[1]万文渊,罗娟△,彭真灵,等.龟甲胶对绝经后骨质疏松大鼠骨代谢的影响及机制研究[J].中国中医骨伤科杂志,2024,32(04):18-23.[doi:10.20085/j.cnki.issn1005-0205.240404]
　WAN Wenyuan,LUO Juan,PENG Zhenling,et al.Efficacy and Mechanism of Tortoise Shell Glue on Bone Metabolism in Postmenopausal Osteoporosis Rats[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2024,32(04):18-23.[doi:10.20085/j.cnki.issn1005-0205.240404]
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龟甲胶对绝经后骨质疏松大鼠骨代谢的影响及机制研究()
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《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第32卷
期数:
2024年04期
页码:
18-23
栏目:
实验研究
出版日期:
2024-04-01

文章信息/Info

Title:
Efficacy and Mechanism of Tortoise Shell Glue on Bone Metabolism in Postmenopausal Osteoporosis Rats
文章编号:
1005-0205(2024)04-0018-06
作者:
万文渊1; 罗娟1△; 彭真灵1; 朱莹莹1; 罗杰1
1株洲市中医伤科医院(湖南 株洲,412000)
Author(s):
WAN Wenyuan1; LUO Juan1△; PENG Zhenling1; ZHU Yingying1; LUO Jie1
1Zhuzhou Traditional Chinese Medicine Traumatology Hospital,Zhuzhou 412000,Hunan China.
关键词:
绝经后骨质疏松;  龟甲胶;  骨密度;  骨代谢
Keywords:
postmenopausal osteoporosis;  tortoise shell glue;  bone mineral density;  bone metabolism
分类号:
R-33
DOI:
10.20085/j.cnki.issn1005-0205.240404
文献标志码:
A
摘要:
目的:观察龟甲胶对绝经后骨质疏松(PMOP)大鼠能量代谢和骨代谢稳态的影响,及对腺苷酸激活蛋白激酶(AMPK)/沉默信息调节因子2相关酶1(SIRT1)通路的调控作用。方法:将60只雌性SD大鼠随机分为假手术组、模型组、阳性对照组、龟甲胶低、中、高剂量组,每组各10只。除假手术组外,其余大鼠采用双侧卵巢摘除法复制绝经后骨质疏松大鼠模型,以尼尔雌醇片作为阳性对照药,于造模完成后给予尼尔雌醇片1 mg/kg灌胃,1次/d,龟甲胶低、中、高剂量组分别给予0.45,0.90,1.80 g/kg灌胃,2次/d,持续治疗8周。称量大鼠子宫质量并计算子宫指数,双能X线骨密度仪测定L4/5及双侧股骨骨密度,显微CT分析骨形态计量学参数,ELISA法实验检测血清骨钙素(BGP)、骨碱性磷酸酶(BALP)、抗酒石酸酸性磷酸酶5b(TRACP-5b)、Ⅰ型胶原交联羧基末端肽(CTX-Ⅰ)含量,Western Blot法及RT-PCR检测骨组织AMPK/SIRT1通路相关蛋白和mRNA表达。结果:与假手术组相比,模型组子宫指数、L4/5及双侧股骨骨密度明显降低,差异有统计学意义(P<0.05); 骨形态计量学参数骨体积分数(BV/TV)、骨小梁数量(Tb.N)、骨小梁厚度(Tb.Th)明显降低,骨小梁间隔(Tb.Sp)明显增高,差异有统计学意义(P<0.05); 血清中血清骨钙素、骨碱性磷酸酶、TRACP-5b、CTX-Ⅰ的水平均明显升高,差异有统计学意义(P<0.05); 骨组织中p-AMPK/AMPK、SIRT1、PGC1-α的蛋白及AMPK、SIRT1、PGC1-α的mRNA水平明显降低,差异有统计学意义(P<0.05)。龟甲胶高、中、低剂量组治疗后,子宫指数、L4/5及双侧股骨骨密度较模型组明显增高,差异有统计学意义(P<0.05); BV/TV、Tb.N、Tb.Th明显升高,Tb.Sp明显降低,差异有统计学意义(P<0.05); 血清中血清骨钙素、骨碱性磷酸酶、TRACP-5b、CTX-Ⅰ的水平均明显降低,差异有统计学意义(P<0.05); 同时明显上调了骨组织中p-AMPK/AMPK、SIRT1、PGC1-α的蛋白及AMPK、SIRT1、PGC1-α的mRNA水平,差异有统计学意义(P<0.05)。结论:龟甲胶对绝经后骨质疏松大鼠具有较好的改善效果,能够保护大鼠骨微结构,调节骨代谢水平,其作用机制可能与激活AMPK/SIRT1通路、提高骨骼肌能量代谢相关。
Abstract:
