首页|Absorbed Bioactive Compounds Replicate Guanxin Ⅱ-Induced Endothelium-Associated in/ex vivo Vasodilation
Absorbed Bioactive Compounds Replicate Guanxin Ⅱ-Induced Endothelium-Associated in/ex vivo Vasodilation
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Objective:To develop an interference-free and rapid method to elucidate Guanxin Ⅱ(GXⅡ)'s representative vasodilator absorbed bioactive compounds(ABCs)among enormous phytochemicals.Methods:The contents of ferulic acid,tanshinol,and hydroxysafflor yellow A(FTA)in GX Ⅱ/rat serum after the oral administration of GX Ⅱ(30 g/kg)were detected using ultra-performance liquid chromatography-mass spectrometry.Totally 18 rats were randomly assigned to the control group(0.9％normal saline),GX Ⅱ(30 g/kg)and FTA(5,28 and 77 mg/kg)by random number table method.Diastolic coronary flow velocity-time integral(VTI),i.e.,coronary flow or coronary flow-mediated dilation(CFMD),and endothelium-intact vascular tension of isolated aortic rings were measured.After 12 h of exposure to blank medium or 0.5 mmol/L H2O2,endothelial cells(ECs)were treated with post-dose GXⅡ of supernatant from deproteinized serum(PGSDS,300 µL PGSDS per 1 mL of culture medium)or FTA(237,1539,and 1510 mg/mL)for 10 min as control,H2O2,PGSDS and FTA groups.Nitric oxide(NO),vascular endothelial growth factor(VEGF),endothelin-1(ET-1),superoxide dismutase(SOD),malondialdehyde(MDA)and phosphorylated phosphoinositide 3 kinase(p-PI3K),phosphorylated protein kinase B(p-AKT),phosphorylated endothelial nitric oxide synthase(p-eNOS)were analyzed.PGSDS was developed as a GX Ⅱ proxy of ex vivo herbal crude extracts.Results:PGSDS effectively eliminates false responses caused by crude GX Ⅱ preparations.When doses equaled the contents in GX Ⅱ/its post-dose serum,FTA accounted for 98.17％of GX Ⅱ-added CFMD and 92.99％of PGSDS-reduced vascular tension.In ECs,FTA/PGSDS was found to have significant antioxidant(lower MDA and higher SOD,P＜0.01)and endothelial function-protective(lower VEGF,ET-1,P＜0.01)effects.The increases in aortic relaxation,endothelial NO levels and phosphorylated PI3K/Akt/eNOS protein induced by FTA/PGSDS were markedly abolished by NG-nitro-L-arginine methyl ester(L-NA,eNOS inhibitor)and wortmannin(PI3K/AKT inhibitor),respectively,indicating an endothelium-dependent vasodilation via the PI3K/AKT-eNOS pathway(P＜0.01).Conclusion:This study provides a strategy for rapidly and precisely elucidating GX Ⅱ's representative in/ex vivo cardioprotective absorbed bioactive compounds(ABCs)-FTA,suggesting its potential in advancing precision ethnomedicine.
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