左归丸拆方通过调节β2-AR介导的OPG/RANKL信号通路提高去卵巢大鼠骨密度  

Effect of Decomposed Zuoguiwan Recipes in Improving BMD of Ovariectomized Rats by Regulating OPG/RANKL Pathway Mediated byβ2AR

作　　者：张明玥 谭峰[1] 叶素敏 柴毅[1] 刘飞祥[1] 麦聪英 李星[1] 徐道明[2] 樊巧玲[1] ZHANG Ming-yue;TAN Feng;YE Su-min;CHAI Yi;LIU Fei-xiang;MAI Cong-ying;LI Xing;XU Dao-ming;FAN Qiao-ling(School of Basic Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China;Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China)

机构地区：[1]南京中医药大学基础医学院,南京210023 [2]南京中医药大学第一附属医院,南京210029

出　　处：《中国实验方剂学杂志》2020年第12期49-55,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81573874,81873229);江苏省高等学校自然科学研究面上项目(18KJB360006);江苏高校优势学科建设工程资助项目(中西医结合);江苏省研究生科研创新计划项目(KYCX19_1283)。

摘　　要：目的:探讨左归丸及其拆方治疗绝经后骨质疏松症(PMOP)的作用机制。方法:去卵巢法建立骨质疏松症模型,将40只SD大鼠平均随机分为假手术组,模型组(0.1 mL·kg^-1·d^-1),17β-雌二醇组(50μg·kg^-1·d^-1)和左归丸补阳药低、高剂量(0.55,1.1 g·kg^-1·d^-1)组。灌胃给药12周后,显微CT(μ-CT)检测右侧远端股骨骨密度(BMD)及显微结构;苏木精-伊红(HE)染色观察大鼠股骨形态学变化;酶联免疫吸附法(ELISA)检测大鼠血清中Ⅰ型胶原交联羧基端肽(β-CTX)和骨特异性碱性磷酸酶(BALP)浓度;实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测大鼠下丘脑β2-肾上腺素能受体(β2AR),胫骨平台骨保护素(OPG)和核转录因子-κB受体激活因子配体(RANKL)蛋白及mRNA表达水平。结果:与假手术组相比,模型组大鼠BMD显著下降(P<0.01),骨组织形态破坏;血清BALP含量降低,β-CTX含量升高(P<0.01);下丘脑β2AR,胫骨RANKL mRNA及蛋白表达量升高(P<0.05,P<0.01),胫骨OPG mRNA及蛋白表达量降低(P<0.05,P<0.01)。治疗后,与模型组比较,各用药组大鼠BMD升高(P<0.01),骨组织形态改善;血清BALP含量升高,β-CTX含量降低(P<0.01);下丘脑β2AR,胫骨RANKL mRNA及蛋白表达降低(P<0.05,P<0.01),胫骨平台OPG mRNA和蛋白表达升高(P<0.05,P<0.01)。结论:左归丸拆方中补阳药能通过调节β2AR介导的OPG/RANKL信号通路提高去卵巢大鼠骨密度,"阳中求阴"配伍特点是左归丸治疗PMOP的关键之一。Objective:To investigate the mechanism of decomposed Zuoguiwan(ZGW)recipes in treating ovariectomized osteoporosis rats.Method:Forty Sprague-Dawley female rats were equally and randomly divided into Sham-operated group,ovariectomized model group,positive group,and low and highdose ZGW groups.After 12 weeks of administration by gavage,the bone mineral density(BMD)of rats’distal femur was measured by micro-CT,the morphology of bone tissue were observed by hematoxylin-eosin staining(HE),β-cross-linked c-telopeptide of typeΙcollagen(β-CTX)and bone-specific alkaline phosphatase(BALP)in serum were detected by enzyme-linked immunosorbent assay(ELISA),and the mRNA and protein expressions ofβ2 AR,OPG and RANKL were evaluated by Western blot analysis and real-time quantitative polymerase chain reaction(PCR).Result:Compared with Sham-operated group,BMD of rats in ovariectomized model group was decreased(P<0.01),morphology of bone tissue was destroyed,serum BALP was reduced,whileβ-CTX was boosted(P<0.01),mRNA and protein expressions of OPG in tibia were reduced,while RANKL were increased,and mRNA and protein expressions ofβ2 AR in the hypothalamus were decreased(P<0.05,P<0.01).Compared with ovariectomized model group,BMDs of rats in low and highdose ZGW groups were increased(P<0.01),morphology of bone tissue was repaired,serum BALP and mRNA and protein expressions of OPG in tibia were up-regulated(P<0.05,P<0.01),whereas serumβ-CTX and mRNA and protein expressions ofβ2 AR in the hypothalamus and RANKL in tibia were down-regulated(P<0.05,P<0.01).Conclusion:Yang-nourishing components in decomposed Zuoguiwan recipes can improve BMD of ovariectomized rats by regulating OPG/RANKL pathway mediated byβ2 AR."Seeking Yin in Yang"is a crucial mechanism of Zuoguiwan in treating ovariectomized osteoporosis in rats.

关 键 词：左归丸 左归丸拆方 绝经后骨质疏松症 阳中求阴 核转录因子-κB受体激活因子配体(RANKL) 骨保护素(OPG) β2-肾上腺能受体(β2AR) 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]