[1]卫朗秋,邹佳盈,龙赵凡,等.基于AMPK/mTOR自噬通路探讨抗疏健骨颗粒改善骨稳态作用机制[J].陕西中医,2026,(4):441-448.[doi:DOI:10.3969/j.issn.1000-7369.2026.04.002]
　WEI Langqiu,ZOU Jiaying,LONG Zhaofan,et al.The mechanism of Kangshu Jiangu granules in improving bone homeostasis based on the AMPK/mTOR autophagy pathway[J].,2026,(4):441-448.[doi:DOI:10.3969/j.issn.1000-7369.2026.04.002]
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基于AMPK/mTOR自噬通路探讨抗疏健骨颗粒改善骨稳态作用机制

《陕西中医》[ISSN:1000-7369/CN:61-1281/TN]

卷:
期数:
2026年4期
页码:
441-448
栏目:
基础研究
出版日期:
2026-04-05

文章信息/Info

Title:
The mechanism of Kangshu Jiangu granules in improving bone homeostasis based on the AMPK/mTOR autophagy pathway
作者:
卫朗秋1; 邹佳盈1; 龙赵凡1; 史激特1; 汪宁茜1; 宋丽1; 史传道1; 2; 刘启玲1; 2
（1.陕西中医药大学，陕西 咸阳 712046；2.环境相关疾病与中医药防控陕西省高等学校重点实验室，陕西 咸阳 712046）
Author(s):
WEI Langqiu1; ZOU Jiaying1; LONG Zhaofan1; SHI Jite1; WANG Ningxi1; SONG Li1; SHI Chuandao1; 2; LIU Qiling1; 2
（1.Shaanxi University of Chinese Medicine,Xianyang 712046,China；2.The Key Laboratory of Environment-Related Diseases and Traditional Chinese Medicine Prevention and Control,Shaanxi Higher Education Institutions,Xianyang 712046,China）
关键词:
骨质疏松; 抗疏健骨颗粒; 自噬; AMPK/mTOR通路; 骨代谢; 成骨细胞
Keywords:
Osteoporosis; Kangshu Jiangu granules; Autophagy; AMPK/mTOR signaling pathway; Bone metabolism; Osteoblast
分类号:
R 681
DOI:
DOI:10.3969/j.issn.1000-7369.2026.04.002
文献标志码:
A
摘要:
目的：研究抗疏健骨颗粒(KSJG)对成骨细胞自噬通路腺苷酸活化蛋白激酶（AMPK）/哺乳动物雷帕霉素靶蛋白（mTOR）的作用，为其在骨质疏松症（OP）早期预防中的应用提供实验依据。方法：通过Label-free非标记定量分析，对对照组、模型组、KSJG组大鼠右侧股骨组织进行蛋白组学测序，进行GO和KEGG通路分析后确定AMPK/mTOR自噬通路；将KSJG含药血清作用于MC3T3-E1成骨细胞，实验分为对照组、KSJG组、Compound C组和Compound C+KSJG组。采用CCK-8法检测法检测细胞活力；Western blot法检测自噬标志性蛋白（LC3、Beclin-1、Atg7、p62）及AMPK/mTOR通路蛋白（AMPK、mTOR和p-mTOR等）水平；MDC荧光染色观察自噬体形成情况，茜素红S染色评价成骨矿化能力。结果：Compound C浓度为4 μmol/L时，p-AMPK水平最低，抑制效果最好（P<0.05），且细胞存活率与对照组比较差异无统计学意义（P>0.05）。KSJG含药血清作用于细胞的最佳浓度和时间为30%和24 h；与对照组相比，Compound C组成骨细胞中AMPK磷酸化水平显著降低，自噬相关蛋白LC3-Ⅱ、Beclin-1、Atg7表达降低，p62、p-mTOR表达升高（均P<0.05），自噬水平受到抑制；KSJG干预后，可逆转上述现象，即成骨细胞中p-AMPK表达明显升高，LC3-Ⅱ、Beclin-1、Atg7表达上调，p62与p-mTOR表达下调（均P<0.05）。与对照组相比，KSJG组自噬体数目增多，而Compound C组和Compound C+KSJG组自噬体数目下降（均P<0.05），且Compound C+KSJG组自噬体数目在Compound C组的基础上进一步下降（P<0.05）。与对照组相比，KSJG组钙盐沉积物升高，而Compound C组和Compound C+KSJG组钙盐沉积物下降（均P<0.05），且Compound+KSJG组钙盐沉积物在Compound组的基础上进一步下降（P<0.05）。结论：KSJG可通过激活AMPK、抑制mTOR磷酸化水平，增强成骨细胞自噬活性，提高成骨矿化能力，进而改善骨代谢失衡状态，是KSJG早期预防骨质疏松的机制之一。
Abstract:
