Objective:To explore the effects of 6-gingerol,asarinin,and deoxyschizandrindthe main components of Zingiber officinale(Willd.)Rosc.(Gan Jiang),Asarum heterotropoides f.var.mandshuricum(Maxim.)(Xi Xin),and Schisandra ...Objective:To explore the effects of 6-gingerol,asarinin,and deoxyschizandrindthe main components of Zingiber officinale(Willd.)Rosc.(Gan Jiang),Asarum heterotropoides f.var.mandshuricum(Maxim.)(Xi Xin),and Schisandra chinensis(Turcz.)Baill.(Wu Wei Zi),respectivelydon an interleukin(IL)-13einduced BEAS-2B cell model in vitro.Methods:The BEAS-2B cell model was established using 25 ng/mL IL-13 combined with 1%fetal bovine serum(FBS)in vitro.Mitoquinone mesylate(Mito-Q)treatment was used as a positive control group,and different concentrations of 6-gingerol,asarinin,and deoxyschizandrin were used to treat the models.The level of reactive oxygen species(ROS)production was detected by flow cytometry.The expression levels of LC3B,Beclin1,adenosine 50-monophosphate(AMP)eactivated protein kinase(AMPK),phosphory-lated-AMPeactivated protein kinase(P-AMPK),dynamin-related protein 1(DRP1),and mitochondrial fusion protein 2(MFN2)were detected by Western blot.Mitochondrial membrane potential(MMP)assay kit with JC-1 was utilized to detect the level of MMP.Results:The BEAS-2B cells exposed to 25 ng/mL IL-13 with 1%FBS showed an increased ROS level and a decreased MMP.6-Gingerol,asarinin,and deoxyschizandrin were able to downregulate ROS level and upregulate the MMP in the BEAS-2B model.Asarinin and deoxyschizandrin reduced the expression of autophagy protein LC3B,while deoxyschizandrin significantly increased the expression of DRP1 in the BEAS-2B model.Conclusion:6-Gingerol,asarinin,and deoxyschizandrin can reduce ROS generation and increase MMP,but have different regulatory effects on the expression of autophagy protein and mitochondrial mitotic protein.The three components have both synergistic and complementary effects in classic medicine compatibility.This study may provide an innovative strategy to reduce the lung inflammation related to IL-13.展开更多
基金This study was supported by the National Natural Science Foundation of China(81403313)the Vertical Development Fund of Beijing University of Chinese Medicine(2019-ZXFZJJ-062).
文摘Objective:To explore the effects of 6-gingerol,asarinin,and deoxyschizandrindthe main components of Zingiber officinale(Willd.)Rosc.(Gan Jiang),Asarum heterotropoides f.var.mandshuricum(Maxim.)(Xi Xin),and Schisandra chinensis(Turcz.)Baill.(Wu Wei Zi),respectivelydon an interleukin(IL)-13einduced BEAS-2B cell model in vitro.Methods:The BEAS-2B cell model was established using 25 ng/mL IL-13 combined with 1%fetal bovine serum(FBS)in vitro.Mitoquinone mesylate(Mito-Q)treatment was used as a positive control group,and different concentrations of 6-gingerol,asarinin,and deoxyschizandrin were used to treat the models.The level of reactive oxygen species(ROS)production was detected by flow cytometry.The expression levels of LC3B,Beclin1,adenosine 50-monophosphate(AMP)eactivated protein kinase(AMPK),phosphory-lated-AMPeactivated protein kinase(P-AMPK),dynamin-related protein 1(DRP1),and mitochondrial fusion protein 2(MFN2)were detected by Western blot.Mitochondrial membrane potential(MMP)assay kit with JC-1 was utilized to detect the level of MMP.Results:The BEAS-2B cells exposed to 25 ng/mL IL-13 with 1%FBS showed an increased ROS level and a decreased MMP.6-Gingerol,asarinin,and deoxyschizandrin were able to downregulate ROS level and upregulate the MMP in the BEAS-2B model.Asarinin and deoxyschizandrin reduced the expression of autophagy protein LC3B,while deoxyschizandrin significantly increased the expression of DRP1 in the BEAS-2B model.Conclusion:6-Gingerol,asarinin,and deoxyschizandrin can reduce ROS generation and increase MMP,but have different regulatory effects on the expression of autophagy protein and mitochondrial mitotic protein.The three components have both synergistic and complementary effects in classic medicine compatibility.This study may provide an innovative strategy to reduce the lung inflammation related to IL-13.
文摘目的基于蛋白组学探讨细辛水提物诱发肝损伤的作用机制。方法30只大鼠随机分为对照组和细辛高、低剂量(12、6 g/kg)组,每组10只,每日ig给药1次,连续28 d。末次给药后采取血清样本和肝脏样本,检测血清丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天冬氨酸氨基转移酶(aspartate aminotransferase,AST)活性;苏木素-伊红(hematoxylineosin,HE)染色法观察肝脏病理组织形态;串联质谱标签定量蛋白质组学法检测肝组织差异表达蛋白;免疫组化法测定大鼠肝脏组织细胞色素P4501A1(cytochrome P4501A1,CYP1A1)、金属硫蛋白-1(metallothionein-1,MT-1)、肌肉特异性烯醇化酶3(recombinant enolase 3,ENO3)、抗原转运蛋白(transporter associated with antigen processing 1,TAP1)蛋白表达。CCK-8法考察细辛水提物对小鼠肝实质AML-12细胞活力的影响,并采用qRT-PCR与Western blotting检测AML-12细胞中CYP1A1、MT-1、ENO3、TAP1蛋白及m RNA表达。结果与对照组比较,细辛组大鼠血清AST、ALT活性均显著升高(P<0.01);肝组织出现明显炎性改变;肝组织中存在179个差异蛋白,主要富集于次级代谢产物的生物合成途径、视黄醇代谢途径、吞噬体等;肝组织CYP1A1、MT-1蛋白表达显著增加(P<0.01),ENO3、TAP1蛋白表达显著减少(P<0.01)。体外实验结果显示,细辛组AML-12细胞CYP1A1、MT-1 m RNA与蛋白表达显著增加(P<0.05、0.01),ENO3、TAP1 m RNA与蛋白表达水平显著降低(P<0.05、0.01)。结论细辛水提物可诱发大鼠肝损伤,引起肝功能异常和肝组织病理改变,肝损伤机制与上调CYP1A1、MT-1 m RNA及蛋白表达及下调ENO3、TAP1 m RNA及蛋白表达有关。
