Study On Optimization Of Niweikang Prescription

Objective: To ameliorate the prescription of “Niweikang”,reduce its side effects on the basis of ensuring the curative effect.And explore the pharmacological effect of the prescription at the cellular level.Methods: 1.The gastritis model were established byethanol and sodium salicylate.Using Xuedan,Astragalus membranaceus,Laolongpi,Zhushaqi,Banzhilian,Haipiaoqiao and normal saline respectively to carry on the mice.By observing the ratio of kaolin to rat food,search for which ingredients caused vomiting.2.Use the original prescription,the original prescription with halved Zhushaqi,the original prescription with halved Zhushaqi and ginger,the original prescription with halved Zhushaqi and orange peel,the original prescription with halved Zhushaqi and Banxia,and normal saline to carry on the mice respectively.Select the prescription with the best anti-vomiting effect.3.Use the original prescription,optimized prescription and normal saline to carry on the mice respectively.Detecting the expressions of COX-2 and TNF-α by immunohistochemistry(IHC)to make a comparison of the effect between "Niweikang" and the optimized prescription.4.MC cells were stimulated by the optimized prescription.Apply the flow cytometry to detect the apoptosis of the cells.Apply real-time fluorescence quantitative PCR(RT-q PCR)to analyze the expressions of C-myc and Cyclin D1 genes in cells.Apply Western Blot to detect the expressions of Bcl-2,GSK-3β,Bax,β-catenin,PTEN,Caspase-3 and p-GSK-3β proteins in cells.Results 1.After 10 days,the mice were obviously in a listless state and lost weight.Through the gastric pathology,submucosal edema,inflammatory cell infiltration,erosion as well as shallow ulcer were obviously observed.The ratio of kaolin to rat food was the highest in the Zhushaqi group.2.The ratio of kaolin to rat food was lowest in the orange peel group and ginger group.After continuous gastric lavage fluid for about 2 weeks,the state and body weight of the mice were basically restored.3.Compared to the model group,the average optical density values of COX-2 and TNF-α in the original Niweikang group and the optimized group were remarkablly reduced(P<0.01).There was no remarkable difference between the original Niiweikang group and the optimized group.4.The optimized Niweikang groupcould promote the apoptosis of MC cells in a concentration dependent manner.The expression levels of C-myc and Cyclin D1 genes in MC cells were decreased after 48 h.Meanwhile,the expressions of Bcl-2,β-catenin and p-GSK-3β proteins in MC cells were significantly decreased and the expressions of Bax,Caspase-3,PTEN and GSK-3β proteins were increased.Conclusions 1.Zhushaqi may cause nausea.So the dose of zhushaqi in the optimized group was halved.2.The mice showed the lowest level of nausea in the orange peel group and ginger group.15 g of Xuedan,10 g of Astragalus membranaceus,10 g of Laolongpi,3 g of Zhushaqi,15 g of Banzhilian,10 g of Haipiaoqiao,6 g of Orange peel and 6 g of Ginger were selected as the components of optimized "Niweikang".3.The Optimized prescription and the original Niweikang prescription could improve the clinical indications of gastritis model in mice.The therapeutic effect of the two prescriptions was similar,and there was no remarkable difference.4.The optimized prescription could promote the apoptosis of MC cells,andthis prescription may inhibit the activation of Wnt/β-catenin signaling pathway effectively.