Background:The purpose of the study was to investigatethe active ingredients and potential biochemicalmechanisms of Simiao Wan(SMW)in obesity-associated insulin resistance.Methods:An integrated network pharmacology me...Background:The purpose of the study was to investigatethe active ingredients and potential biochemicalmechanisms of Simiao Wan(SMW)in obesity-associated insulin resistance.Methods:An integrated network pharmacology method to screen the active compoundsand candidate targets,construct the protein-protein-interaction network,and ingredients-targets-pathways network was constructed for topological analysis to identify core targets and main ingredients.To find the possible signaling pathways,enrichment analysis was performed.Further,a model of insulin resistance in HL-7702 cells was established to verify the impact of SMW and the regulatory processes.Results:An overall of 63 active components and 151 candidate targets were obtained,in which flavonoids were the main ingredients.Enrichment analysis indicated that the PI3K-Akt signaling pathway was the potential pathway regulated by SMW in obesity-associated insulin resistance treatment.The result showed that SMW could significantly ameliorate insulin sensitivity,increase glucose synthesis and glucose utilization and reduce intracellular lipids accumulation in hepatocytes.Also,SMW inhibited diacylglycerols accumulation-induced PKCεactivity and decreased its translocation to the membrane.Conclusion:SMW ameliorated obesity-associated insulin resistance through PKCε/IRS-1/PI3K/Akt signaling axis in hepatocytes,providing a new strategy for metabolic disease treatment.展开更多
Simiao Wan(SMW)is a traditional Chinese formula,including Atractylodis Rhizoma,Achyranthis Bidentatae Radix,Phellodendri Chinensis Cortex and Coicis Semen at the ratio of 1:1:2:2.It can be used to the treatment of dia...Simiao Wan(SMW)is a traditional Chinese formula,including Atractylodis Rhizoma,Achyranthis Bidentatae Radix,Phellodendri Chinensis Cortex and Coicis Semen at the ratio of 1:1:2:2.It can be used to the treatment of diabetes.However,its bioactive compounds and underlying mechanism are unclear.This study was designed to screen the antilipolytic fraction from SMW and investigate its therapeutic mechanisms on hepatic insulin resistance.Different fractions of SMW were prepared by membrane separation combined with macroporous resin and their antilipolytic activity was screened in fasted mice.The effects of 60%ethanol elution(ESMW)on lipolysis were investigated in 3T3-L1 adipocytes stimulated by palmitic acid(PA)and high fat diet(HFD)-fed mice.In our study,ESMW was the bioactive fraction responsible for the antilipolytic activity of SMW and 13 compounds were characterized from ESMW by UHPLC-QTOF-MS/MS.ESMW suppressed protein kinase A(PKA)-hormone-sensitive lipase(HSL)related lipolysis and increased AMP-activated protein kinase(AMPK)phosphorylation in PA challenged 3T3-L1 adipocytes.AMPKa knockdown abolished the inhibitory effects of ESMW on IL-6 and HSL pSer-660,revealing that the antilipolytic and anti-inflammatory activities of ESMW are AMPK dependent.Furthermore,ESMW ameliorated insulin resistance and suppressed lipolysis in HFD-fed mice.It inhibited diacylglycerol accumulation in the liver and inhibited hepatic gluconeogenesis.Conditional medium collected from ESMW-treated 3T3-L1 cells ameliorated insulin action on hepatic gluconeogenesis in liver cells,demonstrating that the antilipolytic activity contributed to ESMW beneficial effects on hepatic glucose production.In conclusion,ESMW,as the antilipolytic fraction of SMW,inhibited PKA-HSL related lipolysis by activating AMPK,inhibiting diacylglycerol(DAG)accumulation in the liver and thereby improving insulin resistance and hepatic gluconeogenesis.展开更多
基金supported by the National Natural Science Foundation of China(81903871)Natural Science Foundation of Jiangsu Province(BK20190565)+1 种基金Fundamental Research Funds for the Central Universities(2632021ZD16)Zhenjiang City 2022 Science and Technology Innovation Fund(SH2022084).
文摘Background:The purpose of the study was to investigatethe active ingredients and potential biochemicalmechanisms of Simiao Wan(SMW)in obesity-associated insulin resistance.Methods:An integrated network pharmacology method to screen the active compoundsand candidate targets,construct the protein-protein-interaction network,and ingredients-targets-pathways network was constructed for topological analysis to identify core targets and main ingredients.To find the possible signaling pathways,enrichment analysis was performed.Further,a model of insulin resistance in HL-7702 cells was established to verify the impact of SMW and the regulatory processes.Results:An overall of 63 active components and 151 candidate targets were obtained,in which flavonoids were the main ingredients.Enrichment analysis indicated that the PI3K-Akt signaling pathway was the potential pathway regulated by SMW in obesity-associated insulin resistance treatment.The result showed that SMW could significantly ameliorate insulin sensitivity,increase glucose synthesis and glucose utilization and reduce intracellular lipids accumulation in hepatocytes.Also,SMW inhibited diacylglycerols accumulation-induced PKCεactivity and decreased its translocation to the membrane.Conclusion:SMW ameliorated obesity-associated insulin resistance through PKCε/IRS-1/PI3K/Akt signaling axis in hepatocytes,providing a new strategy for metabolic disease treatment.
基金supported by the National Key R&D Program of China(Nos.2018YFC1704500 and 2018YFC 1704503)the National Natural Science Foundation of China(Nos.82074080 and 81773877)the Natural Science Foundation of Jiangsu Province(No.BK20181328).
文摘Simiao Wan(SMW)is a traditional Chinese formula,including Atractylodis Rhizoma,Achyranthis Bidentatae Radix,Phellodendri Chinensis Cortex and Coicis Semen at the ratio of 1:1:2:2.It can be used to the treatment of diabetes.However,its bioactive compounds and underlying mechanism are unclear.This study was designed to screen the antilipolytic fraction from SMW and investigate its therapeutic mechanisms on hepatic insulin resistance.Different fractions of SMW were prepared by membrane separation combined with macroporous resin and their antilipolytic activity was screened in fasted mice.The effects of 60%ethanol elution(ESMW)on lipolysis were investigated in 3T3-L1 adipocytes stimulated by palmitic acid(PA)and high fat diet(HFD)-fed mice.In our study,ESMW was the bioactive fraction responsible for the antilipolytic activity of SMW and 13 compounds were characterized from ESMW by UHPLC-QTOF-MS/MS.ESMW suppressed protein kinase A(PKA)-hormone-sensitive lipase(HSL)related lipolysis and increased AMP-activated protein kinase(AMPK)phosphorylation in PA challenged 3T3-L1 adipocytes.AMPKa knockdown abolished the inhibitory effects of ESMW on IL-6 and HSL pSer-660,revealing that the antilipolytic and anti-inflammatory activities of ESMW are AMPK dependent.Furthermore,ESMW ameliorated insulin resistance and suppressed lipolysis in HFD-fed mice.It inhibited diacylglycerol accumulation in the liver and inhibited hepatic gluconeogenesis.Conditional medium collected from ESMW-treated 3T3-L1 cells ameliorated insulin action on hepatic gluconeogenesis in liver cells,demonstrating that the antilipolytic activity contributed to ESMW beneficial effects on hepatic glucose production.In conclusion,ESMW,as the antilipolytic fraction of SMW,inhibited PKA-HSL related lipolysis by activating AMPK,inhibiting diacylglycerol(DAG)accumulation in the liver and thereby improving insulin resistance and hepatic gluconeogenesis.
