松油烯-4-醇通过调控KLF4/NF-κB信号通路抑制高糖诱导的VSMCs增殖作用研究  

Terpinen-4-ol inhibits proliferation of VSMCs exposed to high glucose via regulating KLF4/NF-κB signaling pathway

作　　者：何丽 张林 张菊 蒋鸿 何永祥 冷栋国 巩迎新 杨钉 宋艳 熊传银 张彦燕[2] HE Li;ZHANG Lin;ZHANG Ju;JIANG Hong;HE Yong-xiang;LENG Dong-guo;GONG Ying-xin;YANG Ding;SONG Yan;XIONG Chuan-yin;ZHANG Yan-yan(Qianxi People′s Hospital,Qianxi 551500,China;Key Laboratory of Optimal Utilization of Natural Medicine Resources,Guizhou Medical University,Guiyang 550025,China)

机构地区：[1]黔西市人民医院,贵州黔西551500 [2]贵州医科大学天然药物资源优效利用重点实验室,贵州贵阳550025

出　　处：《中国中药杂志》2023年第9期2530-2537,共8页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(82060775,82260827);贵州省科技支撑计划项目([2020]4Y093);贵州省科学技术基金重点项目([2020]1Z069)。

摘　　要：研究松油烯-4-醇(terpinen-4-ol,T4O)对高糖(HG)诱导的血管平滑肌细胞(VSMCs)增殖的抑制作用,以及对Krüppel样因子4(Krüpple-like factor 4,KLF4)表达和NF-κB信号通路的影响。预先给予T4O预孵育2 h后,加入HG共孵育48 h诱导VSMCs损伤模型。MTT法检测VSMCs增殖;流式细胞仪检测细胞周期;划痕实验检测细胞迁移率;ELISA法检测VSMCs上清液中白细胞介素(IL)-6、肿瘤坏死因子-α(TNF-α)的含量;Western blot检测增殖细胞核抗原PCNA、周期相关蛋白Cyclin D1,以及KLF4和炎症相关蛋白NF-κB p-p65、IL-1β和IL-18的表达水平。siRNA-KLF4转染实现KLF4在VSMCs的沉默表达,观察T4O对HG诱导VSMCs细胞周期及上述蛋白表达水平的影响。结果显示,与HG组比较,不同剂量的T4O干预给药可明显抑制HG诱导的VSMCs增殖,增加G1期细胞百分比、降低S期细胞百分比,抑制细胞迁移能力,下调HG诱导的PCNA和Cyclin D1蛋白表达水平;同时,下调KLF4蛋白表达和NF-κB p-p65/NF-κB p65、IL-1β和IL-18的表达水平,降低炎症因子IL-6和TNF-α的分泌与释放;沉默KLF4可实现KLF4的低表达,与si-NC+HG组比较,siKLF4+HG组可降低S期细胞百分比,增加G1期细胞百分比,下调PCNA、Cyclin D1、KLF4蛋白表达水平及抑制NF-κB信号通路的激活,沉默KLF4与T4O共处理后可进一步促进上述指标的变化。上述结果表明,T4O对HG诱导的VSMCs增殖具有抑制作用,其作用可能与下调KLF4的水平,抑制NF-κB信号的激活有关。This study aimed to observe the effect of terpinen-4-ol(T4O)on the proliferation of vascular smooth muscle cells(VSMCs)exposed to high glucose(HG)and reveal the mechanism via the Krüppel-like factor 4(KLF4)/nuclear factor kappaB(NF-κB)signaling pathway.The VSMCs were first incubated with T4O for 2 h and then cultured with HG for 48 h to establish the model of inflammatory injury.The proliferation,cell cycle,and migration rate of VSMCs were examined by MTT method,flow cytometry,and wound healing assay,respectively.The content of inflammatory cytokines including interleukin(IL)-6 and tumor necrosis factor-alpha(TNF-α)in the supernatant of VSMCs was measured by enzyme-linked immunosorbent assay(ELISA).Western blot was employed to determine the protein levels of proliferating cell nuclear antigen(PCNA),Cyclin D1,KLF4,NF-κB p-p65/NF-κB p65,IL-1β,and IL-18.The KLF4 expression in VSMCs was silenced by the siRNA technology,and then the effects of T4O on the cell cycle and protein expression of the HG-induced VSMCs were observed.The results showed that different doses of T4O inhibited the HG-induced proliferation and migration of VSMCs,increased the percentage of cells in G1 phase,and decreased the percentage of cells in S phase,and down-regulated the protein levels of PCNA and Cyclin D1.In addition,T4O reduced the HG-induced secretion and release of the inflammatory cytokines IL-6 and TNF-αand down-regulated the expression of KLF4,NF-κB p-p65/NF-κB p65,IL-1β,and IL-18.Compared with si-NC+HG,siKLF4+HG increased the percentage of cells in G1 phase,decreased the percentage of cells in S phase,down-regulated the expression of PCNA,Cyclin D1,and KLF4,and inhibited the activation of NF-κB signaling pathway.Notably,the combination of silencing KLF4 with T4O treatment further promoted the changes in the above indicators.The results indicate that T4O may inhibit the HG-induced proliferation and migration of VSMCs by down-regulating the level of KLF4 and inhibiting the activation of NF-κB signaling pathway.

关 键 词：松油烯-4-醇 血管平滑肌细胞 高糖 KLF4 NF-ΚB 

分 类 号：R285[医药卫生—中药学]