不同滋阴补肾法干预去势大鼠破骨通路的机制  

Mechanisms of different yin nourishing and kidney tonifying methods on osteoclastysis pathway in ovariectomized rats

作　　者：黄小彬 葛继荣 李生强 谢丽华 黄景文 何艳艳 薛立鹏 Huang Xiaobin;Ge Jirong;Li Shengqiang;Xie Lihua;Huang Jingwen;He Yanyan;Xue Lipeng(Key Research Laboratory of Osteoporosis Symptom Genomics,Fujian Academy of Traditional Chinese Medicine,Fuzhou 350000,Fujian Province,China;Fujian Key Laboratory of Integrative Medicine for Osteoporosis Prevention and Control(Rehabilitation Hospital of Fujian University of Traditional Chinese Medicine,Fujian Academy of Traditional Chinese Medicine),Fuzhou 350000,Fujian Province,China)

机构地区：[1]福建省中医药科学院骨质疏松证候基因组学重点研究室,福建省福州市350000 [2]福建省中西医结合防治骨质疏松重点实验室(福建省中医药科学院,福建中医药大学附属康复医院),福建省福州市350000

出　　处：《中国组织工程研究》2025年第6期1214-1219,共6页Chinese Journal of Tissue Engineering Research

基　　金：福建省中医药科学院2021年省属公益类科研院所基本科研专项项目(2021R1003007),项目负责人:黄小彬。

摘　　要：背景:六味地黄丸以“三补三泻”为配伍特点,补泻兼施以滋阴补肾;左归丸以“阳中求阴”为配伍特点,以阳促阴补肾。二者同属于滋阴补肾法,其在证候和细胞分子水平等都发挥较好的疗效。目的:观察六味地黄丸和左归丸在骨代谢中的作用,探讨其在破骨通路骨保护素(osteoprotegerin,OPG)/核因子κB受体活化因子配体(receptoractivatorofnuclearfactor-κBligand,RANKL)的作用机制。方法:将32只SD随机大鼠分为模型组、六味地黄丸组、左归丸组、假手术组,每组8只。除假手术组大鼠外,其余大鼠切除双侧卵巢制备骨质疏松模型。术后30 d开始六味地黄丸组大鼠灌胃六味地黄丸1.125 g/(kg·d);左归丸组大鼠灌胃左归丸2.25 g/(kg·d);假手术组和模型组灌胃生理盐水10 mL/(kg·d)。灌胃12周后,取大鼠胫骨检测骨密度,采用ELISA检测血清中雌激素E2、骨碱性磷酸酶、cAMP/cGMP水平,Westernblot法检测股骨中OPG/RANKL的表达情况。结果与结论:①与假手术组相比,模型组骨密度及雌激素E2、骨碱性磷酸酶水平降低(P<0.05),cAMP/cGMP水平升高(P<0.05)。②与模型组相比,六味地黄丸组和左归丸组骨密度显著提高(P<0.05);骨碱性磷酸酶均显著提高(P<0.05);左归丸组cAMP/cGMP水平明显降低(P<0.05)和上调OPG蛋白表达(P<0.05),六味地黄丸组上调OPG蛋白表达和下调RANKL蛋白表达均有作用(P<0.05);两组均不能显著提高雌激素E2水平(P>0.05)。③结果显示,“阳中求阴”的左归丸能够有效提高OPG表达,不能下调RANKL蛋白,而“三补三泻”的六味地黄丸能双向调节OPG和RANKL蛋白的表达,提示了六味地黄丸对抑制破骨功能方面可能比左归丸显著,此结论仍有待进一步的研究加以验证。BACKGROUND:Liuwei Dihuang Wan takes“three tonifying and three reducing effects”as its compatibility feature to nourish yin and tonify the kidneys,while Zuogui Wan takes“seeking yin in yang”as its compatibility feature to nourish yin and tonify the kidneys by promoting yang.Both of them belong to the same method of nourishing yin and tonifying the kidneys,and have better curative effects at the symptomatic and cellular molecular levels.OBJECTIVE:To observe the effects of Liuwei Dihuang Wan and Zuogui Wan in bone metabolism,and to explore their mechanism of action in the osteoprotegerin(OPG)/receptor activator of nuclear factor-κB ligand(RANKL)osteoblastic pathway.METHODS:Thirty-two Sprague-Dawley rats were randomized into model,Liuwei Dihuang Wan,Zuogui Wan,and sham operation group,with eight rats in each group.Osteoporosis models were prepared using removal of both ovaries in the first three groups.Starting at 30 days postoperatively,rats in the Liuwei Dihuang Wan group were gavaged with Liuwei Dihuang Wan 1.125 g/kg/d;rats in the Zuoqui Wan group were gavaged with Zuogui Wan 2.25 g/kg/d;and rats in the sham operation group and the model group were gavaged with saline 10 mL/kg/d.After 12 weeks of gavage,the rat tibia was taken to measure bone mineral density.The serum levels of estrogen,bone alkaline phosphatase,and cAMP/cGMP were measured using ELISA,and the expression of OPG/RANKL in the femur was detected using western blot.RESULTS AND CONCLUSION:Compared with the sham operation group,the model group showed a decrease in bone mineral density and levels of estrogen and bone alkaline phosphatase(P<0.05)and an increase in cAMP/cGMP level(P<0.05).Compared with the model group,the Liuwei Dihuang Wan group and the Zuogui Wan group significantly increased bone mineral density(P<0.05)and bone alkaline phosphatase levels(P<0.05);the Zuogui Wan group significantly decreased cAMP/cGMP levels(P<0.05)and upregulated OPG expression(P<0.05);the Liuwei Dihuang Wan group upregulated OPG expression and downregulated RA

关 键 词：绝经后骨质疏松症 肾阴虚证 六味地黄丸 左归丸 骨保护 OPG RANKL 蛋白印迹 

分 类 号：R496[医药卫生—康复医学] R318[医药卫生—临床医学] R446