Investigation On Compatibility Of Huang-Lian-Wan In Treatment Of Type 2 Diabetes Mellitus Based On Pharmacokinetics

Objectives:Huang-Lian-Wan(HLW), a traditional Chinese medicine prescription used in the treatment of diabetes mellitus in ancient China, was consisted of Rhizoma coptidis and Radix rehmanniae and had been reported to possess hypoglycemic effect. The purpose of this study was to investigate the compability of HLW in the treatment of type 2 diabetes mellitus based on pharmacokinetic characteristics. In this study, we developed HPLC fingerprints of alkaloids and iridoid glycosides contained in HLW, HPLC methods to respectively determine the contents of alkaloids and iridoid glycosides in different HLW extracts, and a LC-MS method to simultaneously determine the contents of 4 alkaloids(berberine, coptisine, palmatine and jatrorrhizine) in rats plasma. Type 2 diabetic model rats were also developed. The compability of HLW in the treatment of type 2 diabetes mellitus was then explored with the results of pharmacokinetics, which could provide experimental basis for the clinic use of HLW in the treatment of type 2 diabetes mellitus. Methods:(1)A HPLC method was used to develop the HPLC fingerprints of alkaloids contained in HLW extracts and the methodology was evaluated.(2)A HPLC method was used to develop the HPLC fingerprints of iridoid glycosides contained in HLW extracts and the methodology was evaluated.(3)A HPLC method was developed to simultaneously determine the contents of 4 alkaloids contained in HLW extracts and the methodology was evaluated.(4)A HPLC method was developed to determine the contents of catalpol contained in HLW extracts and the methodology was evaluated.(5)A LC-MS method was developed to simultaneously determine the contents of 4 alkaloids in rats plasma and the methodology was evaluated.(6)Type 2 diabetic rats were induced by high fat diet in combination with intraperitoneal injection of streptozotocin.(7)The type 2 diabetic rats were divided into HLW extracts group(QF) and Rhizoma coptidis extract group(HL), and then we gave HLW extract(1:1, W/W) and Rhizoma coptidis extract to the 2 groups respectively by intragastric administration. The LC-MS method was used to determine the contents of 4 alkaloids in blood samples and then we got the pharmacokinetic parameters after the plasma concentration datas was processed. Results:(1)HPLC fingerprints of alkaloids contained in HLW extracts was successfully developed.(2)HPLC fingerprints of iridoid glycosides contained in HLW extracts was successfully developed.(3)A HPLC method to simultaneously determine the contents of 4 alkaloids contained in HLW extracts was successfully developed. The contents of berberine, coptisine, jatrorrhizine and palmatine in HL extact were 231.93、39.99、27.95 and 47.32 mg/g, respectively;those in HLW extact(1:1, W/W) were 56.55、8.37、5.01 and 11.7 mg/g, respectively;those in HLW extact(1:4, W/W) were 33.03、4.95、3.13 and 6.40 mg/g, respectively.(4) A HPLC method to determine the contents of catalpol contained in HLW extracts was successfully developed. The content of catalpol in Radix rehmanniae extract was 2.2 mg/g; that in HLW extact(1:1, W/W) was 2.1 mg/g;that in HLW extact(1:4, W/W) was 1.4 mg/g。(5)A LC-MS method to simultaneously determine the contents of 4 alkaloids in rats plasma was successfully developed.(6)Type 2 diabetic rats were successfully induced with the success rate of 73%.(7)The AUC0-24 h of berberine and coptisine in QF were significantly higher than those in HL, while the AUC0-24 h of jatrorrhizine and palmatine in QF had no obvious difference with those in HL; the Cmax of berberine and coptisine in QF were significantly higher than those in HL; the Tmax of coptisine in QF was significantly shorter than that in HL; the Cmax of palmatine in QF were higher than that in HL; the MRT of 4 alkaloids in QF had no obvious difference with those in HL. Conclusions:(1)The HPLC fingerprints of alkaloids contained in HLW extracts showed that berberine, coptisine, palmatine and jatrorrhizine were all found in both Rhizoma coptidis extract and HLW extracts(1:1、1:4), which indicated that there were no losses of major alkaloids after the compability of Rhizoma coptidis and Radix rehmanniae. Moreover, berberine, coptisine, palmatine and jatrorrhizine could be served as the markers in the HPLC fingerprints of alkaloids contained in HLW extracts.(2)The HPLC fingerprints of iridoid glycosides contained in HLW extracts showed that catalpol was found in both Radix rehmanniae extract and HLW extracts(1:1、1:4), which indicated that there were no losses of major iridoid glycosides after the compability of Rhizoma coptidis and Radix rehmanniae. Moreover, catalpol could be served as the marker in the HPLC fingerprints of iridoid glycosides contained in HLW extracts.(3) The HPLC method to simultaneously determine the contents of 4 alkaloids contained in HLW extracts developed in this study was accutate and reliable.(4)The HPLC method to determine the contents of catalpol contained in HLW extracts developed in this study was accutate and reliable.(5) The LC-MS method to simultaneously determine the contents of 4 alkaloids in rats plasma developed in this study was accurate and reliable.(6) Compared to Rhizoma coptidis, the pharmacokinetic characteristics of alkaloids(especially the absorption of berberine and coptisine) in type 2 diabetic rats were significantly altered after the compability of Rhizoma coptidis and Radix rehmanniae. In pharmacology, the alkaloids such as berberine were the effective components in the treatment of type 2 diabetes mellitus, and the increasing of their absorption could enhance the therapeutic effect. Thus the research results of this study indicated that the compatibility of HLW did help to enhance its therapeutic effect in the treatment of type 2 diabetes mellitus.