| objective: To study the effect of Capsaicin(Cap)with different dosage on the gastric motility,transient receptor potential vanilloid1(TRPV1),calaitonin gene related peptide(CGPR)and neurotensin(NT)in gastric anturm and brain tissue,content of motility(MTL)in plasm of rat with chronic stress(CUMS)in different periods and to explore the mechanism of Capsaicin.Methods :(1)The experimental animal grouping: One hundred twenty SD rats randomly selected 20 as the normal control group(Group A),The remaining were established CUMS models.After the success of modeling,the remaining rats were randomly divided into 5 group: blank control group(Group B),solvent control group(Group C),Cap 0.04mg/kg group(Group D),Cap0.4mg/kg group(Group E),Cap 4mg/kg group(Group F),each group had 20 rats.In order to explore the effects of Cap on gastric motility in different periods,the experiment was divided into I,II periods,I period was 1 week,II period was 2 weeks.According to the corresponding periods,Group A,Group B,Group C,Group D,Group E and Group F were randomly divided into 2groups,namely the I periods was divided into Group A1,Group B1,Group C1,Group D1,Group E1,Group F1,the II periods was divided into Group A2,Group B2,Group C2,Group D2,Group E2,Group F2,with 10 rats in each group.(2)The establishment of CUMS rat model: Twenty SD rats were randomly divided into control group and model group,each group of 10.Ratsin model group were established by exerting the factors of fasting,Water deprivation,stimulating tails,horizontal shaking,ice water swimming,hot water swimming and reverse of day and night,prohibited activities and moist litter,adjacent 2 days of stimulation does not repeat,each stimulus repeat 2-3times for 3 weeks.(3)The preparation of Cap: Accurately weighed Cap 100 mg(98.37% Cap powder 101.66 mg),10 mg(98.37% Cap powder 10.17 mg),1mg(98.37% Cap powder 1.02 mg),after dissolving in 1ml of Tween 80 completely,were added to 99 ml normal saline mixed respectively,the concentration of 1mg/ml,0.1mg/ml,0.01mg/ml Cap preparation.Control solvent:1ml of Tween 80 added to 99 ml normal saline mixed.(4)The experimental method of Capsaicin intervention: After the completion of modeling,the rats in each group were given different solutions by intragastric administration,Group A had no treatment,Group B was given distilled water,Group C was given the control solvent,Group D was given Cap 0.04 mg/kg,Group E was given Cap 0.4 mg/kg,Group F was given Cap 4 mg/kg.All rats were routinely fed after administration.The rats in each group were fed by stages,all the rats were fasted for 24 hours,and then lavaged them with phenol red solution(concentration was 50mg/dL)in the second day.All the rats were killed 30 minutes later.(5)Detection index:(1)Collected the gastric contents for testing the gastric emptying rate of phenol red.(2)Collection of heart blood 2ml,After centrifugation at 4?,the supernatant was extracted.(3)The gastric mucosal lesion was observed under microscope and the histopathological scorewas observed(4)TRPV1,CGRP and NT content in gastric anturm and brain were measured by ELISA and immunohistochemical staining.Results: 1.The model validation:(1)General states of the rats:Rats in control group were in good condition,activity sensitive,normal feeding water and urine normal.The mental state of rats in the CUMS model group was poorer than that in the control group,and the rats in the CUMS model group were thin,fatigue,lazy,had a dry stool,a poor appetite and dry stool.