甘草干姜汤对人肾癌细胞及人尿源性干细胞的作用

doi:10.3969/j.issn.1004-616x.2024.05.004

摘要/Abstract

摘要： 目的： 探讨甘草干姜汤(LDGD)对体外培养的人肾癌769-P细胞和人尿源性干细胞(hUSCs)活力和凋亡的作用。方法： 常温离心法从人的新鲜尿液中分离培养hUSCs，观察hUSCs的生长状态并采用MTT法测定其活力；流式细胞术鉴定hUSCs表面标志物的表达。MTT法检测2.5~40 mg/mL LDGD作用769-P细胞和hUSCs 24~72 h后的活力，观察其细胞形态的变化；流式细胞术检测10 mg/mL LDGD诱导769-P细胞和hUSCs凋亡的情况，计算其凋亡率。结果： 通过常温离心法获得细胞形态良好、生长活性较高的hUSCs，hUSCs表达间充质干细胞表面标志物CD44和CD90，不表达内皮细胞表面标志物CD31和造血干细胞表面标志物CD34。5~10 mg/mL LDGD作用24~72 h可抑制769-P细胞活力并呈时间和剂量-效应关系，而对hUSCs活力有促进作用。10 mg/mL LDGD作用48 h后，倒置显微镜下可见769-P细胞发生明显皱缩，体积缩小，细胞折光率下降，而hUSCs形态无明显变化。10 mg/mL LDGD作用769-P细胞48 h后的凋亡率为(11.93±0.51)%，相较对照组显著升高(P<0.01)，而作用hUSCs后的凋亡率为(0.01±0.10)%，与对照组比较差异无统计学意义(P<0.05)。结论： 在本实验条件下，LDGD可抑制体外培养的人肾癌细胞活力，诱导其凋亡；可促进hUSCs的细胞活力，且无明显诱导细胞凋亡作用。LDGD对两种细胞的作用存在明显差异。

关键词: 甘草干姜汤, 肾癌细胞, 人尿源性干细胞, 细胞活力, 凋亡

Abstract: OBJECTIVE： To investigate the effects of licorice and dried ginger decoction (LDGD) on the viability and apoptosis of human renal carcinoma cells (769-P) and human urine-derived stem cells (hUSCs) in vitro. METHODS： hUSCs were isolated and cultured from fresh human urine using room temperature centrifugation. The growth status of hUSCs was observed，and their viability was measured using the MTT assay. Flow cytometry was employed to identify the expression of surface markers on hUSCs. The MTT assay was used to evaluate the effects of 2.5-40 mg/mL LDGD on the viability of 769-P and hUSCs cells after 24-72 hours，compared with the control group，and morphological changes in cells were observed. Flow cytometry was utilized to assess the apoptosis rate of 769-P and hUSCs cells after treatment with 10 mg/mL LDGD，and apoptosis rates were calculated. RESULTS： Cells with good morphology and high growth activity were obtained using room temperature centrifugation. hUSCs expressed mesenchymal stem cell surface markers CD44 and CD90，but did not express endothelial cell marker CD31 or hematopoietic stem cell marker CD34. LDGD at 5-10 mg/mL inhibited the viability of 769-P cells in a time- and concentration-dependent manner after 24-72 hours，while promoting the viability of hUSCs. After 48 hours of treatment with 10 mg/mL LDGD，769-P cells exhibited significant shrinkage，reduced size，and decreased refractive index under an inverted microscope，whereas hUSCs showed no significant morphological changes. The apoptosis rate of 769-P cells was (11.93±0.51)% after 48 hours of treatment with 10 mg/mL LDGD，significantly higher than the control group，while the apoptosis rate of hUSCs was (0.01±0.10)%，showing no significant apoptosis compared to the control. CONCLUSION： Under the conditions of this experiment，LDGD inhibited the viability of human renal carcinoma cells in vitro and induced apoptosis，while promoting the viability of hUSCs without significantly inducing apoptosis. The effects of LDGD on the two types of cells were markedly different.

Key words: licorice and dried ginger decoction, renal carcinoma cells, human urine-derived stem cells, cell viability, apoptosis

中图分类号:

R961

黄吉, 张忆雪, 孙震晓. 甘草干姜汤对人肾癌细胞及人尿源性干细胞的作用[J]. 癌变·畸变·突变, 2024, 36(5): 359-364,390.

HUANG Ji, ZHANG Yixue, SUN Zhenxiao. Effects of licorice and dried ginger decoction on human renal carcinoma cells and human urine-derived stem cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2024, 36(5): 359-364,390.

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