血根碱通过激活分泌型糖蛋白/β-链蛋白信号通路促进脊柱骨折大鼠骨折愈合的实验研究  

Experimental Study of Sanguinarine Promoting Fracture Healing in Spinal Fracture Rats by Activating Secretory Glikoproteinas-3a/β-Catenin Signaling Pathway

作　　者：王慧 潘茂圣 WANG Hui;PAN Maosheng(Department of Orthopedics,Suzhou Xiangcheng People's Hospital,Suzhou 215000,Jiangsu China)

机构地区：[1]苏州市相城人民医院骨科,江苏苏州215000

出　　处：《中国中医骨伤科杂志》2023年第8期13-18,共6页Chinese Journal of Traditional Medical Traumatology & Orthopedics

摘　　要：目的:探讨血根碱(SAG)对脊柱骨折大鼠骨折愈合的影响及其作用机制。方法:选用SD大鼠72只,随机分为6组(12只/组):假手术组、模型组、血根碱低剂量组(10 mg/kg)、血根碱中剂量组(20 mg/kg)、血根碱高剂量组(40 mg/kg)、抑制剂组(Wnt/β-catenin通路抑制剂,Wnt-C59,30 mg/kg)。构建脊柱骨折大鼠模型,造模成功后,灌胃相应药物,1次/d,连续4周。BBB评分法进行神经功能评定;采用微计算机断层扫描技术(Micro CT)观察大鼠骨骼微结构;苏木精-伊红(HE)染色观察大鼠脊髓病理损伤;TUNEL染色观察大鼠骨髓神经细胞凋亡情况;酶联免疫吸附(ELISA)法检测大鼠血清骨钙素(OC)和Ⅰ型胶原交联C-末端肽(CTX-Ⅰ)、骨形态发生蛋白2(BMP-2)的含量变化;Western Blot法检测受损脊髓中Bcl-2相关X蛋白(Bax)、碱性磷酸酶(ALP)及Wnt/β-catenin信号通路蛋白表达。结果:与假手术组比较,模型组大鼠BBB评分,骨密度,BV/TV、Tb.N、Tb.Th、Ct.Th、OC、BMP-2含量,ALP、Wnt-3a、β-catenin蛋白表达水平降低,脊髓病理损伤严重,神经细胞凋亡指数、CTX-Ⅰ含量、Bax、GSK-3β蛋白表达水平升高,差异有统计学意义(P<0.05);与模型组比较,血根碱低、中、高剂量组大鼠BBB评分,骨密度,BV/TV、Tb.N、Tb.Th、Ct.Th、OC、BMP-2含量,ALP、Wnt-3a、β-catenin蛋白表达水平升高,脊髓病理损伤减轻,神经细胞凋亡指数,CTX-Ⅰ含量,Bax、GSK-3β蛋白表达水平降低,差异有统计学意义(P<0.05);与血根碱高剂量组比较,抑制剂组大鼠BBB评分,骨密度,BV/TV、Tb.N、Tb.Th、Ct.Th、OC、BMP-2含量,ALP、Wnt-3a、β-catenin蛋白表达水平降低,脊髓病理损伤严重,神经细胞凋亡指数,CTX-Ⅰ含量,Bax、GSK-3β蛋白表达水平升高,差异有统计学意义(P<0.05)。结论:血根碱可促进成骨细胞的分化、抑制破骨细胞活性,起到促进脊柱骨折大鼠骨折愈合及改善神经损伤的作用,其作用机制可能与激活Wnt/β-catenin信号通路有关。Objective:To explore the effect and potential mechanism of sanguinarine(SAG)on fracture healing in rats with spinal fractures.Methods:72 SD rats were randomly divided into 6 groups,with 12 rats in each group:sham group,model group,SAG low-dose group(10 mg/kg),SAG medium-dose group(20 mg/kg),SAG high-dose group(40 mg/kg),and inhibitor group(Wnt/β-catenin pathway inhibitor,Wnt-C59,30 mg/kg).The spinal fracture rat model was established,and the corresponding drugs were administered once a day for 4 weeks after successful model establishment.The nerve function was evaluated using the Basso,Beattie,and Bresnahan(BBB)scoring method.Micro-CT was utilized to observe the bone microstructure,while hematoxylin-eosin(HE)staining was employed to assess the pathological injury of the spinal cord.TUNEL staining was used to observe the apoptosis of nerve cells in the rat bone marrow.The levels of serum osteocalcin(OC),C-terminal telopeptides of typeⅠcollagen(CTX-Ⅰ),and bone morphogenetic protein 2(BMP-2)were measured using enzyme-linked immunosorbent assay(ELISA).Western Blot analysis was conducted to detect the expression of Bcl-2-associated X protein(Bax),alkaline phosphatase(ALP),and Wnt/β-catenin signal pathway proteins in the injured spinal cord.Results:Compared with the sham group,the model group exhibited decreased BBB scores,bone mineral density(BMD),bone volume fraction(BV/TV),trabecular number(Tb.N),trabecular thickness(Tb.Th),cortical thickness(Ct.Th),OC,BMP-2 contents,ALP,Wnt-3a,andβ-catenin protein expression levels.Additionally,the pathological injury of the spinal cord was severe,and the apoptosis index of nerve cells,CTX-Ⅰcontent,Bax,and GSK-3βprotein expression levels were increased(P<0.05).In contrast,the SAG low,medium,and high-dose groups showed increased BBB scores,BMD,BV/TV,Tb.N,Tb.Th,Ct.Th,OC,BMP-2 contents,ALP,Wnt-3a,andβ-catenin protein expression levels compared to the model group.The pathological injury of the spinal cord was alleviated,and the apoptosis index of nerve cells,CTX-Ⅰcontent,B

关 键 词：血根碱 分泌型糖蛋白/β-链蛋白 脊柱骨折愈合 神经损伤 

分 类 号：R-33[医药卫生]