(2)The gastric emptying rate of phenol red:The emptying rate of gastric red phenol in the normal and CUMS model groups were 80.36%±3.27% and 65.83%±3.90% respectively.The gastric emptying rate in CUMS model group was significantly lower than control group(p<0.05),suggesting that the model was produced successfully.2.Cap intervention results:(1)General states of the rats:No death was observed in all groups,Group A rats were in good condition,activity sensitive,normal feeding water and urine normal.The mental state of rats in the Group B and Group C were poorer,and were thin,fatigue,lazy,had a dry stool,a poor appetite and dry stool.Group D1,Group E1 and Group F1 were the same.The rats in Group D2 the general situation were slightly better,activities slightly slow,thin,feeding is normal and urine normal.Group E2 were the same.The rats in Group F2 in poor condition,and that were thin,fatigue,lazy,but the feeding and the urine was normal.(2)The gastric emptying rate of phenol red: Group A1,B1,C1,D1,E1,F1 was 79.54%±2.98%,69.37%±4.21%,68.15%±3.67%,65.58%±7.50%,66.58%±5.31%,62.52%±5.34% respectively.Group A2,B2,C2,D2,E2,F2 was 78.61%±4.04%,67.14%±3.44%,66.95%±3.93%,92.68%±2.31%,86.24%±2.75%,79.53%±2.24% respectively.In the phase I trial,the emptying rate of gastric phenol red in Group A1 was significantly higher than that in other groups(P<0.05),but there was no significant difference among the other groups(P>0.05).In the phase II study,the emptying rate of gastric red phenol in Group D2,E2 and F2 was significantly higher than Group B2,C2(P<0.05).Group D2 was significantly higher than Group E2(P<0.05).Group E2 was significantly higher than Group F2(P<0.05).At the same dose of Cap,the emptying rate of gastric phenol red in Group D2 was significantly higher than Group D1(P<0.05),Group E2 was significantly higher than Group E1(P<0.05),Group F2 was significantly higher than Group F1(P<0.05).(3)ELISA test results:(1)The expression of TRPV1 in gastric antrum in Group A1,B1,C1,D1,E1,F1 was 63.62±9.45pg/ml ? 269.38±11.07pg/ml ? 274.67±10.92pg/ml ?138.16±12.10pg/ml ? 147.60±15.18pg/ml ? 150.62±11.95pg/ml respectively.Group A2,B2,C2,D2,E2,F2 was 59.42±7.05pg/ml?273.06±13.98pg/ml?271.57±11.59pg/ml?73.89±6.53pg/ml?99.50±5.94pg/ml?130.44±7.63pg/ml respectively.In the phase I trial,the expression of TRPV1 in gastric antrum of Group A1 was significantly lower than that in other groups(P<0.05).Group D1,E1,F1 was significantly lower than Group B1 and C1(P<0.05).The expression of TRPV1 between Group D1,E1 and F1 had no significant differences(P>0.05).In the phase II study,the expression of TRPV1 of Group A2 was significantly lower than that in other groups(P<0.05).Group D2,E2,F2 was significantly lower than Group B2 and C2(P<0.05).And the Group D2 was significantly lower than Group E2(P<0.05),Group E2 was significantly lower than Group F2(P<0.05).At the same dose of Cap,Group D1 was significantly higher than Group D2(P<0.05),Group E1 was significantly higher than Group E2(P<0.05),Group F1 was significantly higher than Group F2(P<0.05).(2)The expression of TRPV1 in hypothalamus in Group A1,B1,C1,D1,E1,F1 was 61.67±6.74pg/ml,284.06±6.88pg/ml,288.01±9.76pg/ml,154.48±8.53pg/ml,159.48±10.14pg/ml,151.77+13.25pg/ml respectively.Group A2,B2,C2,D2,E2,F2 was 64.20±6.14pg/ml,284.50±5.45pg/ml,285.71±8.48pg/ml,78.52±6.31pg/ml,108.21±4.27pg/ml,132.26±4.19pg/ml respectively.In the phase I trial,the expression of TRPV1 in hypothalamus of Group A1 was significantly lower than that in other groups(P<0.05).Group D1,E1,F1 was significantly lower than Group B1 and C1(P<0.05).The expression of TRPV1 between Group D1,E1 and F1 had no significant differences(P>0.05).In the phase II study,the expression of TRPV1 of Group A2 was significantly lower than that in other groups(P<0.05).Group D2,E2,F2 was significantly lower than Group B2 and C2(P<0.05).And the Group D2 was significantly lower than Group E2(P<0.05),Group E2 was significantly lower than Group F2(P<0.05).At the same dose of Cap,Group D1 was significantly higher than Group D2(P<0.05),Group E1 was significantly higher than Group E2(P<0.05),Group F1 was significantly higher than Group F2(P<0.05).