白皮杉醇通过抑制HMGB1/TLR4/NF-κB信号通路减轻多柔比星诱导的H9c2心肌细胞损伤  

Through Inhibition of HMGB1/TLR4/NF-κB Signaling Pathway,Piceatannol Attenuates the Damage Caused to H9c2 Cardiomyocytes by Doxorubicin

作　　者：吴其芮 杨秋实 汪雅玲 贺武斌 王一 黄建华 WU Qirui;YANG Qiushi;WANG Yaling;HE Wubin;WANG Yi;HUANG Jianhua(Key Laboratory of Surgery,The First Affiliated Hospital of Jinzhou Medical University,Jinzhou 121000,China)

机构地区：[1]锦州医科大学附属第一医院外科学重点实验室,辽宁锦州121000

出　　处：《中国现代应用药学》2025年第5期735-742,共8页Chinese Journal of Modern Applied Pharmacy

基　　金：辽宁省兴辽英才科技创新领军人才项目(XLYC1802113)。

摘　　要：目的探讨白皮杉醇对多柔比星(doxorubicin,DOX)引起的H9c2心肌细胞损伤的保护作用及机制。方法分组培养H9c2大鼠心肌细胞后,加入终浓度为1μmol·L^(-1)的DOX和梯度浓度的白皮杉醇处理24h,应用MTT法检测细胞存活率,并筛选出最佳联合给药浓度。采用Hoechst 33342染色及流式细胞术检测H9c2心肌细胞的凋亡情况。采用DCFH-DA染色荧光显微镜照相检测ROS水平。Western blotting检测细胞凋亡蛋白Bcl-2、Bax、Cleaved caspase-3蛋白表达水平,并使用TLR4激动剂脂多糖进一步验证白皮杉醇与HMGB1/TLR4/NF-κB信号通路的关系。结果MTT表明DOX可以导致H9c2心肌细胞活性明显降低,而白皮杉醇可抑制此过程。与正常对照组比较,AnnexinV-FITC/PI染色和Heochest 33342染色表明白皮杉醇可以降低DOX诱导的H9c2心肌细胞的凋亡率。ROS染色结果显示,白皮杉醇对DOX干预后细胞ROS水平的升高有明显降低作用,Westernblotting结果表明白皮杉醇可以一定程度上促进DOX干预后抗凋亡蛋白Bcl-2表达,抑制促凋亡蛋白(Bax、Cleaved caspase-3)的表达,进一步研究发现白皮杉醇能够抑制DOX引起的H9c2心肌细胞内TLR4蛋白水平的升高,而TLR4激动剂脂多糖逆转了这一作用。结论白皮杉醇可能通过抑制HMGB1/TLR4/NF-κB通路改善DOX诱导的H9c2心肌细胞损伤。OBJECTIVE To examine the potential protective effect and underlying mechanism of piceatannol against doxorubicin(DOX)-induced injury in H9c2 cardiomyocytes.METHODS H9c2 cardiomyocytes were cultured in groups and subsequently treated with doxorubicin at a final concentration of 1μmol·L^(-1) and piceatannol at various levels.The cells were incubated for 24 h,and the MTT method was employed to assess the cell survival rate and determine the most effective co-administration concentration.Hoechst 33342 staining and flow cytometry were utilized to detect apoptosis in H9c2 cardiomyocytes.ROS levels were assessed using DCFH-DA staining and fluorescence microscopy.Western blotting was employed to determine the expression levels of apoptotic proteins Bcl-2,Bax,and Cleaved caspase-3.The relationship between piceatannol and the HMGB1/TLR4/NF-κB signaling pathway was further confirmed by using the TLR4 agonist lipopolysaccharide.RESULTS DOX treatment resulted in a significant decrease in H9c2 cardiomyocytes activity,which was inhibited by piceatannol.Annexin V-FITC/PI staining and Hoechst 33342 staining demonstrated that piceatannol reduced the apoptotic rate of DOX-induced H9c2 cardiomyocytes.ROS staining results showed that piceatannol significantly decreased cellular ROS levels.Western blotting analysis revealed that piceatannol increased the expression level of the anti-apoptotic protein Bcl-2 and decreased the expression levels of the pro-apoptotic proteins(Bax,Cleaved caspase-3).Furthermore,it was found that piceatannol inhibited the increase of TLR4 protein level in H9c2 cardiomyocytes induced by DOX,and this effect was reversed by the TLR4 agonist lipopolysaccharide.CONCLUSION Piceatannol may ameliorate doxorubicin-induced H9c2 cardiomyocytes injury by inhibiting the HMGB1/TLR4/NF-κB pathway.

关 键 词：白皮杉醇 多柔比星 H9C2心肌细胞 高迁移率族蛋白B1/Toll样受体4/核因子-κB通路 

分 类 号：R285.5[医药卫生—中药学]