| ObjectiveThe clinical study was conducted to elucidate the clinical efficacy and safety of xiangsha liujunzi in treating Functional dyspepsia(FD)syndrome of spleen deficiency.Based on the established rat model of FD syndrome of spleen deficiency,the mechanism of xiangsha liujunzi was further explored from the perspective of EGC-NGFMethodsPart one:Clinical StudyA total of 60 patients with FD spleen-deficiency syndrome were randomly divided into the treatment group and the control group.The treatment group was treated with xiangsha liujunzi granule,and the control group with placebo for 4 weeks.The main outcome measures include response rate of the clinical global impression scale,and the secondary outcome measures include the the main symptom score,TCM syndrome score and the SF-36 scale.The above outcome measures were observed and recorded,and the adverse events were also evaluated.Part two:Animal ExperimentThe rat model with FD spleen deficiency syndrome was established by iodoacetamide gavage combined with small platform.During the experiment,the general state,body weight,food intake,water intake and grasping ability of the rats were observed.The depression was evaluated by sucrose preference test,and the visceral hypersensitivity was evaluated by VFH fiber test.The degree of spleen deficiency was assessed by the levels of lactic acid,D-xylose and amylase in the serum.HE staining was used to evaluate the pathological changes of gastric antral mucosa.Immunohistochemistry(IHC)was used to detect the mean optical density(MOD)of GFAP,NGF,TRPV1 in the distribution of gastric antrum mucosa.The co-localization of GFAP and NGF was observed by immunofluorescence double-standard method.Western blot was used to detect the expression of GFAP,NGF,TrkA,Akt and TRPV1.ResultsPart one:Clinical Study1.Comparison of clinical global impression scale between the two groupsAfter 4 weeks of treatment,3 cases(11.54%)of dyspepsia symptoms were completely relieved in the treatment group,15 cases(57.69%)were significantly relieved,the response rate was 69.23%.In the control group,one patient had complete remission,accounting for 3.57%.Significant remission was observed in 11 cases(39.29%),the response rate was 42.86%.There was no significant difference in the response rates between the two groups(P>0.05)2.Comparison of main symptom scores between the two groups(1)Total scores of main symptoms:compared with the total scores of main symptoms before treatment,the scores in the two groups were significantly reduced after 2 weeks and 4 weeks of treatment(P<0.01).After 2 weeks of treatment,there was no significant difference between the two groups(P>0.05),After 4 weeks of treatment,the total score of main symptoms in the treatment group was significantly lower than that in the control group(P<0.05).(2)Single score of the main symptom:after 2 weeks of treatment,the scores of post-prandial fullness,discomfort and upper abdominal pain(P<0.01)and upper abdominal burning sensation(P<0.05)in the treatment group were significantly lower than those before treatment.In the control group,the scores of post-meal fullness and discomfort,upper abdominal pain and upper abdominal burning sensation were significantly lower than those before treatment(P<0.01).Compared with the control group,the score of upper abdominal pain in the treatment group were decreased significantly(P<0.05).After 4 weeks of treatment,the scores of post-meal fullness,discomfort and upper abdominal pain(P<0.01),early satiety and upper abdominal burning sensation(P<0.05)in the treatment group were significantly lower than those before treatment.In the control group,the scores of post-meal fullness and discomfort,upper abdominal pain and upper abdominal burning sensation(P<0.01)were significantly lower than those before treatment(P<0.05).Compared with the control group,the scores of post-meal fullness,discomfort and upper abdominal pain in the experimental group were decreased significantly(P<0.01),while there was no significant difference in the scores of other symptoms(P>0.05).3.Comparison of TCM syndrome scores between the two groupsAfter 4 weeks of treatment,the total effective rate of the treatment group was 80.77%,while that of the control group was 57.14%.There was no significant difference in the total effective rate between the two groups(P>0.05).Compared with that before treatment,the total score of TCM syndrome in the two groups decreased significantly after 2 weeks and 4 weeks of treatment(P<0.01).After 2 weeks of treatment,there was no significant difference in the total score of TCM syndrome between the two groups(P>0.05).After 4 weeks of treatment,the total score of TCM syndrome in the treatment group decreased more significantly than that in the control group(P<0.05).The disappearance rate of three TCM syndromes of fullness,epigastric pain and weakness in the treatment group was significantly higher than that in the control group,with statistically significant differences(P<0.01,P<0.05)4.Comparison of SF-36 scale scores between the two groupsAfter 