红景天苷对低氧诱导的大鼠肺动脉平滑肌细胞表型转化的影响  

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

作　　者：毛稼琦 刘川川[1,2,4] 张雨薇[1,2] 张晴晴 刘红 马兰 MAO Jia-qi;LIU Chuan-chuan;ZHANG Yu-wei;ZHANG Qing-qing;LIU Hong;MA Lan(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;the Echinococcosis Key Laboratory of Qinghai University,Xining 810001,China;Department of Respiratory,Affiliated Hospital of Qinghai University,Xining 810001,China)

机构地区：[1]青海大学,青海西宁810001 [2]青海大学医学院高原医学研究中心,青海西宁810001 [3]青海省糖脂代谢疾病防控中医药重点实验室,青海西宁810001 [4]青海大学包虫病重点实验室,青海西宁810001 [5]青海大学附属医院呼吸科,青海西宁810001

出　　处：《中国中药杂志》2022年第4期1024-1030,共7页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(32060207,31560292);青海省自然科学基金项目(2021-ZJ-738)。

摘　　要：探讨红景天苷对低氧引起的大鼠肺动脉平滑肌细胞(pulmonary artery smooth muscle cells,PASMCs)表型转化的影响。采用组织消化法分离大鼠肺动脉,培养PASMCs,用CCK-8(cell counting kit-8)法测定不同浓度红景天苷处理细胞48 h后的A,确定红景天苷作用的适宜浓度范围。将细胞分为常氧对照组、低氧对照组和低氧+红景天苷处理组(40、60、80μg·mL^(-1)),采用实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)检测各组细胞收缩型标志物如α-平滑肌肌动蛋白(α-SMA)、平滑肌22(SM22)、钙结合蛋白(calponin)和合成型标志物波形蛋白(vimentin)的基因表达水平;Western blot检测各组细胞表型标志物和核增殖抗原(PCNA)蛋白表达水平;EdU(5-ethynyl-2′-deoxyuridine)法检测各组细胞增殖情况;Transwell法测定各组细胞迁移情况。结果表明,与常氧对照组相比,低氧可诱导PASMCs收缩型表型标志物mRNA和蛋白表达降低,合成型标志物mRNA和蛋白升高;与低氧对照组相比,红景天苷可以下调PASMCs合成型标志物mRNA和蛋白表达,上调收缩型表型标志物mRNA和蛋白表达。与常氧对照组相比,低氧对照组增殖和迁移能力增加;与低氧对照组相比,红景天苷抑制表型转化后细胞的增殖和迁移能力降低。结果提示,红景天苷可以抑制PASMCs合成型标志物表达,促进收缩型标志物表达,从而抑制低氧引起的PASMCs表型转化;且红景天苷抑制PASMCs增殖和迁移的机制与其抑制了PASMCs的表型转化有关。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 80μg·mL^(-1)).Quantitative real-time PCR(qRT-PCR)was used to detect the mRNA expression of cell contractile markers in each group,such asα-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 migrat

关 键 词：低氧 肺动脉平滑肌细胞 表型转化 红景天苷 

分 类 号：R285.5[医药卫生—中药学]