Background: RenShenJian decoction, a combination of Pueraria lobata (Willd.) Ohwi and Panax ginseng C. A. Mey, has been used in China since the Song Dynasty (960–1279 C.E.) to relieve symptoms of diabetes mellitus. However, the key compounds in RenShenJian that ameliorate insulin resistance remain unclear. This study identified the anti-diabetic compounds in RenShenJian by rescuing the decreased function of adenosine 5'-monophosphate-activated protein kinase (AMPK), sirtuin 3 (SIRT3), or glucose transporter isoform 4 (GLUT4). Methods: After streptozotocin-induced diabetic mice were treated with RenShenJian, fasting blood glucose levels and protein expression of SIRT3, p-AMPK, and AMPK were determined. Compounds from RenShenJian in plasma were monitored using multiple responses by liquid chromatography – mass spectrometry. Additionally, two insulin-resistant cell models were incubated with compounds identified in RenShenJian in the blood. Glucose uptake was determined using the fluorescent analog 2-(N-(7-nitrobenz-2-oxa-1,3-dia-xol-4-yl)amino)-2-deoxyglucose. Protein expression levels of p-AMPK, AMPK, SIRT3, and GLUT4 were detected by western blotting. Results:RenShenJian decreased FBG levels and upregulated SIRT3 expression and AMPK phosphorylation in diabetic mice. Thirteen RenShenJian extracts were identified in the blood, 11 of which increased the ratios of 2-(N-(7-nitrobenz-2-oxa-1,3-dia-xol-4-yl)amino)-2-deoxyglucose uptake in two insulin-resistant cell models. Nine extracts increased AMPK phosphorylation, nine increased SIRT3 expression, and six elevated GLUT4 expression in palmitate-induced HepG2 cells. Five extracts – puerarin, puerarin 6'-O-xyloside, genistein, ginsenoside Rb1, and ginsenoside Rd – simultaneously activated AMPK, SIRT3 and GLUT4. Conclusion: A series of compounds in RenShenJian that target AMPK, SIRT3, and/or GLUT4 was confirmed and indicate the chemical material basis of amelioration of insulin resistance by RenShenJian.

Zhen-Hua Lan;Lan-Fang Tan;Guo-En Wang;Meng-Qiu Tang;Ruo-Hong Wang;Shu-Mei Wang

Guangdong Pharmaceutical University,Guangzhou 510006,China;Key Laboratory of Digital Quality Evaluation of Chinese Materia Medica of State Administration of Traditional Chinese Medicine and Engineering&Technology Research Center for Chinese Materia Medica Quality of Guangdong Province,Guangdong Pharmaceutical University,Guangzhou 510006,China

传统医学研究（英文版）

[1]Zhen-Hua Lan;Lan-Fang Tan;Guo-En Wang;Meng-Qiu Tang;Ruo-Hong Wang;Shu-Mei Wang-.Active compounds in RenShenJian decoction ameliorate insulin resistance in vitro)[J].传统医学研究（英文版）,2022(04):8-16

RenShenJian,nitrobenz,xol,deoxyglucose,xyloside

Active,compounds,decoction,ameliorate,insulin,resistance,vitro,Background,combination,Pueraria,lobata,Willd,Ohwi,Panax,ginseng,Mey,has,been,used,China,since,Song,Dynasty,relieve,symptoms,diabetes,mellitus,However,key,that,remain,unclear,This,study,identified,anti,diabetic,by,rescuing,decreased,function,adenosine,monophosphate,activated,protein,kinase,AMPK,sirtuin,SIRT3,transporter,isoform,GLUT4,Methods,After,streptozotocin,induced,mice,were,treated,fasting,blood,levels,expression,determined,Compounds,from,plasma,monitored,using,multiple,responses,liquid,chromatography,mass,spectrometry,Additionally,two,resistant,models,incubated,Glucose,uptake,was,fluorescent,analog,oxa,amino,Protein,detected,western,blotting,Results,FBG,upregulated,phosphorylation,Thirteen,extracts,which,increased,ratios,Nine,nine,six,elevated,palmitate,HepG2,cells,Five,puerarin,genistein,ginsenoside,Rb1,Rd,simultaneously,Conclusion,series,target,confirmed,indicate,chemical,material,basis,amelioration

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