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氨磷汀通过上调PKM2表达增强肾近端小管上皮细胞抵抗顺铂毒性
Amifostine enhances the resistance of renal proximal tubular epithelial cells to cisplatin by upregulating PKM2
- 陆军军医大学学报 2023年45卷第5期 页码：433-440
- 作者机构：
1.730000 兰州，甘肃省人民医院肾内科
2.730000 兰州，甘肃省人民医院普外科
3.730050 兰州，西宁联勤保障中心药品仪器监督检验站
- 作者简介：
[李小丽，E-mail：shenneike2022@163.com LI Xiaoli, E-mail: shenneike2022@163.com
- 基金信息：国家自然科学基金地区科学基金项目(81660398);甘肃省自然科学基金(21JR7RA625);甘肃省自然科学基金(22JR11RA256)
DOI：10.16016/j.2097-0927.202207159
中图分类号： R322.61;R966;R979.19收稿：2022-07-25，
修回：2022-09-16，
纸质出版：2023-03-15
- 稿件说明：
移动端阅览
薛嵘, 马金刚, 李莹屏, 等. 氨磷汀通过上调PKM2表达增强肾近端小管上皮细胞抵抗顺铂毒性[J]. 陆军军医大学学报, 2023,45(5):433-440.
Rong XUE, Jingang MA, Yingping LI, et al. Amifostine enhances the resistance of renal proximal tubular epithelial cells to cisplatin by upregulating PKM2[J]. Journal of Army Medical University, 2023, 45(5): 433-440.
薛嵘, 马金刚, 李莹屏, 等. 氨磷汀通过上调PKM2表达增强肾近端小管上皮细胞抵抗顺铂毒性[J]. 陆军军医大学学报, 2023,45(5):433-440. DOI： 10.16016/j.2097-0927.202207159.
Rong XUE, Jingang MA, Yingping LI, et al. Amifostine enhances the resistance of renal proximal tubular epithelial cells to cisplatin by upregulating PKM2[J]. Journal of Army Medical University, 2023, 45(5): 433-440. DOI： 10.16016/j.2097-0927.202207159.
摘要
目的
2
探究氨磷汀缓解顺铂肾毒性的分子机制。
方法
2
在肾近端小管上皮细胞系HK-2及原代细胞中，采用CCK-8细胞活性实验检测氨磷汀对顺铂细胞毒性的抑制作用。通过在线数据库tabula-muris分析肾脏组织各细胞亚群
PKM
2的表达。通过流式细胞术检测细胞凋亡比例。通过荧光定量PCR和蛋白免疫印迹实验检测细胞中
PKM
2基因表达。通过小RNA干扰技术敲低细胞中
PKM
2基因表达。
结果
2
氨磷汀显著增强HK-2细胞和原代肾近端小管上皮细胞抵抗顺铂的毒性。在顺铂处理的细胞中，随着氨磷汀浓度的增加，细胞的活力从0.37增加到0.45至0.77(
P
<
0.001)。另外，随着氨磷汀浓度的增加，HK-2细胞中
PKM
2基因表达依次增加，在1 μmol/L浓度时增加到8.76倍(
P
<
0.001)。在HK-2细胞中干扰
PKM
2基因表达，氨磷汀对顺铂的细胞毒性的保护作用显著受到抑制。顺铂诱导的细胞凋亡比例、标志蛋白Cleaved-PARP和Cleaved-caspase3的表达被氨磷汀显著抑制。单细胞数据分析的结果表明，相对于肾脏组织其他细胞亚群，
PKM
2在肾近端小管上皮细胞中表达最低。
结论
2
氨磷汀通过上调肾近端小管上皮细胞
PKM
2的表达抑制顺铂诱导的细胞凋亡，从而缓解顺铂的细胞毒性。
Abstract
Objective
2
To explore the molecular mechanism of amifostine in relieving cisplatin nephrotoxicity.
Methods
2
CCK-8 assay was used to detect the inhibitory effect of amifostine on cisplatin cytotoxicity in HK-2 and primary renal proximal tubular epithelial cells. The expression of
PKM
2 was analyzed by online database tabula muris. The proportion of apoptotic cells was detected by flow cytometry. The expression of
PKM
2 gene was detected by fluorescence quantitative PCR and Western blotting.
PKM
2
gene expression was knocked down by small RNA interference.
Results
2
Amifostine significantly enhanced the resistance of renal proximal tubular epithelial cells to cisplatin toxicity. In cisplatin treated cells
with the increase of the concentration of amifostine
the cell viability was increased from 0.37 to 0.45 to 0.77 (
P
<
0.001). In addition
with the increase of amifostine concentration
PKM
2 gene expression in HK-2 cells was increased in turn. The expression of
PKM
2 gene was increased to 8.76 times higher at 1 μmol/L of amifostine (
P
<
0.001).
PKM
2 gene expression in HK-2 cells was increased with the increase of amifostine concentration. When
PKM
2 gene expression was depleted in HK-2 cells
and the protective effect of amifostine on cisplatin cytotoxicity was significantly inhibited. The proportion of apoptotic cells was reduced and the levels of apoptotic markers Cleaved-PARP and Cleaved-Caspase3 induced by cisplatin were significantly inhibited by amifostine. Single-cell data analysis indicated that the expression of
PKM
2 in renal proximal tubular epithelial cells was the lowest compared with other cell subsets in renal tissue.
Conclusion
2
Amifostine inhibits cell apoptosis induced by cisplatin by up-regulating
PKM
2 in renal proximal tubular epithelial cells
and thus alleviates the cytotoxicity of cisplatin.
关键词
Keywords
references
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