电针干预去卵巢骨质疏松模型大鼠骨组织Runx2启动子甲基化水平的变化  

Runx2 promoter methylation level changes in bone tissue of rats with ovariectomized osteoporosis treated by electroacupuncture

作　　者：陈赵 莫雨晴 唐宏宇[4] Chen Zhao;Mo Yuqing;Tang Hongyu(Guangzhou University of Chinese Medicine,Guangzhou 510405,Guangdong Province,China;Zhaoqing First People's Hospital,Zhaoqing 526000,Guangdong Province,China;First Clinical School of Guangzhou University of Chinese Medicine,Guangzhou 510405,Guangdong Province,China;The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,Guangdong Province,China)

机构地区：[1]广州中医药大学,广东省广州市510405 [2]肇庆市第一人民医院,广东省肇庆市526000 [3]广州中医药大学第一临床医学院,广东省广州市510405 [4]广州中医药大学第一附属医院,广东省广州市510405

出　　处：《中国组织工程研究》2021年第29期4644-4649,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81774339),项目参与人:唐宏宇;广东省中医药管理局资助项目(20191099),项目负责人:唐宏宇。

摘　　要：背景:肾俞、足三里为针刺治疗骨质疏松症的常用穴位,但其具体机制尚不明确。目的:基于骨组织Runt相关转录因子2启动子甲基化水平,探究电针肾俞、足三里等穴位对去卵巢骨质疏松症大鼠的疗效机制。方法:将50只SD大鼠随机分为假手术组、模型组、电针穴位组、电针非穴位组、雌二醇组,每组10只。假手术组以外的4组大鼠采用去卵巢法建立骨质疏松症模型,电针穴位组选取肾俞、足三里穴位进行电针治疗,电针非穴位组选取非穴位处进行干预,其余3组不进行电针干预;雌二醇组给予雌二醇片(0.09 mg/kg)灌胃治疗,假手术组和模型组均灌胃10 mL/kg的生理盐水。干预12周后,测定各组大鼠右侧下肢股骨及椎骨骨密度、股骨生物力学性能、血清骨钙与Ⅰ型原胶原N端前肽表达水平、骨组织骨桥蛋白与骨唾液蛋白表达水平、骨组织Runt相关转录因子2启动子甲基化表达水平。结果与结论:①与假手术组相比,模型组大鼠股骨、椎骨骨密度明显下降(P<0.05);股骨最大载荷、刚度及弹性模量、血清骨钙、Ⅰ型原胶原N端前肽水平均明显下降(P<0.01);骨组织中骨桥蛋白和骨唾液蛋白、Runt相关转录因子2启动子CpG1和CpG2、CpG3和CpG4.5的表达明显上升(P<0.01);②与模型组相比,电针穴位组和雌二醇组对上述各指标改善明显(P<0.01);而电针非穴位组骨组织中Runt相关转录因子2启动子CpG1、CpG2、CpG3、CpG4.5的表达下降不明显(P>0.05),此外其余上述指标均有所改善(P<0.05);③与雌二醇组相比,电针穴位组对血清Ⅰ型原胶原N端前肽水平的上调较为明显(P<0.05),而其余指标差异无显著性意义(P>0.05);④提示电针肾俞、足三里穴位可通过降低骨组织Runt相关转录因子2启动子甲基化水平,增强骨密度,改善股骨的生物力学性能等,从而有效防治骨质疏松症。BACKGROUND:Shenshu and Zusanli are commonly used acupuncture acupoints in the treatment of osteoporosis;however,the specific mechanism is still unclear.OBJECTIVE:To explore the therapeutic mechanism of electroacupuncture at Shenshu and Zusanli acupoints in ovariectomized rats with osteoporosis based on the methylation level of Runx2 promoter in bone tissue.METHODS:Fifty Sprague-Dawley rats were randomly divided into sham operation group,model group,electroacupuncture acupoint group,electroacupuncture non-acupoint group and estradiol group(n=10 per group).The model of osteoporosis was established by ovariectomy in all the groups except for sham operation group.Shenshu and Zusanli acupoints were selected for electroacupuncture treatment in the electroacupuncture acupoint group.Nonacupoint electroacupuncture was selected for intervention in the electroacupuncture non-acupoint group,while the other three groups did not receive electroacupuncture intervention.The estradiol group was given estradiol tablets(0.09 mg/kg)by gavage,and the sham operation group and the model group were given 10 mL/kg normal saline.Femoral and vertebral bone mineral density,biomechanical properties of the femur,serum bone calcium,type I tropocollagen n-terminal propeptide(P1NP),bone tissue osteopontin and bone sialoprotein levels,and Runx2 promoter methylation level in bone tissue were determined at 12 weeks after intervention.RESULTS AND CONCLUSION:Bone mineral density levels of the femur and vertebrae decreased significantly in the model group compared with the sham operation group(P<0.05).The maximum load,stiffness and elastic modulus of the femur,serum calcium,and P1NP levels were significantly decreased in the model group compared with the sham operation group(P<0.01).The osteopontin and bone sialoprotein levels and expressions of Runx2 promoters CpG1,CpG2,CpG3 and CpG4.5 in bone tissue were significantly increased in the model group compared with the sham operation group(P<0.01).Compared with the model group,electroacupuncture at acupo

关 键 词：电针 骨质疏松 表观遗传 甲基化 生物力学 大鼠 Runx2启动子 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]