Therapeutic Effect of JXYG on Sjogren’s Syndrome and Immune Regulation on Tfh/Tfr

Abstract

Background: Sjogren’s syndrome (SS) is a common autoimmune disease, which mainly invades glands and belongs to global disease, seriously affecting the quality of life of patients. The purpose of this study is to observe the therapeutic effect of Jinxueyuan Granules (JXYG) on SS and the regulatory effect of JXYG on follicular helper T (Treg) cells/follicular regulatory T cell (Tfh/Tfr) immune balance through animal experimental research. Methods: Statistical method was used to analyze the changes in water intake of NOD (non obese diabetic) mice after JXYG treatment. Hematoxylin and eosin (H&E) staining was used to observe the pathological morphology of the submandibular gland of NOD mice. Flow cytometry was used to analyze the effect of JXYG on the changes of Tfh cells and Tfr cells in the peripheral blood of NOD mice. Immunohistochemistry was used to analyze the effects of blood source on the expressions of CD4 (cluster of differentiation 4), CXCR5 (C-X-C chemokine receptor type 5), FOXP3 (fork head box P3), Bcl-6 (B-cell lymphoma 6), and Blimp-1 (B lymphocyte-induced maturation protein-1) in NOD mice’s submandibular gland and western blot was applied to analyze the expression of Bcl-6, Blimp-1, STAT3 (signal transducer and activator of transcription 3), and p-STAT3 (phosphorylated-signal transducer and activator of transcription 3) in submandibular glands of NOD mice in each group. Results: The amount of drinking water and the degree of lymphocyte infiltration in the submandibular gland of NOD mice could be reduced after the intervention of JXYG, which was closer to the normal level, and the effect of the high-dose group was more significant than that of the low-dose group. JXYG could promote the decrease of Tfh/CD4⁺ T, Tfr/CD4⁺ T, and Tfh/Tfr ratio in the peripheral blood of NOD mice, but there is no significant difference. JXYG could remarkably reduce the expression levels of Bcl-6, Blimp1, CD4, and p-STAT3, and increase those of FOXP3 in the submandibular gland of NOD mice. Conclusions: The above results suggested that JXYG may play a regulatory role in Tfh/Tfr immune balance by regulating the above molecular pathways. This study investigated the possible mechanisms underlying the treatment of SS with JXYG from a cellular and molecular perspective, thereby elucidating the pathogenesis of SS and providing an evidence-based foundation for the clinical treatment of dryness syndrome with the use of Chinese herbal compounds.