吉林大学学报(医学版) ›› 2024, Vol. 50 ›› Issue (1): 120-127.doi: 10.13481/j.1671-587X.20240115

• 基础研究 • 上一篇    

鹿茸多肽对骨质疏松模型大鼠的改善作用及对SIRT1/FOXO1信号通路的影响

迟雪婷1,黄晓巍1,2,陈芳园1,周高峰1,王晋冀1,律广富3,林喆1(),龚庆4()   

  1. 1.长春中医药大学药学院临床药学与中药药理教研室,吉林 长春 130117
    2.长春中医药大学东北亚中医药研究院基础研究所,吉林 长春 130117
    3.长春中医药大学 吉林省人参科学研究院中药药理组,吉林 长春 130117
    4.长春中医药大学附属医院骨科中心,吉林 长春 130021
  • 收稿日期:2023-03-02 出版日期:2024-01-28 发布日期:2024-01-31
  • 通讯作者: 林喆,龚庆 E-mail:linzhe1228@163.com;272632635@qq.com
  • 作者简介:迟雪婷(1999-),女,吉林省松原市人,在读硕士研究生,主要从事心血管和内分泌药理学方面的研究。
  • 基金资助:
    吉林省科技厅发展计划项目(20210101200JC);长春中医药大学大学生创新训练基金项目(202210199049);吉林省发改委创新能力建设项目(2021C011)

Improvement effect of velvet antler polypeptide in osteoporosis model rats and its effect on SIRT1/FOXO1 signaling pathway

Xueting CHI1,Xiaowei HUANG1,2,Fangyuan CHEN1,Gaofeng ZHOU1,Jinji WANG1,Guangfu LYU3,Zhe LIN1(),Qing GONG4()   

  1. 1.Department of Clinical Pharmacy and Pharmacology of Chinese Medicine,School of Pharmaceutical Sciences,Changchun University of Chinese Medicine,Changchun 130117,China
    2.Basic Research Institute,Northeast Asia Institute of Traditional Chinese Medicine,Changchun University of Traditional Chinese Medicine,Changchun 130117,China
    3.Department of Pharmacology of Traditional Chinese Medicine,Jilin Ginseng Academy,Changchun University of Traditional Chinese Medicine,Changchun 130117,China
    4.Orthopedic Center,Affiliated Hospital,Changchun University of Chinese Medicine,Changchun 130021,China
  • Received:2023-03-02 Online:2024-01-28 Published:2024-01-31
  • Contact: Zhe LIN,Qing GONG E-mail:linzhe1228@163.com;272632635@qq.com

摘要:

目的 探讨鹿茸多肽(VAP)对骨质疏松(OP)模型大鼠的保护作用,并阐明其可能的作用机制。 方法 60只12周龄SD大鼠随机分为对照组、模型组、阳性药组(1 mg·kg-1·d-1阿仑膦酸钠灌胃)、低剂量(100 mg·kg-1·d-1)VAP组、中剂量(200 mg·kg-1·d-1)VAP组和高剂量(300 mg·kg-1·d-1)VAP组,每组10只。除对照组外,其余各组大鼠肌肉注射地塞米松(2 mg·kg-1)复制OP大鼠模型,对照组大鼠肌肉注射等体积生理盐水,每周2次,连续11周。双能X射线骨密度仪检测各组大鼠股骨骨密度(BMD),酶联免疫吸附试验(ELISA)法检测各组大鼠血清中血钙(Ca2+)、血磷(P)、骨保护素(OPG)、碱性磷酸酶(ALP)和骨钙素(OCN)水平,生化法检测各组大鼠血清中丙二醛(MDA)水平和超氧化物歧化酶(SOD)活性,HE染色观察各组大鼠骨组织病理形态表现,Western blotting法检测各组大鼠骨组织中沉默信息调节因子1(SIRT1)、过氧化氢酶(CAT)、Runt相关转录因子2(RUNX2)和叉头框蛋白O1(FOXO1)蛋白表达水平。 结果 与对照组比较,模型组大鼠股骨BMD明显降低(P<0.05);与模型组比较,阳性药组、中剂量VAP组和高剂量VAP组大鼠股骨BMD明显升高(P<0.05或P<0.01)。与对照组比较,模型组大鼠血清中Ca2+、P和OPG水平及SOD活性明显降低(P<0.05),ALP、OCN和MDA水平明显升高(P<0.05);与模型组比较,低剂量VAP大鼠血清中OPG水平明显升高(P<0.05),阳性药组、中剂量VAP组和高剂量VAP组大鼠血清中Ca2+、P和OPG水平及SOD活性明显升高(P<0.05或P<0.01),阳性药组和各剂量VAP组大鼠血清中ALP、OCN和MDA水平明显降低(P<0.05或 P<0.01)。HE染色,与对照组比较,模型组大鼠骨组织中骨细胞数量减少且排列混乱,骨小梁纤细,出现大片断裂,髓腔扩大;与模型组比较,阳性药组、中剂量VAP组和高剂量VAP组大鼠骨组织中骨小梁粗壮,排列紧密。Western blotting法,与对照组比较,模型组大鼠骨组织中SIRT1、CAT、RUNX2和FOXO1蛋白表达水平明显降低(P<0.05);与模型组比较,阳性药组、中剂量VAP组和高剂量VAP组大鼠骨组织中SIRT1、CAT、RUNX2和FOXO1蛋白表达水平明显升高(P<0.05或P<0.01)。 结论 VAP对OP模型大鼠具有保护作用,其作用机制可能与介导SIRT1/FOXO1信号通路抗氧化应激作用有关。

