摘要
目的探讨人参皂苷Rb1(Ginsenoside Rb1,Rb1)对PDGF-BB诱导的大鼠主动脉平滑肌细胞(Rat aortic smooth muscle cells,RASMCs)增殖的影响,并探讨可能的机制。方法免疫荧光法检测RASMCs的α-SM-actin。MTT法检测PDGF-BB(0、2、4、8、16、32 ng/mL组)诱导的RASMCs增殖效应和Rb1(0、20、40、80μg/mL组)对其增殖的抑制作用。AnnexinV/PI法检测Rb1对细胞凋亡的影响。流式细胞术检测Rb1对细胞周期的影响。Western blot检测细胞PCNA、CDK2、CyclinE、P53和P21的表达。采用颈动脉球囊损伤术建造颈动脉内膜增生大鼠模型。将大鼠分成4组:正常组、损伤非治疗组、低剂量治疗组和高剂量治疗组。HE染色检测并计算颈动脉的内膜和管腔面积。免疫组化分析4组大鼠颈动脉内膜PCNA、P53和P21的表达。结果 RASMCs的纯度达(95±1.4)%。PDGF-BB诱导RASMCs增殖具有剂量时间依赖性。Rb1抑制PDGF-BB诱导的RASMCs增殖具有剂量依赖性并能够抑制其PCNA的蛋白表达。Rb1对PDGF-BB诱导增殖的RASMCs的凋亡无明显作用(P>0.05),但是能够减少S期细胞,阻滞细胞于G0/G1期。Rb1能够上调RASMCs的P53和P21的表达,而下调CDK2和CyclinE表达。球囊损伤组大鼠的颈动脉内膜比其他各组厚(P<0.05),管腔较其他组窄(P<0.05)。Rb1治疗后大鼠颈动脉内膜变薄,管腔变窄,PCNA的表达显著减少,而P21和P53表达增加。结论 Rb1能够在体外抑制PDGF-BB诱导的RASMCs增殖,体内同样能够抑制VSMCs的增殖。这与Rb1上调VSMCs的P53和P21表达明显相关。
Objective To investigate the effect of ginsenoside Rb1 on PDGF-BB induced proliferation of rat aortic smooth muscle cells（RASMCs）,and to explore the potential mechanism.Methods Immunofluorescence was used to detect α-SM-actin in RASMCs.MTT assay was applied to analyze the effects of proliferation induced by PDGF-BB （0,2,4,8,16,32 ng/mL） and the inhibitory effects of Rb1 （0,20,40,80 μg/mL） on the proliferation in RASMCs.The apoptosis induced by Rb1 was examined by AnnexinV/PI assay.The changes of cell cycle were detected by flow cytometry.Western blot was used to analyze the expression of PCNA,CDK2,CyclinE,P53 and P21.The rat carotid artery balloon-injured model was used as the practical model.The rats were divided into 4 groups：normal group,non-treated group,low dose-treated group and high dose-treated group.HE staining was used to detect and calculate the carotid artery intimal area and lumen area.Immunohistochemical assays were applied to analyze the expression of PCNA,P53 and P21 in rat carotid artery intima of four groups.Results The purity of RASMCs was （95 ± 1.4） ％.PDGF-BB induced proliferation of RASMCs was in a dose-time dependent manner.Inhibitory effects of Rb1 on PDGFBB induced proliferation of RASMCs was in a dose-dependent manner,and the expression of PCNA was inhibited by Rb1.The effect of Rb1 on apoptosis of PDGF-BB-induced RASMCs was not significant（P ＞ 0.05）.However,Rb1 could reduce the proportion of S phase cells and made cells arrested at G0/G1 phase.Rb1 could up-regulate the expression of P53 and P21 in RASMCs,and down-regulate the expression of CDK2 and CyclinE.Carotid artery intima of balloon-injured group was thicker（P ＜ 0.05）,as well as the lumen was narrower than other groups （P ＜ 0.05）.The intima was thinner and the lumen was narrower after treating with Rb1 in rats.The expression of PCNA significantly decreased,while the expression of P21 and P53 increased.Conclusion Rb1 could inhibit the PDGF-BB-induced proliferation of RASMCs in vitro.Similarly
出处
《实用药物与临床》
CAS
2014年第12期1516-1522,共7页
Practical Pharmacy and Clinical Remedies
基金
2012年浙江省医药卫生科技计划立项项目(2012KYA155)
