文献类型：期刊文献

中文题名：固本增骨颗粒对骨质疏松大鼠模型Wnt/β-catenin信号通路的研究

英文题名：Effect of Gubenzenggu Granule on Wnt/β-catenin signaling pathway in osteoporosis rat model

作者：齐雅茜[1];宁浩驰[1];潘静[1];赵瑞[1];雷新宇[1];宋敏[1];张浩令[2];张忠文[3];王薇[4];宋志靖[1,5]

第一作者：齐雅茜

机构：[1]甘肃中医药大学中医临床学院,甘肃兰州730000;[2]马来西亚理科大学高级医学与牙科研究所,马来西亚槟城132003;[3]甘肃中医药大学公共卫生学院,甘肃兰州730000;[4]甘肃中医药大学针灸推拿学院,甘肃兰州730000;[5]教育部敦煌医学与转化重点实验室,甘肃兰州730000

第一机构：甘肃中医药大学中医临床学院

年份：2024

卷号：30

期号：1

起止页码：37

中文期刊名：中国骨质疏松杂志

外文期刊名：Chinese Journal of Osteoporosis

收录：CSTPCD;;北大核心:【北大核心2023】；CSCD:【CSCD2023_2024】；

基金：国家自然科学基金(81960877);甘肃省高等学校创新基金(2021A-076);甘肃省科技计划(创新基地和人才计划)项目(21JR7RA561);敦煌医学与转化教育部重点实验室开放基金项目(DHYX20-16);甘肃省中医药研究中心专项开放课题(zyzx-2020-zx10);甘肃省自然基金(21JR1RA267);甘肃省教育科技创新项目(2022A-067);甘肃省高等学校创新基金(2023A-088);甘肃省自然科学基金(22JR5RA582)。

语种：中文

中文关键词：Wnt/β-catenin信号通路;骨质疏松症;固本增骨颗粒;成骨作用

外文关键词：Wnt/β-catenin signaling pathway;osteoporosis;Gubenzenggu granules;osteogenesis

摘要：目的研究固本增骨颗粒防治骨质疏松症的作用机制,揭示固本增骨颗粒通过Wnt/β-catenin信号通路抑制骨质疏松的分子机制。方法随机将66只大鼠分为假手术组13只和造模组53只,造模组采取背部双侧卵巢切除法制备模型,假手术组不摘除卵巢,造膜8周后将每组进行称重,并随机选取3只进行骨密度检测,待模型鉴定成功后,将造模组随机分为模型组、固本增骨颗粒高、中、低剂量组和阳性药物组,每组10只,固本增骨颗粒高、中、低剂量组分别按24.4、12.2、6.1 g/kg不同浓度进行灌胃,阳性药物组给予0.09 mg/kg浓度的雌二醇进行灌胃,假手术组及模型组给予生理盐水灌胃,各组大鼠的每日灌胃体积均为2 mL。每周记录各组大鼠的行为,给药干预12周后将大鼠处死股骨取材,进行骨组织形态学HE染色,通过IHC检测股骨组织中Wnt7b、TCF3、GSK3β蛋白含量。运用RT-PCR和WB测定股骨组织中Wnt7b、TCF3、GSK3βmRNA及其蛋白含量的表达。结果HE染色显示假手术组骨结构正常,小梁骨厚度均匀,结构完整,小梁连接良好,脂肪细胞和骨细胞形态清晰;模型组股骨骨小梁变细不规则、网状结构杂乱无章,部分骨小梁明显细长或断裂,两者连接不完全,固本增骨颗粒干预后,固本增骨颗粒治疗组大鼠骨小梁结构逐渐趋向成熟,厚度有所增加,逐渐均匀化,骨细胞增多,骨小梁与周围组织连接紧密,连续性逐渐变好,整体结构逐渐转好。与假手术组相比,模型组大鼠股骨组织中Wnt7b、TCF3表达量显著降低(P<0.01),GSK3β表达量显著升高(P<0.01);与模型组比较,阳性药物组、固本增骨颗粒高、中剂量组Wnt7b、TCF3表达量均升高(P<0.05),GSK3β表达量均下降(P<0.05)。结论固本增骨颗粒可通过调控Wnt/β-catenin信号通路,上调Wnt7b、TCF3的表达以促进骨形成,下调GSK3β的表达防止骨过度吸收。
Objective To study the mechanism of Gubenzenggu granules in preventing and treating osteoporosis,and to reveal the molecular mechanism of Gubenzenggu granules inhibiting osteoporosis through Wnt/β-catenin signaling pathway.Methods A total of 66 rats were randomly divided into sham operation group(13 rats)and modeling group(53 rats).The modeling group underwent bilateral back ovariectomy to prepare the model,while the sham operation group did not remove the ovaries.Each group was weighed 8 weeks after membrane construction,and 3 rats were randomly selected for bone mineral density detection.The modeling group was randomly divided into model group,Gubenzenggu granule high,medium and low dose groups and positive drug group,with 10 in each group.Gubenzenggu granule was given gavage at different concentrations of 24.4 g/kg,12.2 g/kg and 6.1 g/kg at high,medium and low doses,respectively.The positive drug group was given estradiol at 0.09 mg/kg concentration by gavage,and the sham operation group and model group were given normal saline gavage.The daily gavage volume of rats was 2 ml.The behavior of rats in each group was recorded every week.After 12 weeks of intervention,the femurs were sacrificed,and bone morphology was stained with HE,and the protein contents of Wnt7b,TCF3 and GSK3βin the femur tissues were detected by IHC.The mRNA and protein levels of Wnt7b,TCF3 and GSK3βin femur tissues were determined by RT-PCR and WB.Results HE staining showed that the bone structure of the sham operation group was normal,the trabecular bone thickness was uniform,the structure was complete,the trabecular connection was good,and the morphology of fat cells and bone cells was clear.In the model group,bone trabeculae of femur became thiner and irregular with a disorganized mesh structure,and some bone trabeculae were obviously elongated or fractured,with incomplete connection between the two.After the intervention of Gubenzenggu Granule,the bone trabecular structure of rats in the Gubenzenggu granule treatment group gradually tended to mature,the thickness increased,and the bone cells increased.The bone trabeculae were closely connected to the surrounding tissues,and the continuity gradually improved.The overall structure gradually improved.Compared with sham operation group,the expressions of Wnt7b and TCF3 in femur tissue of model group were significantly decreased(P<0.01),while the expression of GSK3βwas significantly increased(P<0.01).Compared with model group,the expression levels of Wnt7b and TCF3 in positive drug group,Gubenzenggu granule high dose and medium dose groups were increased(P<0.05),and the expression levels of GSK3βwere decreased(P<0.05).Conclusion Gubenzenggu granules can regulate the Wnt/β-catenin signaling pathway,up-regulate the expression of Wnt7b and TCF3 to promote bone formation,down-regulate the expression of GSK3βto prevent bone over-absorption.