丹参酮Ⅰ在肾脏缺血再灌注损伤中的保护作用研究

Protective effect of Tanshinone Ⅰ against renal ischemia/reperfusion injury

目的:探讨丹参酮Ⅰ(T-Ⅰ)在肾脏缺血再灌注损伤(RIRI)中的保护作用。方法:(1)体内实验:将雄性ICR小鼠随机分为假手术(sham)组、sham+T-Ⅰ组、缺血再灌注(IR)组、IR+T-Ⅰ组4组,术前每组腹腔注射等量T-Ⅰ或玉米油2周,左肾蒂夹闭50 min诱导RIRI,收集术后48 h血清及肾脏标本,比较各组之间的血清肌酐(Cr)水平、尿素氮(BUN)水平、肾脏组织病理学评分、肾脏组织凋亡比例及肾脏组织内超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量。(2)体外实验:用过氧化氢处理人肾小管上皮细胞(HK-2)建立氧化应激模型,将HK-2分为对照(C)组、T-Ⅰ组、H2O2组、H2O2+T-Ⅰ组4组;用T-Ⅰ或其溶剂处理24 h,然后用过氧化氢处理2 h,比较各组细胞活力、细胞凋亡及细胞内活性氧水平。结果:(1)体内实验:与IR组相比,IR+T-Ⅰ组血清Cr、BUN水平显著降低,肾组织内SOD活力升高而MDA含量显著下降,肾脏组织凋亡比例显著降低,且IR+T-Ⅰ组肾脏组织学病理评分较IR组显著降低;(2)体外实验:经过T-Ⅰ预处理,可以显著降低过氧化氢处理后的细胞内活性氧水平,增强细胞活力,减少细胞凋亡。结论:T-Ⅰ通过抗氧化活性在RIRI中起到保护作用。

Objective： To investigate the protective effect of Tanshinone Ⅰ（T-Ⅰ） on renal ischemia/reperfusion injury（RIRI）. Methods：（1） In vivo study： the male ICR mice were divided into 4 groups： sham group,sham ＋T-Ⅰ group,IR group and IR ＋ T-Ⅰ group. All the mice received T-Ⅰ or corn oil via intraperitoneal injection for 2 weeks before surgery. Left renal pedicle was clamped with vascular clamp for 50 min to induce ischemia reperfusion injury. Serum samples and kidney tissue were collected after 48 h. Blood ureanitrogen（BUN）,creatinine（Cr）,superoxideismutase（SOD）,malondialdehyde（MDA） and kidney histopathology score were compared among the groups.（2） In vitro study： human tubular epithelial cells（HK-2） were treated with H2 O2 to induce oxidative stress injury,and HK-2 were divided into 4 group,i. e.,control（C） group,H2 O2 group,T-Ⅰ group and H2 O2＋ T-Ⅰgroup. Control（C） group and H2 O2 group were pretreated with solvent while T-Ⅰ group and H2 O2＋ T-Ⅰ group were pretreated with T-Ⅰ for 24 h followed by exposure to H2 O2 for another 2 h. Cell viability and reactive oxygen species were compared among all groups. Results：（1） In vivo study： serum BUN and Cr in IR ＋ T-Ⅰ group were significantly decreased compared with those in the IR group. Level of SOD was significantly higher while level of MDA was significantly lower in IR ＋ T-Ⅰ group compared with those in IR group; histological score in IR ＋ T-Ⅰgroup was significantly reduced compared with that in IR group.（2） In vitro study： T-Ⅰ attenuated hydrogen peroxide-induced viability decrease and apoptosis of HK-2 cells,in addition,T-Ⅰ reduced the generation of ROS induced by hydrogen peroxide in HK-2 cells. Conclusion： T-Ⅰ plays a protective role in renal ischemia-reperfusion injury via antioxidant activity.