The Effect And Mechanism Of Capsaicin Prevented Gastric Mucosal Injury By Indomethacin

objective:The study of capsaicin(CAP)on the effect and mechanism of indomethacin induced gastric mucosal injury in different period.Methods:We pretreated the rats by given CAP1mg/kg/d 2weeks or 4weeks,and then given the rats with indomethacin 40mg/kg 1 time caused gastric mucosal injury model,observed the effect of CAP on indomethacin induced gastric mucosal injury in rats.The detailed methods as follows.1.Grouping method:80 SD rats were randomly divided into 8 groups with 10 rats in each group.The experiment was completed in two phases,and the?period was 2 weeks,the?period was 4 weeks.Including group Al(control 1 group),group A2(control 2 group),group B1(model 1 group),group B2(model 2 group),group C1(CAP 1 group),group C2(CAP 2 group),group D1(CAP+indomethacin 1 group)and group D2(CAP + indomethacin 2 group).I phase:group A1 was given normal saline 10ml/kg/d by intragastric administration.2 weeks later the rats were given normal saline 10ml/kg 1times by intragastric administration.Group B1 was givennormal saline 10ml/kg/d by intragastric administration.2 weeks later the rats were given indomethacin 40mg/kg 1 time by intragastric administration.Group Cl was given CAP lmg/kg/d by intragastric administration.2 weeks later the rats were given normal saline 10ml/kg 1 times by intragastric administration.Group D1 was given CAP 1mg/kg/d by intragastric administration.2 weeks later the rats were given indomethacin 40mg/kg 1 time by intragastric administration.?phase:group A2 was given normal saline 10ml/kg/d by intragastric administration.4 weeks later the rats were given normal saline 10ml/kg 1 times by intragastric administration.Group B2 was given normal saline 10ml/kg/d by intragastric administration.4 weeks later the rats were given indomethacin 40mg/kg 1 time by intragastric administration.Group C2 was given CAP lmg/kg/d by intragastric administration.4 weeks later the rats were given normal saline 10ml/kg1 times by intragastric administration.Group D2 was given CAP 1mg/kg/d by intragastric administeration.4 weeks later the rats were given indomethacin 40mg/kg 1 time by intragastric administration.2.CAP intervention method:in the experiment,the rats of group C1,group C2,group D1 and group D2 were given CAP lmg/kg intragastric administration at 8 am in each day.After the end,the rats were given free to eat and drink.3.Indomethacin model:after the rats of group Bl,group B2,group D1 and group D2 last irrigation the rats were fasted 24h,can not help but water,and then given them indomethacin 40mg/kg 1 time by intragastric administration.4.Detection index and method:after the rats were killed,gastric juice was collected by acidbase titration method to determine the total acidity of gastric acid.The gastric mucosa injury index wascounted according to the Guth standard.The gastric antrum tissue were trimmed inthe size of 0.5×0.5cm,4 pieces intotal.Taken 1 piece for HE staining.To score the degree of of gastric mucosa pathological injury according to Masuda standards.And used immunohistochemical method to detect the expression of capsaicin receptor(TRPV1),calcitoningene related peptide(CGRP)in gastric mucosa,taken 1 piece to homogenated to detect the malondialdehyde(MDA)by the method of thiobarbituric acid(TBA),taken 1 piece to homogenated to detect the superoxide dismutase(SOD)by the method of nitroblue tetrazolium(NBT),taken 1 piece to homogenatedto detect the prostacyclin(PGI2)by the methodof elisa.Results:1.Animal condition:in the experiment,each group was in good spirits,normal eating,moved easily,nomal bowels and water,no bitingeach other.There were no death in each group.2.Morphological changes of gastric mucosa and mucosal injury score in rats under naked eye:In the group A1,the