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在体外,降钙素基因相关肽通过抑制p38丝裂原活化蛋白激酶途径保护高糖刺激的膀胱平滑肌细胞。

CGRP protects bladder smooth muscle cells stimulated by high glucose through inhibiting p38 MAPK pathway in vitro.

作者信息

Xue Jun, Liu Yadong, Zhang Sichong, Ding Liucheng, Shen Baixin, Shao Yunpeng, Wei Zhongqing

机构信息

Department of Urology, The Second Affiliated Hospital of Nanjing Medical University, 121 Jiangjiayuan Road, Nanjing, 210011, Jiangsu, China.

Department of Urology, The Third People's Hospital of Yancheng, Yancheng, 224001, Jiangsu, China.

出版信息

Sci Rep. 2021 Apr 7;11(1):7643. doi: 10.1038/s41598-021-87140-y.

Abstract

This study aimed to explore the effect of calcitonin gene-related peptide (CGRP) on bladder smooth muscle cells (BSMCs) under high glucose (HG) treatment in vitro. BSMCs from Sprague-Dawley rat bladders were cultured and passaged in vitro. The third-generation cells were cultured and divided into control group, HG group, HG + CGRP group, HG + CGRP + asiatic acid (AA, p-p38 activator) group, CGRP group, AA group, HG + CGRP + CGRP-8-37 (CGRP receptor antagonist) group and HG + LY2228820 (p38 MAPK inhibitor) group. The cell viability, apoptosis, malondialdehyde (MDA) and superoxide dismutase (SOD) levels of BSMCs were observed by the relevant detection kits. The expressions of α-SM-actin, p38 and p-p38 were detected by qRT-PCR or Western blot analysis. Compared with the control group, the cell viability, SOD and α-SM-actin levels of BSMCs were decreased and apoptotic cells, MDA and p-p38 levels were increased after HG treatment, while these changes could be partly reversed when BSMCs were treated with HG and CGRP or LY2228820 together. Moreover, AA or CGRP-8-37 could suppress the effect of CGRP on BSMCs under HG condition. Our data indicate that CGRP protects BSMCs from oxidative stress induced by HG in vitro, and inhibit the α-SM-actin expression decrease through inhibiting the intracellular p38 MAPK signaling pathway.

摘要

本研究旨在探讨体外高糖(HG)处理下降钙素基因相关肽(CGRP)对膀胱平滑肌细胞(BSMCs)的影响。从Sprague-Dawley大鼠膀胱中分离出BSMCs并进行体外培养和传代。将第三代细胞培养后分为对照组、HG组、HG + CGRP组、HG + CGRP + 积雪草苷(AA,p-p38激活剂)组、CGRP组、AA组、HG + CGRP + CGRP-8-37(CGRP受体拮抗剂)组和HG + LY2228820(p38丝裂原活化蛋白激酶抑制剂)组。通过相关检测试剂盒观察BSMCs的细胞活力、凋亡、丙二醛(MDA)和超氧化物歧化酶(SOD)水平。采用qRT-PCR或蛋白质免疫印迹分析检测α-SM-肌动蛋白、p38和p-p38的表达。与对照组相比,HG处理后BSMCs的细胞活力、SOD和α-SM-肌动蛋白水平降低,凋亡细胞、MDA和p-p38水平升高,而当BSMCs同时用HG和CGRP或LY2228820处理时,这些变化可部分逆转。此外,AA或CGRP-8-37可抑制HG条件下CGRP对BSMCs的作用。我们的数据表明,CGRP在体外可保护BSMCs免受HG诱导的氧化应激,并通过抑制细胞内p38丝裂原活化蛋白激酶信号通路来抑制α-SM-肌动蛋白表达的降低。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0393/8027675/b0f754f7669e/41598_2021_87140_Fig1_HTML.jpg

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