伏立诺他对干燥综合征大鼠免疫功能的影响

Effects of vorinostat on immune function in rats with Sjögren syndrome

目的探讨伏立诺他(SAHA)调节白细胞介素6(IL-6)/信号转导及转录活化因子3(STAT3)信号通路对干燥综合征(SS)大鼠免疫功能的影响。方法采用弗氏完全/不完全佐剂免疫诱导法构建SS大鼠模型,并将造模成功的大鼠分为模型组,SAHA低、中、高剂量组(25、50、100 mg/kg)和SAHA高剂量+IL-6/STAT3信号通路激活剂组(100 mg/kg SAHA+0.05 mg/kg重组IL-6蛋白),每组10只;另选10只健康大鼠为对照组。各组大鼠腹腔注射相应药液/生理盐水,每天1次,连续2周。检测各组大鼠的饮水量(1 d)、唾液流量(10 min)和脾脏指数、颌下腺指数,观察其颌下腺组织病理变化,检测其血清干扰素γ(IFN-γ)、IL-17、IL-10水平和外周血T淋巴细胞亚群分化情况,并检测其颌下腺组织中IL-6/STAT3信号通路相关蛋白的表达情况。结果与对照组比较,模型组大鼠颌下腺组织可见明显的淋巴细胞局灶性浸润,腺泡细胞明显减少;其饮水量,脾脏指数,颌下腺组织病理评分,血清IFN-γ、IL-17水平,外周血辅助性T细胞1(Th1)、Th17比例,以及颌下腺组织中IL-6蛋白的表达水平和STAT3蛋白的磷酸化水平均显著升高(P<0.05);唾液流量、颌下腺指数、血清IL-10水平和外周血调节性T细胞(Treg)比例均显著降低(P<0.05)。与模型组比较,SAHA各剂量组大鼠颌下腺组织病理损伤均有所减轻,各定量指标均显著改善,且有剂量依赖性(P<0.05);重组IL-6蛋白可逆转SAHA对大鼠上述指标的改善作用(P<0.05)。结论SAHA可减轻SS大鼠的颌下腺损伤,调节其免疫功能,上述作用可能与抑制IL-6/STAT3信号通路有关。

OBJECTIVE To investigate the effects of vorinostat(SAHA)on immune function in rats with Sjögren syndrome(SS)by regulating interleukin-6(IL-6)/signal transducer and activator of transcription 3(STAT3).METHODS SS model was constructed by complete/incomplete Freund’s adjuvant immune induction,and the rats with successful modeling were divided into model group,SAHA low-dose,medium-dose and high-dose groups(25,50,100 mg/kg),and high-dose SAHA+IL-6/STAT3 signaling pathway activator group(100 mg/kg SAHA+0.05 mg/kg recombinant IL-6 protein),with 10 rats in each group.Another 10 healthy rats were collected as the control group.Rats in each group were intraperitoneally injected with the corresponding drug/normal saline,once a day,for consecutive 2 weeks.The water intake(1 d)and saliva flow(10 min)of rats in each group were measured,and the spleen index and submandibular gland index were measured;pathological changes in submandibular gland tissue were observed.The serum levels of interferon-γ(IFN-γ),IL-17 and IL-10 were detected,and the differentiation of T lymphocyte subgroups in peripheral blood was also detected;the expressions of IL-6/STAT3 signaling pathway-related proteins in submandibular gland were detected.RESULTS Compared with the control group,obvious focal infiltration of lymphocytes was observed in the submandibular gland tissue of rats in the model group,and the number of acinar cells was decreased significantly;the water intake,spleen index,pathological score of submandibular gland,serum levels of IFN-γand IL-17,and the ratios of helper T cell 1(Th1)and Th17 as well as the protein expression of IL-6 and the protein phosphorylation of STAT3 in submandibular gland were increased obviously(P<0.05);the saliva flow,submandibular gland index,serum level of IL-10 and regulatory T cell(Treg)ratio were obviously decreased(P<0.05).Compared with the model group,the damage to the submandibular gland in SAHA groups was relieved,the quantitative indexes were obviously improved in dose-dependent manner(P<0.05);recombinant IL-6 protein could reverse the improvement effect of SAHA on above indexes of rats(P<0.05).CONCLUSIONS SAHA can relieve the injury of submandibular gland and regulate its immune function in SS rats,which may be related to inhibiting the IL-6/STAT3 signaling pathway.