gastric mucosa was in the color of orange,with smooth surface,no congestion and edema,no damage,no hemorrhageic effusion.In the group A2,the gastric mucosa was in the color of orange,with smooth surface,no congestion and edema,no damage,no hemorrhageic effusion.In the group Bl,the gastric mucosal was congestion,edema.And there was a large number of point bleeding,linear bleeding or strip oferosion,the surface of the gastric mucosa attached to the blood crust,the gastric juice was in the color of coffee,partial visibl blood effusion.In the group B2,the gastric mucosal was congestion,edema.And there was a large number of point bleeding,linear bleeding or strip oferosion,the surfaceof the gastric mucosa attached to the blood crust,the gastric juice was in the color of coffee,partial visibl blood effusion.In the group C1,the gastric mucosa was in the color of orange,with smooth surface,no congestionand edema,no damage,no hemorrhagic effusion.In the group C2,the gastricmucosa was in the color oforange,with smooth surface,no congestionand edema,no damage,no hemorrhagic effusion.In the group D1,the gastric mucosal was congestion,edema,the surface of the gastric mucosa attached to the blood crust,partial visible theerosionof the focal point,partial visible the erosion of the lesion,and the gastric juice was in the color of coffee.In the group D2,gastric mucosal injury was significantly reduced,there was a little bleeding point and scattered in the point of erosion,gastric juice was in the color of light coffee color,partly in the color of coffee.Gastricmucosal injury score:group A1:0.0±0.0,group A2:0.0±0.0,group B1:39.90±5.70,group B2:40.15±5.32,group C1:0.0±0.0,group C2:0.0±0.0,group D1:35.5±3.78,group D2:22.5±5.04.Among them,the groupB1 was higher than group A1(P<0.05)and group C1(P<0.05);the group B2 was higher than group A2(P<0.05),group C2(P<0.05)and group D2(P<0.05);the group D1 was higher than group A1(P<0.05),group C1(P<0.05)and group D2(P<0.05);the group D2 was higher than group A2(P<0.05)and group C2(P<0.05).3.Histopathological changes of gastric antrum inrats and integration of mucosal pathological changes:in the group A1,the gastric antrum mucosal epithelial cells were neatly arranged,no glandular structure disorder.In the group A2,the gastric antrum mucosal epithelial cellswere neatly arranged,no glandular structure disorder.In the group Bl,the gastric antrum mucosal was congestion,hemorrhage,edema,epithelial cell loss,and can visible glandular structure disorder.In the group B2,the gastric antrum mucosal was congestion,hemorrhage,edema,epithelial cell loss,and can visible glandular structure disorder.Inthe group Cl,the gastric antrum mucosal epithelial cells arranged in order,some of the submucosal blood vessels were visible,and there was no structural disorder.In the group C2,the gastric antrum mucosal epithelial cells arrangedin order,some of the submucosal blood vessels were visible,and there wasno structural disorder.In the group D1,the gastric antrum mucosal was congestion,hemorrhage,edema,epithelial cells loss,and some can visible glandular structure disorder.In the group D2,the gastric antrum mucosal was congestion,edema,can visible epithelial cells loss,no glandular structure disorder.The pathological tissue injury scores of gastric antrum mucosa:group A1:0.0±0.0,group A2:0.0±0.0,group B1:5.50±2.12,groupB2:5.45±2.07,groupC1:0.0±0.0,groupC2:0.0±0.0,group D1:5.20±2.25,group D2:2.13±0.99.Among them,the integral of group B1 was higher than that of group A1(P<0.05)and group C1(P<0.05).The integral of group B2 was higher than that of group A2(P<0.05),group C2(P<0.05)ang group D2(P<0.05).The integral of group D1 was higher tha n that of group A1(P<0.05),group C1(P<0.05)and group D2(<0.05).The integral of group D2 was higher than that of group A2(P<0.05)a nd group