H(+)/Cl(‑) exchange transporter 7 promotes lysosomal acidification‑mediated autophagy in mouse cardiomyocytes.

Jiezhi Lin, Jinyu Wei, Yanling Lv, Xingyue Zhang, Ruo Fan Yi, Chen Dai, Qiong Zhang, Jiezhi Jia, Dongxia Zhang, Yuesheng Huang

Molecular medicine reports 2021 Mar

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Autophagy protects cardiomyocytes in various pathological and physiological conditions; however, the molecular mechanisms underlying its influence and the promotion of autophagic clearance are not completely understood. The present study aimed to explore the role of H(+)/Cl(‑) exchange transporter 7 (CLC‑7) in cardiomyocyte autophagy. In this study, rapamycin was used to induce autophagy in mouse cardiomyocytes, and the changes in CLC‑7 were investigated. The expression levels of CLC‑7 and autophagy‑related proteins, such as microtubule associated protein 1 light chain 3, autophagy related 5 and Beclin 1, were detected using western blotting or immunofluorescence. Autolysosomes were observed and analyzed using transmission electron microscopy and immunofluorescence following CLC‑7 silencing with small interfering RNAs. Cellular viability was assessed using Cell Counting Kit‑8 and lactate dehydrogenase assays. Lysosomal acidification was measured using an acidification indicator. Increased CLC‑7 co‑localization with lysosomes was identified during autophagy. CLC‑7 knockdown weakened the acidification of lysosomes, which are the terminal compartments of autophagy flux, and consequently impaired autophagy flux, ultimately resulting in cell injury. Collectively, the present study demonstrated that in cardiomyocytes, CLC‑7 may contribute to autophagy via regulation of lysosomal acidification. These findings provide novel insights into the role of CLC‑7 in autophagy and cytoprotection.

Citation

Jiezhi Lin, Jinyu Wei, Yanling Lv, Xingyue Zhang, Ruo Fan Yi, Chen Dai, Qiong Zhang, Jiezhi Jia, Dongxia Zhang, Yuesheng Huang. H(+)/Cl(‑) exchange transporter 7 promotes lysosomal acidification‑mediated autophagy in mouse cardiomyocytes. Molecular medicine reports. 2021 Mar;23(3)