Objective:To investigate the effect of Kangshu Jiangu granules on the AMPK/mTOR mediated autophagy pathway in osteoblasts,and to provide experimental evidence for its application in the early prevention of osteoporosis (OP).Methods:Using label-free quantitative proteomics analysis on the right femoral tissues of rats from the control group,model group,and Kangshu Jiangu granules (KSJG) group,we determined the AMPK/mTOR autophagy pathway to be the key regulatory pathway following GO and KEGG pathway enrichment analyses.MC3T3-E1 osteoblasts were treated with KSJG medicated serum and divided into four groups:Control group,KSJG group,Compound C group and Compound C+KSJG group.Cell viability was assessed using the CCK-8 assay.The expression of autophagy-related proteins (LC3,Beclin-1,Atg7,and p62) and AMPK/mTOR pathway-related proteins (AMPK,mTOR,and p-mTOR) were determined by Western blot analysis.Autophagosome formation was observed by MDC fluorescence staining,and osteogenic mineralization was evaluated using Alizarin Red S staining.Results:Compound C exhibited the strongest inhibitory effect on p-AMPK expression at 4 μmol/L (P<0.05)，and without affecting cell viability (P>0.05).The optimal concentration and intervention time of KSJG medicated serum were determined to be 30% and 24 h,respectively.Compared with the control group,Compound C treatment significantly reduced the expression of p-AMPK and downregulated the expression of LC3-Ⅱ,Beclin-1,and Atg7,while upregulated the expression of p62 and p-mTOR (all P<0.05),indicating autophagic activity was suppressed.KSJG intervention markedly reversed these changes,the expression of p-AMPK,LC3-Ⅱ,Beclin-1 and Atg7 increased,p62 and p-mTOR decreased (all P<0.05).Compared with the control group,the number of autophagosomes was significantly increased in the KSJG group,whereas it was decreased in the Compound C group and the Compound C+KSJG group (P<0.05).In addition,calcium deposition was significantly increased in the KSJG group,while decreased in the Compound C group and the Compound C+KSJG group (all P<0.05),and was further reduced in the Compound C+KSJG group compared with the Compound C group (P<0.05).Conclusion:Kangshu Jiangu granules can enhance the autophagic activity of osteoblasts and improve osteogenic mineralization capacity by activating AMPK and inhibiting the phosphorylation level of mTOR,thereby ameliorating the imbalance of bone metabolism,which is one of the mechanisms underlying its efficacy in the early prevention of osteoporosis.

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备注/Memo

备注/Memo:
陕西省自然科学基础研究计划项目（2024JC-YBMS-729）；陕西省疾病预防控制局公共卫生科研创新团队项目（202512）；陕西省中医药管理局中医药科技创新计划项目（2025-CXTD-04）；陕西省教育厅一般项目（24JK0399）；陕西省咸阳市优秀创新团队计划项目（L2024-CXNL-KJRCTD-KJTD-0014）；陕西省秦创原中医药产业创新聚集区项目（L2024-QCY-ZYYJJQ-X138，X183）
更新日期/Last Update: 2026-04-05