C2(<0.05).4.Total acidity of gastric acid(mmol/L):group A1:41.90±1.87,group A2:42.20±1.96,group B1:48.15±1.88,group B2:48.42±1.93,g roup C1:38.2±2.09,group C2:34.16±2.36,group D1:45.13±2.21,group D2:41.86±2.07.Among them,the integral of group A1 was higher than that ofg roup C1(P<0.05).The integral of group A2 was higher than that of gro up C2(P<0.05).The integralof group B1 was higher than thatof groupA 1(P<0.05),group C1(P<0.05),group D1(P<0.05).The integralof grou p B2 was higher than that of group A2(P<0.05),group C2(P<0.05),gr oup D2(P<0.05).The integral of group C1 was higher than that of grou p C2(P<0.05).The integral of group D1 was higher than that of group A1(P<0.05),group C1(P<0.05),group D2(<0.05).The integral of grou p D2 was higher than that of group C2(P<0.05).5.MDA content in ga stric antrum tissue(nmol/mgprot):group A1:1.51±0.07,group A2:1.52±0.08,groupB1:3.88±0.17,group B2:3.91±0.21,group C1:1.49±0.06,group C2:1.47±0.08,group D1:3.78±0.19,group D2:2.75±0.23.Among them,the integral ofgr oup B1 was higher than that of group A1(P<0.05)and group C1(P<0.05).The integral of group B2 was higher than that of group A2(P<0.05),group C2(P<0.05)and group D2(P<0.05).The integral of group D 1 washigher than that of group A1(P<0.05),group C1(P<0.05)and gr oup D2(P<0.05).The integral of group D2 was higher than that of gro up A2(P<0.05)and group C2(P<0.05).6.SOD activity in gastric antr um tissue(u/mg prot):group A1:151.05±16.76,group A2:150.54±15.85,gr oup B1:100.48±9.69,group B2:101.48±9.85,group C1:148.58±16.82,groupC 2:155.57±13.18,group D1:113.86±11.63,group D2:123.54±9.93.Among them,the integral of group B1 was lower than that of group A1(P<0.05),group C1(P<0.05)and group D1(P<0.05).The integral of group B2 was lower than that of group A2(P<0.05),group C2(P<0.05)and group D2(P<0.05).The integral of group D1 was lower than that of group A1(P<0.05)and group C1(P<0.05).The integral of group D2 was lower than that of group A2(P<0.05)and group C2(P<0.05).7.PGI2 content in gastric antrum tissue(ng/L):group A1:172.16±19.82,group A2:170.72±18.72,group B1:113.74±17.99,group B2:110.95±16.58,group C1:225.14±16.96,g roup C2:259.49±19.48,group D1:132.55±16.05,group D2:163.11±23.32.Among them,the integral of group A1 was lower than that of group C1(P<0.05).The integral of group A2 was lower than that of group C2(P<0.05).The integral of group B1 was lower than that of group A1(P<0.05),group C1(P<0.05)and group D1(P<0.05).The integral of group B2 was lower than that of group A2(P<0.05),group C2(P<0.05)and group D2(P<0.05).The integral of group C1 was lower than that of group C2(P<0.05).The integral of group D1 was lower than that of group A1(P<0.05),group C1(P<0.05),group D2(P<0.05).The integral of group D2 was lower than that of group C2(P<0.05).8.The expression of TRPV1 and CGRP in gastric antrum mucosa of rats and the scores:the positive products of TRPV1 and CGRP of gastric antrum mucosa in rats were expressed in all groups,and the products were mainly located in the cytoplasm of the cells.(1)The expression of TRPV1 in gastric antrum mucosa of rats:group A1:The color of the positive products were light yellow or yellow,and the distribution was sparse or medium.Group A2:The coloro f the positive products were light yellow or yellow,and the distribution w as sparse or medium.Group B1:The color of the positive products were li ght yellow,and the distribution was sparse or medium.Group B2:The colo r of the positive products were light yellow,and the distribution was spar se or medium.Group C1:The color of the positive products were yellow,t he distribution was mediumor dense,and the partial distribution was spars e.Group C2:The color of the positive products were yellow or brown yell ow,and the distribution was very denseor especially dense,partial distribut ion was