Objective:To observe the efficacy of tortoise shell glue on energy metabolism and bone metabolism homeostasis in postmenopausal osteoporosis(PMOP)rats,as well as its regulatory efficacy on the AMPK/SIRT1 pathway.Methods:60 female SD rats were randomly divided into a sham surgery group,a model group,a positive control group,a high,medium and low dose group of tortoise shell glue,with 10 rats in each group.In addition to the sham surgery group,the other rats used bilateral ovariectomy to duplicate the PMOP model.Nilestriol tablets were used as a positive control drug,1 mg/kg of nilestriol tablets were administered by gavage once a day after the completion of modeling.The low,medium,and high dose groups of tortoise shell glue were given 0.45,0.90,1.80 g/kg by gavage twice a day for 8 weeks.The weight of the rat uterus was weighed and the uterine index was calculated.Dual energy X-ray bone mineral density(BMD)instrument was used to measure the BMD of L4/5 and bilateral femurs.Bone morphometric parameters were analyzed by micro CT.The serum levels of osteocalcin(BGP),bone alkaline phosphatase(BALP),anti tartaric acid phosphatase 5b(TRACP-5b),and type Ⅰ collagen cross-linked carboxyl terminal peptide(CTX-Ⅰ)were detected by ELISA.Western Blot and RT-PCR were used to detect the expression of AMPK/SIRT1 pathway related proteins and mRNA in bone tissue.Results:The uterine index,L4/5 and bilateral femoral BMD in the model group were significantly lower compared with the sham surgery group(P<0.05).The bone metrology parameters,bone volume fraction(BV/TV),trabecular number(Tb.N),trabecular thickness(Tb.Th),decreased significantly,and trabecular septa(Tb.Sp)were increased significantly(P<0.05).The levels of BGP,BALP,TRACP-5b and CTX-Ⅰ in serum were significantly increased(P<0.05).The protein levels of p-AMPK/AMPK,SIRT1,PGC1-α and the mRNA levels of AMPK,SIRT1,PGC1-α in bone tissue were significant reduced(P<0.05).The uterine index,L4/5 and bilateral femoral BMD were significantly increased compared to the model group after treatment with high,medium,and low doses of tortoise shell glue(P<0.05).BV/TV,Tb.N,Tb.Th were significantly increased,while Tb.Sp was significantly decreased(P<0.05).The levels of BGP,BALP,TRACP-5b and CTX-Ⅰ in serum were significantly decreased(P<0.05).The protein levels of p-AMPK/AMPK,SIRT1,PGC1-α and the mRNA levels of AMPK,SIRT1,PGC1-α in bone tissue were simultaneously significantly upregulated(P<0.05).Conclusion:Tortoise shell glue has a good improvement efficacy on PMOP rats,which can protect the bone microstructure and regulate bone metabolism levels,its mechanism of action may be related to activating the AMPK/SIRT1 pathway and improving skeletal muscle energy metabolism.

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备注/Memo

备注/Memo:
基金项目:湖南省中医药管理局株洲市联合中医药科研课题(E2023016)
△通信作者 E-mail:zyj135920@163.com
更新日期/Last Update: 2024-04-15