(3)The expression of CGRP in gastric antrum in Group A1,B1,C1,D1,E1,F1 was 80.16±7.43pg/ml,218.66±10.24pg/ml,224.13±11.60pg/ml,110.37±8.90pg/ml,164.97±14.52pg/ml,197.67±10.04pg/ml respectively,Group A2,B2,C2,D2,E2,F2 was 84.01±6.58pg/ml,220.79±9.71pg/ml,223.04±12.01pg/ml,110.79±8.74pg/ml,143.21±9.20pg/ml,182.93±9.31pg/ml respectively.In the phase I trial,the expression of CGRP in gastric antrum of Group A1 was significantly lower than that in other groups(P<0.05).Group D1,E1,F1 was significantly lower than Group B1 and C1(P<0.05).But Group D1 was significantly lower than Group E1(P<0.05),Group E1 was significantly lower than Group F1(P<0.05).In the phase II study,the expression of CGRP of Group A2 was significantly lower than that in other groups(P<0.05).Group D2,E2,F2 was significantly lower than Group B2 and C2(P<0.05).And Group D2 was significantly lower than Group E2(P<0.05),Group E2 was significantly lower than Group F2(P<0.05).At the same dose of Cap,Group E1 was significantly higher than Group E2(P<0.05),Group F1 was significantly higher than Group F2(P<0.05).(4)The expression of CGRP in hypothalamus in Group A1,B1,C1,D1,E1,F1 was 78.14±9.34pg/ml,219.64±9.28pg/ml,227.55±6.56pg/ml,109.88±7.29pg/ml,174.64±8.23pg/ml,206.78±6.81pg/ml respectively,Group A2,B2,C2,D2,E2,F2 was 82.69±7.22pg/ml,224.64±9.66pg/ml,232.55±4.97pg/ml,106.07±8.42pg/ml,156.79±5.39pg/ml,176.41±8.60pg/ml respectively.In the phase I trial,the expression of CGRP in hypothalamus of Group A1 was significantly lower than that in other groups(P<0.05).Group D1,E1,F1 was significantly lower than Group B1 and C1(P<0.05).But Group D1 was significantly lower than Group E1(P<0.05),Group E1 was significantly lower than Group F1(P<0.05).In the phase II study,the expression of CGRP of Group A2 was significantly lower than that in other groups(P<0.05).Group D2,E2,F2 was significantly lower than Group B2 and C2(P<0.05).And Group D2 was significantly lower than Group E2(P<0.05),Group E2 was significantly lower than Group F2(P<0.05).At the same dose of Cap,Group E1 was significantly higher than Group E2(P<0.05),Group F1 was significantly higher than Group F2(P<0.05).(5)The expression of NT in gastric antrum in Group A1,B1,C1,D1,E1,F1 was 78.35±7.73pg/ml,247.40±8.25pg/ml,256.21±9.36pg/ml,108.72±10.18pg/ml,46.03±12.65pg/ml,170.56±8.08pg/ml respectively.Group A2,B2,C2,D2,E2,F2 was82.08±8.09pg/ml,243.80±6.44pg/ml,251.41±7.33pg/ml,107.93±8.43pg/ml,136.13±7.73pg/ml,174.40±7.04pg/ml respectively.In the phase I trial,the expression of NT in gastric antrum of Group A1 was significantly lower than that in other groups(P<0.05).Group D1,E1,F1 was significantly lower than Group B1 and C1(P<0.05).But Group D1 was significantly lower than Group E1(P<0.05),Group E1 was significantly lower than Group F1(P<0.05).In the phase II study,the expression of NT of Group A2 was significantly lower than that in other groups(P<0.05).Group D2,E2,F2 was significantly lower than Group B2 and C2(P<0.05).And Group D2 was significantly lower than Group E2(P<0.05),Group E2 was significantly lower than Group F2(P<0.05).