硫酸羟氯喹治疗NOD小鼠干燥综合征疗效及对IL-1β和TNF-α产生的影响

Effect of Hydroxychloroquine on IL-1β and TNF-α in non-obese diabetic mice with Sjgren's syndrome

目的观察免疫抑制剂硫酸羟氯喹对非肥胖型糖尿病(NOD)小鼠干燥综合征的疗效及其对白细胞介素(IL)-1β和肿瘤坏死因子(TNF)-α产生的影响。方法8周龄雌性NOD小鼠20只,随机分为两组,每组10只,9周龄时开始给药。羟氯喹(HCQ)组动物给予硫酸羟氯喹溶液0．4ml灌胃,对照组给予蒸馏水0．4ml灌胃。定期观察每组的唾液流率。小鼠在15周末和20周末分别取左、右侧颌下腺组织,并断尾取血。用苏木素-伊红(HE)染色评价颌下腺组织的病理学改变。用免疫组织化学检查评价IL-1β和TNF-α在颌下腺组织的蛋白质表达情况。用反转录聚合酶链反应(RT-PCR)技术测定颌下腺组织中IL-1β和TNF-α的mRNA含量。用酶联免疫吸附试验(ELISA)检测外周血血清IL-1β和TNF-α的含量。结果HCQ组与对照组相比,唾液流率明显增加,颌下腺组织淋巴细胞浸润程度明显减轻,IL-1β和TNF-α的蛋白质表达显著降低,IL-1βmRNA和TNF-αmRNA的表达明显下降,外周血血清的IL-1β和TNF-α含量明显降低。结论硫酸羟氯喹对NOD小鼠干燥综合征疗效显著,其作用机制可能与其明显下调NOD小鼠颌下腺中IL-1β和TNF-α的基因表达及蛋白质表达有关。

Objective To investigate the effect of hydroxychloroquine （HCQ） in treating non-obese diabetic mice （NOD mice） with Sjogren＇s syndrome and its mechanism. Methods Twenty female 8-weeks-old female NOD mice were randomly divided into 2 groups. One week later, HCQ and normal saline were administered intragastricly in HCQ group and control group retrospectively. The changes of salivary flow rate were observed. At the age of 15 weeks and 20 weeks, bilateral submandibular glands were resected. The histopathological change was examined by HE staining. The mRNA expression of IL-1β and TNF-α was measured by re- verse transcription-polymerase chain reaction （RT-PCR）. The protein expression of IL-1β and TNF-α was de- tected by immuohistochemical method. Results Compared with the control group, the mice in HCQ group showed that： ①salivary flow rate was increased significantly;②histopathological change was significantly ameliorated, ③The protein expression of IL-1β and TNF-α was significantly decreased;④the expression of IL-1β mRNA and TNF-α mRNA was significantly down-regulated. Conclusion HCQ can ameliorate Sjogren＇s syndrome on NOD mice effectively. The down-regulation of gene expression and protein synthesis may play an important role in the treatment.