摘要
目的探讨温肾通络止痛方对老年性骨质疏松(SOP)小鼠模型的干预作用和机制。方法将60只6月龄雄性C57BL/6J小鼠随机分为对照组、模型组、复方低、中、高剂量组及阿仑膦酸钠组,每组10只。采用腹腔注射D-半乳糖(每天150 mg/kg,8周)制备SOP小鼠模型,8周后复方低、中、高剂量组每天灌胃0.105、0.210、0.420 g/10 kg温肾通络止痛方,对照组及模型组灌胃等体积生理盐水,阿仑膦酸钠组每周灌胃1.53 mg/kg阿仑膦酸钠,共12周。酶联免疫吸附分析(ELISA)检测血清中骨保护素(OPG)和β-骨胶原交联(β-CTX)的水平,采用苏木素-伊红(HE)染色法观察胫骨组织病理学变化,micro-CT检测小鼠股骨骨密度(BMD)、骨体积分数(BV/TV)、骨小梁(TB)数、TB厚度和TB分离度;Western Blot法检测小鼠股骨骨髓中p16、p53、哺乳动物ATG6同源蛋白(Beclin-1)、微管相关蛋白1轻链3-Ⅱ(LC3-Ⅱ)、磷酸化腺苷酸活化蛋白激酶(p-AMPK)、AMPK、磷酸化哺乳动物雷帕霉菌靶蛋白(p-m TOR)、m TOR的蛋白表达水平。结果与对照组比较,模型组小鼠血清中OPG水平显著下降,β-CTX水平升高(P<0.05,P<0.01);小鼠TB数量减少,间隙增宽变细多断裂;Micro-CT扫描结果显示,模型组小鼠股骨组织中TB数量、TB厚度、BMD、BV/TV、TB模式因子均显著降低(P<0.05,P<0.01),TB分离度显著升高(P<0.01);模型组小鼠股骨中的衰老相关标记蛋白p16、p53、p-mTOR/mTOR的比值表达均显著升高(P<0.05),自噬相关蛋白Beclin-1、LC3-Ⅱ、p-AMPK/AMPK比值下降均降低(P<0.01)。与模型组比较,复方中、高剂量组及阿仑膦酸钠组SOP模型小鼠血清中OPG水平增高,β-CTX水平下降(P<0.05,P<0.01);股骨中的衰老相关标记蛋白p16、p53蛋白表达明显降低(P<0.05,P<0.01);Beclin-1、LC3-Ⅱ蛋白表达显著升高(P<0.05,P<0.01);复方高剂量组及阿仑膦酸钠组TB数量、TB厚度、BMD、BV/TV、TB模式因子均显著增加(P<0.05),TB分离度显著降低(P<0.05)。p-AMPK/AMPK�
Objective To study the intervention effects and mechanism of Wenshen Tongluo Zhitong Recipe(WTZR)on senile osteoporosis(SOP)model mice.Methods Sixty 6-month-old male C57BL/6J mice were randomly divided into six groups,i.e.,control group,model group,low-dose WTZR group,medium-dose WTZR group,high-dose WTZR group,and Alendronate Sodium group,10 mice in each group.The SOP mouse model was established by intraperitoneal injection of D-galactose(150 mg/kg per day for 8 weeks).After 8 weeks,mice in low,medium,and high dose WTZR groups were orally administered with 0.105,0.210,and 0.420 g/10 kg WTZR by gastrogavage every day.Mice in the control group and model group were administered with equal volume of physiological saline by gastrogavage.Mice in Alendronate Sodium group were administered with 1.530mg/kg Alendronate Sodium by gastrogavage every week.All administration lasted for a total of 12 weeks.Enzyme-linked immunosorbent assay(ELISA)was used to measure the serum levels of bone calmodulin-associated protein(OPG)and β-bone collagen cross-linked(β-CTX).Histopathological changes of the tibia were observed by HE staining.micro-CT was used to detect bone mineral density(BMD),bone volume fraction(BV/TV),trabecular(TB)number,TB thickness,and TB separation.Western Blot was used to detect the expression levels of p16,p53,Beclin-1,and microtubule-associated protein 1 in mouse femur bone marrow,as well as the levels of microtubule associated protein light chain 3-Ⅱ(LC3-Ⅱ),phosphorylated adenylate activated protein kinase(p-AMPK),AMP-activated protein kinase(AMPK),phosphorylated mammalian target protein of Rapa mold(p-mTOR),and mTOR protein.Results Compared with the control group,the serum level of OPG significantly decreased,while the level of β-CTX increased(P<0.05,P<0.01).The TB number decreased,TB thickness widened,the gap widened,narrowed,and broke more frequently.micro-CT scan results showed a significant reduction in the number of TB,TB thickness,BMD,BV/TV,and TB pattern factor in the femur tissue of the model gr
作者
牛园园
张天驰
李沐哲
郭杨
王礼宁
尹恒
关钛元
马勇
NIU Yuan-yuan;ZHANG Tian-chi;LI Mu-zhe;GUO Yang;WANG Li-ning;YIN Heng;GUAN Tai-yuan;MA Yong(Laboratory for Novel Repairing and Reconstructive Technology of Orthopedics and Traumatology,Nanjing University of Traditional and Western Medicine,Nanjing(210000);School of Integrated Chinese and Western Medicine,School of TCM,Nanjing University of Traditional and Western Medicine,Nanjing(210000);Wuxi Affiliated Hospital of Nanjing University of Traditional Chinese Medicine,Jiangsu Province Traditional Chinese Medicine Retrogressive Osteoarthritis Clinical Medical Innovation Center,Jiangsu(214000))
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2024年第1期84-90,共7页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然科学基金面上项目(No.82174411)
江苏省自然科学青年基金项目(No.BK20220470)
江苏省中医退行性骨关节病临床医学创新中心资助项目(No.苏中医科教[2021]4号)。
