右归饮抑制大鼠破骨细胞作用的分子机制研究

Molecular mechanism of Youguiyin inhibit the activity on the osteoclasts in rats

目的探讨右归饮抑制大鼠破骨细胞作用的分子机制。方法 105只Ⅱ型胶原诱导的类风湿关节炎(CIA)模型大鼠分为7组:3个剂量右归饮组、甲氨蝶呤组、阳性与阴性病毒组、模型组,每组15只;另取15只正常大鼠作为空白组。高中低3个剂量右归饮组分别给与相当于人临床剂量的22.5,15,7.5倍的量,每周3次;甲氨蝶呤组,按照1μg·g-1体重给药,每周1次;阳性病毒组,双膝各关节注射2.5×106TU Anx A2与Atp6i慢病毒,阴性病毒组,同法注射阴性慢病毒,均给药8周。观察给药前后各组血清抗酒石酸酸性磷酸酶-5b(TRACP-5b)、白细胞介素-1β(IL-1β)含量变化、膝关节骨密度,钼靶X线下观察大鼠膝关节骨皮质的变化。结果 8周后,右归饮、甲氨蝶呤、Anx A2和Atp6i慢病毒(阳性病毒)均能显著下调IL-1β和TRACP-5b表达(P<0.05);右归饮高剂量组的骨密度明显优于中、低剂量组(P<0.05);右归饮组,大鼠膝关节骨皮质较完整,骨量分布较均匀,且高剂量组较中低2个剂量组更明显。结论在类风湿关节炎骨破坏过程中,右归饮可能通过调控Anx A2、IL-1β基因来抑制破骨细胞骨吸收活性,从而抑制类风湿关节炎的膝关节骨破坏。

Objective To evaluate the molecular mechanism of You-guiyin inhibit the activity of rat osteoclasts through inhibiting Annexin A 2 （ AnxA2） and Atp61.Methods One hundred and five rats of collagen-induced arthritis （ CIA） were divided in to 7 groups：high, moderate and low doses Youguiyin groups , methotrexate group , lentiviruses （ ＋） group, lentiviruses （ -） group and model control group.And 15 rats without any treatment were included as blank group.Rats in the Yougui-yin groups were given Youguiyin intragastric.Rats in the methotrexate group were given methotrexate 1 μg· g -1 intragastric.Rats in the lenti-viruses （＋） group were given 2.5 ×10 6 TU AnxA2 and Atp6 i lentiviru-ses double knee joints injection.Rats in the lentiviruses （-） group were given lentiviruses double knee joints injection.At the time of 8 weeks af-ter administration the rats were put to death.The serum level of tartrate resistant acid phosphatase -5b（TRACP-5b）, the interleukin -1 beta （ IL-1β） , and knee joint bone density were evaluated.The change of the knee joint bone cortex in rats was evaluated by molybdenum target X-ray before and after administration.Results After 8 weeks, the IL-1βand TRACP -5b levels in CIA model group were much higher than that of blank group （P〈0.05）.IL-1βand TRACP-5b levels in Youguiyin, methotrexate, and lentiviruses （ ＋） groups were much lower ＆ nbsp;than that of CIA model group （ P〈0.05 ）.The bone density in the Youguiyin and methotrexate groups were lower than the blank group （ P〈0.05 ） but significant improve in the high dosages Youguiyin group than that of moderate and low dosage groups （ P〈0.05 ） .After administration with Youguiyin , bone cortex is complete , bone mass distribution is uni-form according to X-ray in the high dosage group compared with the moderate and low groups.Conclusion In the process of osteoclasia in rats with rheumatoid arthritis , Youguiyin can inhibit the osteoclasia effects by regulate and con-trol the AnxA2 and IL-1βexpression.