Journal of Shandong University (Health Sciences) ›› 2023, Vol. 61 ›› Issue (4): 1-9.doi: 10.6040/j.issn.1671-7554.0.2022.1226

• 基础医学 •    

ACE2 gene protects against renal ischemia-reperfusion injury by regulating the Nrf2/HO-1 signaling pathway

ZHANG Jiaying, SU Rongyun, WANG Yinghui, WANG Honggang, LIU Gang   

  1. Department of Nephrology, The Second Hospital of Shandong University, Jinan 250033, Shandong, China
  • Published:2023-04-11

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Abstract: Objective To investigate the effects of angiotensin-converting enzyme 2(ACE2)on the oxidative stress, inflammation, apoptosis and the nuclear factor E2-related factor 2(Nrf2)/ heme oxygenase 1(HO-1)signaling pathway in renal tubular epithelial cells(HK-2)induced by hypoxia/reoxygenation(H/R). Methods HK-2 cells were transfected with ACE2 lentivirus, and divided into the control group, H/R group, H/R-NC group, and H/R-ACE2 group. After H/R treatment, cell viability was measured with CCK-8 assay; the levels of inflammatory factors including interleukin-6(IL-6), tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)were measured with ELISA and RT-PCR; superoxide dismutase(SOD)and malondialdehyde(MDA)levels were measured with colorimetric assay; protein levels of Caspase-3, Bcl-2, Bax, Nrf2, and HO-1 were measured with Western blotting. After ML385 and SnPPIX were used to inhibit the Nrf2/HO-1 pathway, changes in the expressions of Caspase-3, Bcl-2, Bax, Nrf2 and HO-1 were detected with Western blotting, and changes in SOD and MDA levels were detected with colorimetry. Results Compared with the control group, the H/R group showed lower cell viability(t=7.58, P<0.001), higher expression levels of MDA, IL-1β, IL-6, TNF-α, Caspase-3 and Bax(tMDA=11.08, PMDA<0.001; tPCR-IL-6=5.82, PPCR-IL6<0.001; tPCR-TNF-α=7.69, PPCR-TNF-α<0.001; tPCR-IL-1β=4.80, PPCR-IL-1β=0.001; tELISA-IL-6=34.11, PELISA-IL-6<0.001; tELISA-TNF-α=14.12, PELISA-TNF-α<0.001; tELISA-IL-1β=9.63, PELISA-IL-1β<0.001; tCaspase-3=2.73, PCaspase-3=0.026; tBax=27.75, PBax<0.001), but lower levels of SOD, Bcl-2 and ACE2(tSOD=7.74, PSOD<0.001; tBcl-2=75.49, PBcl-2<0.001; tACE2=11.41, PACE2<0.001). Compared with the H/R group, the H/R-ACE2 group had higher cell viability(t=3.61, P=0.002), lower levels of MDA, IL-1β, IL-6, TNF-α, Caspase-3 and Bax(tMDA=6.15, PMDA<0.001; tPCR-IL-6=3.34, PPCR-IL6=0.006; tPCR-TNF-α=3.65, PPCR-TNF-α=0.007; tPCR-IL-1β=4.06, PPCR-IL-1β=0.004; tELISA-IL-6=14.62, PELISA-IL-6<0.001; tELISA-TNF-α=10.42, PELISA-TNF-α<0.001; tELISA-IL-1β=8.65, PELISA-IL-1β<0.001; tCaspase-3=3.74, PCaspase-3=0.006; tBax=30.52, PBax<0.001), higher levels of SOD, Bcl-2, ACE2, Nrf2, and HO-1(tSOD=3.58, PSOD=0.007; tBcl-2=63.86, PBcl-2<0.001; tACE2=58.72, PACE2<0.001; tNrf2=44.55, PNrf2<0.001; tHO-1=14.19, PHO-1<0.001). However, ML385 and SnPPIX inhibited the protective effects of ACE2 gene overexpression on HK-2 cells under H/R(FBax=11.02, PBax=0.003; FBcl-2=21.48, PBcl-2<0.001; FCaspase-3=20.80, PCaspase-3<0.001; FSOD=133.49, PSOD<0.001; FMDA=14.06, PMDA=0.001). Conclusion ACE2 inhibits the oxidative stress, regulates inflammation, and ameliorates apoptosis in HK-2 cells under H/R, and the Nrf2/HO-1 signaling pathway may play an important role in this progress.

Key words: Angiotensin-converting enzyme 2, Ischemia-reperfusion injury, Oxidative stress, Apoptosis, Signaling pathway

CLC Number: 

  • R574
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