玉女煎对棕榈酸诱导的L细胞损伤的影响

Effect of Yunvjian on L cell injury induced by palmitic acid

目的:研究玉女煎对棕榈酸(PA)诱导的肠L细胞损伤的影响。方法:(1)将不同浓度(1.5、3和6 g/L)玉女煎溶液分别与二苯代苦味肼基自由基(DPPH·)溶液(100 mg/L)混合并避光反应30 min,采用全波长酶标仪检测吸光度的变化,评价玉女煎对DPPH·的清除作用。(2)将小鼠肠内分泌细胞系GLUTag分为空白对照组、PA(400μmol/L)组、PA+玉女煎(1.5 g/L)组、PA+玉女煎(3 g/L)组和PA+玉女煎(6 g/L)组,通过Hoechst 33342细胞核染色实验和MTT实验评价玉女煎对PA诱导的细胞凋亡的影响。(3)将GLUTag细胞分为空白对照组、PA组和PA+玉女煎(6 g/L)组,采用免疫荧光染色检测细胞内胰高血糖素样肽1(GLP-1)表达水平,通过2´,7´-二氯荧光素二乙酸酯(DCFH-DA)荧光探针检测细胞内活性氧(ROS)水平,通过Western blot实验检测超氧化物歧化酶1(SOD1)表达水平。结果:(1)玉女煎在体外可显著清除DPPH·(P<0.01)。(2)与PA组比较,玉女煎能以浓度依赖性方式抑制PA诱导的GLUTag细胞活力降低,在6 g/L时效果最明显(P<0.01)。(3)与PA组比较,6 g/L玉女煎能显著恢复GLUTag细胞的GLP-1(P<0.05)和SOD1表达水平(P<0.01),降低ROS水平(P<0.01)。结论:玉女煎可能通过降低细胞内氧化应激水平抑制PA诱导的GLUTag细胞损伤。

ATM:To explore the impact of Yunvjian on palmitic acid(PA)-induced injury of intestinal L cells.METHODS:(1)Various concentrations(1.5,3 and 6 g/L)of Yunvjian solution and 1,1-diphenyl-2-picrylhydrazyl(DPPH·)solution(100 mg/L)were incubated in darkness for 30 min.A full-wavelength microplate reader was employed to measure the absorbance,thereby assessing the scavenging effect of Yunvjian on DPPH·.(2)Mouse intestinal endocrine cell line GLUTag was divided into control group,PA group,PA+Yunvjian(1.5 g/L)group,PA+Yunvjian(3 g/L)group,and PA+Yunvjian(6 g/L)group.The influence of Yunvjian on PA-induced apoptosis was ascertained through Hoechst 33342 nuclear staining and MTT assay.(3)The GLUTag cells were divided into control group,PA group,and PA+Yunvjian(6 g/L)group.The expression of glucagon-like peptide-1(GLP-1)was quantified by immunofluorescence staining.The level of reactive oxygen species(ROS)was determined using 2',7'-dichlorofluorescein diacetate(DCFHDA)fluorescent probe,and superoxide dismutase 1(SOD1)protein level was analyzed by Western blot.RESULTS:(1)Yunvjian exhibited significant DPPH·scavenging activity in vitro(P<0.01).(2)Compared with PA group,Yunvjian markedly attenuated PA-induced decrease in GLUTag cell viability in a concentration-dependent manner,with the most pronounced effect observed at 6 g/L(P<0.01).(3)Compared with PA group,6 g/L Yunvjian significantly enhanced the expression of GLP-1(P<0.05)and SOD1(P<0.01),but reduced ROS level(P<0.01)in GLUTag cells.CONCLUSION:Yunvjian potentially mitigates PA-induced injury in GLUTag cells by diminishing intracellular oxidative stress.