Zengyetang Regulates The Signal Pathway Of Aquaporin In The Treatment Of Sjogren's Syndrome Model

Sjogren's syndrome(SS)is a kind of chronic autoimmune disease,patients with dry mouth,dry eye symptoms as the initial presentation,often with multiple organ damage,due to infringement of lacrimal gland and salivary gland exocrine gland to secrete decrease.In this study,Zengye Tang of "Wenbing Tiao bian" obtain better efficacy,which was used in clinical treatment of SS.Therefore,a series of experimental studies were carried out to explore the treatment mechanism.The preliminary results showed that Zengye Tang could reduced the daily water intake of SS mice,increased the rate of saliva flow,and significantly alleviated the symptoms of dry mouth in SS model mice.Zengye Tang increased defecation frequency,reduced the lymphocyte infiltration of submandibulargland,lung,and intestinal epithelial cells,inhibiting inflammatory cytokines IL-6,IFN-gamma mRNA gene transcription;Immunohistochemical experiments found that Zengye Tang can promote the expression of AQP1 and AQP5 in submandibular glands,lung epithelial cells,and intestinal epithelial cells,and the expression rules of AQP1 and AQP5 is different,These results suggest that Zengye Tang may promote glandular secretion and increase the water transport rate of various tissues by reducing the inflammatory response and up-regulating the expression of aquaporin,thereby alleviating the overall dryness of the animal model of SS.Therefore,we carried out the research on the signal transduction pathway through which Zengye Tang regulates aquaporins and treats Sjogren's syndrome.Objective:This project intends to explore the molecular mechanism of Zengye Tang promoting the expression of aquaporin in the treatment of Sjogren's syndrome,and to test and verify the effect of Zengye Tang on the key proteins of the signaling pathway related to aquaporin and Sjogren's syndrome at the gene level.To clarify the signal transduction process of Zengye Tang regulating the expression of aquaporin,and to clarify the mechanism of "Increasing water" in the treatment of SS.Methods:This topic research observed the Zengye Tang for SS model of phenotype observation,and the impact on the AQPs expression of the glands,clear liquid soup increasing overall treatment of SS model.Therefore,the colonic epithelial cells of primary mice and the lung epithelial cells of primary mice were cultured to observe the effect of LPS on the expression of AQP1 and AQP5 at different concentrations,and the optimal time point and cells with obvious changes were selected.JoinZengyeTang to intervene,differences in protein enrichment of GO and in protein KEGG pathway enrichment analysis,on the basis of the result analysis of the influence of LPS and Zengye Tang intervened the expression of key protein signaling pathways,and Western blotting protein imprinting on the result,FAS relating to aquaporins and sjogren's syndrome,JAK1,JAK2,AKt-STAT,the nf-kappa B p65,p38 lightning,p42,phosphorylation p42,phosphorylated p38 lightning,TLR4 verifies the change of the key protein.Results:1.According to the GO enrichment analysis results and the literature review results,it was found that Zengye Tang mainly affected pancreatic kinin,phenylalanine,cytosine,growth hormone,tyrosine,glutathione,ABC transporter and other factors in cells,and combined with these factors and the signal pathways involved in sain syndrome,the signal transduction system related to these factors was intended to be studied.2.Compared with the control group,LPS can significantly promote the expression of FAS protein(p<0.05),which is enhanced with the extension of action time and the increase of concentration.Zengye Tang had an obvious inhibitory effect on FAS,and the longer the action time,the more obvious the inhibitory effect of Zengye Tang on FAS.The expression of JAK1 and JAK2 was significantly enhanced at low concentration of LPS for 48 hours(p<0.05),and the inhibition effect of Zengyeng Tang on JAK1 and JAK2 was the most obvious at 40%concentration at 24 hours(p<0.05).LPS significantly enhanced the expression of NF-kB p65 at 48h(p<0.05).LPS significantly enhanced the expression of p38 and p-p38 at 48h(p<0.05),but the effect on P42 was not significantly different from that of the control group(p>0.05).After 24 hours,Zengye Tang had the most significant inhibitory effect on p38 and p-p38(p<0.05).The expression of p-p42 was significantly inhibited at 40%concentration for 24 hours,compared with the control group(p<0.01).Compared with the control group,LPS increased the expression of TLR4 most significantly at 10%concentration for 48h(p<0.05).The inhibitory effect of Zengyeng Tang on TLR4 was significant after 24 hours(p<0.05).Conclusion:Zengye Tang has better clinical curative effect for autoimmune diseases sjogren's syndrome,previous studies have found that it can adjust the aquaporins of submandibular gland,alveolar epithelial cells and intestinal epithelial cells,have the effect of moisture transport barriers and relieve dryness syndrome.This project is in order to explore its action mechanism by studied the signal path,through GO and WB,We have found the following conclusions.The effect of Zengye Tang on the key proteins of several signaling pathways was analyzed,Zengye Tang first inhibits TLR-4,TLR-4 as a membrane surface inflammatory substances recognition receptors,After receiving the inflammatory substances stimulation,to transmit signals into the cell membrane,activate p38MAPK pathway,activating the nf-kappa B signaling pathways downstream,and p38 lightning from within the nucleus outgoing will touch the STAT1,STAT1 is an important transcription factor of JAK-STAT.Zengye Tang inhibit p38 lightning,the NF-kappa B and JAK1 in order to control the progress of the inflammation.At the same time,FAS was significantly inhibited by Zengye Tang,and the longer the time,the more significant the inhibition effect was,and FAS can affected downstream NF-kappa B and ERK,ERK is an important branch of MAPK,which can down-regulated AQP5.These pathways are closely related to Sjogren's syndrome,which can not only induce inflammation,but also affect the expression of aquaporins.Zengye Tang has an inhibitory effect on the key proteins in the above pathways,which can reduce inflammation and indirectly promote the upregulation of aquaporins,thus alleviating the clinical manifestations of Sjogren's syndrome.