摘要
目的:探究人参皂苷CK(CK)对血小板衍生生长因子-BB(PDGF-BB)诱导肺动脉平滑肌细胞(PASMC)的迁移和胶原沉积的作用以及相关的机制。方法:以体外培养的大鼠PASMC为研究对象,PDGF-BB为诱导剂,CK为干预剂。实验分为三组(每组每次实验有5个负孔,独立重复3次即每组n=3):采用CCK-8法检测对照组(不做处理)、模型组(给予PDGF-BB 20 mg/L)、干预组(同时给予PDGF-BB 20 mg/L+CK 5μmol/L)细胞增殖情况;细胞划痕实验检测细胞迁移;实时荧光定量PCR检测Ⅰ型胶原蛋白(COLⅠ)和Ⅲ型胶原蛋白(COLⅢ)信使RNA(mRNA)的相对表达;酶联免疫吸附(ELISA)法检测细胞培养液中COLⅠ和COLⅢ蛋白的表达;蛋白免疫印迹(Western blot)法检测COLⅠ、COLⅢ和β-连环蛋白(β-catenin)的相对表达。结果(:1)与对照组比较,模型组中PDGF-BB显著诱导了PASMC的增殖;与对照组比较,模型组PASMC在12 h、24 h的迁移率[(37%vs.26%)、(68%vs.51%)]均呈显著升高,差异均有统计学意义(P均<0.01)。PASMC培养液中,与对照组比较,模型组培养液中COLⅠ蛋白含量升高约3.5倍和COLⅢ蛋白含量升高2.5倍(P<0.01),同时PASMC中COLⅠ、COLⅢ、β-catenin的m RNA和蛋白表达水平均明显升高(P均<0.01)。(2)与模型组比较,干预组中CK抑制了PDGF-BB诱导的PASMC增殖;与模型组比较,干预组12 h、24 h迁移率分别为[(16%vs.37%)、(48%vs.68%)]较模型组均明显降低(P均<0.01),PASMC培养液中COLⅠ蛋白含量减少了45%和COLⅢ减少了56%(P<0.01),同时PASMC中COLⅠ、COLⅢ和β-catenin的m RNA和蛋白表达水平均明显降低(P均<0.01),差异均有统计学意义。结论:CK明显抑制了PDGF-BB诱导PASMC的增殖、迁移以及胶原沉积,这表明CK可能对胶原沉积类疾病有抑制作用。
Objectives:To investigate the effects of ginsenoside compound K(CK)on the proliferation,migration,and collagen deposition of platelet-derived growth factor-BB(PDGF-BB)-induced pulmonary artery smooth muscle cells(PASMCs)and the related mechanisms.Methods:In vitro cultured rat PASMCs were used,PDGF-BB was used as the inducer,CK was used as the intervention agent.PASMCs were divided into 3 groups:control group,model group(treated with PDGF-BB 20 mg/L),intervention group(treated with PDGF-BB 20 mg/L and CK 5μmol/L).Cell proliferation was detected by CCK-8 assay.Cell migration was detected by cell scratch assay.RT-qPCR was used to detect the mRNA expression of collagen type I(COL I)and collagen typeⅢ(COLⅢ);ELISA was used to detect protein secretion of COL I and COLⅢ.Western blot was used to detect the protein expression of COL I,COLⅢ,andβ-catenin.Results:Compared with the control group,PDGF-BB significantly induced PASMCs proliferation in the model group.The mobility at 12 h(37%vs.26%,P<0.01)and 24 h(68%vs.51%,P<0.01)was significantly enhanced in the model group than in the control group.The protein contents of COL I and COLⅢin culture medium increased by about 3.5 fold and 2.5 fold in the model group as compared to control group(P<0.01),and the mRNA and protein expression levels of COL I,COLⅢ,andβ-catenin were significantly upregulated in model group than in control group(P<0.01).Compared with the model group,PDGF-BB-induced PASMCs proliferation,and enhanced mobility(at 12 h:16%vs.37%,P<0.01;at 24 h:48%vs.68%,P<0.01)were significantly inhibited in the intervention group.The protein contents of COL I,COLⅢin culture medium were also significantly decreased by 45%and 56%in the intervention group as compared to the model group(both P<0.01),and the mRNA and protein expression levels of COL I,COLⅢ,andβ-catenin in PASMCs were also significantly decreased in the intervention group than in the model group(all P<0.01).Conclusions:Ginsenoside compound K could significantly inhibit PDGF-BB-induced PASMCs pro
作者
刘桃
朱路
李刚
贾鹏
谢亮
刘瀚旻
刘斌
LIU Tao;ZHU Lu;LI Gang;JIA Peng;XIE Liang;LIU Hanmin;LIU Bin(Sichuan Clinical Research Center for Birth Defects,Department of Pediatrics,The Affiliated Hospital of Southwest Medical University,Luzhou 646000,Sichuan,China)
出处
《中国循环杂志》
CSCD
北大核心
2021年第12期1229-1236,共8页
Chinese Circulation Journal
基金
四川省科技厅泸州市西南医科大学科研基金(Z1432)资助。
关键词
人参皂苷CK
肺动脉平滑肌细胞
迁移
胶原蛋白
胶原沉积
ginsenoside compound k
pulmonary arterial smooth muscle cells
migration
collagen
collagen deposition
