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目的:观察三七总皂苷(PNS)对顺铂(DDP)损伤的人肾近曲小管上皮细胞(HK-2)增殖和氧化指标的影响。方法:体外培养HK-2细胞株,使其细胞数约为1×10~6个/ml,接种于96孔培养板,分为空白组、模型组、阳性药物组和PNS高、中、低剂量组6组。加入终浓度为6.25μg.L~(-1)DDP 20μl建立DDP损伤HK-2细胞模型,各组分别给予生理盐水、氨磷汀和不同浓度的PNS处理48 h。MTT法测定细胞存活率;比色法测定乳酸脱氢酶(LDH)活性;裂解细胞,取上清液采用紫外分光光度法监测细胞内谷胱甘肽(GSH)含量,黄嘌呤氧化酶法测定超氧化物歧化酶(SOD)活性,TBA法测定MDA含量;荧光素DCFH-DA探针法测定细胞内ROS含量。结果:与空白组比较,模型组的细胞存活率、细胞悬液中的SOD酶活性和GSH-PX含量减低,细胞培养液中的LDH活性、细胞悬液中MDA含量和HK-2细胞内ROS水平显著升高,差异均具有显著统计学差异(P<0.05);与模型组比较,阳性组和PNS高、中、低剂量组的细胞存活率、细胞悬液中的SOD酶活性和GSH-PX含量明显升高,细胞培养液中的LDH活性、细胞悬液中MDA含量和细胞内ROS水平显著降低,且作用均有浓度依赖性,差异均具有显著统计学差异(P<0.05)。结论:PNS能促进DDP损伤HK-2细胞的增殖,降低培养液中LDH水平和细胞内的ROS水平,改善其细胞内氧化水平,对DDP损伤的HK-2细胞具有保护作用。其保护机制可能与其抗氧化作用有关。
Objective: To observe the effects of Panax Notoginseng Saponins (PNS) on the proliferation and oxidative indexes of human renal proximal tubule epithelial cells (HK-2) induced by cisplatin (DDP). Methods: HK-2 cells were cultured in vitro, and the number of cells was about 1 × 10 ~ 6 cells / ml. The cells were inoculated into 96-well plates and divided into blank group, model group, positive drug group and high, Dose group 6 groups. DDP-induced HK-2 cell model was established by adding 20μl DDP at the final concentration of 6.25μg.L-1, and each group was given physiological saline, amifostine and PNS at different concentrations for 48h. Cell viability was measured by MTT assay. Lactate dehydrogenase (LDH) activity was assayed by colorimetric assay. Cells were lysed and the supernatant was assayed for glutathione (GSH) by UV spectrophotometry. Xanthine oxidase assay The activity of superoxide dismutase (SOD) and the content of MDA were determined by TBA method. The intracellular ROS content was determined by fluorescein DCFH-DA probe method. Results: Compared with the blank group, the cell viability, SOD activity and GSH-PX content in the cell suspension decreased, the activity of LDH in the cell culture medium, the content of MDA in the cell suspension and the level of ROS in HK-2 cells (P <0.05). Compared with the model group, the cell viability, the activity of SOD in cell suspension and the activity of GSH- PX content was significantly increased, LDH activity in cell culture medium, MDA content in cell suspension and intracellular ROS levels were significantly decreased, and the effect was concentration-dependent, the differences were statistically significant (P <0.05). CONCLUSION: PNS can promote the proliferation of HK-2 cells induced by DDP, decrease the levels of LDH and intracellular ROS in the medium, and improve the intracellular oxidative potential of HK-2 cells. Its protective mechanism may be related to its antioxidant effect.