4 weeks of treatment,compared with those before treatment,the scores in terms of general health(GH),vitality(VT)(P<0.01)and body pain(BP),the role of emotion(RE),mental health(MH)(P<0.05)were significantly increased in the treatment group,and the scores in terms of the general health(GH),vitality(VT)(P<0.01),and mental health(MH)(P<0.05)were significantly improved in the control group.After 4 weeks of treatment,the scores in terms of GH,BP,VT,RE and MH in the treatment group was higher than that in the control group(P<0.05).5.Adverse eventsThere were no serious adverse events in the two groups during the treatment,and no obvious abnormal changes were observed in blood routine,liver function(ALT,AST),kidney function(BUN,Cr),urine routine,stool routine and electrocardiogramPart two:Animal Experiment1.General conditionsCompared with the normal control(NC)group,the weight,food-intake,water-intake and grasping ability of rats in the model group were significantly decreased(P<0.01).Compared with the model group,the body weight and food-intake of rats in each medication group were increased to different degrees(P<0.05,P<0.01).The water-intake in the mosapride and high-dose group was significantly increased(P<0.01).Rats in the TCM medium-dose group(P<0.01),the TCM high-dose group(P<0.05)and the mosapride group(P<0.05)showed significantly higher grasping power.2.Sucrose preference testWhen the model was finished,there was no significant difference between the two groups in terms of sucrose consumption and sucrose preference rate(P>0.05).3.VFH fiber testCompared with the NC group,the pain threshold of rats in the model group was significantly decreased(P<0.05).Compared with the model group,The pain threshold of rats in low dose group(P<0.05),medium dose group,high dose group and mosapride group(P<0.01)were significantly increased.4.Amylase activity and the levels of D-xylose and lactic acid in the serumCompared with the NC group,the serum amylase activity and D-xylose content of rats in the model group were significantly decreased(P<0.05),while the lactate content was significantly increased(P<0.05).Compared with the model group,the serum amylase activity and D-xylose content in each group significantly increased(P<0.01),and the serum lactic acid content in each group significantly decreased(P<0.01).5.HE staining resultsThe structure of gastric antral mucosa of rats in each group was complete and clear,with normal cell morphology.No obvious infiltration of inflammatory cells,deep damage or glandular atrophy were observed.6.Immunohistochemical resultsPositive stainings of GFAP,NGF and TRPV1 were observed in the mucosal lamina propria of gastric antrum in rats.Compared with the NC group,the MOD of GFAP,NGF and TRPV1 in gastric antral mucosa of rats in the model group was significantly increased(P<0.01,p<0.05).Compared with the model group,the MOD of GFAP and NGF in gastric antrum mucosa of each medication group was decreased to different degrees(P<0.01,P<0.05),and the MOD of TRPV1 in gastric antrum mucosa of the medium-dose group was significantly decreased(P<0.05).7.Double immunofluorescence resultsGFAP staining and NGF staining were co-located in the mucosa lamina propria of gastric antrum in each group.Compared with the NC group,the yellow fluorescence produced by GFAP-NGF co-localization in the model group was significantly increased.Compared with the model group,the co-location of GFAP-NGF in each drug group was decreased to different degrees.8.Western blotCompared with the NC group,the expressions of GFAP,TrkA,Akt(P<0.01)and NGF,TRPV1(P<0.05)in the model group were significantly up-regulated.Compared with the model group,the expressions of GFAP in each medication group were significantly decreased(P<0.01),the expressions of NGF in the medium-dose and high-dose group were significantly decreased(P<0.05),the expressions of TrkA,TRPV1 in each TCM group were significantly decreased(P<0.01),the expressions of Akt in the low-dose and medium-dose group(P<0.01)and in the high-dose group(P<0.05)were significantly decreasedConclusions1.The symptoms of postprandial distress syndrome,abdominal pain were significantly alleviated with the treatment of xiangsha liujunzi granule,which can also reduce the total score of syndromes,thus significantly improve the quality of life.Moreover,There was no serious adverse events2.The rat established by the method of iodoacetamide gavage combined with small platform can be considered as an ideal model used to study the visceral hypersensitivity of FD with spleen deficiency syndrome.The co-localization of GFAP and NGF was found in the model rats.The expressions of GFAP,NGF,TrkA,Akt and TRPV1 in the model rats were significantly up-regulated.Xiangsha liujunzi decoction could reduce the expressions of the above proteins,especially in the medium-dose group.It is assumed that xiangsha liujunzi decoction may alleviate visceral hypersensitivity by regulating EGC-NGF network and its downstream PI3K/Akt-TRPV1 pathway. |
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