Yu Yun-fei, Xu Hong-guang, Wang Hong, Zhang Wei, Xiong Shou-liang, Zhang Min
Department of Orthopedic Surgery, Yijishan Hospital, Wannan Medical College, Wuhu 241001, China.
Department of Orthopedic Surgery, Yijishan Hospital, Wannan Medical College, Wuhu 241001, China. Email:
Zhonghua Yi Xue Za Zhi. 2013 Dec 3;93(45):3632-5.
To explore the expression and significance of autophagy in endplate cartilage of rats during aging process.
The end-plate chondrocytes were isolated from 3, 6 and 12-month SD rats respectively. And the natural culture and rapamycin groups were assigned. Alizarin red staining was used to observe the morphological changes of cells. And RT-PCR was employed to detect the expressions of type II collagen, proteoglycan, SOX-9 and matrix metalloproteinase (MMP-13). The expressions of Beclin-I and LC3 were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The rate of autophagy was observed by monodansylcadaverine (MDC) staining and methyl thiazolyl tetrazolium (MTT) for cell survival rate.
Alizarin red staining showed that cells might reflect the process of intervertebral disc degeneration. The expressions of polysaccharide, Sox-9, type II collagen, Beclin-1 and LC3 in endplate chondrocytes significantly decreased with advancing age (P < 0.05). The incidence of autophagy significantly decreased (P < 0.05). The cell viability of each group significantly decreased (P < 0.05). Compared with control group, the cell viability of rapamycin group significantly increased (P < 0.05).
During aging process, the expressions of autophagy related-gene LC3 and Beclin-1 significantly decrease with the reduced activity of end-plate chondrocyte. And autophagy activity may be correlated with the development and degeneration of intervertebral disc.
探讨自噬在大鼠终板软骨衰老过程中的表达及意义。
分别从3、6和12月龄的SD大鼠分离终板软骨细胞，分为自然培养组和雷帕霉素组。采用茜素红染色观察细胞形态变化，采用RT-PCR检测Ⅱ型胶原、蛋白聚糖、SOX-9和基质金属蛋白酶（MMP-13）的表达。采用逆转录-聚合酶链反应（RT-PCR）和蛋白质印迹法检测Beclin-Ⅰ和LC3的表达。采用单丹磺酰尸胺（MDC）染色观察自噬率，采用甲基噻唑基四氮唑（MTT）检测细胞存活率。
茜素红染色显示细胞可反映椎间盘退变过程。随着年龄增长，终板软骨细胞中多糖、Sox-9、Ⅱ型胶原、Beclin-1和LC3的表达显著降低（P<0.05）。自噬发生率显著降低（P<0.05）。各组细胞活力显著降低（P<0.05）。与对照组相比，雷帕霉素组细胞活力显著升高（P<0.05）。
在衰老过程中，自噬相关基因LC3和Beclin-1的表达随着终板软骨细胞活性降低而显著下降。自噬活性可能与椎间盘的发育和退变相关。
