OBJECTIVE To investigate the compatibility of the prescription Rhodiolarosea,Cordycepsmilitaris,and Rhubarb root.METHODS The DIO mice accompanying with insulin resistance,hypercholesterolemia and obesity were induced ...OBJECTIVE To investigate the compatibility of the prescription Rhodiolarosea,Cordycepsmilitaris,and Rhubarb root.METHODS The DIO mice accompanying with insulin resistance,hypercholesterolemia and obesity were induced by high fat diet(HFD)in C57BL/6mice.For evaluating the insulin sensitivity in vivo,the values of serum insulin,HOMA-IR,and the glucose infuse rate(GIR)in hyperinsulinemiceuglycemic clamp test were determined;the time-dependent blood glucose changes were measured after insulin loading in insulin tolerance test(ITT),or after glucose loading in glucose tolerance test(GTT),respectively.Recombinant human protein tyrosine phosphatase 1B(PTP1B)was overexpressed by hGST-PTP1BBL21 E.coli and purified by GST affinity chromatography.The activities of PTP1 Band glutamine:fructose 6-phosphate amidotransferase(GFAT)were measured by the pNPP and GDH method,respectively.The activity ofα-glucosidase was determined with the substrates of p-nitrophenyl-alpha-D-glucopyranoside(pNPG).The IC50 value was calculated using the software GraphPad Prism 5.RESULTS 1st,the standard for the raw materials and for the extraction process of Rhodiolarosea,Cordycepsmilitaris,and Rhubarbroot are formulated according to the inhibition on PTP1 B,α-glucosidase,and GFAT in vitro,and the improvement on impaired glucose tolerance(IGT),hypercholesterolemia,and obesity in vivo,respectively.2nd,the composition was determined by the synergistic effects of Rhodiolarosea,Cordycepsmilitaris,and Rhubarb root on insulin resistance in DIO mice.3rd,in accordance with the IC50 values from the L9(34)and L8(27)orthogonal experiments targeting on PTP1 B and α-glucosidase,respectively,the best rate of Rhodiolarosea∶Cordycepsmilitaris∶Rhubarbroot was selected.4th,the effective dose for the treatment of metabolic syndrome was assessed based on the effects on IGT,hypercholesterolemia,obesity,insulin resistance,respectively,in DIO mice.CONCLUSION The prescription compatibility of Rhodiolarosea,Cordycepsmilitaris,and Rhubarbroot with the rate of 20∶1∶1was formed to treat the metabolic syndrome,such as IGT,hypercholesterolemia,and obesity,with the effective dose of 200 mg·kg-1 by the major mechanism of the improvement on insulin sensitivity.Monarch drug,Rhodiola,can clear away the lung-heat,tonify Qi,resolve stasis and nourish the heart.Adjuvant drug,Cordycepsmilitaris,can tonify the lung Qi and the kidney essence,strong waist and knee,compatibility with Rhodiola to enhance the function of invigorating lung and kidney.Assistant drug,Rhubarb,can clear heat,detoxify,remove blood stasis.These three herbal are compatible to show the effects of tonifying Qi,nourishing essence,cleaning heat,reducing phlegm and resolving masses for the treatment of metabolic syndrome.展开更多
基金The project supported by National Major Special Project on New Drug Innovation of China(2009ZX09103-4322012ZX09103-101-063+1 种基金2012ZX09301002-004)We also appreciate the support of Novo Nordisk Union Diabetes Research Talent Fund
文摘OBJECTIVE To investigate the compatibility of the prescription Rhodiolarosea,Cordycepsmilitaris,and Rhubarb root.METHODS The DIO mice accompanying with insulin resistance,hypercholesterolemia and obesity were induced by high fat diet(HFD)in C57BL/6mice.For evaluating the insulin sensitivity in vivo,the values of serum insulin,HOMA-IR,and the glucose infuse rate(GIR)in hyperinsulinemiceuglycemic clamp test were determined;the time-dependent blood glucose changes were measured after insulin loading in insulin tolerance test(ITT),or after glucose loading in glucose tolerance test(GTT),respectively.Recombinant human protein tyrosine phosphatase 1B(PTP1B)was overexpressed by hGST-PTP1BBL21 E.coli and purified by GST affinity chromatography.The activities of PTP1 Band glutamine:fructose 6-phosphate amidotransferase(GFAT)were measured by the pNPP and GDH method,respectively.The activity ofα-glucosidase was determined with the substrates of p-nitrophenyl-alpha-D-glucopyranoside(pNPG).The IC50 value was calculated using the software GraphPad Prism 5.RESULTS 1st,the standard for the raw materials and for the extraction process of Rhodiolarosea,Cordycepsmilitaris,and Rhubarbroot are formulated according to the inhibition on PTP1 B,α-glucosidase,and GFAT in vitro,and the improvement on impaired glucose tolerance(IGT),hypercholesterolemia,and obesity in vivo,respectively.2nd,the composition was determined by the synergistic effects of Rhodiolarosea,Cordycepsmilitaris,and Rhubarb root on insulin resistance in DIO mice.3rd,in accordance with the IC50 values from the L9(34)and L8(27)orthogonal experiments targeting on PTP1 B and α-glucosidase,respectively,the best rate of Rhodiolarosea∶Cordycepsmilitaris∶Rhubarbroot was selected.4th,the effective dose for the treatment of metabolic syndrome was assessed based on the effects on IGT,hypercholesterolemia,obesity,insulin resistance,respectively,in DIO mice.CONCLUSION The prescription compatibility of Rhodiolarosea,Cordycepsmilitaris,and Rhubarbroot with the rate of 20∶1∶1was formed to treat the metabolic syndrome,such as IGT,hypercholesterolemia,and obesity,with the effective dose of 200 mg·kg-1 by the major mechanism of the improvement on insulin sensitivity.Monarch drug,Rhodiola,can clear away the lung-heat,tonify Qi,resolve stasis and nourish the heart.Adjuvant drug,Cordycepsmilitaris,can tonify the lung Qi and the kidney essence,strong waist and knee,compatibility with Rhodiola to enhance the function of invigorating lung and kidney.Assistant drug,Rhubarb,can clear heat,detoxify,remove blood stasis.These three herbal are compatible to show the effects of tonifying Qi,nourishing essence,cleaning heat,reducing phlegm and resolving masses for the treatment of metabolic syndrome.
