摘要
目的在体外水平探讨健脾益气利水汤对小鼠肾上皮细胞及肿瘤细胞的细胞毒性缓解作用,并从OCT-MATE转运系统探索其调控机制。方法通过MTT法检测不同给药组健脾益气利水汤对顺铂诱导的原代肾上皮细胞和肿瘤细胞的细胞毒性作用的影响,再通过ICP-MS检测不同给药组对顺铂诱导的原代肾上皮细胞和HEK293肾癌细胞内Pt含量的影响,最后通过RT-PCR和Western Blot检测不同浓度健脾益气利水汤对顺铂诱导的原代肾上皮细胞内OCT-1/2、MATE-2基因及蛋白表达变化。结果健脾益气利水汤能有效降低12.9μmol/L顺铂所致对体外小鼠肾上皮细胞的细胞毒性作用,其中预防组和同时组与模型组比较有显著性差异(P<0.05);同时发现健脾益气利水汤24h内对6.25~50μmol/L顺铂诱导的食管癌细胞EC109;大肠癌细胞HP-29;鼻咽癌细胞HK-1;肾癌细胞HEK293;肺癌细胞A549;胃癌细胞MKN-45的细胞毒性作用均无显著性差异(P>0.05)。ICP-MS检测发现,预防给药组和同时给药组能显著降低原代肾上皮细胞内Pt的含量(P<0.05),而在HEK293细胞内,各给药组细胞内Pt含量与模型组之间均无显著差异(P>0.05)。RT-PCR和Western Blot检测均发现,健脾益气利水汤显著降低顺铂诱导的小鼠肾上皮细胞内OCT-2表达水平(P<0.05),而OCT-1和MATE-2无明显差异(P>0.05)。结论健脾益气利水汤通过抑制OCT-2表达从而抑制顺铂进入肾上皮细胞,具有预防降低顺铂肾细胞毒性作用,同时对顺铂进入肿瘤细胞发挥抗肿瘤活性无显著影响。
Objective To investigate the cytotoxic effect of Jianpi Yiqi Lishui Tang on mouse renal epithelial cells and tumor cells in vitro,and to explore its regulatory mechanism from OCT-MATE transporters.Methods The effects of Jianpi Yiqi Lishui Tang on the cytotoxic effects of cisplatin induced primary renal epithelial cells and tumor cells were detected by MTT method.Then,the effects of different groups on the content of Pt in the primary renal epithelial cells and HEK293 renal cancer cells were detected by ICP-MS.Finally,the changes of OCT-1/2,MATE-2 gene and protein expression in primary renal epithelial cells induced by cisplatin were detected by RT-PCR and Western blot.Results The study found that Jianpi Yiqi Lishui Tang can effectively reduce the cytotoxic effects of 12.9μmol/L cisplatin on renal epithelial cells in vitro were significantly different between the prevention and simultaneous administration groups and the model group(P<0.05).There was no significant difference in the cytotoxic effect of Jianpi Yiqi Lishui Tang within 24 hours on EC109,HP-29,HK-1,HEK293,A549,MKN-45 tumor cells induced by cisplatin(P>0.05).ICP-MS showed that the content of Pt in primary renal epithelial cells was significantly reduced in the prevention and simultaneous administration groups(P<0.05),but in HEK293 cells,there was no significant difference(P>0.05).RT-PCR and Western blot showed that the expression of OCT-2 in cisplatin induced renal epithelial cells was significantly reduced by Jianpi Yiqi Lishui Tang(P<0.05),but OCT-1 and MATE-2 were not significantly different(P>0.05).Conclusion Jianpi Yiqi Lishui Tang can inhibit cisplatin from entering renal epithelial cells by inhibiting the expression of OCT-2,which can prevent and reduce the toxicity of cisplatin renal cells,and has no significant effect on the antitumor activity of cisplatin into tumor cells.
作者
周丽
翁纯雪
邱煌沛
高奥
林晓辉
贺龙刚
ZHOU Li;WENG Chun-xue;QIU Huang-pei;GAO Ao;LIN Xiao-hui;HE Long-gang(Fujian University of Traditional Chinese medicine,Fuzhou Fujian 350122,China)
出处
《时珍国医国药》
CAS
CSCD
北大核心
2022年第5期1106-1110,共5页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金(81302873)
福建省自然科学基金(2019J01340,2021J01927)
福建中医药大学校管课题重点项目(X2020013-重点)。