| 基于Akt/mTOR信号通路探讨补肾活血方对去卵巢大鼠软骨细胞自噬的影响 |
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Received:September 20, 2022
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| 作者 | Author | 单位 | Unit | E-Mail |
| 陶帅 |
TAO Shuai |
南京中医药大学泰州附属医院, 江苏 泰州 225300 |
Taizhou Hospital of TCM Affiliated to Nanjing University of Chinese Medicine, Taizhou 225300, Jiangsu, China |
taoshuai130@163.com |
| 姜宏 |
JIANG Hong |
南京中医药大学苏州附属医院, 江苏 苏州 215009 |
Suzhou Hospital of TCM Affiliated to Nanjing University of Chinese Medicine, Suzhou 215009, Jiangsu, China |
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| 周海燕 |
ZHOU Hai-yan |
南京中医药大学翰林学院, 江苏 泰州 225300 |
Hanlin College of Nanjing Univerisity of Chinese Medicine, Taizhou 225300, Jiangsu, China |
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| 期刊信息：《中国骨伤》2024年37卷，第2期，第196-206页 |
| DOI：10.12200/j.issn.1003-0034.20220321 |
| 基金项目:泰州市科技支撑计划（社会发展）项目（编号：TS201909） |
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目的：研究补肾活血方是否可以通过调控Akt/mTOR信号通路促进去卵巢大鼠软骨细胞自噬反应来保护关节软骨。
方法：选取30只SPF级12周龄雌性SD大鼠，体重（247.0±7.0） g，先随机选择6只为空白对照组，剩余大鼠采用卵巢切除术结合右膝关节腔注射碘乙酸钠（monosodium iodoacetate，MIA）建立绝经后膝骨关节炎模型，随机分为模型组、BSHXR-L组、BSHXR-M组、BSHXR-H组，每组6只。运用肉眼观察评分、番红O-固绿染色、免疫组化等方法明确补肾活血方对大鼠关节软骨损伤的保护作用，Western-blot检测自噬相关蛋白的表达，qPCR检测Akt、mTOR及下游自噬基因的相对表达。
结果：造模后BSHXR （L、M、H）各组均可减轻软骨组织形态学损伤，免疫组化示Collagen-Ⅱ、Aggrecan表达逐渐升高，基质金属蛋白酶-13（matrix metallopriteinase-13，MMP-13）表达逐渐降低，其中BSHXR-M组和BSHXR-H组与模型组相比差异均有统计学意义（P<0.05）。Western-blot结果示BSHXR （L、M、H）各组自噬通路蛋白p-Akt/Akt、p-mTOR/mTOR受到抑制，下游蛋白Beclin-1、LC3Ⅱ表达逐渐升高，而p62逐渐降低，呈剂量效应。qPCR结果示BSHXR （L、M、H）各组均可促进Beclin-1、LC3ⅡmRNA的相对表达，抑制p62、Akt、mTOR mRNA的相对表达，与模型组相比差异有统计学意义（P<0.05）。
结论：补肾活血方可以通过抑制Akt/mTOR信号通路增强软骨细胞自噬反应，进而发挥保护软骨的作用。 |
| [关键词]:补肾活血方 绝经 膝骨关节炎 软骨细胞自噬 Akt/mTOR信号通路 |
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| Effect of Bushen Huoxue(补肾活血) recipe on autophagy of ovariectomized rat chondrocytes based on Akt/mTOR signaling pathway |
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Abstract:
Objective To investigate whether Bushen Huoxue recipe can protect articular cartilage by regulating Akt/mTOR signaling pathway to promote the autophagy of chondrocytes in ovariectomized rats.
Methods Among 30 SPF 12-week-old female SD rats weighing (247.0±7.0) g, 6 were randomly selected as the blank control group, and the remaining rats were randomly divided into model group, BSHXR-L group, BSHXR-M group and BSHXR-H group, with 6 rats in each group. The protective effect of Bushen Huoxue recipe on articular cartilage injury in rats was determined by visual observation score, muscovine O-solid green staining and immunohistochemistry. The expression of autophagy related proteins was detected by Western-blot, and the relative expression of Akt, mTOR and downstream autophagy genes was detected by qPCR.
Results After modeling, BSHXR (L, M, H) groups could alleviate the histological damage of cartilage. Immunohistochemistry showed that the expression of Collagen-Ⅱand Aggrecan gradually increased, and the expression of MMP-13 gradually decreased, and the differences between BSHXR-M and BSHXR-H groups and model group were statistically significant (P<0.05). The results of Western-blot showed that the autophagy pathway proteins p-Akt/Akt and p-mTOR/mTOR were inhibited in the BSHXR(L, M, H) groups, and the expressions of downstream proteins Beclin-1 and LC3Ⅱwere gradually increased, while p62 was gradually decreased, showing a dose effect. QPCR results showed that BSHXR(L, M, H) groups could promote the relative expression of Beclin-1 and LC3ⅡmRNA, and inhibit the relative expression of p62, Akt, mTOR mRNA, and the differences were statistically significant compared with model group (P<0.05).
Conclusion Bushen Huoxue recipe can enhance the cartilage autophagy response by inhibiting the Akt/mTOR signaling pathway, and then protect the cartilage. |
| KEYWORDS:Bushen Huoxue recipe Menopause Knee osteoarthritis Chondrocyte autophagy Akt/mTOR signaling pathway |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 陶帅,姜宏,周海燕.基于Akt/mTOR信号通路探讨补肾活血方对去卵巢大鼠软骨细胞自噬的影响[J].中国骨伤,2024,37(2):196~206 |
| 英文格式: | TAO Shuai,JIANG Hong,ZHOU Hai-yan.Effect of Bushen Huoxue(补肾活血) recipe on autophagy of ovariectomized rat chondrocytes based on Akt/mTOR signaling pathway[J].zhongguo gu shang / China J Orthop Trauma ,2024,37(2):196~206 |
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