Wang Ying, Zhou Chun, Gao Hui, Li Cuixian, Li Dong, Liu Peiqing, Huang Min, Shen Xiaoyan, Liu Liang
Department of Pharmacology, School of Pharmacy, Fudan University, Shanghai, China; Laboratory of Pharmacology and Toxicology, School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou, China; Department of Pharmacy, Huashan Hospital, Fudan University, Shanghai, China.
Laboratory of Pharmacology and Toxicology, School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou, China.
Biochem Pharmacol. 2017 Aug 15;138:119-129. doi: 10.1016/j.bcp.2017.05.006. Epub 2017 May 16.
The balance between T helper 17 (Th17) cells and regulatory T (Treg) cells, plays a critical role in rheumatoid arthritis (RA). The differentiation of Th17 cells requires the activation of STAT3, which determines the balance of Th17/Treg. Here, we investigated the therapeutic effect of Cryptotanshinone (CTS) on collagen induced mouse arthritis and explored the underlying mechanisms.
Arthritis was induced in DBA/1 mice with bovine collagen type II and complete Freund's adjuvant. CTS was given at 20mgkgd or 60mgkgd by gavage for 6weeks. The immuno-inflammation and joint destruction were evaluated and the balance of Th17/Treg was determined. STAT3 acetylation and phosphorylation were detected by western blotting, and the involvement of p300 was investigated by siRNA and plasmid overexpression.
CTS at a dose of 60mgkgd ameliorated the inflammation and joint destruction in CIA mice. It improved Th17/Treg imbalance, and inhibited both acetylation and phosphorylation of STAT3. CTS reduced p300 expression and its binding to STAT3, but increased phosphorylated AMPK. Knockdown of p300 mimicked the inhibitory effect of CTS on STAT3 acetylation and phosphorylation, which could be partially rescued by overexpression of p300-WT, but not p300-dominant negative (DN) construct.
Our study suggested that the anti-arthritis effects of CTS were attained through suppression of p300-mediated STAT3 acetylation. Our data suggest that CTS might be a potential immune modulator for RA treatment.
辅助性T细胞17（Th17）与调节性T（Treg）细胞之间的平衡在类风湿性关节炎（RA）中起关键作用。Th17细胞的分化需要信号转导和转录激活因子3（STAT3）的激活，而这决定了Th17/Treg的平衡。在此，我们研究了隐丹参酮（CTS）对胶原诱导的小鼠关节炎的治疗作用，并探讨了其潜在机制。
用牛II型胶原和完全弗氏佐剂诱导DBA/1小鼠患关节炎。通过灌胃给予CTS，剂量为20mg/kg/d或60mg/kg/d，持续6周。评估免疫炎症和关节破坏情况，并确定Th17/Treg的平衡。通过蛋白质免疫印迹法检测STAT3的乙酰化和磷酸化，并通过小干扰RNA（siRNA）和质粒过表达研究p300的作用。
剂量为60mg/kg/d的CTS改善了胶原诱导的关节炎（CIA）小鼠的炎症和关节破坏。它改善了Th17/Treg失衡，并抑制了STAT3的乙酰化和磷酸化。CTS降低了p300的表达及其与STAT3的结合，但增加了磷酸化的腺苷酸活化蛋白激酶（AMPK）。敲低p300模拟了CTS对STAT3乙酰化和磷酸化的抑制作用，过表达野生型p300（p300-WT）可部分挽救这种作用，但过表达显性负性p300（p300-DN）构建体则不能。
我们的研究表明，CTS的抗关节炎作用是通过抑制p300介导的STAT3乙酰化实现的。我们的数据表明，CTS可能是一种用于RA治疗的潜在免疫调节剂。
