Extraction And Purification Process Optimization And Effect Of Urine Protein On Rats With Nephrotic Syndrome Of Total Flavonoids From Penthorum Chinense Pursh

Objective: To optimize extraction and purification process of total flavonoids from Penthorum chinense Pursh.(Stems,leaves and flowers),determined the content of six flavonoids in the stem and studied the effect on urinary protein in rats with adriamycin-induced nephrotic syndrome.It laid the foundation for the study of the effect of total flavonoids on other kidney diseases and the development of new drugs for the treatment of nephrotic syndrome and other nephropathy.Methods : The content of total flavonoids was determined by ultraviolet spectrophotometry.The pinocembrin was used as reference substance.The content of total flavonoids was used as index,the extraction process was optimized by orthogonal experiment based on single test.The polyamide was used to compare the adsorption of total flavonoids from Penthorum chinense Pursh..by polyamide mesh,times,pH,sample concentration,sample loading,water dosage,elution solvent pH,flow rate,ethanol concentration and ethanol dosage.Rate,desorption rate,determine the best purification and extraction process.Under the optimal extraction conditions and determined six flavonoids in the stems of Penthorum chinense pursh by HPLC.Sixty male Sprague-Dawley rats were randomly selected from the control group,and the other 52 were injected with doxorubicin(5 mg/kg twice)to make a model.After the 21-day model was successfully prepared,the remaining rats were randomly divided into four groups: group B(model group),group C(positive control group),group D(high-dose group of total flavonoids from Penthorum chinense Pursh.),E Group(low-dose group of total flavonoids from Penthorum chinense Pursh.),with 13 rats in each group.The adult clinical dose of total flavonoids of C.chinensis was 0.016-0.032 g/(kg·d).According to the dosage method of adult and rat,the high and low dose groups were given 0.2 g/(Kg·d),0.1 g/(kg·d),positive control group of prednisone acetate tablets,mixed with distilled water into a suspension,the dose is 6 mg/(kg·d),blank control group and model group irrigation.The same volume of distilled water in the stomach was continuously administered for 4 weeks,urine was collected every 2 weeks,and the volume of urine was recorded,and the change in the urine protein level of the rats 24 hours was observed.After 4 weeks of administration,the rats were sacrificed and serum was taken.The kidney tissues were fixed with 10% formalin and stained with HE and Masson respectively.The changes of renal tissue were observed under light microscope.Results: The best extraction process was as follows: the powder of Penthorum chinense pursh.was extracted for 1.5 h with 6 times of 55% ethanol by reflux for 3 times.The best purification process: the concentration is 5.0 mg.mL-1,the volume is 3.5 BV,the pH of the water solution is 3,the water amount is 25 BV,and the solution is eluted with a 10 BV volume of 75% ethanol with a pH of 8.The purified total flavonoids was 77.88%.Under this condition,the content of total flavonoids was the highest from Penthorum chinense Pursh.The content of quercetin and apigenin in the six flavonoids in the stem is relatively high.The total flavonoids from Penthorum chinense Pursh.can significantly reduce the urine protein in rats with nephrotic syndrome.