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桑色素调节NLRP3/Caspase-1通路改善大鼠实验性自身免疫性甲状腺炎的研究

Morin Improves Experimental Autoimmune Thyroiditis in Rats via NLRP3/Caspase-1 Pathway

    摘要
  • 摘要:
      目的  研究桑色素调节Nod样受体（NLR）家族中含有pyrin结构域的蛋白3（NLRP3）/半胱氨酸天冬氨酸蛋白酶1（Caspase-1）通路对大鼠实验性自身免疫性甲状腺炎的影响。
      方法  将大鼠随机分为6组：对照组，实验性自身免疫性甲状腺炎组（EAT），桑色素低、中、高剂量组（建模后每日灌胃桑色素50、100、200 mg/kg，连续6周）和雷公藤多苷组（LGT，建模后每日灌胃雷公藤多苷6.25 mg/kg，连续6周）。除对照组以外，其余各组均采用皮下注射0.1 mL含猪甲状腺球蛋白的不完全弗氏佐剂建立实验性自身免疫性甲状腺炎大鼠模型，放射免疫分析法检测血清中甲状腺球蛋白（Tg-Ab）、甲状腺过氧化物酶抗体（TPO-Ab）、三碘甲腺原氨酸（T3）和四碘甲腺原氨酸（T4）水平；逆转录-聚合酶链反应检测白细胞介素（IL）-17、IL-4、γ干扰素（INF-γ）的mRNA水平；试剂盒检测血清中蛋白质羰基含量、8-羟基脱氧鸟苷（8-OHdG）水平、丙二醛（MDA）活力；蛋白免疫印迹检测NLRP3、凋亡z相关斑点样蛋白（ASC）、Caspase-1蛋白表达。
      结果  与EAT组相比，桑色素低、中、高剂量组和LGT组血清中TPO-Ab、Tg-Ab、T3和T4水平降低（P<0.01），IL-17、INF-γ的mRNA水平上调（P<0.01），IL-4的mRNA水平上调（P<0.01），蛋白质羟基含量降低（P<0.01），MDA活性降低（P<0.01），8-OHdG水平降低（P<0.01），NLRP3、ASC、Caspase-1蛋白水平降低（P<0.01），桑色素低、中剂量组上述指标与LGT组相比差异有统计学意义（P<0.05），桑色素高剂量组和LGT组相比上述指标无明显差异，桑色素低、中、高剂量组间相比上述指标差异有统计学意义（P<0.05）。
      结论  桑色素通过调节NLRP3/Caspase-1通路能改善大鼠实验性自身免疫性甲状腺炎。

     

    Abstract:
      Objective  To investigate the effects of morin-regulated NLRP3/Caspase-1 pathway on experimental autoimmune thyroiditis in rats.
      Methods   The rats were randomly assigned to 6 groups: control group, experimental autoimmune thyroiditis group (EAT), low-, medium- and high-dose morin groups (post-modeling gavage of 50, 100 and 200 mg/kg morin hydrate per day for 6 weeks) and tripterygium wilfordii polyglycosides group (LGT group, post-modeling gavage of 6.25 mg/kg tripterygium wilfordii polyglycosidesper day for 6 weeks). Except for the control group, the rat model of experimental autoimmune thyroiditis was established by subcutaneous injection of 0.1 mL incomplete Freund’s adjuvant containing porcine thyroglobulin. The levels of serum thyroglobulin (TgAb), thyroid peroxidase antibody (TPOAb), triiodothyronine (T3) and tetraiodothyronine (T4) in serum were detected by radioimmunoassay. The mRNA levels of interleukin-17 (IL-17), interleukin-4 (IL-4) and interferon γ (INF-γ) were detected by reverse transcription-polymerase chain reaction. The levels of serum protein carbonyl content, 8-hydroxydeoxyguanosine (8-OHdG), and malondialdehyde (MDA) activity were checked with test kits. Expressions of NLRP3, apoptosis-related speck-like protein (ASC), and Caspase-1 were detected by Western blot.
      Results   Compared with the EAT group, serum levels of TPOAb, TgAb, T3, and T4 in low-, medium- and high-dose Morin groups and LGT group were reduced (P<0.01) and the mRNA levels of IL-17, INF-γ and IL-4 were increased (P<0.01), the protein hydroxyl content, MDA activity, and 8-OHdG levels were reduced (P<0.01). The levels of NLRP3, ASC and Caspase-1 were reduced (P<0.01), the levels of 8-OHdG were significantly reduced (P<0.01), and the levels of NLRP3, ASC and Caspase-1 were significantly reduced (P<0.01). There were statistically significant differences between the data from the low-dose and the medium-dose Morin groups and the data of the LGT group (P<0.05), while data from the high-dose Morin group showed no significant difference compared with the data of the LGT group. Data from low-, medium- and high-dose Morin groups showed no statistically significant differences (P<0.05).
      Conclusion   The findings suggest that Morin improved experimental autoimmune thyroiditis in rats through regulating NLRP3/Caspase-1 pathway.

     

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