| Objective: To study the effects and mechanism of Weile capsule(WL)on chronic gastric ulcer induced by acetic acid in rats,The Maximum Feasible Dose(MFD)acute toxicity test,the Salmonella typhimurium reverse mutation test(Ames test),the Mammalian erythrocyte micronucleus test and the Sperm deformity test were used to investigate the potential acutetoxicity and genotoxicity of WL.Methods: In the test of the effects of WL on chronic gastric ulcer induced by acetic acid in rats,after 5 days of adaptation,sixty male Spraque-Dawley(SD)rats were randomly divided into six groups of ten rats each,the rats were fasted,but allowed water,for 14 hours to 16 hours prior to the surgery.Rats in Sham group were subserosal injected of 0.9 % saline(0.1 mL/rat),rats in other groups were subserosal injected of acetic acid(20 %;0.1 mL/rat).From day 1 after surgery,0.5 % Carboxy methylcellulose(CMC;Sham and Model groups),Omeprazole(OMRZ;4.2 mg/kg),WL(189,378,756 mg/kg)were administered orally 10 mL/kg once per day for ten days after the acetic acid injection.On day 10 one hour after treatment,the blood samples were collected and the stomachs were removed.The levels of prostaglandin E2(PGE2)in the serum and stomach and epidermal growth factor(EGF)in the stomach were determined by the double-antibody sandwich enzyme-linked immunosorbent assay(ELISA),the levels of Superoxide dismutase(SOD)and Malondialdehyde(MDA)in the serum and stomach were determined by biochemical estimation and histopathological changes were evaluated by hematoxylin and eosin(HE)staining and then examined by the light microscopy.In the acute toxicity test,after 3 days of adaptation,forty SPF KunMing mice were randomly divided into two groups: WL group(84 g/kg)and Control group of twenty mice each,10 males and 10 females per group,the mice were fasted,but allowed water,for 14 hours to 16 hours prior to the treatment.0.5% CMC(Control group),WL(84 g/kg)were administered orally with 40 mL/kg thrice in 24 hours with the interval 4-5 hours.Acute toxicity was observed and recorded after treatment for 14 days.In the Ames test,strains TA97,TA98,TA100,TA102 and TA1535 were used.In the condition of presence or absence of an exogenous metabolic activation system(S9).All the five strains were made into bacterial suspension,and were exposed to WL at doses of 5000,1000,200,40,8 ?g/plate,Control group and Positive groups were prepared at the same time.Three plates per strain were used each dose in parallel,and the test was repeated twice.The bacterium numbers were recorded after incubated at 37? for 48 h.In the Mammalian erythrocyte micronucleus test,after 3 days of adaptation,thirty SPF KunMing male mice were randomly divided into five groups of six mice each.The mice were fasted,but allowed water,for 14 hours to 16 hours prior to the treatment.0.5 % CMC(Control group),Cyclophosphamide(CP;40 mg/kg),WL(3500,7000,14000 mg/kg)were administered orally 20 mL/kg once.The numbers of micronucleus and the micronucleus rate were detested on 24 hours and 48 hours later of three samples each point.In the Sperm deformity test,after 3 days of adaptation,twenty-five SPF KunMing male mice were randomly divided into five groups of five mice each.The mice were fasted,but allowed water,for 14 hours to 16 hours prior to the treatment.0.5% CMC(Control group),Cyclophosphamide(CP;40 mg/kg),WL(3500,7000,14000 mg/kg)were administered orally 20 mL/kg once per day for five days.On day 35,the numbers and rates of abnormal sperm of all type was observed and recorded.Results:In the test of the effects of WL on chronic gastric ulcer induced by acetic acid in rats,after treated with WL for ten days,compared with the Model group,(1)The Bodyweight was significantly increased(P<0.05).(2)The Ulcer indexes and Ulcer scores were significantly decreased(P<0.05),the ulcer inhibition rates in WL groups(3500,7000,14000 mg/kg)were 22.38 %,30.23 %,41.86 %.(3)The levels of PGE2 in the serum and stomach and EGF in the stomach were significantly increased(P<0.05).(4)The levels of SOD in the serum were significantly increased(P<0.05),and the levels of SOD in the stomach were increased,the levels of MDA in the serum and stomach were decreased,but no significantly(P>0.05).(5)Stomach pathological injury were alleviated,the inflammation and oedema in submucosal layer were decreased.In the acute toxicity test,the result showed no sign of toxicity,no adverse effects were observed,no statistically significant differences of body weight were observed(P>0.05),behavioral analysis such as tremors,convulsions,salivation,diarrhea,lethargy,sleep and coma were no observed,and no pathologic changes of organs were observed by naked eye.In the Ames test,in the condition of presence or absence of S9,compared with the Control group,the result showed no statistically significant differences of the bacterium numbers(P>0.05),and the bacterium numbers of all the strains were no more than twice the spontaneous colonies for each dosage group.But the bacterium numbers of all the strains in the Positive groups were far more than twice the spontaneous colonies,and were statistical significantly increased(P<0.05).In the Mammalian erythrocyte micronucleus test,compared with the Control group,there were no statistically significant differences of the micronucleus rates(P>0.05)in each dosage group on 24 hs and 48 hs,the micronucleus rates were between 1.3 ‰~4.0 ‰.But the micronucleus rates in CP group were statistical significantly increased(P<0.05)on 24 hs and 48 hs,the micronucleus rates were 29.5 ‰,23.2 ‰ respectively.In the sperm deformity test,compared with the Control group,there were no statistically significant differences of the sperm abnormality rates in each dosage group(P>0.05),the sperm abnormality rates were between 14.6 ‰~ 16.6 ‰.The main types of abnormal sperm were hookless,amorphous,fat head,banana,tail fold,but there were no statistically significant differences of the sperm abnormality rates of particular type in each dosage group(P>0.05).The sperm abnormality rates in CP group were statistical significantly increased(P<0.05),the sperm abnormality rates were 69.6 ‰,and the the sperm abnormality rates of the five types were statistical significantly increased(P<0.05).Conclusions:Weile capsule can alleviate the stomach pathological injury in submucosal layer,we speculate that the mechanism of Weile capsule stomach protection effects on chronic gastric ulcer rats induced by acetic acid,is that the up-regulation of PGE2,EGF,and the increase of anti-oxidation activity.No acutetoxicity and genotoxicity of WL was observed under there test condition,no mutagenicity was observed of WL. |
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