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红景天苷抑制缺氧诱导的大鼠肺动脉平滑肌细胞表型转化

[Salidroside inhibits phenotypic transformation of rat pulmonary artery smooth muscle cells induced by hypoxia].

作者信息

Mao Jia-Qi, Liu Chuan-Chuan, Zhang Yu-Wei, Zhang Qing-Qing, Liu Hong, Ma Lan

机构信息

Qinghai University Xining 810001, China Research Center for High Altitude Medicine, Medical School, Qinghai University Xining 810001, China Qinghai Provincial Key Laboratory of Traditional Chinese Medicine Research for Glucolipid Metabolic Diseases Xining 810001, China.

Qinghai University Xining 810001, China Research Center for High Altitude Medicine, Medical School, Qinghai University Xining 810001, China the Echinococcosis Key Laboratory of Qinghai University Xining 810001, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2022 Feb;47(4):1024-1030. doi: 10.19540/j.cnki.cjcmm.20211103.706.

Abstract

This study investigated the effect of salidroside on phenotypic transformation of rat pulmonary artery smooth muscle cells(PASMCs) induced by hypoxia. Rat pulmonary arteries were isolated by tissue digestion and PASMCs were cultured. The OD values of cells treated with salidroside at different concentrations for 48 hours were measured by cell counting kit-8(CCK-8) to determine the appropriate concentration range of salidroside. The cells were divided into a normal(normoxia) group, a model(hypoxia) group, and three hypoxia + salidroside groups(40, 60, and 80 μg·mL~(-1)). Quantitative real-time PCR(qRT-PCR) was used to detect the mRNA expression of cell contractile markers in each group, such as α-smooth muscle actin(α-SMA), smooth muscle 22(SM22), and calcium-binding protein(calponin), and synthetic marker vimentin. The expression levels of cell phenotypic markers and proliferating cell nuclear antigen(PCNA) were detected by Western blot. The proliferation of cells in each group was detected by the 5-ethynyl-2'-deoxyuridine(EdU) assay. Cell migration was measured by Transwell assay. As revealed by results, compared with the normal group, the model group showed decreased mRNA and protein expression of contractile phenotypic markers of PASMCs and increased mRNA and protein expression of synthetic markers. Compared with the conditions in the model group, salidroside could down-regulate the mRNA and protein expression of synthetic markers in PASMCs and up-regulated the mRNA and protein expression of contractile phenotypic markers. Compared with the normal group, the model group showed potentiated proliferation and migration. Compared with the model group, the hypoxia + salidroside groups showed blunted proliferation and migration of cells after phenotypic transformation. The results suggest that salidroside can inhibit the expression of synthetic markers in PASMCs and promote the expression of contractile markers to inhibit the hypoxia-induced phenotypic transformation of PASMCs. The mechanism of salidroside in inhibiting the proliferation and migration of PASMCs is related to the inhibition of the phenotypic transformation of PASMCs.

摘要

本研究探讨了红景天苷对缺氧诱导的大鼠肺动脉平滑肌细胞(PASMCs)表型转化的影响。通过组织消化分离大鼠肺动脉并培养PASMCs。用细胞计数试剂盒-8(CCK-8)测定不同浓度红景天苷处理48小时的细胞的OD值,以确定红景天苷的合适浓度范围。将细胞分为正常(常氧)组、模型(缺氧)组和三个缺氧+红景天苷组(40、60和80μg·mL⁻¹)。采用定量实时聚合酶链反应(qRT-PCR)检测各组细胞收缩标志物如α-平滑肌肌动蛋白(α-SMA)、平滑肌22(SM22)和钙结合蛋白(钙调蛋白)以及合成标志物波形蛋白的mRNA表达。通过蛋白质免疫印迹法检测细胞表型标志物和增殖细胞核抗原(PCNA)的表达水平。采用5-乙炔基-2'-脱氧尿苷(EdU)检测法检测各组细胞的增殖情况。通过Transwell检测法测量细胞迁移。结果显示,与正常组相比,模型组PASMCs收缩表型标志物的mRNA和蛋白表达降低,合成标志物的mRNA和蛋白表达增加。与模型组相比,红景天苷可下调PASMCs中合成标志物的mRNA和蛋白表达,并上调收缩表型标志物的mRNA和蛋白表达。与正常组相比,模型组细胞增殖和迁移增强。与模型组相比,缺氧+红景天苷组细胞表型转化后增殖和迁移减弱。结果表明,红景天苷可抑制PASMCs中合成标志物的表达,促进收缩标志物的表达,从而抑制缺氧诱导的PASMCs表型转化。红景天苷抑制PASMCs增殖和迁移的机制与抑制PASMCs的表型转化有关。

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