槲皮素联合山柰酚抑制TNF信号通路保护大鼠背根神经节细胞免受高糖损伤  

Quercetin Combined with Kaempferol Protects Rat Dorsal Root Ganglion Cells from Hyperglycemic Damage by Inhibiting TNF Signaling Pathway

作　　者：卯明艳 余炫颉 张丽君[4] MAO Ming-yan(Liupanshui Vocational and Technical College,Liupanshui 553000,Guizhou,China)

机构地区：[1]六盘水职业技术学院,贵州六盘水553000 [2]湖北科技学院医学部药学院 [3]咸宁市中心医院介入科 [4]湖北科技学院医学部基础学院,湖北咸宁437100

出　　处：《牡丹江医学院学报》2024年第1期16-21,15,共7页Journal of Mudanjiang Medical University

基　　金：湖北省重点研发计划(大健康领域支持地方专项)(2022BCE051);国家自然科学基金面上项目(81571664)。

摘　　要：目的 探究槲皮素(Quercetin, Que)联合山柰酚(Kaempferol, Kae)对高糖(high glucose, HG)引起大鼠背根神经节(dorsal root ganglion, DRG)细胞氧化损伤及凋亡的保护作用。方法 通过体外培养DRG细胞,CCK-8检测细胞活力,确定糖尿病周围神经病变(diabetic peripheral neuropathy, DPN)体外模型建设条件、Que、Kae和二者组合的最佳给药浓度。DRG细胞分为5组(Con、HG、HG+Que、HG+Kae、HG+Que+Kae),ROS Assay Kit试剂盒和细胞凋亡荧光Hoechst 33342/PI双染试剂盒检测单药或组合药对HG诱导细胞产生ROS的影响和细胞过量凋亡的抑制作用;蛋白印迹法检测细胞死亡受体通路关键蛋白TNFR1、FADD、Caspase-8和Caspase-3的表达水平。结果 HG浓度达到85μM,作用时间24 h,细胞活力降至62%左右,低于对照组,差异有统计学意义(P<0.05);与HG组相比,20μM的Que、5μM的Kae和10μM的Que加5μM的Kae对HG环境中DRG细胞保护性最好,细胞活力最高,差异均有统计学意义(P<0.05);单药或组合药均能降低细胞内ROS水平和抑制DRG细胞过量凋亡,具有抗氧化和抗凋亡作用,组合药优于单药;单药组或组合药组TNFR1、FADD、Caspase-8和Caspase-3的表达水平均低于HG组,其中组合组蛋白水平表达最低,差异有统计学意义(P<0.05),虽然HG+Que组有降低趋势,但是无统计学意义(P>0.05)。结论 Que联合Kae通过抗氧化和抑制细胞死亡受体通路,降低DRG细胞凋亡量,延缓DPN的进展。Objective To investigate the protective effect of quercetin(Que)combined with kaempferol(Kae)on dorsal root ganglion(DRG)cells of rats in high glucose(HG)environment through anti-oxidation and inhibition of apoptosis.Methods DRG cells were cultured in vitro and cell viability was detected by CCK-8 to determine the conditions for the construction of diabetic peripheral neuropathy(DPN)model in vitro and the optimal concentration of Que,Kae,and their combination.DRG cells were divided into 5 groups(Con,HG,HG+Que,HG+Kae,HG+Que+Kae).The effect of single or combination drugs on HG-induced ROS production and the inhibition of excessive apoptosis by ROS assay kit and apoptosis fluorescence Hoechst 33342/PI double-staining kit.Results The cell viability decreased to about 62%with 85Mm HG and the treatment time was 24 h,which was lower than the control group(P<0.05).Compared with the HG group,20μM Que,5μM Kae,and 10μM Que plus 5μM Kae had the best protective effect on DRG cells in the HG environment,and the cell viability was the highest,and the differences were statistically significant(P<0.05).Both single and combination drugs can reduce intracellular ROS levels inhibit excessive apoptosis of DRG cells,and have antioxidant and anti-apoptotic effects.In addition,combination drugs are superior to single drugs.The expression levels of TNFR1,FADD,Caspase-8,and Caspase-3 in the monotherapy group or combination group were lower than those in the HG group,and the expression levels of combination histone were the lowest,the difference was statistically significant(P>0.05).Although there was a decreasing trend in HG+Que group,there was no statistical significance.Conclusion Que combined with Kae can reduce the amount of apoptosis of DRG-producing cells and delay the progression of DPN through anti-oxidation and inhibition of the cell death receptor pathway.

关 键 词：糖尿病病周围神经病变 槲皮素 山柰酚 氧化损伤 TNF信号通路 

分 类 号：R285.5[医药卫生—中药学]