关键词: 骨质疏松症, 地塞米松, 鹿茸多肽, 氧化应激, 沉默信息调节因子1, 叉头框蛋白O1, 信号通路

Abstract:

Objective To discuss the protective effect of velvet antler peptide (VAP) in the osteoporosis (OP) model rats,and to clarify the possible mechanism. Methods Sixty 12-week-old SD rats were randomly divided into control group, model group, positive drug group (treated with 1 mg·kg-1·d-1 of alendronate sodium by gavage), low dose of VAP group (treated with 100 mg·kg-1·d-1 VAP), medium dose of VAP group (treated with 200 mg·kg-1·d-1 VAP), and high dose of VAP group (treated with 300 mg·kg-1·d-1 VAP), and there were ten rats in each group. Except for control group,the rats in the other groups were injected with dexamethasone (2 mg·kg-1) to replicate the OP rat model, while the rats in control group were injected with the equivalent volume of saline twice a week for 11 consecutive weeks. Dual-energy X-ray absorptiometry was used to detect the bone mineral density (BMD) of femur tissue of the rats in various groups;enzyme-linked immunosorbent assay (ELISA) method was used to detect the levels of serum calcium (Ca2+), phosphate (P), osteoprotegerin (OPG), alkaline phosphatase (ALP), and osteocalcin (OCN)in serum of the rats in various groups; biochemical method was used to detect the malondialdehyde (MDA) level and superoxide dismutase (SOD) activity in serum of the rats in various groups; HE staining was used to observe the pathomorphology of bone tissue of the rats in various groups; Western blotting method was used to detect the expression levels of silent information regulator 1 (SIRT1), catalase (CAT), Runt-related transcription factor 2 (RUNX2), and forkhead box protein O1 (FOXO1) proteins in bone tissue of the rats in various groups. Results Compared with control group, the BMD of femoral tissue of the rats in model group was decreased (P<0.05); compared with model group, the BMD of femur tissue of the rats in positive drug group, medium dose of VAP group, and high dose of VAP group were increased (P<0.05 or P<0.01). Compared with control group, the levels of Ca2+, P, OPG, and SOD activities in serum of the rats in model group were decreased (P<0.05), and the levels of ALP, OCN, and MDA were increased (P<0.05); compared with model group, the level of OPG in serum of the rats in low dose of VAP group was significantly increased(P<0.05),the levels of Ca2+, P, OPG, and activities of SOD in serum of the rats in positive drug group, medium dose of VAP group, and high dose of VAP group were significantly increased (P<0.05 or P<0.01), and the levels of ALP, OCN, and MDA in serum of the rats in positive drug group and different doses of VAP groups were decreased (P<0.05 or P<0.01). The HE staining results showed that compared with control group, the rats in model group had fewer bone cells and disordered arrangements in the bone tissue, thinner bone trabeculae with large fractures, and an expanded marrow cavity; compared with model group, the rats in positive drug group, medium dose of VAP group, and high dose of VAP group had thicker bone trabeculae arranged more tightly. The Western blotting results showed that compared with control group, the expression levels of SIRT1, CAT, RUNX2, and FOXO1 proteins in bone tissue of the rats in model group were decreased (P<0.05); compared with model group, the expression levels of SIRT1, CAT, RUNX2, and FOXO1 proteins in bone tissue of the rats in positive drug group, medium dose of VAP group, and high dose of VAP group were significantly increased (P<0.05 or P<0.01). Conclusion VAP has the protective effect against OP in the rats, and its mechanism may be related to mediating the antioxidant stress action through the SIRT1/FOXO1 signaling pathway.

Key words: Osteoporosis, Dexamethasone, Velvet antler polypeptide, Oxidative stress, Silent information regulator 1, Forkhead box protein O1, Signaling pathway

中图分类号: 

  • R285.5