Protective effects of obestatin are mediated by proliferative rather than anti-apoptotic properties in MES23.5 dopaminergic cells
首發時間:2013-11-04
Abstract:In this paper Objective Obestatin is an endogenous peptide sharing the same peptide precursor with ghrelin. We aimed to investigate whether and how obestatin could exert neuroprotective effects on MES23.5 dopaminergic cells against 1-methyl-4-phenylpyridinium (MPP+)-induced neurotoxicity. Methods MES23.5 cells were pretreated with obestatin (10-13 ~ 10-6 mol/L) for 20 min prior to 200 μmol/L MPP+ incubation for 12 or 24 hrs, or treated with different doses of obestatin (10-13 ~ 10-7 mol/L) for 0, 6, 12, 24 hrs, respectively. Methyl thiazolyl tetrazolium (MTT) assay was used to measure cell viability. Flow cytometry was used to measure the caspase-3 activity and the mitochondrial transmembrane potential. Proliferating cell nuclear antigen (PCNA) protein levels were determined by western blotting. Results Obestatin (10-13 ~ 10-7 mol/L) pretreatment could block or even reverse MPP+-induced reduction of cell viability in MES23.5 cells. Solely obestatin treatment increased cell viability. However, it had no effects on MPP+-induced mitochondrial transmembrane potential collapse and caspase-3 activation. Elevated PCNA levels were observed with 10-7, 10-9, 10-11 and 10-13 mol/L obestatin treatment for 12 hrs. Conclusion The protective effects of obestatin against MPP+ in MES23.5 cells were due to its proliferative rather than anti-apoptotic effects.
keywords: Obestatin MPP+ Proliferation Apoptosis Dopaminergic cells
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Obestatin在多巴胺能MES23.5細胞系中通過增殖而非抗凋亡發揮保護作用
摘要:目的 探討obestatin(與ghrelin來自同一前體)對MPP+誘導的MES23.5多巴胺能細胞損傷的保護作用及其機制。方法Obestatin (10-13 ~ 10-6 mol/L)預處理20分鐘后,與200 μmol/L MPP+共孵育MES23.5細胞12或24小時,或obestatin (10-13 ~ 10-7 mol/L)單獨孵育0, 6,12,24小時。使用MTT法檢測細胞存活率,流式細胞儀檢測線粒體膜電位和caspase-3的活性的變化,免疫印跡方法檢測PCNA蛋白表達。結果 MPP+導致MES23.5多巴胺能細胞存活率降低,Obestatin(10-13 ~ 10-7 mol/L)預處理能夠拮抗這一作用,但不能拮抗由MPP+導致的線粒體跨膜電位差的降低和caspase-3的激活。Obestatin(10-13 ~ 10-7 mol/L)單獨作用于MES23.5細胞,能夠使細胞的存活率顯著增加。Obestatin(10-7,10-9,10-11,10-13 mol/L)處理細胞12小時后PCNA蛋白表達增多。結論 在MPP+處理的MES23.5多巴胺能細胞上,obestatin的保護作用歸因于它的細胞增殖作用,而非抗凋亡作用。。
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Obestatin在多巴胺能MES23.5細胞系中通過增殖而非抗凋亡發揮保護作用
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