摘要
目的：研究冬虫夏草提取液（Cordyceps sinensis,C.sinensis）对血管紧张素Ⅱ（AngⅡ）诱导的肾小管上皮细胞凋亡的保护机制。方法：C.sinensis（0,5,10,20,40 mg/L）与10-8mol/L AngⅡ共同孵育NRK-52E 24,48,72 h,了解C.sinensis对细胞增殖的影响,确定最佳干预浓度;取AngⅡ（0,10-12,10-10,10-8,10-6mol/L）与NRK-52E共同培育24 h及10-8mol/L AngⅡ与NRK-52E共同培养24,48,72 h,观察AngⅡ对NRK-52E凋亡的影响,确定AngⅡ最佳干预浓度、时间后设立对照组,AngⅡ（10-8mol/L）组,AngⅡ（10-8mol/L）＋C.sinensis（40mg/L）组,AngⅡ（10-8mol/L）＋福辛普利（10-5mol/L）组和AngⅡ（10-8mol/L）＋C.sinensis（40 mg/L）＋福辛普利（10-5mol/L）组,培养24 h后进行实验。MTT法检测不同浓度C.sinensis（0,5,10,20,40 mg/L）对NRK-52E作用24,48,72 h后的增殖情况,流式细胞仪Annexin V/PI双染检测凋亡率,比色法测定caspase-3酶活性。结果：C.sinensis（10~40 mg/L）在一定范围内可呈浓度依赖地促进培养的肾小管上皮细胞增殖（P〈0.05）;AngⅡ呈浓度（10-10~10-6mol/L）和时间（12~24 h）依赖地诱导NRK-52E凋亡率增加,caspase-3酶活性增加（P〈0.05）。C.sinensis（10~40 mg/L）能部分地抑制AngⅡ诱导的NRK-52E凋亡和降低caspase-3酶活性（P〈0.01）,与福辛普利对细胞凋亡的抑制无明显区别,二者联合用药效果与单用效果差异无统计学意义（P〉0.05）。结论：C.sinensis对AngⅡ诱导的肾小管上皮细胞凋亡具有一定抑制作用,可能与抑制NRK-52E中caspase-3的激活有关。
Objective： To investigate the mechanism of the protective effect of Cordyceps sinensis（C.sinensis） on the apoptosis of cultured NRK-52E induced by angiotension Ⅱ（AngⅡ）. Methods： NRK-52E cells were incubated with C.sinensis（0,5,10,20,and 40 mg/L） and 10-8 mol/L AngⅡ for 24,48,72 h.The optimal concentration of C.sinensis was selected.Either NRK-52E cells were incubated with different doses of AngⅡ（0,10-12,10-10,10-8,and 10-6 mol/L） for 24 h,or with 10-8 mol/L AngⅡ for 24,48,and 72 h,to observe the effect of AngⅡ on the apoptosis of NRK-52E cells.The optimal concentration and time of AngⅡ were selected.In another experiment cells were divided into 5 groups： a control,AngⅡ（10-8 mol/L）,AngⅡ（10-8 mol/L）＋ C.sinensis（40 mg/L）,Ang Ⅱ（10-8 mol/L）＋ fosinopril（10-5 mol/L）,and Ang Ⅱ（10-8 mol/L）＋ fosinopril（10-5 mol/L）＋C.sinensis（40 mg/L）.MTT assay was used to test the changes in the proliferation of NRK-52E cultured with different concentration of C.sinensis for 24,48,72 h.The Annecxin V-FITC and PI stainings were applied to detect the apoptosis rate induced by AngⅡ by flow cytometer（FCM） and to determine the effects of C.sinensis.The activity of caspase-3 was assayed by spectrophotometry. Results： Certain concentrations of C.sinensis（10-40 mg/L） promoted the proliferation of NRK-52E cells inhibited by AngⅡ（P0.05）.AngⅡ induced the apoptosis of NRK-52E in a dose and time-dependent manner,accompanied with increased activity of caspase-3（P0.05）.C.sinensis partially suppressed the apoptosis of NRK-52E induced by AngⅡ,and declined the activity of caspase-3（P0.01）.No significant difference was shown as between the fosinopril group and the fosinopril＋C.sinensis group（P0.05）. Conclusion： C.sinensis can suppress the apoptosis of NRK-52E by AngⅡ,and the protective effect of C.sinensis may be inhibiting the activation of caspase-3 during the AngⅡ-induced apoptosis of NRK-52E.
出处
《中南大学学报（医学版）》
CAS
CSCD
北大核心
2012年第1期67-72,共6页
Journal of Central South University ：Medical Science
