益母草醇提物调控Rho A/ROCK1信号通路对血管平滑肌细胞表型转化的影响OA
Effects of Alcohol Extracts of Leonuri Herba on Phenotypic Transformation of Vascular Smooth Muscle Cells via RhoA/ROCK1 Signaling Pathway
目的:研究益母草醇提物对血小板衍生因子(PDGF-BB)诱导的血管平滑肌细胞(VSMCs)增殖、迁移及表型转化的影响及其对Rho A/ROCK 信号通路的作用.方法:建立PDGF-BB 诱导的原代大鼠血管平滑肌细胞模型,CCK-8 法筛选益母草醇提物对 VSMCs 的安全剂量;Brdu 试验检测益母草醇提物对 VSMCs 增殖的影响;通过划痕试验和 Transwell 法检测 VSMCs 的迁移和侵袭能力;流式细胞术、PI 染色法检测益母草醇提物对 VSMCs周期分布的影响;通过免疫荧光染色观察益母草醇提物对 VSMCs 骨架重塑的影响;Western blot 法检测 VSMCs 收缩和合成表型转化及 Rho A/ROCK 信号通路相关蛋白的表达.结果:与空白对照组相比,模型对照组中 VSMCs细胞迁移能力增强(P<0.01),细胞阻滞在 S 期(P<0.01),VSMCs 收缩型标记物 Calponin、SM22α 的表达明显下调(P<0.05),VSMCs 合成型标记物 OPN、PCNA、Rho A、ROCK、JNK 的磷酸化表达明显上调(P<0.05 或P<0.01);与模型对照组相比,益母草醇提物能有效抑制 PDGF-BB 诱导的 VSMCs 细胞的增殖、迁移及表型转化(P<0.01);益母草醇提物25、50、100 μg/mL 组平滑肌细胞 Calponin、SM22α、α-SMA 蛋白表达明显上调(P<0.05或0.01),OPN、PCNA、Rho A、ROCK1、JNK 的磷酸化表达明显下调(P<0.05 或 0.01).结论:益母草醇提物通过调控 Rho A/ROCK1 信号通路,抑制JNK 磷酸化,进而抑制PDGF-BB 诱导的原代大鼠VSMCs 增殖、迁移及向合成表型的转化,改善血管重塑.
Objective:To investigate the effects of alcohol extracts of Leonuri Herba on the proliferation,migration,and phenotypic transformation of vascular smooth muscle cells(VSMCs)induced by platelet-derived growth factor-BB(PDGF-BB),and its mechanism based on the Ras homolog gene-Rho-related coiled-coil kinase(RhoA/ROCK)signa-ling pathway.Methods:A primary rat VSMC model induced by PDGF-BB was established.The CCK-8 assay was used to determine the safe dose of alcohol extracts of Leonuri Herba for VSMCs.The BrdU assay was used to assess the effect of alcohol extracts of Leonuri Herba on VSMC proliferation.Cell migration and invasion abilities were detected by scratch and Transwell assays.Flow cytometry combined with PI staining was employed to detect the effect of alcohol extracts of Leonuri Herba on the cycle distribution of VSMCs.Immunofluorescence staining was used to investigate the effect of al-cohol extracts of Leonuri Herba on cytoskeletal remodeling in VSMCs.Western blot was utilized to detect the phenotypic transformation of VSMCs from contractile to synthetic phenotype markers,as well as proteins related to the RhoA/ROCK signaling pathway.Results:Compared with the blank control group,VSMCs in the model control group exhibited en-hanced migration ability(P<0.01)and significant S-phase arrest(P<0.01).The expression of contractile phenotype markers calponin and SM22α was significantly downregulated(P<0.05),while the phosphorylation levels of synthetic phenotype markers OPN,PCNA,RHOA,ROCK,and JNK were significantly upregulated(P<0.05 or P<0.01).Com-pared with the model control group,alcohol extracts of Leonuri Herba significantly inhibited PDGF-BB-induced VSMC proliferation,migration,and phenotypic transformation(P<0.01).Treatment with alcohol extracts of Leonuri Herba at 25,50,and 100 μg/mL significantly upregulated the expression of calponin,SM22α,and α-SMA(P<0.05 or P<0.01)and downregulated the phosphorylation levels of OPN,PCNA,RHOA,ROCK1,and JNK(P<0.05 or P<0.01).Conclu-sion:Alcohol extracts of Leonuri Herba may improve vascular remodeling by inhibiting the proliferation,migration,and transformation of primary rat VSMCs to the synthetic phenotype induced by PDGF-BB,through regulation of the RhoA/ROCK1 signaling pathway and suppression of JNK phosphorylation.
宋柯妍;刘金杰;李梓宁;冉怡;张晗;李霄
天津中医药大学中医药研究院,天津 301617||组分中药国家重点实验室,天津 301617||方剂学教育部重点实验室,天津 301617天津中医药大学中医药研究院,天津 301617||组分中药国家重点实验室,天津 301617||方剂学教育部重点实验室,天津 301617翼城县人民医院,临汾 043500天津中医药大学中西医结合学院,天津 301617天津中医药大学中医药研究院,天津 301617||组分中药国家重点实验室,天津 301617||方剂学教育部重点实验室,天津 301617天津中医药大学中医药研究院,天津 301617||组分中药国家重点实验室,天津 301617||方剂学教育部重点实验室,天津 301617
益母草血管平滑肌细胞增殖迁移表型转化
Leonuri HerbaVascular smooth muscle cellsProliferationMigrationPhenotypic transformation
《中药药理与临床》 2026 (5)
91-99,9
天津市科技计划项目(编号:24ZYJDSS00320)现代中药新质生产力科技创新工程专项(编号:24ZXZKSY00010)组分中药国家重点实验室资助课题(编号:CBQM2022106).
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