medium.Group D1:The color of the positive products were light yellow or yellow,and the distribution was medium or dense,and the parti al distribution was sparse.Group D2:The color of the positive products w ere yellow or brown yellow,and the distribution was medium or dense,pa rtial distribution was very dense.The TRPV1 scores of gastric antrummuco sa in rats:group A1:1.38±0.52,group A2:1.36±0.48,group B1:1.33±0.50,grou p B2:1.35±0.53,group C1:2.25±0.71,group C2:3.38±0.74,group D1:2.13±0.64,group D2:3.25±0.71.Among them,the TRPV1 scores of group A1 was lo wer than that of group C1(P<0.05)and group D1(P<0.05).TheTRPV1 scores of group A2 was lower than that of group C2(P<0.05)and gr oup D2(P<0.05).The TRPV1 scores of group B1 was lower than that of group C1(P<0.05)and group D1(P<0.05).The TRPV1 scores of gro up B2 was lower than that of group C2(P<0.05)and group D2(P<0.05).The TRPV1 scores of group C1 was lower than that of group C2(P<0.05).The TRPV1 scores of group D1 was lower than that of groupD2(P<0.05).(2)The expression of CGRP in gastric antrum mucosa of rats:group A1:The color of the positive products were light yellow,and the di stribution was sparse or medium.Group A2:The color of the positive prod ucts were light yellow,and the distribution was sparse or medium.Group B1:The color of the positive products were light yellow,and the distributi on was sparse or medium.Group B2:The color of the positive products were light yellow,and the distribution was sparse or medium.Group C1:Th e color of the positive products were light yellow or yellow,and the distr ibution was sparse,medium or dense.Group C2:The color of the positive products were yellow,partly visible brown,and the distribution was mediu m,dense or especially dense.Group D1:The color of the positive products were light yellow,and the distribution was sparse,medium or dense.Grou p D2:The color of the positive products were light yellow or yellow,part ly visible brown,and the distribution was medium,dense or especially dens e.TheCGRP scores of gastric mucosal in rats:group A1:1.30±0.76,group A 2:1.32±0.75,group B1:1.20±0.42,group B2:1.23±0.45,group C1:2.0±0.67,gro up C2:3.1±0.74,group D1:1.90±0.74,group D2:2.9±0.73.Among them,the CGRP scores of group A1 was lower than that of group C1(P<0.05)a nd group D1(P<0.05).The CGRP scores of group A2 was lower than t hat of group C2(P<0.05)and group D2(P<0.05).The CGRP scores o f group B1 was lower than that of group C1(P<0.05)and group D1(P<0.05).The CGRP scores of group B2 was lower than that of groupC2(P<0.05)and group D2(P<0.05).The CGRP scores of group C1 was lower than that of group C2(P<0.05).The CGRP scores of group D1w as lower than that of group D2(P<0.05).(3)Correlation analysis of ex pression of TRPV1 and CGRP in rat gastric antrum mucosa:pearson co rrelation analysis was used to detect the relativity of TRPV1 and CGRP in gastricantrum mucosa.The results showed that the correlation coefficient of TRPV1 and CGRP was 0.998.There was apositive correlation between the expression of TRPV1 and CGRP in the gastric antrum mucosaoconclusion:1.There was no damage to the general morphology and histology of gastric mucosa in rats by intragastric CAP 1mg/kg/d for 2 weeks and 4 weeks.2.It could prevent that indomethacin induced gastric mucosal injury in rats by pretreated with CAP 1mg/kg/d for 4weeks.3.It could decrease the total acidity of gastric acid in rats by intragastric CAP 1 mg/kg/d for 2 weeks and 4 weeks.4.It could increase the expression of TRPV1 and CGRP in rats' gastric antrum mucosa,PGI2 content in gastric antrum tissues,the SOD activity of gastric antrum tissues in indomethacin model by intragastric CAP lmg/kg/d for 2 weeks and 4 weeks.It could decrease the content of MDA in gastric antrum tissues in indomethacin model by intragastric CAP 1mg/kg/d for 4 weeks.