茵陈对阿霉素诱导的NRK-52e细胞凋亡的保护作用研究  

Protective Effect of Yinchen(Artemisia Capillaris Thunb.) on Adriamycin-induced Apoptosis of NRK-52e

作　　者：黄国顺 雍晨 王晓芳 陈玮 姚敏[1] 束洋 李一卓[1] 周恩超[1] HUANG Guoshun;YONG Chen;WANG Xiaofang;CHEN Wei;YAO Min;SHU Yang;LI Yizhuo;ZHOU Enchao(Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,Jiangsu,China;Key Laboratory for Metabolic Diseases in Chinese Medicine,Nanjing University of Chinese Medicine,Nanjing 210029,Jiangsu,China)

机构地区：[1]南京中医药大学附属医院,江苏南京210029 [2]南京中医药学大学代谢病中医研究重点实验室,江苏南京210029

出　　处：《中华中医药学刊》2022年第10期171-174,I0025-I0028,共8页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金(81873270);江苏省“六大人才高峰”高层次人才项目(WSN-012);江苏省中医院“高峰学术”人才项目(y2018rc17);江苏省研究生科研创新计划(KYCX1456);南京市科技发展计划(202002051)。

摘　　要：目的 探究茵陈水提物冻干粉(freeze-dried powder of Artemisia Capillaris Thunb. water extract, WAC)对阿霉素(adriamycin, ADR)诱导的大鼠近端肾小管上皮细胞(NRK-52e)损伤的保护作用。方法 ADR(1μg·mL^(-1))体外诱导NRK-52e损伤,通过加入不同质量浓度的WAC(1、2、4 mg·mL^(-1))作用后,分别采用形态学观察、细胞增殖毒性检测法(CCK-8法)检测WAC对细胞的保护作用,通过脱氧核糖核苷酸末端转移酶(TUNEL)染色法以及蛋白免疫印迹法(Western Blot)检测细胞凋亡的表达水平。结果 与空白组比较,给予ADR干预后,NRK-52e细胞数目明显减少,间隙明显增大,形态皱缩变形,同时漂浮在培养液中的损伤细胞明显增加,细胞活性明显降低(P<0.01),TUNEL法检测凋亡率明显增加(P<0.01),能诱导相关凋亡蛋白Bcl-2/Bax表达明显降低(P<0.01)、Cleaved Caspase-3/Caspase-3显著提高(P<0.01),并且伴随一定的剂量依赖性。与模型组相比,在加入不同质量浓度的WAC(1、2、4 mg·mL^(-1))后,能减弱ADR(质量浓度为1μg·mL^(-1))诱导的细胞形态改变,细胞活性显著提高(P<0.05,P<0.01),TUNEL法检测凋亡率明显降低(P<0.01),检测凋亡相关蛋白Bcl-2/Bax表达明显升高(P<0.01)、Cleaved Caspase-3/Caspase-3表达明显降低(P<0.01),均呈现一定的剂量依赖性。结论 WAC能抑制ADR诱导的肾小管细胞凋亡而发挥保护作用,作用机制可能与线粒体途径有关。Objective To observe the protective effect of freeze-dried powder of Yinchen(Artemisia Capillaris Thunb.) water extract(WAC) on the adriamycin(ADR)-induced injury of rat proximal renal tubular epithelium(NRK-52 e). Methods ADR(1 μg·mL^(-1)) induced NRK-52 e injury in vitro. After adding WAC with different mass concentrations(1,2,4 mg·mL^(-1)), the protective effect of WAC on cells was detected by morphological observation and cell proliferation toxicity test(CCK-8 method). The expression level of apoptosis was detected by deoxyribonucleotide terminal transferase(TUNEL) staining and Western Blot. Results After administration of adriamycin intervention, compared with those of the blank group, the number of NRK-52 E cells decreased significantly, the gap increased significantly, the morphology shrunk and deformed, the injured cells floating in the culture medium increased significantly, the cell activity decreased significantly(P<0.01), the apoptosis rate detected by TUNEL method increased significantly(P<0.01), and the expression of related apoptosis protein Bcl-2/Bax decreased significantly(P<0.01). Cleared Caspase-3/Caspase-3 was significantly increased(P<0.01), and accompanied by a certain dose-dependent. Compared with those of the model group, the addition of WAC with different mass concentrations(1,2,4 mg·mL^(-1)) can weaken ADR-induced(mass concentration was 1 μg·mL^(-1)) cell morphological changes, cell activity increased significantly(P<0.05,P<0.01). The apoptosis rate decreased significantly(P<0.01), the expression of apoptosis related protein Bcl-2/Bax increased significantly(P<0.01), and the expression of cleaved Caspase-3/Caspase-3 decreased significantly(P<0.01). Conclusion WAC can inhibit ADR-induced apoptosis of renal tubule cells and plays a protective role possibly through the mitochondrial pathway.

关 键 词：茵陈 阿霉素 肾小管上皮细胞 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]