摘要
目的探讨补肾益精方(BSYJF)通过骨髓神经元性凋亡囊泡对老年性骨质疏松(OP)小鼠的作用研究。方法取年轻和老年男性骨折患者骨髓,将两组骨髓培养上清液处理骨髓间充质干细胞(BMSCs),形态学检测碱性磷酸酶(ALP)、茜素红(ARS)和油红O染色,评价BMSCs成骨成脂分化能力,蛋白质组学分析骨髓上清液中神经元凋亡囊泡标志物L1-细胞粘附分子(L1CAM)、血小板反应蛋白(TH)、补体C3(C3b)的蛋白表达水平。取30只雄性C57BL/6J小鼠,随机分为空白组、造模组、BSYJF干预组,每组10只。造模及BSYJF干预组采用腹腔注射D-半乳糖(150 mg/kg·d^(-1),8周)制备OP小鼠模型,8周后BSYJF干预组每天灌胃0.40 g/10 kg BSYJF药液,采用等容生理盐水给空白组及造模组灌胃,共12周。检测血清保护素(OPG)、β-骨胶原交联(β-CTx)吸光度值,股骨做Micro CT扫描和骨组织形态学检测(HE染色)分析,实时荧光定量聚合酶链式反应分析(qPCR)神经元凋亡囊泡标志物表达水平。结果与年轻组比较,老年来源的骨髓上清液处理BMSCs ALP和ARS阳性染色明显减少,而脂肪细胞的油红O染色明显增强;老年组骨髓中神经元源性凋亡囊泡标志物L1CAM、TH、C3b表达明显增高。动物实验结果显示:与造模组比较,BSYJF组明显改善SOP小鼠血清OPG、β-CTx表达水平及骨组织形态;BSYJF组骨密度(BMD)、骨小梁数目(Tb.N)、骨小梁厚度(Tb.Th)、骨体积分数(BV/TV)均明显增加(P<0.05),骨小梁分离度(Tb.Sp)明显降低(P<0.05);BSYJF组则明显下调神经元凋亡囊泡标志物L1CAM、TH、C3b(P<0.01)。结论BSYJF能够提高SOP小鼠的BMD,促进其骨形成,改善OP,其机制可能与抑制神经元性凋亡囊泡有关。
Objective To investigate the effect of Bu Shen Yi Jing Fang(BSYJF)on senile osteoporosis(SOP)mice through bone marrow neuronal apoptosis vesicles.Methods Bone marrow samples were collected from young and elderly male fracture patients.Two groups of bone marrow culture supernatants were used to treat bone marrow mesenchymal stem cells(BMSCs).Morphological analysis was performed on alkaline phosphatase(ALP),alizarin red(ARS),and oil red O staining to evaluate the osteogenic and adipogenic differentiation ability of BMSCs.Proteomic analysis was performed on the protein expression levels of neuronal apoptosis vesicle markers L1 cell adhesion molecule(L1CAM),platelet reactive protein(TH),and complement C3(C3b)in the bone marrow supernatants.Thirty male C57BL/6J mice were randomly divided into a blank group,a modeling group,and a BSYJF intervention group,with 10 mice in each group.The SOP mouse model was prepared by intraperitoneal injection of D-galactose(150 mg/kg/d,8 weeks)for modeling and BSYJF intervention group.After 8 weeks,the BSYJF intervention group was orally administered with 0.40 g/10 kg BSYJF solution per day,and the blank group and modeling group were orally administered with equal volume physiological saline for 12 weeks.Detect the absorbance values of serum protective protein(OPG)andβ-collagen cross-linking(β-CTx),perform micro CT scanning and bone tissue morphology analysis(HE staining)on the femur,and analyze the expression levels of neuronal apoptosis vesicle markers using real-time fluorescence quantitative pdymerase chain reaction(qPCR).Results Compared with the younger group,the treatment of BMSCs with bone marrow supernatant from the elderly significantly reduced ALP and ARS positive staining,while the oil red O staining of adipocytes was significantly enhanced;The expression of neuronal apoptosis vesicle markers L1CAM,TH,and C3b in the bone marrow of the elderly group was significantly increased.The animal experiment results showed that compared with the modeling group,the BSYJF group significantly improved the serum OPG,β-CTx expression levels,and bone tissue morphology of SOP mice;The bone mineral density(BMD),number of trabeculae(Tb.N),thickness of trabeculae(Tb.Th),and bone volume fraction(BV/TV)in the BSYJF group were significantly increased(P<0.05),while the degree of trabecular separation(Tb.Sp)was significantly decreased(P<0.05);The BSYJF group significantly downregulated the neuronal apoptosis vesicle markers L1CAM,TH,and C3b(P<0.01).Conclusion The formula of tonifying kidney and nourishing essence can increase bone density,promote bone formation,and improve osteoporosis in SOP mice.Its mechanism may be related to the inhibition of neuronal apoptotic vesicles.
作者
叶俊星
李霞
杨晓斐
程飞
石树文
冯政
袁凤来
YE Junxing;LI Xia;YANG Xiaofei;CHENG Fei;SHI Shuwen;FENG Zheng;YUAN Fenglai(Department of Orthopedics,Affiliated Hospital of Jiangnan University,Wuxi 214000;Department of Pharmacy,Affiliated Hospital of Jiangnan University,Wuxi 214000;Department of Orthopedics,Jiangyin Second People's Hospital,Wuxi 214401;Institute of Integrated Traditional Chinese and Western Medicine,Affiliated Hospital of Jiangnan University,Wuxi 214000)
出处
《颈腰痛杂志》
2025年第5期838-843,共6页
The Journal of Cervicodynia and Lumbodynia
基金
江苏省中医药科技发展计划项目(MS2023069)
无锡市科技创新资金(Y20232018)
江南大学附属医院临床科研项目(LCYJ202307)。
关键词
骨质疏松
补肾益精方
骨髓
老年男性
小鼠
神经源性凋亡囊泡
osteoporosis
Bu Shen Yi Jing Fang
bone marrow
elderly males
mouse
neurogenic apoptotic vesicles
