Study On The Mechanism Of Kangfuxin Solution On Treating Gastric Ulcer

Objective:Kangfuxin Solution?KFXS?is a Chinese patent drug separated and refined from blattidae America ethanol extract of Periplaneta dried worms.KFXS is effective in treating gastric ulcer?GU?,but its mechanism is not clear.This study aims to explore the mechanism of KFXS on treating experimental GU in order to provide basis for its clinical application.Methods:1.Rats were administered for 7 days by KFXS at doses of 20ml/kg,10ml/kg and 5ml/kg.Then the acute GU model of rats was established by using the method of bounding in cold water.Ulcer index was measured after the experiment on restraint water-immersion stress.And gastric histology was observed by H.E staining.The expressions of TFF2 and bFGF in gastric tissue were determined by immunohistochemistry and serum contents of 5-HT,TNF-?,MTL,and GAS were determined by Elisa assay kits.The expression of apoptosis related proteins in gastric tissue was determined by Western blot.2.Chronic GU model was established by acetic acid.Rats were administered for 14 days with KFXS in 20ml/kg,10ml/kg and5ml/kg.The effect of KFXS on gastric mucosal morphology in rats of chronic GU was observed by direct observation and HE staining.The expression of TFF2 and bFGF in gastric tissue was determined by immunohistochemistry.T-SOD,NO and MDA levels in serum,and VEGF as well as PGE₂ contents in gastric tissue were determined by Elisa analysis.3.Gastric mucosal epithelial cells GES-1 were cultured in vitro.Injuried gastric mucosal epithelial cell model was prepared with nonsteroidal antiinflammatory drugs NS398.Effect of different concentration of KFXS on the proliferation of GES-1 was determined by MTT assay.To observe the effect of KFXS on the morphological changes of GES-1,Hoechst/PI staining was used to detect the occurrence of apoptosis.The levels of Erk1/2,pErk1/2 and apoptosis related proteins were detected by Western blot.Results:1.For the acute GU rats,spread ulcerous spot and impared ulcer-like changes were observed in gastric mucosa in the model group.The results of immunohistochemistry showed that the content of TFF2 and bFGF in gastric tissue was increased after modeling.The results of Western blot showed that the content of Bcl-2 protein in gastric tissue of model group was decreased,and the content of Bax and Caspase-3 protein increased.Compared with the model group,the levels of 5-HT and TNF-?in serum of rats in KFXS groups were significantly lower,and the contents of GAS and MTL were increased.The expression of Bcl-2 protein was increased and the expression of Bax and Caspase-3 were decreased in all drug administration groups.2.For chronic GU rats,a large scale of ulcerous spot and impared ulcer-like changes by pathological assay were observed in gastric mucosa for rats in model group.Compared with the control group,the positive expression of TFF2 and bFGF in the other groups increased,and the difference of KFXS in each dose group of positive expression was significant?P<0.05 or P<0.01?.Compared with model group,KFXS significantly decreased the MDA and EGF level,and increased the NO,T-SOD levels in serum,and elevated VEGF as well as PGE₂ levels in gastric tissue.The KFXS in low dose group increased the content of VEGF has significant difference.3.MTT results showed that KFXS diluted 10 times on the function of GES-1 48h does not inhibit cell growth.When the reaction time increases to 72h,regardless of KFXS concentration is too high or too low will inhibit GES-1 growth.KFXS diluted 80 times after administration of 24h can repair cell damage caused by NSS398.After Hoechst/PI staining,KFXS was observed by High Content Analysis can reduce cell damage induced by NS398,and the decrease of apoptosis and death cells.Western blot results showed that compared with the control group,the ratio of Bcl-2 and Bax in the NS398 group decreased,and the difference was significant.Compared with NS398 group,the ratios of NS398+KFXS group were higher,and the difference is significant?P<0.01?.Conclusions:1.KFXS is effective on both acute and chronic GU,involved in regulating gastrointestinal hormone and repairing damaged gastric mucosa.2.The mechanisms of KFXS on acute and chronic GU were not the same.To the former one,the mechanism is mainly concerned with the regulation of gastrointestinal hormones and regulation between mucosal defense factor and damage factor balance and anti-apoptosis of gastric mucosal cells.Antioxidant injury is one of the mechanisms of chronic GU.3.KFXS protects human gastric mucosal epithelial cell injury caused by NSAIDs with the potential mechanism of regulating Erk1/2signaling pathway.