ca^2＋ content were measured. RESULTS： All 108 rats were involved in the result analysis. （1）The SOD activity of the PNS group was higher than that of the I/R group, while the MDA level was lower than that of the I/R group. （2）The plasma Scr and BUN levels of PNS group were lower than those of the I/R group, while the CGRP level was higher than that of the I/R group. （3）Additionalty, injuries of the mitochondria were greatly alleviated in the PNS group under electron microscope, and the ridge was lightly decreased and regularly arranged. No mitochondrial disaggregation was found. （4）The activities of mitochondrial COX and SDH in the PNS group were higher than those of the I/R group, while the plasma Ca^2＋ level was greatly lower than the I/R group. CONCLUSION： PNS could protect kidney from I/R injuries, which may be related to the elimination of free radicals, increase in COX and SDH activities, elevation of CGRP level, decrease in renal calcium overload and alleviation of mitochondrial injury.