Calcitonin gene-related peptide alleviates hyperoxia-induced human alveolar cell injury via the CGRPR/TRPV1/Ca2<SUP>+</SUP> axis

作者全名：Li, Jun-Hui; Wan, Han-Xing; Wu, Li-Hong; Fang, Fang; Wang, Jian-Xin; Dong, Hui; Xu, Feng

作者地址：[Li, Jun-Hui; Wan, Han-Xing; Wu, Li-Hong; Fang, Fang; Dong, Hui; Xu, Feng] Chongqing Med Univ, Childrens Hosp, Dept Pediat Intens Care Unit, Natl Clin Res Ctr Child Hlth & Disorders,Minist Ed, 136 Zhongshan Second Rd, Chongqing 400037, Peoples R China; [Wang, Jian-Xin; Dong, Hui] Qingdao Univ, Med Coll, Sch Pharm, Dept Pharmacol, 1 Ningde Rd, Qingdao 266073, Shandong, Peoples R China

通信作者：Dong, H; Xu, F (通讯作者)，Chongqing Med Univ, Childrens Hosp, Dept Pediat Intens Care Unit, Natl Clin Res Ctr Child Hlth & Disorders,Minist Ed, 136 Zhongshan Second Rd, Chongqing 400037, Peoples R China.; Dong, H (通讯作者)，Qingdao Univ, Med Coll, Sch Pharm, Dept Pharmacol, 1 Ningde Rd, Qingdao 266073, Shandong, Peoples R China.

来源：MOLECULAR MEDICINE REPORTS

ESI学科分类：CLINICAL MEDICINE

WOS号：WOS:001219621100001

JCR分区：Q2

影响因子：3.4

年份：2024

卷号：30

期号：1

开始页：　

结束页：　

文献类型：Article

关键词：hyperoxia; A549 cells; TRPV1; proliferation; apoptosis

摘要：Although exogenous calcitonin gene-related peptide (CGRP) protects against hyperoxia-induced lung injury (HILI), the underlying mechanisms remain unclear. The present study attempted to elucidate the molecular mechanism by which CGRP protects against hyperoxia-induced alveolar cell injury. Human alveolar A549 cells were treated with 95% hyperoxia to establish a hyperoxic cell injury model. ELISA was performed to detect the CGRP secretion. Immunofluorescence, quantitative (q)PCR, and western blotting were used to detect the expression and localization of CGRP receptor (CGRPR) and transient receptor potential vanilloid 1 (TRPV1). Cell counting kit-8 and flow cytometry were used to examine the proliferation and apoptosis of treated cells. Digital calcium imaging and patch clamp were used to analyze the changes in intracellular Ca2(+) signaling and membrane currents induced by CGRP in A549 cells. The mRNA and protein expression levels of Cyclin D1, proliferating cell nuclear antigen (PCNA), Bcl-2 and Bax were detected by qPCR and western blotting. The expression levels of CGRPR and TRPV1 in A549 cells were significantly downregulated by hyperoxic treatment, but there was no significant difference in CGRP release between cells cultured under normal air and hyperoxic conditions. CGRP promoted cell proliferation and inhibited apoptosis in hyperoxia, but selective inhibitors of CGRPR and TRPV1 channels could effectively attenuate these effects; TRPV1 knockdown also attenuated this effect. CGRP induced Ca2(+) entry via the TRPV1 channels and enhanced the membrane non-selective currents through TRPV1 channels. The CGRP-induced increase in intracellular Ca2(+) was reduced by inhibiting the phospholipase C (PLC)/protein kinase C (PKC) pathway. Moreover, PLC and PKC inhibitors attenuated the effects of CGRP in promoting cell proliferation and inhibiting apoptosis. In conclusion, exogenous CGRP acted by inversely regulating the function of TRPV1 channels in alveolar cells. Importantly, CGRP protected alveolar cells from hyperoxia-induced injury via the CGRPR/TRPV1/Ca2(+ )axis, which may be a potential target for the prevention and treatment of the HILI.

基金机构：National Natural Science Foundation of China

基金资助正文：Not applicable.