Slit3/Robo1通路探讨健骨颗粒对大鼠骨量丢失模型的影响  

Effect of Strong-bone Granules on rats with a bone loss model by modulating the Slit3/Robo1 pathway

作　　者：陈赛楠 黄云梅[1,2] 周芬 王志强 魏振朴 林燕萍[1,2] CHEN Sainan;HUANG Yunmei;ZHOU Fen;WANG Zhiqiang;WEI Zhenpu;LIN Yanping(Academy of Integrative Medicine,College of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Fujian Provincial Key Laboratory of Integrative Medicine on Geriatrics,Fuzhou 350122,China;College of Acupuncture and Moxibustion,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Key Research Section of Syndrome of Osteoporosis Genomics,Fujian Academy of Chinese Medical Sciences,Fuzhou 350003,China;Fujian Key Laboratory of Integrated Traditional Chinese and Western Medicine for Osteoporosis(Fujian Academy of Chinese Medical Sciences,Rehabilitation Hospital Affiliated to Fujian University of TCM),Fuzhou 350003,China)

机构地区：[1]福建中医药大学中西医结合学院中西医结合研究院,福建福州350122 [2]福建省中西医结合老年性疾病重点实验室,福建福州350122 [3]福建中医药大学针灸学院,福建福州350122 [4]福建省中医药科学院骨质疏松证候基因组学研究室,福建福州350003 [5]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),福建福州350003

出　　处：《中国骨质疏松杂志》2025年第3期313-320,共8页Chinese Journal of Osteoporosis

基　　金：国家自然科学基金资助项目(81904228、81574003);福建省自然科学基金项目(2023J01336);福建省财政专项资助项目(X2019002-财政专项);福建省属公益类科研院所基本科研专项(2022R1003003)。

摘　　要：目的观察健骨颗粒对去卵巢快速骨量丢失模型大鼠H型血管和Slit3/Robo1通路的影响,探讨其补虚化瘀的潜在作用机制。方法采用双侧卵巢切除术制备SD大鼠模型,灌胃干预6周,micro-CT观察胫骨近端微观结构,免疫荧光染色观察H型血管,ELISA检测血清CTX和BALP,Real-time PCR检测股骨Slit3和Robo1 mRNA的表达水平,免疫组织化学染色观察Slit3蛋白的表达水平,Western Blot检测Robo1蛋白的表达水平。结果模型组骨密度、骨小梁体积分数、骨小梁数量和骨小梁厚度均显著降低(P<0.01)和骨小梁分离度显著增大(P<0.001),H型血管阳性面积占比显著降低(P<0.001),Slit3 mRNA和蛋白表达水平显著升高(P<0.01),Robo1 mRNA和蛋白表达水平未见显著变化。与模型组相比,健骨颗粒组骨密度升高(P<0.05)和骨小梁分离度降低(P<0.001),H型血管阳性面积占比升高(P<0.05),血清CTX水平显著降低(P<0.001)、BALP水平略降低,Slit3 mRNA表达略下调和蛋白平均光密度值显著下降(P<0.05),Robo1 mRNA和蛋白表达水平显著升高(P<0.01)。结论健骨颗粒可通过Slit3/Robo1信号通路促进H型血管新生,提高骨量和改善骨微结构发挥抗骨质疏松的作用。Objective To investigate the impact of Strong-bong Granules on type H vessels and the Slit3/Robo1 pathway in ovariectomized rats with rapid bone loss,with the aim of elucidating its potential mechanism in promoting angiogenesis and osteogenesis.Methods A rat model was established through bilateral ovariectomy and oral administration of Strong-bone Granules for a duration of 6 weeks.The microstructure of the proximal tibia was examined using micro-CT.Immunofluorescence staining was employed to observe type H vessels.Serum CTX and BALP levels were measured using ELISA.The expression levels of Slit3 and Robo1 mRNA in the femur were assessed with real-time PCR.The immunohistochemistry staining was employed to observe the expression levels of Slit3 protein.Western blotting was used to detect the expression level of Robo1 protein.Results The model group exhibited a significant reduction in bone mineral density(BMD),bone volume fraction(BV/TV),trabecular number(Tb.N),and trabecular thickness(Tb.Th,P<0.01),accompanied by a significant increase in trabecular separation(Tb.Sp,P<0.001).Additionally,there was a notable decrease in the proportion of type H vessels positive area(P<0.001),along with a significant increase in the expression levels of Slit3 mRNA and protein(P<0.01).Conversely,no significant changes were observed in the expression levels of Robo1 mRNA and protein.Compared to the model group,the Strong-bone Granule group exhibited a statistically significant increase in BMD(P<0.05)and a significant decrease in Tb.Sp(P<0.001).Additionally,there was a significant increase in the proportion of type H vessels positive area(P<0.05),a significant decrease in serum CTX content(P<0.001),a slight decrease in BALP content,a slight down-regulation of Slit3 mRNA expression,a significant decrease in average protein optical density(P<0.05),and a significant increase in Robo1 mRNA and protein expression levels(P<0.01).Conclusion The administration of Strong-bone Granules has been found to stimulate type H vessels,to enhance bone

关 键 词：绝经后骨质疏松症 健骨颗粒 H型血管 Slit3/Robo1信号通路 

分 类 号：R965[医药卫生—药理学]