Department of Biotechnology, Bharathiar University, Coimbatore, Tamil Nadu, 641046, India.
Centre for Animal Research, Training and Services (CAReTS), Central Inter-Disciplinary Research Facility (CIDRF), Mahatma Gandhi Medical College and Research Institute (MGMCRI) Campus, Puducherry, 607403, India.
Mol Cell Biochem. 2018 Mar;440(1-2):199-208. doi: 10.1007/s11010-017-3167-x. Epub 2017 Aug 29.
Osteoblast differentiation is an essential event in the developmental process, which is favoured by the production of extra cellular matrix proteins and various enzymes including discrete ones like acetylcholinesterase (AChE). Despite the fact that AChE facilitates osteoblast differentiation, the significance of its catalytic functions [esterase and aryl acylamidase (AAA) activities] in the process is yet to be ascertained. In this context, SaOS-2 cell line was used in the present study to implicate the catalytic activities of AChE in process of osteoblast differentiation and mineralization. During differentiation, it was found that the activity of both esterase and AAA increased 1.13 and 1.46 folds respectively, signifying the involvement of catalytic activities of AChE in the process. Inhibition of both the catalytic activities of AChE with edrophonium significantly reduced the amount of mineralization by decreasing the alkaline phosphatase (ALP) activity and expression of differentiation-related genes such as RUNX-2, COL1A, ALP, OC, and OP significantly (p < 0.05). Inhibition of esterase activity without altering the AAA activity using gallamine significantly increased the level ALP activity and expression of differentiation-associated genes (p < 0.05), thus favouring mineralization. Therefore, this study concludes and confirms that the AAA activity of AChE is actively involved in the process of osteoblast differentiation and mineralization.
成骨细胞分化是发育过程中的一个重要事件，它受到细胞外基质蛋白和各种酶的产生的促进，包括离散的酶如乙酰胆碱酯酶（AChE）。尽管 AChE 促进成骨细胞分化，但它的催化功能（酯酶和芳基酰基酰胺酶[AAA]活性）在该过程中的意义尚未确定。在这方面，本研究使用 SaOS-2 细胞系来暗示 AChE 的催化活性在成骨细胞分化和矿化过程中的作用。在分化过程中，发现酯酶和 AAA 的活性分别增加了 1.13 和 1.46 倍，表明 AChE 的催化活性参与了该过程。用依酚氯铵抑制 AChE 的两种催化活性显著降低了矿化量，通过降低碱性磷酸酶（ALP）活性和显著降低分化相关基因如 RUNX-2、COL1A、ALP、OC 和 OP 的表达来减少矿化量（p < 0.05）。用加兰他敏抑制酯酶活性而不改变 AAA 活性显著增加了 ALP 活性和分化相关基因的表达（p < 0.05），从而促进矿化。因此，本研究得出并证实 AChE 的 AAA 活性积极参与成骨细胞分化和矿化过程。
