Combined With Network Pharmacology To Explore The Effect And Mechanism Of Jiawei Lianmei Yin Sjogren's Syndrome Model In Mice

Background and objective: Sjogren's syndrome(SS)is a chronic autoimmune disease characterized by invasion of exocrine glands.The main symptoms of early onset are xerophthalmia,fatigue and arthralgia.In view of the main demands of SS patients to improve dryness,fatigue,muscle pain,modern medicine has no specific treatment,only symptomatic treatment,symptomatic treatment drugs can not prevent the progress of the disease.Traditional Chinese medicine in the treatment of Sjogren's syndrome can effectively improve the early symptoms of patients with less side effects,so traditional Chinese medicine in the treatment of Sjogren's syndrome has been more and more recognized and concerned.Jiawei Lianmei Yin is a clinical effective prescription for Sjogren's syndrome by Professor Gao Yongxiang of Chengdu University of traditional Chinese medicine,but its therapeutic characteristics and mechanism are still unclear.Therefore,in order to verify the therapeutic effect of Jiawei Lianmei Yin on SS,to explore the characteristics and mechanism of Jiawei Lianmei Yin,and to provide laboratory basis for clinical rational application of Jiawei Lianmei Yin in the treatment of SS,we designed this experiment..Methods: 1.Tcmsp,drugbank,geo,CNKI and other databases were used to obtain the active ingredients of Jiawei Lianmei Yin and the differential genes between Sjogren's syndrome and healthy people.The active ingredients of Jiawei Lianmei Yin and drug disease interaction targets were analyzed.According to the drug disease interaction targets,the protein protection network was constructed,and go and KEGG were analyzed;2.NOD / Ltj mice were used as Sjogren's syndrome model,C57 / bl-6mice were used as blank control group,hydroxychloroquine sulfate was used as positive control drug,and different doses of Jiawei Lianmeiyin concentrate were used to intervene the mice.The general situation,salivary flow,organ index and submandibular gland HE staining of mice in each group were observed.The expression of IL-17,IL-6,TNF-? and IL-23 in serum was detected by ELISA,and the expression of IL-17,IL-6 and TNF-? in submandibular gland was detected by immunohistochemistry;3.The number of Th17 in spleen was detected by flow cytometry,and the expression of ror ? T,I-? B,NF-? B p65,p-p65 and AQP5 in submandibular gland was detected by Western blot.Results:1.According to the results of network pharmacology and literature analysis,we speculate that quercetin,isoliquiritigenin,apigenin,glycine and kaempferol may be the important components of Jiawei Lianmei Yin in the treatment of SS.These ingredients have therapeutic effects on autoimmune diseases mainly through antioxidant,anti-inflammatory,immune regulation,organ protection and other biological pathways;and through the influence of Th1,Th2,Th17,reduce IL-1,IL-6,TNF-?,il17 and other inflammatory factors,play a therapeutic role in the treatment of SS.Furthermore,it is possible to regulate Th17 cells to affect IL-17 and TNF pathways.Based on this,we propose a further working hypothesis that "Jiawei Lianmei Yin may affect IKB / nf-kb-aqp5 pathway by regulating Th17 cells to achieve the purpose of treating SS".2.Jiawei Lianmei Yin can reduce the number of lesions in submandibular gland of mice,compared with the model group,with statistical significance(P < 0.05);at the same time,the drug can reduce the amount of Yining water of mice;inhibit the expression of serum IL-17,IL-6,IL-23,TNF-?,with statistical significance(P <0.05).The expression of pro-inflammatory factors IL-17,IL-6,IL-23 and TNF-? in submandibular gland tissue was down regulated,and there was statistical significance(P < 0.05).There was a positive correlation between the improvement of each index and the drug concentration,and there was a significant difference between the high-dose group of Jiawei Lianmei Yin and hydroxychloroquine(P < 0.01).3.Western blot was used to detect the expression of ROR?t,I?B,NF-? B p65,p-p65 and AQP5 in submandibular gland,and flow cytometry was used to detect the number of Th17 cells in spleen.It was confirmed that Jiawei Lianmeiyin could inhibit the differentiation of Th17 in spleen,inhibit the expression of ROR?t,I?B,NF-? B p65,p-p65 in submandibular gland,and up regulate the expression of AQP5(P < 0.05).Conclusion: 1.Jiawei Lianmei Yin can protect submandibular gland tissue structure,improve salivary flow,improve early xerostomia of SS,and delay the progress of the disease;2.The mechanism of Jiawei Lianmei Yin may be to inhibit the differentiation of Th17 cells,reduce the expression of related inflammatory factors(IL-6,IL-17,TNF-?,etc.),thus inhibit the activation of NF-?B signaling pathway,and up regulate the expression of AQP5;3.Jiawei Lianmei Yin can inhibit the differentiation of s-17 cells,and reduce the expression of related inflammatory factors(IL-6,IL-17,TNF-?,etc.)The intervention effect of early symptoms of SS is better than that of hydroxychloroquine,which has a certain inhibitory effect on the development of SS,and can be further studied as an early intervention scheme of SS.