维持性血液透析病人外周血单核细胞凋亡与线粒体异常的关系

Reduction of mitochondrial transmembrane potential constitutes an early event of peripheral blood monocytes apoptosis in hemodialysis patients

目的探讨维持性血液透析（ＨＤ）病人外周血单核细胞凋亡与线粒体异常的关系。方法２０例ＨＤ病人体外培养的外周血单核细胞以流式细胞术通过荧光素标记的ＡｎｎｅｘｉｎＶ检测早期凋亡细胞的膜表面磷酯酰丝氨酸残基（ＰＳ），线粒体跨膜潜能（ΔΨｍ）通过线粒体探针ＤｉＯＣ６（３）分析。线粒体渗透转移（ＰＴ）肿胀通过分光光度计记录。结果ＨＤ病人体外培养的外周血单核细胞在２４、７２小时自发凋亡（４１．４±１２．２）％，（２２．０±３．０）％，明显超过正常对照组（２０．６±８．８）％，（９．８±２．１）％；新鲜分离的单核细胞在病人组和正常对照组细胞凋亡无差异，然而在病人组ΔΨｍ降低达（４３±９）％。ＨＤ病人新鲜分离单核细胞ＰＴ肿胀高达（８０±６）％明显超过正常对照组（５±３）％。结论线粒体肿胀和ΔΨｍ降低是导致ＨＤ病人外周血单核细胞凋亡的原因。

Objective To investigate the relationship between monocyte apoptosis and mitochondria function in hemodialysis (HD) patients. Methods Fluorescein labeled Annexin V was used to detect the phospha tidyserine (PS) of apoptosis cells, and mitochondrial transmembrane potential was studied by 3,3′ dihexyloxacarbocyanine iodide, DiOC 6 (3), and analyzed by flow cytometry. Swelling of mitochondrial permeability transition (PT) was recorded by spectrophotometer.Results Spontaneous apoptosis of cultured monocytes increased at 24, 72 hours in HD patients (41 4±12 2)%、(22 0±3 0)% as compared with controls (20 6±8 8)%、(9 8±2 1)%. Freshly isolated monocytes in HD patients had little apoptosis, but mitochondrial transmembrane potential was reduced to half of disruption (43±9)%. As to swelling of PT, those freshly isolated monocytes mitochondria in HD patients experienced overwhelming opening of PT, (80±5 6)%.Conclusion Swelling of PT and breakdown of mitochondrial transmembrane potential constitute the early events of monocytes apoptosis in HD patients, which could contribute to the impaired monocytes function.