|
| 温肾通络止痛方经AMPK/mTOR信号通路调控自噬改善BMSCs细胞衰老 |
| Experimental study of Wenshen Tongluo Zhitong Recipe regulating autophagy through AMPK/mTOR signaling pathway to improve BMSCs senescence |
| |
| DOI：10.3969/j.issn.1006-7108.2025.10.004 |
| 中文关键词: 老年性骨质疏松 温肾通络止痛方 骨髓间充质干细胞 自噬 衰老 |
| 英文关键词:Wenshen Tongluo Zhtong Decoction can inhibit BMSCs senescence by promoting autophagy through AMPK/mTOR signaling pathway. |
| 基金项目:国家自然科学基金面上项目（82074458、82174411）；江苏省自然科学基金项目（BK20221351、BK20220470）；江苏省中医退行性骨关节病临床医学创新中心资助项目(苏中医科教〔2021〕4号），国家中医药管理局高水平中医药重点学科建设项目资助（国中医药人教函〔2023〕85号），江苏省研究生科研创新计划（KYCX24_2224） |
|
| 摘要点击次数: 1 |
| 全文下载次数: 0 |
| 中文摘要: |
| 探讨温肾通络止痛方（Wenshen Tongluo Zhitong Recipe，WTZR)经AMP依赖的蛋白激酶（Adenosine 5‘-monophosphate (AMP)-activated protein kinase，AMPK）/哺乳动物雷帕霉菌靶蛋白(mammalian target of rapamycin, mTOR)信号通路调控自噬改善骨髓间充质干细胞（Bone marrow mesenchymal stromal cells，BMSCs）衰老的机制研究。方法：利用D-半乳糖制备衰老BMSCs模型并随机分为对照组、模型组、WTZR低剂量组、WTZR中剂量组、WTZR高剂量组。β-半乳糖苷酶染色检测检测WTZR干预后衰老BMSCs的衰老情况、Western blot与qPCR检测WTZR干预后衰老BMSCs中p16、p21和p53等衰老标志物的变化。ALP 染色、茜素红染色及油红O染色观察WTZR干预后BMSCs成骨成脂分化的变化。Western blot及免疫荧光检测自噬相关指标微管相关蛋白 1A/1B-轻链3B（Microtubule-Associated Protein 1 Light Chain 3，LC3B)和Beclin1的蛋白表达，评估WTZR对BMSCs自噬的改变。Western blot检测mTOR、p-mTOR、AMPK及p-AMPK的蛋白表达，检验WTZR对AMPK/mTOR信号通路的影响。结果：与模型组相比，WTZR干预后，BMSCs衰老细胞数量降低，BMSCs中p16、p21和p53蛋白及mRNA含量显著减少（P<0.05，P<0.01，P<0.001，P<0.0001），WTZR干预后ALP阳性细胞数、茜素红阳性面积增加,油红 O 染色镜下脂滴数量减少。此外，WTZR可显著增加衰老BMSCs中自噬相关蛋白的含量（P<0.01，P<0.0001）、同时温肾通络止痛方可显著提高p-AMPK/AMPK比值（P<0.05），并降低p-mTOR/mTOR 的比值（P<0.0001）。结论：WTZR可能通过AMPK/mTOR促进自噬从而发挥缓解BMSCs衰老的效果。 |
| 英文摘要: |
| To study the intervention effects of Wenshen Tongluo Zhitong Recipe(WTZR) on Adenosine 5‘-monophosphate (AMP)-activated protein kinase（AMPK）/mammalian target of rapamycin(mTOR) signaling pathways in regulating autophagy to improve the senescence of Bone marrow mesenchymal stromal cells(BMSCs).
Methods: The senescent BMSCs model was established using D-galactose and subsequently divided into five groups: control, model, low-dose drug-treated serum, middle-dose drug-treated serum,and high-dose drug-treated serum.β-galactosidase staining qPCR , and WB were used to assess the levels of senescence markers p16, p21 and p53. WB and immunofluorescence were used to assess the levels of of microtubule-Associated Protein 1 Light Chain 3（LC3B） and Beclin1. Western blot was used to assess the levels of mTOR, p-mTOR, AMPK and p-AMPK in AMPK/mTOR signaling pathway.
Results：In comparison to the model group, the intervention of WTZR significantly reduced the number of positive cells associated with aging. The levels of p16, p21, and p53 proteins and mRNA were also significantly diminished (P < 0.05, P < 0.01, P < 0.001, P < 0.0001), after intervention ALP positive cell number and alizarin red positive area increase, oil red O staining microscopic fewer lipid droplets. Additionally, the WTZR effectively promoted autophagy-related protein expression in BMSCs induced by D-galactose galactose (P < 0.01, P < 0.0001) and improved the P-AMPK/AMPK ratio (P < 0.05), while reducing the ratio of P-mTOR/mTOR (P < 0.0001).
Conclusions : Wenshen Tongluo Zhtong Decoction can inhibit BMSCs senescence by promoting autophagy through AMPK/mTOR signaling pathway. |
| 查看全文 查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|