Lu Wei-Na, Zheng Fen-Ping, Lai Dong-Wu, Li Hong
Department of Endocrinology, Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University, Hangzhou, 310016, China.
Department of Cardiovascular, Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University, Hangzhou, 310016, China.
Chin J Integr Med. 2016 Aug;22(8):611-8. doi: 10.1007/s11655-015-2050-4. Epub 2015 Apr 6.
To investigate the effect of Xuezhikang (, XZK) on renal cell apoptosis in diabetic rats and the possible mechanism.
Sixty-six rats were randomly divided into 3 groups: the normal, model and XZK groups. In each group, the rats were further randomly divided into 3-month and 6-month subgroups, respectively. Diabetes of rats was induced by a single intraperitoneal injection of 1% streptozocin at 60 mg/kg body weight. Rats in the XZK group received gastric perfusion of XZK (1200 mg/kg body weight) everyday for 3 or 6 months, while rats in the normal and model groups received equal volume of saline. Twenty-four hours' urine was collected for urinary albumin excretion (UAE) measurement. Periodic acid-Schiff (PAS) and Masson's trichrome staining were used for saccharides and collagen detection. Cell apoptosis of renal cortex was investigated by TdT-mediated dUTP nick end labeling (TUNEL) staining. Bax and Bcl-2 expressions were detected by immunohistochemistry and Western blot, respectively. Cytochrome C (Cyt C) and caspase-9 concentration were detected by Western blot.
Compared with the model group, XZK treatment could significantly decrease the kidney hypertrophy index, 24 h UAE, renal cell apoptosis, cytoplasmic Cyt C level and active caspase-9 level, as well as suppress the increment of Bax and up-regulate the expression of Bcl-2, leading to the suppression of Bax/Bcl-2 ratio at 3 and 6 months (P<0.05 or P<0.01). Moreover, XZK treatment could alleviate the deposition of PAS-stained saccharides and Masson's trichromestained collagen to different extent.
Renal cell apoptosis was observed in diabetic kidney, in which mitochondrial apoptotic pathway might be involved. XZK treatment could attenuate pathological changes in diabetic kidney and reduce renal cell apoptosis, probably via the suppression of Bax/Bcl-2 ratio, which lead to inhibition of Cyt C release and following caspase-9 activation.
探讨血脂康对糖尿病大鼠肾细胞凋亡的影响及其可能机制。
66只大鼠随机分为3组：正常组、模型组和血脂康组。每组大鼠再随机分为3个月和6个月亚组。通过腹腔注射1%链脲佐菌素（60 mg/kg体重）诱导大鼠糖尿病。血脂康组大鼠每天灌胃给予血脂康（1200 mg/kg体重），持续3或6个月，而正常组和模型组大鼠给予等量生理盐水。收集24小时尿液测定尿白蛋白排泄量（UAE）。采用过碘酸-希夫（PAS）染色和马松三色染色检测糖类和胶原蛋白。通过末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）染色研究肾皮质细胞凋亡。分别采用免疫组化和蛋白质印迹法检测Bax和Bcl-2表达。采用蛋白质印迹法检测细胞色素C（Cyt C）和半胱天冬酶-9浓度。
与模型组相比，血脂康治疗可显著降低肾脏肥大指数、24小时UAE、肾细胞凋亡、细胞质Cyt C水平和活性半胱天冬酶-9水平，抑制Bax增加并上调Bcl-2表达，导致3个月和6个月时Bax/Bcl-2比值降低（P<0.05或P<0.01）。此外，血脂康治疗可不同程度减轻PAS染色糖类和马松三色染色胶原蛋白的沉积。
糖尿病肾病中观察到肾细胞凋亡，可能涉及线粒体凋亡途径。血脂康治疗可减轻糖尿病肾病的病理改变，减少肾细胞凋亡，可能是通过抑制Bax/Bcl-2比值，从而抑制Cyt C释放及随后的半胱天冬酶-9激活。
