The Effect Of Linggui Zhugan Decoction On The Expression Of AQP4 Protein In The Cochlea Of ​​Guinea Pigs With Meniere's Disease Through The P38MAPK Signaling Pathway

Purpose:In this experiment,we observed the effect of Ling-Gui-Zhu-Gan decoction on labyrintine hydrops in guinea pigs with Meniere’s disease.Four proteins,P38 MAPK,p-P38 MAPK,ATF-2 and AQP4,were selected as experimental observation indexes.We used morphological,genetic and molecular biological aspects to observe the differences of expression of the above indicators among blank group,model group,Ling-Gui group and Ling-Gui+alpha-LA group.To explore the possible mechanism of Ling-Gui-Zhu-Gan decoction in the treatment of guinea pig membranous labyrinth hydrops.To provide a new idea for better clinical application of Ling-Gui-Zhu-Gan decoctionin the treatment of Meniere’s disease in the future.Material and method:36 adult healthy red eye guinea pigs were randomly divided into three groups by random number table method.Each group was randomly divided into blank group,model group,Ling-Gui group and Ling-Gui+alpha-LA group.Except the blank group,the other three groups were induced by intraperitoneal injection of desmopressin acetate,6μg/kg/d,continuous injection for 10 days.After successful modeling,guinea pigs in Ling-Gui group were given Ling-Gui-Zhu-Gan decoction 2.79g/kg daily by gastric perfusion.In Ling-Gui+alpha-LA group,alpha-LA was given to guinea pigs 46.5mg/kg daily,and Ling-Gui-Zhu-Gan decoction 2.79g/kg daily by gastric perfusion,too.The blank group was fed with the same amount of saline.The model group was fed normally,and the samples were taken after 7 days.HE staining was used to observe cochlear hydrops in guinea pigs.The expression of P38 MAPK,p-P38 MAPK,ATF-2 and AQP4 proteins in the cochlea of guinea pigs was observed by immunohistochemistry.The gene expression levels of P38 MAPK,p-P38 MAPK,ATF-2 and AQP4 in the cochlea of guinea pigs were observed by fluorescence quantitative PCR.Western-Blot was used to observe the expression levels of P38 MAPK,p-P38 MAPK,ATF-2 and AQP4 proteins in the cochlea of guinea pigs in each group.We use SPSS software to analyze the experimental results and explore the possible mechanism of Ling-Gui-Zhu-Gan decoction in treating Meniere’s disease.Results:1.The total water accumulation rate of model group was 100%,which was significantly different from that of blank group(P<0.01).2.The expression of P38 MAPK,p-P38 MAPK and ATF-2 in the model group was up-regulated,which was significantly different from that in the blank group(P<0.01);the expression level in Ling-Gui group was lower than that in model group,and there was significant difference between Ling-Gui group and model group(P<0.01).3.The expression of AQP4 protein was down-regulated in the model group,and there was significant difference between the model group and the blank group(P<0.01);the expression of AQP4 protein in Ling-Gui group was up-regulated compared with the model group,and there was significant difference between Ling-Gui group and the model group(P<0.01).Conclusion:1.Ling-Gui-Zhu-Gan decoction can alleviate membranous labyrinth hydrops in guinea pigs to some extent.2.The early hydrocele of guinea pig’s membranous labyrinth caused by deaminapressin acetate may be related to the up-regulation of AQP4 protein expression.3.Ling-Gui-Zhu-Gan decoction can up-regulate the expression of AQP4 protein in the cochlea of guinea pigs.4.The up-regulation of AQP4 expression in inner ear of guinea pigs after intraperitoneal injection of vasopressin acetate may be one of the mechanisms of vasopressin-induced hydrops of membranous labyrinth.5.Ling-Gui-Zhu-Gan decoctionin the treatment of Meniere’s disease may be through the P38 MAPK signaling pathway and then up-regulate AQP4 protein.6.The early expression of AQP4 protein in guinea pig model of membranous labyrinthine hydronephrosis may be negatively related to the activation of P38 MAPK signaling pathway.