左、右归丸对去卵巢大鼠骨髓单核/巨噬细胞破骨分化的影响  

Effects of Zuogui Pills(左归丸)and Yougui Pills(右归丸)on Osteoclast Differentiation of Bone Marrow-Derived Macrophage in Ovariectomized Rats

作　　者：冯秀芝[1] 吴继雷[2] 药晓雨 贾连群[1] 任艳玲[1] FENG Xiuzhi;WU Jilei;YAO Xiaoyu;JIA Lianqun;REN Yanling(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847 [2]辽宁中医药大学附属医院,辽宁沈阳110032

出　　处：《中华中医药学刊》2023年第4期72-76,I0014,共6页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82174260,82074145,81774185);辽宁中医药大学自然科学项目(2021LZY011)。

摘　　要：目的基于AMPK/mTOR信号通路研究左、右归丸对去卵巢大鼠骨髓单核/巨噬细胞(bone marrow-derived macrophage,BMMs)破骨分化的作用及机制。方法将60只雌性大鼠随机分为6组,分别为空白组(KB)、假手术组(SHAM)、模型组(OVX)、补佳乐组(BJL)、右归丸组(YGW)和左归丸组(ZGW)。KB组不做处置,SHAM组模拟卵巢切除手术过程,后4组均将双侧卵巢切除。各组分别灌胃蒸馏水(SHAM、OVX)、补佳乐(BJL)、右归丸(YGW)和左归丸(ZGW)。12周后取大鼠股骨和胫骨骨髓,用差时贴壁法获得大鼠BMMs并进行破骨分化诱导。用抗酒石酸酸性磷酸酶活性检测试剂盒检测TRAP活性;qrt-PCR法检测TRAP基因的表达;Western blotting法检测TRAP、RANKL、AMPKɑ、p-AMPKɑ、mTORC1、p-mTORC1蛋白的表达。结果左、右归丸能显著降低去卵巢大鼠BMMs破骨诱导后TRAP活性,降低TRAP基因和蛋白的表达,降低与RANK结合的RANKL蛋白的表达。与KB组比较,OVX组AMPKɑ蛋白磷酸化水平降低,mTORC1蛋白磷酸化水平升高;3个用药组皆能对抗上述改变,其中左归丸优于右归丸。结论左、右归丸均能通过AMPK/mTOR信号通路抑制去卵巢大鼠BMMs的破骨分化。Objective To study the effects and mechanism of Zuogui Pills(左归丸)and Yougui Pills(右归丸)on the osteoclast differentiation of bone marrow-derived macrophage(BMMs)in ovariectomized rats through the AMPK/mTOR signaling pathway.Methods Sixty female rats were randomly divided into 6 groups,which were blank control group(KB),sham operation group(SHAM),model group(OVX),Estradiol Valerate Tablets group(BJL),Yougui Pills group(YGW)and Zuogui Pills group(ZGW).The KB group did not do any treatment,the SHAM group only simulated the process of oophorectomy,and the last 4 groups all underwent oophorectomy.Then groups were gavaged with distilled water(SHAM,OVX),Estradiol Valerate Tablets(BJL),Yougui Pills(YGW)and Zuogui Pills(ZGW).After 12 weeks,the bone marrow tissues of rat femur and tibia were harvested,and the rat BMMs were obtained by the differential time adherence method,and osteoclast differentiation was induced.The TRAP activity was detected by the tartrate-resistant acid phosphatase(TRAP)activity detection kit.The expression of TRAP gene was detected by qrt-PCR.The protein expressions of TRAP,RANKL,AMPKɑ,p-AMPKɑ,mTORC1 and p-mTORC1 were detected by Western blotting.Results Zuogui Pills and Yougui Pills can significantly reduce the activity of TRAP,the expressions of TRAP gene and protein and the expression of RANKL protein combined with RANK after osteoclast induction of BMMs in ovariectomized rats.Compared with that of the KB group,the phosphorylation level of AMPKɑprotein in the OVX group was decreased,and the phosphorylation level of mTORC1 protein was increased.All the three drug groups could resist the above changes,and Zuogui Pills are better than Yougui Pills.Conclusion Both Zuogui Pills and Yougui Pills can inhibit the osteoclast differentiation of BMMs in ovariectomized rats through the AMPK/mTOR signaling pathway.

关 键 词：左归丸 右归丸 骨髓单核/巨噬细胞 破骨分化 AMPK/mTOR信号通路 

分 类 号：R285.5[医药卫生—中药学]