Liu Xi-de, Feng Ying-Ying, Cai Long, Zhou Hong-Juan, Ye Li-Hong, Yu Jian-Ming, Wang Yun-Qing, Du Jing, Yang Meng-Xia
Zhongguo Zhong Xi Yi Jie He Za Zhi. 2016 Oct;36(10):1219-1223.
Objective To observe the effect of Wenhua Juanbi Recipe (WJR) on the expres- sions of DNA methyltransferases (DNMTs) in peripheral blood mononuclear cells (PBMCs) of collagen- inducing arthritis (CIA) , and to study its mechanism for treating CIA. Methods Totally 90 Wistar rats were randomly divided into the model group (n =80) and the normal control group (n = 10). Rats of the model group were injected with type II collagen of bovine (BC II) emulsion from the tail to establish CIA model. Successfully modeled 50 CIA rats were randomly divided into five groups, i.e., the model group, the methotrexate (MTX) group, the low dose WJR group, the middle dose WJR group, the high dose WJR group, 10 in each group. Rats in the model group were administered with normal saline by gastrogavage, once per day. Rats in low, middle, and high dose WJR groups were administered with WJR by gas- trogavage at the daily dose of 22. 9, 45. 8, 68. 7 g/kg, respectively (once per day). Rats in the MTX group were administered with MTX suspension (0.78 mg/kg) by gastrogavage, once per week for 30 successive days. The paw swelling was evaluated using volume method (draining volume). PBMCs were extrac- ted from each group after intervention. mRNA expression levels of DNMTs (DNMT1 , DNMT3a, DNMT3b) were detected by real-time quantitative PCR. Results Compared with the normal group, the paws were obviously swollen in the model group (P <0. 01). Compared with the model group, swollen paws were obviously alleviated in low, middle, and high dose WJR groups, and the MTX group (P <0.01). Compared with before treatment in the same group, swollen paws were obviously alleviated in low, middle, and high dose WJR groups, and the MTX group (P <0. 01 ). Compared with the normal group, expression levels of DNMT1, DNMT3a, and DNMT3b in PBMCs were obviously lowered in the model group (P <0.01). Compared with the model group, expression levels of DNMT1 , DNMT3a, DNMT3b in PBMCs were obviously elevated in low, middle, and high dose WJR groups, and the MTX group (all P <0. 01). There was no sig- nificant difference in expression levels of DNMT1, DNMT3a, or DNMT3b in PBMCs among low, middle, and high dose WJR groups (P>0.05). Conclusions Expression levels of DNMTs in PBMCs of CIA rats decreased. WJR up-regulated the expression level of DNMTs in PBMCs of CIA rats in no obvious dose de- pendent way. One of WJR's mechanisms for treating CIA might be up-regulating expression levels of DN- MTs, and adjusting the state of DNA methylation.
目的 观察文化蠲痹方（WJR）对胶原诱导性关节炎（CIA）大鼠外周血单个核细胞（PBMCs）中DNA甲基转移酶（DNMTs）表达的影响，探讨其治疗CIA的作用机制。方法 将90只Wistar大鼠随机分为模型组（n = 80）和正常对照组（n = 10）。模型组大鼠经尾根部注射牛Ⅱ型胶原（BCⅡ）乳剂制备CIA模型。将成功造模的50只CIA大鼠随机分为5组，即模型组、甲氨蝶呤（MTX）组、WJR低剂量组、WJR中剂量组、WJR高剂量组，每组10只。模型组大鼠每日灌胃给予生理盐水；WJR低、中、高剂量组大鼠分别每日灌胃给予WJR，剂量分别为22.9、45.8、68.7 g/kg；MTX组大鼠每周1次连续30 d灌胃给予MTX混悬液（0.78 mg/kg）。采用容积法（排水体积）评估 paw肿胀情况。干预后从每组大鼠中提取PBMCs。采用实时定量PCR检测DNMTs（DNMT1、DNMT3a、DNMT3b）的mRNA表达水平。结果 与正常组比较，模型组大鼠 paw明显肿胀（P < 0.01）。与模型组比较，WJR低、中、高剂量组及MTX组大鼠 paw肿胀明显减轻（P < 0.01）。与同组治疗前比较，WJR低、中、高剂量组及MTX组大鼠 paw肿胀明显减轻（P < 0.01）。与正常组比较，模型组PBMCs中DNMT1、DNMT3a、DNMT3b的表达水平明显降低（P < 0.01）。与模型组比较，WJR低、中、高剂量组及MTX组PBMCs中DNMT1、DNMT3a、DNMT3b的表达水平明显升高（均P < 0.01）。WJR低、中、高剂量组PBMCs中DNMT1、DNMT3a或DNMT3b的表达水平比较，差异无统计学意义（P > 0.05）。结论 CIA大鼠PBMCs中DNMTs表达水平降低。WJR可上调CIA大鼠PBMCs中DNMTs的表达水平，且无明显剂量依赖性。WJR治疗CIA的机制之一可能是上调DNMTs表达水平，调节DNA甲基化状态。
