The Preliminary Pharmacological Study On Fujian Zexie Terpenoids Components "Clearing Damp And Promoting Diuresis"

Druggability refers to the drug has a certain pharmacological activity,but also includes its pharmacokinetic properties and toxic side effects.Preliminary pharmacodynamic studies are the foundation,early safety evaluation is an important part and the pharmacokinetic study is an indicator in the process of its research.The“Structural Composition of Components theory”(SCCT)simplifies the study of traditional Chinese medicines(TCM)into representative “components” and effectively reflects the role of multi-components and multi-targets on the basis of the overall concept of Chinese medicine.Druggability evaluation is the core and key issue in the research of multi-fractions TCM.Multi-fractions TCM pay attention to the pre-treatment of Chinese herbal medicines/decoction pieces,which is also an important way to study the pre-formulation of formulations.The essence of the component structure of traditional Chinese medicine is the pharmacodynamics,metabolism and safety of multi-component interactions with specific structural proportions.Zexie has the functions of clearing damp and promoting diuresis and use for the treatment of water metabolism disorders.Studies have shown that some terpenes play a role both as an effect and toxic ingredients.Our research group had optimized the preparation of Zexie Terpeneoids Components(ZTCs)in the early stage.In this paper,the pharmacodynamics,pharmacokinetics and safety of ZTCs were evaluated in order to provide reference for the study of Chinese herbal medicines.The main research contents are as follows:1.The efficacy study of Fujian ZTCs "clearing damp and promoting diuresis"This section is based on the previous work basis,select authentic Fujian Zexie and use AB-8 macroporous resin column for separation and preparation of terpenoids from Zexie,then accomplish content determination and analysis validation by HPLC method.The water load model was established in vivo by intragastric administration of distilled water in rats and the changes of urine volume,urinary electrolytes content(Na+,K+ and Cl-)and renal medulla water AQP2 channel protein expression level were investigated.Human renal tubular epithelial cells HK-2 model was used to investigate the expression of AQP2 in vitro.Western blot,Real-time PCR and immunohistochemistry methods were all used to test the expression of AQP2.The results showed that ZTCs could increase urine output of rats,promote the discharge of electrolytes in urine,and significantly down-regulate the expression of the target protein AQP2,like furosemide.The results showed that the ZTCs had a certain diuretic effect,and its diuretic effect is related to the down regulation of AQP2 protein in the kidney.2.Pharmacokinetic study of Fujian ZTCsAccording to the pre-foundation and related literatures,three major leaching terpenes alisol B,23-acetyl alisol B,24-acetyl alisol A were confirmed.The pharmacokinetic characteristics of the rats were examined by time-blood concentration method and UPLC-MRM-MS/MS was used to detect the drug concentration of plasma in rats at different time points after oral administration in rats.The pharmacokinetic software was used to calculate the pharmacokinetic parameters of the three components.According to the area under the curve AUC integrated component pharmacokinetic model,the pharmacokinetic parameters of the components were further fitted,the absorbed terpenoid components were better in vivo absorption,and the half-life was shorter.It provides ideas for the study of the pharmacokinetics of traditional Chinese medicines.3.Early safety evaluation of Fujian ZTCsThis section studies the long-term nephrotoxicity of terpenoids in female rats administered about 12 weeks,determine the terpenoid-induced nephrotoxicity.That is,administration of SD rat from the equivalent low dose of Liu Wei Di Huang Wan to 40 times the clinical dose of ZTCs.Urine p H,urobilinogen,bilirubin,ketone body,occult blood,urine protein,nitrite,p H,glucose and other indicators were determined by Urine test strip;Urinary biochemical indicators like Urine NAG,UP and serum biochemical indexes BUN,Scr,β2-MG were detected by automatic biochemical analyzer.In addition,the expressions of kidney-related protein Kim-1,Clusterin,Cystatin-c and NGAL were detected by RT-PCR and immunohistochemistry methods.HE staining was to investigate the long-term administration of rat kidney and assess its safety.The results showed that long-term and high-dose(about 12 weeks,40-fold dose)administration of ZTCs did not damage the rat kidney.In summary,under the guidance of the “SCCT”,our study choose authentic Fujian Zexie to conduct a preliminary exploration of its druggability.From the preparation of ZTCs to the evaluation of the efficacy in vivo and in vitro,it was concluded that ZTCs have a certain "clearing damp and promoting diuresis" effect.Then the pharmacokinetics and safety of the rats were investigated in vivo.The evaluation of partial druggability of ZTCs fractions was completed.It is of great significance in the study of multi-fractions TCM.