摘要
目的探讨柴胡桂枝汤加减(CGD)调节Wnt/β-连环蛋白(β-catenin)信号通路对骨质疏松大鼠的改善作用。方法将SD大鼠分组为Model组、CK组、高剂量CGD组(CGD-H组,20 g/kg)、低剂量CGD组(CGD-L组,5 g/kg)、戊酸雌二醇组(EV组,9 mg/kg)、DKK-1组(Wnt/β-catenin通路抑制剂,100 mg/kg)和CGD-H+DKK-1组(20 g/kg+100 mg/kg),每组12只。除CK组外,其他组大鼠均通过灌胃70 mg/kg维甲酸的方式构建骨质疏松症(OP)大鼠模型,CK组大鼠灌胃等量的生理盐水。造模4周开始给予相应的药物干预4周。ELISA法检测大鼠血清中Ⅰ型胶-原C端肽(CTX-Ⅰ)、骨钙素(BGP)水平;观察股骨微观结构相关指标骨体积分数、骨小梁厚度、骨密度、骨小梁数量的变化;HE染色检测大鼠股骨病理变化;碱性磷酸酶(ALP)钙-钴染色检测大鼠股骨中成骨细胞活性;抗酒石酸酸性磷酸酶(TRAP)染色检测大鼠股骨组织中破骨细胞活性;Western blot检测各组大鼠股骨组织中Wnt3a、β-catenin蛋白。结果与CK组比较,Model组大鼠股骨组织病理损伤严重,CTX-Ⅰ水平、破骨细胞活性升高,BGP水平、骨体积分数、骨小梁厚度、骨密度、骨小梁数量、成骨细胞活性、Wnt3a、β-catenin蛋白表达降低(P<0.05);与Model组比较,CGD-L组、CGD-H组、EV组大鼠股骨组织病理损伤减轻,CTX-Ⅰ水平、破骨细胞活性降低,BGP水平、骨体积分数、骨小梁厚度、骨密度、骨小梁数量、成骨细胞活性、Wnt3a、β-catenin蛋白表达升高,而DKK-1组对应指标变化趋势与上述相反(P<0.05);与CGD-H组比较,CGD-H+DKK-1组大鼠股骨组织病理损伤严重,CTX-Ⅰ水平、破骨细胞活性升高,BGP水平、骨体积分数、骨小梁厚度、骨密度、骨小梁数量、成骨细胞活性、Wnt3a、β-catenin蛋白表达降低(P<0.05)。结论CGD可能通过激活Wnt/β-Catenin通路改善大鼠OP。
Objective To investigate the improving effect of modified Chaihu Guizhi Decoction(CGD)on osteoporosis rats by regulating the Wnt/β-catenin signaling pathway.Methods SD rats were assigned to Model,CK,high-dose CGD(CGD-H;20 g/kg),low-dose CGD(CGD-L;5 g/kg),and estradiol valerate(EV;9 mg/kg),Wnt/β-catenin pathway inhibitor(DKK-1,100 mg/kg),and CGD-H+DKK-1(20 g/kg+100 mg/kg)groups with 12 rats per group.Except for those in the CK group,rats were administered 70 mg/kg retinoic acid to establish the osteoporosis(OP)model,and rats in the CK group were administered the same amount of normal saline.From week 4 of modeling,the corresponding drug was administered for 4 weeks.ELISA were applied to measure serum levels of collagen type I C-terminal peptide(CTX-Ⅰ)and osteocalcin(BGP).Changes in the bone volume fraction,bone trabecular thickness,bone mineral density,and bone trabecular number were observed.HE staining was applied to assess pathological changes in the rat femur.Alkaline phosphatase(ALP)calcium-cobalt staining was applied to detect osteoblast activity in the rat femur.Osteoclast activity in rat femur tissue was detected by tartrate-resistant acid phosphatase(TRAP)staining.Western blot was applied to detect Wnt3a andβ-catenin proteins in femoral tissue.Results Compared with the CK group,femoral tissue of the Model group had severe pathological damage,the CTX-Ⅰlevel and osteoclast activity were increased,and the BGP level,bone volume fraction,bone trabecular thickness,bone mineral density,bone trabecular number,osteoblast activity,and Wnt3a andβ-catenin protein expression were decreased(P<005).Compared with the Model group,pathological damage of the femur in CGD-L,CGD-H,and EV groups was alleviated,the CTX-Ⅰlevel and osteoclast activity were decreased,the BGP level,bone volume fraction,trabecular thickness,bone mineral density,bone trabecular number,osteoblast activity,and Wnt3a andβ-catenin protein expression were increased,and the opposite trends of the corresponding indicators were observed in the DKK
作者
郭中华
史栋梁
曹玉举
王云飞
杨少祥
秦合伟
张仲博
任博文
周松林
王俊杰
王莉莎
万小冠
GUO Zhonghua;SHI Dongliang;CAO Yuju;WANG Yunfei;YANG Shaoxiang;QIN Hewei;ZHANG Zhongbo;REN Bowen;ZHOU Songlin;WANG Junjie;WANG Lisha;WAN Xiaoguan(Department 1 of Bone Diseases,Henan Hospital of Traditional Chinese Medicine,Zhengzhou 450053;China.2.Department of Orthopaedics,Zhengzhou Traditional Chinese Medicine Orthopedics Hospital,Zhengzhou 450016;.3.Department of Rehabilitation,Henan Hospital of Traditional Chinese Medicine,Zhengzhou 450053;.4.Department of Pain,Henan Hospital of Traditional Chinese Medicine,Zhengzhou 450053;.5.Department of Orthopaedics,Zhengzhou Ninth People’s Hospital,Zhengzhou 450053)
出处
《中国比较医学杂志》
CAS
北大核心
2023年第10期15-22,共8页
Chinese Journal of Comparative Medicine
基金
2021年河南省中医药拔尖人才培养项目资助(豫卫中医函(2021)15号)
河南省中医药科学研究专项课题(2021JDZX2121)。
