Effects Of Cistanche Enzymatic Extract On HKC Cells And Chronic Renal Failure Rats

Objective: Preliminary results of enzyme-assisted extraction of Cistanche showed that this method can significantly improve the benzene content of cistanche glycosides and polysaccharides. Therefore, impact studies on the basis of cstanche extract from HKC cells and chronic renal failure rats can provide theoretical basis for cistanche hydrolysis assisted extraction.Methods: The experiment about enzyme-assisted extraction of Cistanche, through single factor and process parameter optimization and other experiments to determine the optimal enzyme-assisted extraction of Cistanche process parameters. The experiment about cells,cultured normal human embryonic kidney epithelial cell line HKC cells, with the cell toxicity tests to investigate HKC cells after doxorubicin(Doxourbicin) injury, and using cistanche enzyme extract of different concentrations of n-butanol extract in 24 h, 48 h, 72 h affect their survival. After damage by doxorubicin using Hoechst 33258 staining with cistanche enzyme extract of different concentrations of n-butanol extraction treated cells after 24 h to oberseve morphological changes of HKC cells. Through the application of flow cytometry, cistanche enzyme extract of different concentrations of n-butanol extraction affecting the role of the cell cycle distribution after 24 h HKC damage by doxorubicinto was oberserved. The animal experiment, the model of chronic renal failure were made by using adenine rats with gavage,the successful model group were treated with medication, after treatment of 28 days, rats were proceeded serum biochemical test and blood test, then general and kidney tissue changeso were bserved.Results: The experiment of enzyme-assisted extraction of Cistanche finalized optimal parameters for solid-liquid ratio of 1:8, reaction temperature of 55 ℃, enzymatic hydrolysis time of 2.5 h, enzyme dosage of 0.1%. Benzene glycosides content of enzyme-assisted extraction increased by 64% compared with traditional water extraction. The cell experiment,MTT results showed that different concentrations of n-butanol extract of cistanche had significant proliferation to all of damaged HKC cells, which increased the survival rate, and the concentration is important. The different concentrations of n-butanol extract of Cistanche cure the damaged HKC cells after 24 h, apoptosis was significantly reduced. Flow cytometryresults showed that the n-butanol extract of cistanche affect the cycle distribution of damaged cell. When increased the concentrations of n-butanol extract, the percentage of G0/G1 phase cells increased, while G2/M phase percentage gradually decreased. Animal experiments section, compared with the control group, the serum creatinine(Scr) and blood urea nitrogen(BUN) rats of treatment group decreased, red blood cell count(RBC), hemoglobin(HGB) and hematocrit(HCT) rats increased, renal index decreased, pathological examination was normal,and the aqueous layer treatment group did not change.Conclusion: Enzyme-assisted extraction of Cistanche can significantly improve the dissolution rate of effective component of benzene glycosides. The cell experiments and animal experiments indicated that chronic renal failure cistanche had therapeutic effect, and the effective parts of n-butanol extraction played a major role in treatment.