| Objective:To study the relationship between TRPV1 protein and related inflammatory factors and GERD,and the intervention effect of Heweijiangni Decoction on GERD.Methods:Experiment 1:Ninety-six male SD rats were injected intraperitoneally with ovalbumin(OVA)and perfused with acid to replicate the NERD rat model.They were randomly divided into blank group,sham-operated group,model group,low-dose Hewei Jiangni Decoction group,medium-dose Hewei Jiangni Decoction group,high-dose Hewei Jiangni Decoction group,omeprazole group and SB705498 group.The treatment group was given corresponding drugs for 14 days,and the general situation and ultrastructural changes of esophagus of rats in each group were observed.Experiment 2:Ninety-six male SD rats were injected intraperitoneally with ovalbumin(OVA)and perfused with acid to replicate the NERD rat model.They were randomly divided into blank group,sham-operated group,model group,low-dose Hewei Jiangni Fang group,medium-dose Hewei Jiangni Fang group,high-dose Hewei Jiangni Fang group,omeprazole group and SB705498 group.The treatment group was given corresponding drugs for 14 days.The changes of esophageal epithelial structure in model group and blank group were observed under light microscope.The expression and distribution of TRPV1,MC degranulation,PAR2,SP and CGRP were detected by qPCR,immunohistochemistry,Western blot and ELISA.Experiment 3:84 male SD rats were used to establish EE rat models by pyloric hemisection combined with partial cardiotomy.They were randomly divided into sham operation group,model group,Hewei Jiangni Decoction high dose group,Hewei Jiangni Decoction media dose group,Hewei Jiangni Decoction low dose group and omeprazole group.The treatment group was given corresponding drugs for 30 days.The general situation,lower esophageal pH value and esophageal ultrastructure were observed.The expressions of occludin,TRPV1 and CGRP were detected by qPCR,immunohistochemistry and Western blot.And distribution changes.Result:Experiment 1:Compared with the blank group,the weight of the model group had statistical difference(P<0.05),but there was no statistical difference between the omeprazole group and the model group(P>0.05).The weight of the other groups was higher than that of the model group(P<0.05).The DIS of model group was significantly wider than that of blank group(P<0.05),and the other groups(except omeprazole group)were significantly smaller than that of model group(P<0.05).There was no significant difference between the two groups.In model group(b)and omeprazole group(d),the desmosome structure was obviously destroyed and the tension filament was shortened with the increase of intercellular space(P<0.05).The other groups were similar to the blank group,and there was no obvious abnormality in desmosome.Mitochondrial morphology was highly fragmented in model group.Mitochondrial morphology of control group,sham operation group,omeprazole group and high dose group of Hewei Jiangni Fang was regular and cristae was clear.Experiment 2:Compared with the blank group,the gap of esophageal cells was significantly widened under light microscope.The expression of TRPV1 in the model group was significantly up-regulated(P<0.05),and significantly down-regulated compared with the groups of Weijiangni Decoction and SB705498 group(P<0.05).The expression of SP and CGRP in esophagus and peripheral serum was significantly increased in model group(P<0.05),and significantly decreased in model group(P<0.05)as compared with each dose group of Weijiangni Decoction and SB705498 group(P<0.05).The distribution of MC degranulation in model group and omeprazole group increased significantly(P<0.05),and the distribution of other groups was better than that in model group(P<0.05).Experiment 3:Compared with the blank group,the weight of the model group was significantly different(P<0.05),and the weight of the other groups was increased(P<0.05).The cell gap distance in model group was significantly wider than that in sham-operated group(P<0.05),while that in other groups was significantly smaller than that in model group(P<0.05).In the model group,desmosome structure was obviously destroyed and the tension filament was shortened with the increase of intercellular space(P<0.05).No significant abnormalities were found in the other groups.Mitochondrial morphology was highly fragmented in model group.Mitochondria in high dose group,low dose group and omeprazole group had regular morphology and clear cristae.The expression of occludin in the model group was significantly decreased(P<0.05),and the expression of occludin in the model group was significantly higher than that in the treatment group and the model group(P<0.05).The expression of TRPV1 in the model group was significantly up-regulated(P<0.05)and down-regulated(P<0.05)compared with the dose groups of Weijiangni Decoction.The abnormal expression of CGRP increased in the model group(P<0.05),and decreased significantly in the high dose and medium dose groups(P<0.05).Conclusion:1.The DIS of esophageal mucosa cells in NERD rats was significantly widened,and destructive desmosome structure was severely damaged,the number of mitochondria was significantly reduced.Mitochondrial morphology was highly fragmented.2.The esophageal visceral hypersensitivity was observed in NERD rats.The expression of TRPV1,MC,PAR2,CGRP,SP and other related inflammatory factors were increased,forming a cascade reaction.3.Hewei Jiangni Decoction can improve DIS,prevent desmosome damage,protect esophageal mucosa,prevent mitochondrial structural fragmentation,improve the expression and distribution of TRPV1 and related inflammatory factors,and alleviate esophageal visceral hypersensitivity in NERD rats.Medium and high doses were better than omeprazole.4.The changes of esophageal ultrastructure in EE rats were more serious than those in NERD rats,and the expression of tight junction protein occludin was significantly down-regulated.5.The increased expression of TRP V1 protein and abnormal distribution of inflammatory factors such as CGRP were observed in EE rats,suggesting that the inflammatory response of EE lesions was related to TRPV1.6.Hewei Jiangni Decoction can significantly improve esophageal mucosal erosion,reduce DIS value,prevent desmosome structure damage,protect mitochondrial structure,and up-regulate occludin protein,protect esophageal mucosa.Reduce the expression of TRPV1,improve the abnormal distribution of CGRP and alleviate local esophageal inflammation.The effect of high dose group was the best,similar to omeprazole.This study is based on TRPV1,one of the cold and heat sensitive channels in modern molecular biology,to study the mechanism of Hewei Jiangni Decoction,which have strong characteristics of traditional Chinese medicine by regulating and regulating visceral hypersensitive NERD.In the future,we can further study the specific mechanism of Hewei Jiangni Decoction by disassembling prescriptions,and study the causal relationship between this recipe and TRPV1 by using gene mouse technology. |
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