苜蓿素对LPS诱导HUVECs炎症反应的消除作用  

Roles of tricin in eliminating LPS-induced inflammatory response in HUVECs

作　　者：郭海海 范玉雪 郭炎峰 陈喜宏 蒋林峰 玉永雄[1] GUO Haihai;FAN Yuxue;GUO Yanfeng;CHEN Xihong;JIANG Linfeng;YU Yongxiong(College of Animal Science and Technology,Southwest University,Chongqing 404100,China;Chongqing Animal Husbandry Extension Station,Chongqing 404100)

机构地区：[1]西南大学动物科学技术学院,重庆404100 [2]重庆市畜牧推广总站,重庆404100

出　　处：《甘肃农业大学学报》2024年第3期1-9,共9页Journal of Gansu Agricultural University

基　　金：西南山地生态循环农业国家级培育基地项目,国家重点基础研究发展规划项项目(2014CB138701)。

摘　　要：【目的】研究苜蓿素对脂多糖(LPS)诱导的人脐静脉内皮细胞(HUVECs)炎症损伤的保护作用。【方法】以HUVECs为研究对象,用MTT法确定苜蓿素的处理浓度,用酶联免疫分析法确定LPS建模浓度以及检测HUVECs炎症相关因子(TNF-α、iNOS、COX-2、IL-1、IL-6)表达,用Western-blot法测定NF-κB和MAPK信号通路蛋白(β-Actin、P65、IKB、P-P65、P-IKB、P38、JNK、ERK1/2、P-P38、P-JNK、P-ERK1/2)表达。【结果】不同浓度的LPS均能促进TNF-α的表达(P<0.01),其中LPS浓度在2.5~7.5μg/mL时TNF-α的表达量最高,选用2.5μg/mL作为建模浓度,在该浓度下LPS促进炎症相关因子和炎症相关通路蛋白表达(P<0.01),而作为阳性对照辛伐他汀能抑制LPS诱导的炎症相关因子和炎症相关通路蛋白表达(P<0.01);苜蓿素浓度在3μg/mL之内对HUVECs活性无显著差异,因而选用3、2、1μg/mL作为苜蓿素高、中、低剂量组;苜蓿素高、中、低剂量组均能下调LPS诱导的炎症相关因子和炎症相关通路蛋白表达(P<0.01),而且苜蓿素对炎症相关因子的抑制效果弱于辛伐他汀,而对炎症相关通路蛋白的抑制效果强于辛伐他汀,各炎症相关因子表达随苜蓿素浓度升高而降低(P<0.01),炎症相关通路蛋白表达情况也有这个下降趋势。【结论】苜蓿素能够通过抑制NF-κB和MAPK信号通路消除LPS对HUVECs诱导的炎症损伤。【Objective】 This study aimed to evaluate the protective effect of tricin on inflammatory injury in human umbilical vein endothelial cells(HUVECs) induced by lipopolysaccharide(LPS).【Method】 HUVECs were used as the research subject.The MTT method was employed to determine the optimal concentration of tricin for treatment.ELISA was utilized to determine the appropriate LPS concentration for modeling and to assess the expression of inflammation-related factors(TNF-α,iNOS,COX-2,IL-1,IL-6)in HUVECs.Western blot analysis was performed to detect the protein expression of NF-κB and MAPK signal pathways(β-Actin,P65,IKB,P-P65,P-IKB,P38,JNK,ERK1/2,P-P38,P-JNK,P-ERK1/2).【Result】 Different concentrations of LPS were found to promote the expression of TNF-α(P<0.01),with the highest expression observed at LPS concentrations between 2.5 μg/mL and 7.5 μg/mL.Therefore,2.5 μg/mL was chosen as the modeling concentration.At this concentration,LPS induced the upregulation of inflammation-related factors and pathway proteins(P<0.01),while simvastatin,used as the positive control,inhibited the expression of these factors and proteins induced by LPS(P<0.01).HUVECs exhibited no significant difference in activity within tricin concentrations up to 3 μg/ml;therefore,tricin high(3 μg/mL),medium(2 μg/mL),and low(1 μg/mL) dosage groups were selected.Tricin in these dosage groups effectively downregulated the expression of inflammation-related factors and pathway proteins induced by LPS(P<0.01).Tricin showed a weaker inhibitory effect on inflammation-related factors compared to simvastatin,while its inhibitory effect on inflammation-related pathway proteins was stronger than that of simvastatin.The expression of inflammation-related factors decreased with increasing tricin concentration(P<0.01),and a similar downward trend was observed for inflammation-related pathway proteins.【Conclusion】 The findings indicated that tricin can attenuate LPS-induced inflammatory damage in HUVECs by inhibiting the NF-κB and MAPK signa

关 键 词：苜蓿素 人脐静脉内皮细胞(HUVECs) 炎症 脂多糖(LPS) 

分 类 号：S816[农业科学—饲料科学]