Experimental Study Of Jiawei Lianmeiyin Regulating Tfh Cells And Improving SS Secretion Of Submandibular Gland Ectopic Germinal Center In NOD Mice

Background and objective:The Jiawei Lianmeiyin is a clinical effective prescription for the treatment of Sjogren’s syndrome by Professor Gao Yongxiang of Chengdu University of Traditional Chinese Medicine,which is quite effective in improving the symptoms of dry mouth caused by SS.Recent studies have found that follicular helper T cells（TFH）are closely related to the formation and maintenance of Abnormal Germinal centers（GC）.The proliferation of TFH cells makes B lymphocytes clone and secrete a large number of immunoglobulins in the exocrine GC proliferate.This excessive abnormal immune response damages the exocrine glands^[1],resulting in dry mouth and dry eyes.Multiple studies have confirmed that TFH cells are involved in the pathogenesis of SS and the injury of the exocrine glands.By consulting the literature,we put forward the hypothesis that whether Jiawei Lianmeiyin can improve the symptoms of dry mouth in early SS by intervention in TFH cells,reducing the protein expressions of TFH cell-specific molecules Bcl-6 and CXCR5 and the level of its effector IL-21.In this study,NOD/LTJ mice are used as the disease model of Sjogren’s syndrome to explore the efficacy and functional characteristics of Jiawei Lianmeiyin in the treatment of SS,and to explore the possible mechanism of action of Jiawei Lianmeiyin on TFH cells,and to provide new ideas for the treatment of SS.Experiment content:Seventy NOD/LTJ mice were randomly divided into 5 groups:model group,Jiawei Lianmeiyin high-dose,medium-dose,low-dose groups and hydroxychloroquine group.Fourteen C57BL/6J mice were used as blank control group.During the process of experiment we need to observe its body weight,water intake,food intake,saliva secretion,fur gloss and defecation.The effectiveness of Jiawei Lianmeiyin is verified by detecting the amount of saliva secretion and water intake.After 4 weeks,extracting blood from its eyeball,dissecting its bilateral submandibular gland and organ index calculation,HE staining to observe the pathological morphology,ELISA method for the determination of serum level of IL-21,immunohistochemistry,immunofluorescence detection of submandibular gland of the Bcl-6,CXCR5 protein expression,fluid cytology sample determination of peripheral blood CD4⁺CXCR5⁺T cells accounts for the proportion of CD4⁺T cells,using SPSS26.0 analysis differences between groups of experimental data.Experimental results:1)Jiawei Lianmeiyin can improve the saliva secretion and food intake of NOD/LTJ mice,reducing the water intake and the number of lymphocyte infiltration lesions in the submandibular gland of NOD/LTJ mice.2)Jiawei Lianmeiyin can reduce the level of IL-21 in the peripheral blood of NOD/LTJ mice.3)Jiawei Lianmeiyin can down-regulate the protein expressions of Bcl-6 and CXCR5 in submandibular gland of NOD/LTJ mice.4)The ratio of CD4⁺CXCR5⁺T/CD4⁺T cells in the peripheral blood of NOD/LTJ mice was decreased by Jiawei Lianmeiyin,but the difference was not statistically significant compared with the model group（P>0.05）.Summary:Jiawei Lianmeiyin can improve the saliva volume and water intake of NOD/LTJ mice,alleviating its dry mouth symptoms by decreasing the concentration of serum inflammatory factor IL-21,down-regulating the protein expressions of Bcl-6 and CXCR5 in submandibular gland.