红茂草异紫堇碱对LPS诱导小鼠RAW 264.7细胞炎症因子的影响  

Effects of Isocorydine Alkaloid in Dicranostigma leptopodum (Maxim.) Fedde(DLF)on Inflammatory Factors in RAW264. 7 Cells Stimulated by LPS

作　　者：王霞 王廷璞 袁毅君 高义霞 王鹏 赵强 赵仁生 WANG Xia;WANG Ting-pu;YUAN Yi-jun(College of Bioengineering and Technology,Tianshui Normal University,Tianshui,Gansu 741001)

机构地区：[1]天水师范学院生物工程与技术学院

出　　处：《安徽农业科学》2020年第4期178-180,183,共4页Journal of Anhui Agricultural Sciences

基　　金：国家自然科学基金委员会地区科学基金项目(31860707);甘肃省教育厅科技成果转化项目(2017D-24)

摘　　要：[目的]研究红茂草异紫堇碱对细菌脂多糖(lipopolysaccharide,LPS)诱导RAW 264.7细胞释放炎性因子的影响。[方法]用CCK-8试剂盒方法检测不同剂量红茂草异紫堇碱(1、5、15、25、50、100、200、300和400μg/mL)对RAW 264.7细胞的毒性作用;用硝酸酶还原法测定不同浓度红茂草异紫堇碱(50、100、200和400μg/mL)条件下细胞培养液中NO的浓度;用ELISA法测定添加不同浓度红茂草异紫堇碱(100、200和400μg/mL)干预4 h,再加入LPS(0.1μg/mL)孵育20 h后细胞培养液中IL-1β、IL-6、IL-10、PEG-2和TNF-α的含量,通过上述方法测定的结果来评价红茂草异紫堇碱的抗炎作用。[结果]红茂草异紫堇碱浓度在15、25μg/mL时,可显著促进RAW 264.7细胞增殖(P≤0.05);并且在红茂草异紫堇碱存在条件下,促炎因子IL-1β、IL-6、PEG-2和TNF-α释放量都有不程度的降低,而抗炎因子IL-10和NO的释放量有不同程度的升高。[结论]红茂草异紫堇碱能够抑制促炎因子IL-1β、IL-6、PEG-2和TNF-α的释放;另一方面能够促进抗炎因子IL-10和NO的释放来抑制LPS诱导的RAW 264.7细胞炎症反应。[Objective]To study the effects of isocorydine alkaloid in Dicranostigma leptopodum(Maxim.)Fedde(DLF)on secretion of inflammatory factors in RAW 264.7 cell stimulated by lipopolysaccharide,LPS.[Method]Toxic effect on RAW 264.7 cell with different doses of isocorydine alkaloid in DLF(1,5,15,25,50,100,200,300 and 400μg/mL)were detected by CCK 8 kit method.The concentration of NO in cell medium added different doses of isocorydine alkaloid in DLF(50,100,200 and 400μg/mL)were detected by the enzyme nitrate reduction method.After pretreated with various concentration of isocorydine alkaloid in DLF(100,200 and 400μg/mL)4 h.The cell were exposed to 0.1μg/mL LPS for 20 h.The activity of interleukin 6(IL-6)、10(IL-10)and 1β(IL-1β),PEG-2 and tumor necrosis factorα(TNF-α)as well as NO by ELISA and an Nitrate acid reductase.The anti-inflammatory effect was evaluated by above method.[Result]Concentration of isocorydine alkaloid in DLF were 15 and 25μg/mL,which could promote RAW 264.7 cells proliferation significantly(P≤0.05).Isocorydine alkaloid in DLF increased the secretion of IL-10 and NO as well as reduced IL-1β,IL-6,PEG-2 and TNF-αin LPS-induced RAW 264.7 cells.[Conclusion]The anti-inflammatory effectory of isocorydine alkaloid in LDF may be mediated in part of by promoting IL-10 and NO and inhibiting the production of IL-1β,IL-6,PEG-2 and TNF-α.

关 键 词：红茂草异紫堇碱 RAW 264.7细胞 LPS 细胞炎症因子 

分 类 号：R285.5[医药卫生—中药学]