Roxadustat attenuates heat stress-induced apoptosis and senescence in renal tubular epithelial cells by upregulating HIF-1α

Yongwei SONG; Ling WANG; Wenting CHEN; Mingyang ZHANG; Xuesen YANG; Huanzi DAI

doi:10.16016/j.2097-0927.202312123

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Roxadustat attenuates heat stress-induced apoptosis and senescence in renal tubular epithelial cells by upregulating HIF-1α

Basic Medicine | 更新时间：2026-01-09

- Roxadustat attenuates heat stress-induced apoptosis and senescence in renal tubular epithelial cells by upregulating HIF-1α
- Journal of Army Medical University Vol. 46, Issue 10, Pages: 1092-1099(2024)
- 作者机构：
  
  1.400042 重庆，陆军特色医学中心风湿免疫科
  2.400038 重庆，陆军军医大学(第三军医大学)军事预防医学系热带医学教研室
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of Chongqing(CSTB2023NSCQ-MSX1101)
- DOI：10.16016/j.2097-0927.202312123
  CLC： R594.11;R692.9;R977.6
- Received：25 December 2023，
  
  Revised：2024-03-07，
  
  Published：30 May 2024
- 稿件说明：
移动端阅览
Yongwei SONG, Ling WANG, Wenting CHEN, et al. Roxadustat attenuates heat stress-induced apoptosis and senescence in renal tubular epithelial cells by upregulating HIF-1α[J]. Journal of Army Medical University, 2024, 46(10): 1092-1099.
DOI：

Yongwei SONG, Ling WANG, Wenting CHEN, et al. Roxadustat attenuates heat stress-induced apoptosis and senescence in renal tubular epithelial cells by upregulating HIF-1α[J]. Journal of Army Medical University, 2024, 46(10): 1092-1099. DOI： 10.16016/j.2097-0927.202312123.

摘要

目的

探讨罗沙司他对热打击诱导的肾小管上皮细胞(HK-2细胞)凋亡与衰老的影响及其机制。

方法

体外培养HK-2细胞，用不同浓度(10、20、30、40、50 μmol/L)的罗沙司他处理24 h，通过CCK-8法确定罗沙司他的最佳干预浓度。将HK-2细胞分为4组(

=3): 对照组、罗沙司他组(30 μmol/L、24 h)、热打击组(43 ℃、2 h)、热打击+罗沙司他组(30 μmol/L罗沙司他处理24 h后热打击2 h)。通过CCK-8检测细胞活力；Western blot检测缺氧诱导因子-1α(hypoxia-inducible factor-1，HIF-1α)、Cleaved Caspase-3

、p16、p21蛋白表达量；免疫荧光检测HIF-1α分布；细胞衰老β-半乳糖苷酶染色试剂盒检测SA-β-Gal活性；TUNEL染色检测细胞凋亡情况。

结果

用30 μmol/L罗沙司他处理的细胞活力最高。与对照组比较，热打击组细胞活力显著降低(

0.05)；HIF-1α、Cleaved Caspase-3、p16、p21蛋白表达量显著增加(

0.05)；SA-β-Gal活性显著增加(

0.05)；TUNEL阳性细胞占比显著增加(

0.05)。与热打击组比较，热打击+罗沙司他组Cleaved Caspase-3、p16、p21蛋白表达量显著降低(

0.05)；SA-β-Gal活性显著降低[(65.44±5.00)%

(77.15±2.61)%，

0.05

]

；TUNEL阳性细胞占比显著降低[(16.73±2.20)%

(46.40±13.87)%，

0.05

]

，细胞活力显著增加[(86.33±4.51)%

(66.33±8.50)%，

0.05

]

；HIF-1α蛋白表达量进一步增加(

0.05)。免疫荧光结果显示HIF-1α主要分布在细胞核及细胞核周围。

结论

罗沙司他通过上调HIF-1α减轻热打击诱导的肾小管上皮细胞凋亡和衰老。

Abstract

Objective

To investigate the effect and underlying mechanism of roxadustat on apoptosis and senescence of renal tubular epithelial cell line HK-2 induced by heat stress.

Methods

After HK-2 cells were treated with different concentrations of roxadustat (10

40 and 50 μmol/L) for 24 h

CCK-8 assay was used to determine the optimal intervention concentration of roxadustat. HK-2 cells were divided into 4 groups (

=3): control group

roxadustat group (30 μmol/L

24 h)

heat-stress group (43 ℃

2 h)

and heat-stress+roxadustat group (30 μmol/L roxadustat treatmnet for 24 h followed by heat-stress 2 h). Cell viability was detected by CCK-8 assay. Expression of hypoxia-inducible factor-1α (HIF-1α)

Cleaved Caspase-3

p16 and p21 at protein level was detected by Western blotting. Immunofluorescence assay was employed to observe the distribution of HIF-1α. β-galactosidase staining kit was utilized to detect SA-β-Gal activity. TUNEL staining was used to measure cell apoptosis.

Results

The highest

cell viability was observed in the cells after 30 μmol/L roxadustat treatment. Heat stress resulted in a significant decrease in cell viability (

0.05)

elevated protein levels of HIF-1α

Cleaved Caspase-3

p16 and p21(

0.05)

enhanced SA-β-Gal activity (

0.05) and increased percentage of TUNEL-positive cells (

0.05) when compared with the cells in the control group. In comparison with the heat-stress group

the heat-stress+roxadustat group showed significant decrease in the protein levels of Cleaved Caspase-3

p16 and p21 (

0.05)

reduced activity of SA-β-Gal [(65.44±5.00)%

(77.15±2.61)%

0.05

]

and decreased percentage of TUNEL-positive cells [(16.73±2.20)%

(46.40±13.87)%

0.05

]

but increase in cell viability [(86.33±4.51)%

(66.33±8.50)%

0.05

]

as well as HIF-1α protein expression (

0.05). Furthermore

immunofluorescence assay showed that HIF-1α was mainly distributed in the nucleus and perinucleus.

Conclusion

Roxadustat attenuates heat stress-induced apoptosis and senescence of renal tubular epithelial cells by upregulating HIF-1α.

关键词

Keywords

references

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