TLR9依赖的p38MAPK信号通路对鼠原发性舍格伦综合征的影响

Inhibition of TLR9-dependent p38MAPK Signaling Pathway Attenuates the Pathogenesis of primary Sjgren's Syndrome in Mouse

目的:在动物模型NOD(非肥胖型糖尿病)鼠中,观察研究Toll样受体9(Toll Like Receptor 9,TLR9)依赖的p38MAPK信号通路在原发性舍格伦综合征发病机制中的作用。方法:实验选取5周龄的雌性NOD小鼠,分别给予3种抑制剂:ODN2088、VX-792、羟氯喹。利用流式细胞学技术检测小鼠外周血淋巴细胞的情况。利用免疫组化检测小鼠下颌下腺TLR9及p-p38 MAPK的表达情况。利用酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)检测小鼠外周血中血浆抗体的表达。利用Tunel方法检测小鼠下颌下腺腺上皮细胞的凋亡。同时,观察小鼠刺激性唾液流率的改变以及下颌下腺病理学改变。结果:只有ODN2088组的NOD小鼠的唾液流率显著增加。在所有被给予羟氯喹的NOD鼠和未接受治疗的NOD鼠中,下颌下腺均有淋巴细胞浸润灶的出现。但在ODN2088组中,只有1只NOD小鼠出现下颌下腺的淋巴细胞浸润灶。在VX-702组中,所有NOD小鼠均未发现淋巴细胞浸润灶。所有实验组的外周血淋巴细胞的数目显著减少。ODN2088组NOD小鼠的抗SSA/Ro抗体和抗SSB/La抗体的浓度是所有实验组中最低的。结论:TLR9依赖的p38MAPK信号通路的抑制,能一定程度上减轻原发性舍格伦综合征动物模型NOD鼠的临床表现。

Objective： To observe whether the inhibition of TLR9-dependent p38 MAPK pathway are associated with at-tenuation of symptoms and signs in the pathogenesis of primary Sj？gren＆#39;s syndrome （pSS）. Methods： NOD/Ltj mice were selected as a model of pSS and Balb/c mice were used as control group. NOD/Ltj mice were treated by 3 inhibitors：ODN2088, VX-702, and hydroxychloroquine respectively. The changes of stimulated salivary flow rate was measured and the histopathology of submandibular gland was evaluated by HE stain. Peripheral blood mononuclear cells （PBMCs） de-rived from NOD and Balb/c mice were analysis by flow cytometry. The plasma derived from peripheral blood of NOD and Balb/c mice was analyzed by ELISA. Tunel method to detect epithelial cell apoptosis was performed on submandibular gland sections in NOD mice and Balb/c mice. Results： Compared with NOD/Ltj mice, only the salivary secretion of NOD/Ltj mice given ODN2088 was significantly increased. Periductal inflammatory cell foci were observed in the submandibularglands in the whole NOD mice given hydroxychloroquine, 1/5 of the NOD mice given ODN2088 and all of the NOD mice without any treatment. There were no lymphocytes foci in the NOD mice received VX-702. The number of lymphocytes was extremely low in NOD mice given VX-702, the NOD mice given ODN2088 and NOD mice given hydroxychloroquine. Titers of anti-SSA/Ro and anti-SSB/La in plasma of NOD mice given ODN2088 were the lowest among all groups. Conclusion：Inhibition of TLR9-dependent p38MAPK signaling pathway could alleviate the clinical symptoms of pSS to a certain extent. In addition, The activation of TLR9-dependent p38MAPK signaling pathway palys an important role in the patho-genesis of pSS in NOD/Ltj mice.