(6)The expression of NT in hypothalamus in Group A1,B1,C1,D1,E1,F1 was91.59±7.79pg/ml,258.62±6.42pg/ml,261.42±8.08pg/ml,107.29±4.73pg/ml,134.40±8.74pg/ml,183.06±6.11pg/ml respectively.Group A2,B2,C2,D2,E2,F2 was 91.09±6.28pg/ml,262.89±6.37pg/ml,264.68±7.38pg/ml,111.38±2.94pg/ml,146.48±4.19pg/ml,190.67±3.75pg/ml respectively.In the phase I trial,the expression of NT in hypothalamus of Group A1 was significantly lower than that in other groups(P<0.05).Group D1,E1,F1 was significantly lower than Group B1 and C1(P<0.05).But Group D1 was significantly lower than Group E1(P<0.05),Group E1 was significantly lower than Group F1(P<0.05).In the phase II study,the expression of NT of Group A2 was significantly lower than that in other groups(P<0.05).Group D2,E2,F2 was significantly lower than Group B2 and C2(P<0.05).And Group D2 was significantly lower than Group E2(P<0.05),Group E2 was significantly lower than Group F2(P<0.05).(4)The plasma MTL results in Group A1,B1,C1,D1,E1,F1 was 169.44±8.07pg/ml,71.73±8.54pg/ml,73.30±9.67pg/ml,115.46±9.96pg/ml,114.64±10.61pg/ml,105.27±9.23pg/ml respectively.Group A2,B2,C2,D2,E2,F2 was 172.68±7.49pg/ml,75.48±10.03pg/ml,69.22±8.92pg/ml,152.93±5.66pg/ml,135.67±11.92pg/ml,130.34±12.84pg/ml respectively.In the phase I trial,the plasma MTL of Group A1 was significantly higher than that in other groups(P<0.05).Group D1,E1,F1 was significantly higher than Group B1 and C1(P<0.05).And the plasma MTL of Group D1 and E1 were significantly higher than Group F1(P<0.05),the plasma MTL between Group D1 and Group E1 had no significant differences(P>0.05).In the phase IIstudy,the plasma MTL of Group A2 was significantly higher than that in other groups(P<0.05).Group D2,E2,F2 was significantly higher than Group B2 and C2(P<0.05).And the plasma MTL of Group D2 was significantly higher than Group E2 and F2,the plasma MTL between Group E2 and Group F2 had no significant differences(P>0.05).At the same dose of Cap,Group D1 was significantly lower than Group D2(P<0.05),Group E1 was significantly lower than Group E2(P<0.05),Group F1 was significantly lower than Group F2(P<0.05).(5)The histopathological examination results: The gastric mucosal histopathological damage integral(Masuda standard method)in Group A1,B1,C1,D1,E1,F1 was 0.5±0.71,4.7±1.50,4.3±1.33,4.2±1.03,3.8±0.92,4.0±1.41 respectively,Group A2,B2,C2,D2,E2,F2 was 0.3±0.48,4.1±1.45,4.5±1.35,2.1±0.74,2.5±0.85,2.4±0.84 respectively,In the phase I trial,the gastric mucosa injury score of Group A1 was significantly lower than that in other groups(P<0.05).And there was no significant difference among the other groups(P>0.05).In the phase II study,the integral of Group A2 was significantly lower than that in other groups(P<0.05).Group D2,E2,F2 was significantly lower than Group B2 and C2(P<0.05),but Group D2,E2,F2 had no significant differences(P>0.05).At the same dose of Cap,Group D1 was significantly higher than Group D2(P<0.05),Group E1 was significantly higher than Group E2(P<0.05),Group F1 was significantly higher than Group F2(P<0.05).Conclusion:(1)CUMS method could establish an animal model of gastric motility disorder,and the establishment of CUMS model can increasethe expression of TRPV1.(2)Intragastric of 0.04mg/(kg·d)to 4mg/(kg·d)Cap 1week had no effect on gastric motility in CUMS rats.(3)Intragastric of0.04mg/(kg·d)to 4mg/(kg·d)Cap 2 weeks the gastric motility in rats with CUMS significantly promoted,0.04mg/(kg·d)was the best.(4)Short-term(1and 2 weeks)intragastric different doses of Cap can down-regulation the TRPV1 expression which is up-regulated in CUMS rats,and regulate the expression of CGRP and NT,promote the release of MTL.(5)Cap of0.04mg/kg-4mg/kg can protect the gastric mucosa of rats with CUMS. |
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