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Effects of Hypoxia on the Apoptosis and Autophagy of the Goat Temporomandibular Joint Disc Cells after Serum Deprivation
Zhang Fang1, Bao Guangjie1,2,3*, Tang Yuyao1, Liu Lin2,3, Bao Shanying1, Kang Hong1*
1Institute of Stomatology, Lanzhou University, Lanzhou 730000, China; 2Key Laboratory of Stomatology of State Ethnic Affairs Commission, Northwest Minzu University, Lanzhou 730030, China; 3Key Laboratory of Oral Diseases of Gansu Province, Northwest Minzu University, Lanzhou 730030, China
Abstract: In this work， they isolated and cultured temporomandibular joint disc (TMJ disc) cells of goat to p2 generation in vitro. The morphological changes of cells were observed by Hematoxylin-Eosin (HE) staining， Hoechst 33258 and dansylcadaverine (MDC) fluorescence staining were used to observe whether autophagy or apoptosis existed after the serum deprivation. After 0 h， 12 h， 24 h， 36 h， 48 h and 60 h of serum deprivation， apoptosis rate and autophagy rate of cells were detected by flow cytometry and Real-time PCR， respectively. Changes of apoptosis was observed after applying 3-methyladenine (3-MA)， an autophagy inhibitor. The changes of apoptosis and autophagy with serum (10% FBS) or serum deprivation (0% FBS) were detected under the conditions of oxygen (21% O2) or hypoxia (2% O2). The results showed that the rate of apoptosis increased gradually with the prolongation of the time， and the autophagy first increased and then decreased after the serum deprivation. When the autophagy induced by serum deprivation was inhibited by 3-MA， the rate of apoptosis of cells increased obviously. Autophagy could inhibit the apoptosis induced by serum deprivation， which showed that autophagy played an important role in cell survival. Compared with normoxia-cultured cells， the apoptosis rate and autophagy rate of the cells decreased under hypoxia condition. Hypoxia reduced the apoptosis of cells caused by long-term serum deprivation by reducing excessive autophagy， which was more conducive to cell survival than normoxia.
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