柴胡桂枝汤加减通过调节Wnt/ β-catenin 信号通路对骨质疏松大鼠的改善作用及机制研究
作者:
作者单位:

1.河南省中医院骨病一科,郑州 450053;2.郑州中医骨伤病医院骨科,郑州 450016;3.河南省中医院康复科,郑州 450053;4.河南省中医院疼痛科,郑州 450053;5.郑州市第九人民医院骨科,郑州 450053

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R-33


Improving effect and mechanism of modified Chaihu Guizhi Decoction on osteoporosis rats by regulating the Wnt / β-catenin signaling pathway
Author:
Affiliation:

1.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

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    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。

    Abstract:

    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 and β-catenin proteins in femoral tissue. Results Compared with the CK group, femoral tissue of the Model group had severe pathological damage, the CTX-Ⅰ level 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 and β-catenin protein expression were decreased (P< 0. 05). Compared with the Model group, pathological damage of the femur in CGD-L, CGD-H, and EV groups was alleviated, the CTX-Ⅰ level and osteoclast activity were decreased, the BGP level, bone volume fraction, trabecular thickness, bone mineral density, bone trabecular number, osteoblast activity, and Wnt3a and β-catenin protein expression were increased, and the opposite trends of the corresponding indicators were observed in the DKK-1 group (P<0. 05). Compared with the CGD-H group, femoral tissue of the CGD-H+DKK-1 group was severely damaged, the CTX-Ⅰ level 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 and β-catenin protein expression were decreased (P<0. 05). Conclusions CGD may improve OP in rats by activating the Wnt/ β-catenin pathway.

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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.柴胡桂枝汤加减通过调节Wnt/ β-catenin 信号通路对骨质疏松大鼠的改善作用及机制研究[J].中国比较医学杂志,2023,33(10):15~22.

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  • 收稿日期:2022-11-17
  • 在线发布日期: 2023-12-29
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