摘要
目的研究苦丁茶皂苷D对氧化低密度脂蛋白(ox-LDL)诱导的RAW264.7细胞内脂质积累和脂多糖(LPS)诱导的RAW264.7细胞炎症的影响。方法ox-LDL诱导RAW264.7细胞形成泡沫细胞,油红染色观察细胞内脂滴聚集情况,酶法测定细胞内总胆固醇(TC)水平;RT-qPCR和Western blot分别检测清道夫受体CD36和SR-A1,ATP结合转运蛋白A1和G1(ABCA1和ABCG1)的基因和蛋白表达。LPS诱导RAW 264.7细胞炎症,ELISA和RT-qPCR检测白介素-6(IL-6)、白介素-1β(IL-1β)、单核细胞趋化蛋白-1(MCP-1)和肿瘤坏死因子-α(TNF-α)的表达;Western blot检测mTOR和p-mTOR的蛋白表达。结果细胞经苦丁茶皂苷D处理后,与模型组相比,高剂量降低了细胞内TC含量,下调了SR-A1的基因和蛋白表达;同时,苦丁茶皂苷D降低了LPS诱导的IL-6、IL-1β、MCP-1表达,下调了p-mTOR的蛋白表达。结论苦丁茶皂苷D可能通过下调SR-A1的基因和蛋白表达降低细胞内TC含量,可能通过mTOR通路发挥抗炎作用。
Aim To investigate the effects of Kudinoside D on lipid accumulation induced by oxidized low density lipoprotein(ox-LDL)and inflammation induced by lipopolysaccharide(LPS)in RAW264.7 cells.Methods Foam cells were established by incubating the RAW264.7 cells with ox-LDL.The concentration of lipid droplets in the cells was observed by oil red staining,and the level of total cholesterol(TC)in cells was measured by enzyme method.The gene and protein expressions of scavenger receptors CD36 and SR-A1,ATP binding cassette transporters A1 and G1(ABCA1 and ABCG1)were detected by RT-qPCR and Western blot,respectively.The expressions of interleukin-6(IL-6),interleukin-1β(IL-1β),monocyte chemoattractant protein-1(MCP-1)and tumor necrosis factor-α(TNF-α)were detected by ELISA and RT-qPCR.The protein expressions of mTOR and p-mTOR were detected by Western blot.Results Compared with model group,the high dose of Kudinoside D decreased the content of TC and down-regulated the gene and protein expression of SR-A induced by ox-LDL.Meanwhile Kudinoside D also decreased the levels of IL-6,IL-1βand MCP-1 and down-regulated the protein expression of p-mTOR induced by LPS.Conclusions Kudinoside D may reduce the intracellular TC content by down-regulating the gene and protein expression of SR-A1.Kudinoside D may play an anti-inflammatory role through mTOR pathway.
作者
白春艳
孟达
张雅琼
顾雯
张加余
周倩
饶高雄
车彦云
BAI Chun-yan;MENG Da;ZHANG Ya-qiong;GU Wen;ZHANG Jia-yu;ZHOU Qian;RAO Gao-xiong;CHE Yan-yun(Engineering Laboratory for National Healthcare Theories and Products of Yunnan Province,Yunnan University of Chinese Medicine,Kunming 650500,China;School of Chinese Materia Medica and Yunnan Key Laboratory of Southern Medicinal Resource,Yunnan University of Chinese Medicine,Kunming 650500,China;School of Pharmacy,Binzhou Medical University,Yantai,Shandong 264003,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2021年第8期1092-1098,共7页
Chinese Pharmacological Bulletin
基金
云南省科技厅-云南中医学院应用基础研究联合专项资金项目[No 2018FF001(-020)]
国家自然科学基金地区科学基金项目(No 81560643)。
作者简介
白春艳(1995-),女,硕士生,药物化学专业,E-mail:fw1191172410@163.com;通信作者:车彦云(1985-),女,博士,副教授,硕士生导师,从事中药药效物质基础及作用机制研究,E-mail:checpu@163.com。
