2022, Vol. 26 ›› Issue (35): 5665-5668

Melatonin effect on the activity of mouse ATDC5 cells

Sun Jinpeng, Liu Jun, Bai Yunfeng, Hua Feng, Wang Haoran, Zheng Hongrui, Wu Tao   

Abstract: BACKGROUND: Human primary chondrocytes extracted from adolescent iliac cartilage are currently used in the study of adolescent idiopathic scoliosis. However, research on adolescent idiopathic scoliosis is largely limited due to parental concerns and ethical restrictions. Therefore, it is of great scientific and social significance to find a kind of cells that can replace human primary chondrocytes in the study of adolescent idiopathic scoliosis.
OBJECTIVE: To study the activity, receptor and signaling pathway changes of ATDC5 cells stimulated by melatonin, and to explore whether ATDC5 cells can be used to replace human chondrocytes in comparison with the response of human chondrocytes under melatonin stimulation.
METHODS: (1) ATDC5 cells were seeded into 96-well plates and stimulated by melatonin at different concentrations (10-5, 10-6, 10-7, 10-8, 10-9, 10-10, 10-11, 10-12 mol/L). Cells without melatonin stimulation were used as the control. After 72 hours of culture, the proliferation rate of ATDC5 cells was detected by MTT method. (2) ATDC5 cells were seeded into 96-well plates. The control group was not stimulated with melatonin, and the experimental group was stimulated with 10-5 mol/L melatonin. After culture for 24 hours, the protein expression of melatonin receptor and AMPK pathway was detected by western blot assay.
RESULTS AND CONCLUSION: After melatonin stimulation, the proliferative rate of ATDC5 cells was increased with melatonin concentration increasing. After melatonin stimulation, the protein expressions of type II collagen, p-AMPK and melatonin receptor 1B in the experimental group were higher than those in the control group (P < 0.05), while there was no significant difference in the protein expressions of AMPK and melatonin receptor 1A between the two groups (P > 0.05). Therefore, melatonin can promote the proliferation of mouse ATDC5 cells, and the proliferation rate is similar to that of human chondrocytes stimulated by melatonin at the same concentration, suggesting there is a possibility that mouse chondrocyte ATDC5 can be a substitute of human chondrocytes.
Key words: adolescent, idiopathic scoliosis, melatonin, chondrocyte activity, melatonin receptor, AMPK, ATDC5 cell