南五味子二氯甲烷部位化学成分结构鉴定及对HepG2和INS-1细胞的影响

Structural identification of chemical constituents in the dichloromethane part of Schisandra sphenanthera and their effects on HepG2 and INS-1 cells

目的探讨南五味子二氯甲烷部位(SDP)对HepG2细胞胰岛素抵抗和INS-1细胞高糖损伤的影响。方法采用1×10^(-6) mol·L^(-1)胰岛素建立HepG2细胞胰岛素抵抗和30 mmol·L^(-1)葡萄糖建立INS-1细胞高糖损伤模型,并使用不同浓度的南五味子二氯甲烷部位进行干预。检测HepG2细胞中三酰甘油(TG)、丙二醛(MDA)、超氧化物歧化酶(SOD)、细胞内活性氧(ROS)、腺苷三磷酸水平(ATP)、葡萄糖水平;检测INS-1细胞中MDA、SOD、过氧化物歧化酶(GSH)、ROS、基础葡萄糖(BIS)和高糖(GSIS)水平;并采用UPLC-MS/MS法分析SDP中的化学成分。结果经胰岛素诱导48 h的HepG2细胞,模型组TG、MDA、ROS葡萄糖升高(P<0.01),SOD、ATP降低,经SDP干预后,TG、MDA、ROS均有所回调,SOD、ATP显著升高;经高糖诱导48 h的INS-1细胞,模型组MDA、ROS升高(P<0.01),SOD、GSH-Px、BIS、GSIS降低(P<0.01),SDP干预后MDA、ROS回调(P<0.01),SOD、GSH-Px、BIS、GSIS上升(P<0.01)。从SDP样品中共成功鉴定出1824个化学成分,其中134个木脂素类、283个酚酸类、283个香豆素等其他类化合物,而木脂素类化合物相对含量最高。结论SDP能改善HepG2细胞胰岛素抵抗和INS-1细胞高糖损伤,分析了SDP化学成分,为防治T2DM提供实验依据。

Objective To determine the effect of dichloromethane part(SDP)of Schisandra sphenanthera on insulin resistance in HepG2 cells and hyperglycaemic injury in INS-1 cells.Methods HepG2 cell insulin resistance model was established with 1×10^(-6) mol·L^(-1) insulin and INS-1 cell hyperglycaemic injury model with 30 mmol·L^(-1) glucose.Different concentrations of SDP were used for the intervention.Triglyceride(TG),malondialdehyde(MDA),superoxide dismutase(SOD),intracellular reactive oxygen species(ROS),adenosine triphosphate level(ATP),and glucose expression levels were detected in HepG2 cells.Levels of MDA,SOD,peroxisomal dismutase(GSH),ROS,basal glucose(BIS),and hyperglycemic(GSIS)expressions were detected in INS-1 cells.Components in SDP were analyzed by UPLC-MS/MS.Results In HepG2 cells induced by insulin for 48 h,TG,MDA and ROS glucose were significantly increased(P<0.01)and SOD and ATP were decreased in the Model group.After the SDP intervention,and SOD and ATP were increased.In INS-1 cells induced by high glucose for 48 h,both MDA and ROS were increased(P<0.01)while SOD,GSH-Px,BIS and GSIS were decreased(P<0.01)in the model group.The MDA and ROS were decreased(P<0.01),and SOD,GSH-Px,BIS and GSIS were increased(P<0.01)after SDP intervention.Totally 1824 chemical constituents were successfully identified from the SDP samples,including 134 lignans,283 phenolic acids,283 coumarins and other compounds,with the highest relative content of lignans.Conclusion SDP improves insulin resistance in HepG2 cells and high glucose injury in INS-1 cells.The chemical compositions of SDP are analyzed to provide reference for the prevention and treatment of type 2 diabetes mellitus.