摘要
目的:探索补肾活血方调节自噬对去势骨质疏松小鼠H型血管及骨形成的作用。方法:将C57BL/6J雌鼠随机分为假手术组、骨质疏松组和补肾活血方组,补肾活血方组在去势骨质疏松造模成功后以补肾活血方灌胃,假手术组及骨质疏松组以等量生理盐水灌胃,1次/d,连续给药8周。采用Micro-CT检测骨组织形态,免疫荧光检测EMCN、CD31共表达情况,免疫组化检测骨组织LC3和P62表达水平,荧光定量PCR检测骨组织内RUNX2、COL1A1、LC3及P62基因表达水平,Western Blot法检测骨组织内RUNX2、COL1A1、LC3及P62蛋白表达水平。结果:与假手术组相比,骨质疏松组骨组织BV/TV、Tb.N、Tb.Th明显降低,差异有统计学意义(P<0.001);EMCN和CD31共表达面积减少,差异有统计学意义(P<0.001);LC3阳性表达减少,而P62阳性表达增加,RUNX2、COL1A1和LC3基因及蛋白表达水平降低,差异有统计学意义(P<0.001);而P62基因及蛋白表达水平增高,差异有统计学意义(P<0.001)。与骨质疏松组相比,补肾活血方组骨组织BV/TV、Tb.N、Tb.Th升高,差异有统计学意义(P<0.001);EMCN和CD31共表达面积增加,差异有统计学意义(P<0.001);LC3阳性表达增加,而P62阳性表达减少,RUNX2、COL1A1和LC3基因及蛋白表达水平增高,差异有统计学意义(P<0.001);而P62基因及蛋白表达水平降低,差异有统计学意义(P<0.001)。结论:补肾活血方能够提高绝经后骨质疏松小鼠骨组织内自噬水平,增加骨组织内H型血管形成,促进骨组织的形成,起到治疗绝经后骨质疏松的作用。
Objective:To investigate the effects of Bushen Huoxue formula on type H vessels and bone formation through autophagy modulation in ovariectomized osteoporotic mice.Methods:C57BL/6J female mice were randomly divided into sham operation group,osteoporosis group,and Bushen Huoxue formula group.After successful modeling of ovariectomy induced osteoporosis,the Bushen Huoxue formula group received oral gavage with Bushen Huoxue formula.The sham operation group and osteoporosis group received oral gavage with an equal volume of physiological saline.Administer once a day for 8 consecutive weeks.Micro-CT was used to detect bone tissue morphology.Immunofluorescence was used to detect the co-expression of EMCN and CD31.Immunohistochemistry was used to detect the expression levels of LC3 and P62 in bone tissue.Fluorescence quantitative PCR was used to detect the expression levels of RUNX2,COL1A1,LC3 and P62 genes in bone tissue.Western Blot was used to detect the protein expression levels of RUNX2,COL1A1,LC3 and P62 in bone tissue.Results:Compared with sham operation group,the bone tissue BV/TV,Tb.N,Tb.Th in the osteoporosis group were significantly reduced(P<0.001),the number of osteoclasts was increased(P<0.001),the co-expression area of EMCN and CD31 was reduced(P<0.001),the positive expression of LC3 was reduced,and the positive expression of P62 was increased.The expression levels of RUNX2,COL1A1,LC3 genes and proteins were reduced(P<0.001),while the expression levels of P62 genes and proteins were increased(P<0.001).Compared with osteoporosis group,the bone tissue BV/TV,Tb.N,Tb.Th in the Bushen Huoxue formula group increased(P<0.001),the co-expression area of EMCN and CD31 increased(P<0.001),the positive expression of LC3 increased,while the positive expression of P62 decreased.The expression levels of RUNX2,COL1A1,LC3 genes and proteins increased(P<0.001),while the expression levels of P62 genes and proteins decreased(P<0.001).Conclusion:The Bushen Huoxue formula can ameliorate postmenopausal osteoporosis through upregulating autophagy activity in bone tissue,promoting angiogenesis of type H vessels,and enhancing osteogenic formation.
作者
胡劲涛
阮立奇
钱剑胜
王振威
倪月明
胡淼锋
HU Jintao;RUAN Liqi;QIAN Jiansheng;WANG Zhenwei;NI Yueming;HU Miaofeng(Hangzhou Traditional Chinese Medicine Hospital Affiliated to Zhejiang Chinese Medical University,Hangzhou 310007,China;Jiangnan Hospital Affiliated to Zhejiang Chinese Medical University,Hangzhou 311201,China;Hangzhou Fuyang Hospital of Traditional Chinese Medicine Orthopedics and Traumatology,Hangzhou 311400,China)
出处
《中国中医骨伤科杂志》
2025年第8期18-23,共6页
Chinese Journal of Traditional Medical Traumatology & Orthopedics
基金
浙江省中医药科技计划项目(2022ZB232,2024ZR119)
杭州市医药卫生科技项目(A20220507,20220919Y045)
浙江省医药卫生科技计划项目(2023RC068)。
关键词
绝经后骨质疏松
补肾活血方
自噬
H型血管
骨形成
postmenopausal osteoporosis
Bushen Huoxue formula
autophagy
type H vessels
bone formation
