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外周血单核细胞表达PEDF在2型糖尿病视网膜病变中的意义[J]. 中山大学学报(医学科学版), 2015,36(1).
Level of PEDF Increased in Monocytes in Diabetic Retinopathy[J]. Journal of Sun Yat-sen University (Medical Sciences), 2015, 36(1).
【目的】 色素上皮衍生因子(PEDF)通过抗血管增生、抗炎、抗氧化应激等影响糖尿病视网膜病变(DR)发生发展,而单核/巨噬细胞(CD14+)与慢性炎症关系密切。本研究拟通过观察外周血CD14+细胞中PEDF的表达水平,为进一步探讨DR发病机制提供线索。【方法】 选取2011年10月至2013年4月在我院内分泌科住院确诊为2型糖尿病患者(DM组)108例,分为无糖尿病视网膜病变组(NDR组)52例和糖尿病视网膜病变组(DR组)56例,同期健康体检者作为对照组(NC组)52例。收集患者外周血白细胞及其分类细胞计数相关指标;密度梯度离心法收集患者PBMC,免疫磁珠分离法提取CD14+ 细胞;剩余PBMC提取总蛋白,western blot 比较总PBMC(tPBMC)与分离掉CD14+ 细胞的PBMC[PBMC(exCD14+)]中PEDF蛋白表达水平。 【结果】 DR组和NDR组单核细胞绝对值较NC组升高(P < 0.05),但DR组和NDR组间单核细胞绝对值的差异无统计学意义(P > 0.05);DR组淋巴细胞绝对值较NDR组明显减低(P < 0.05);与tPBMC相比,PBMC(exCD14+)中PEDF蛋白表达水平明显下降(P < 0.05)。【结论】 DR患者外周血单核细胞数量增加。单核细胞是PBMC中表达PEDF的主要细胞。
【Objective】 Pigment epithelium derived factor (PEDF) has been widely implicated in Diabetic retinopathy (DR) via inhibition of angiogenesis and inflammation.Monocyte/macrophage (CD14+ cell) is closely associated with chronic inflammation. The present study is aimed to investigate whether PEDF is mainly expressed in monocytes in DR patients. 【Methods】 Type 2 diabetic patients served as DM group (n = 108), which were sub-divided into non-diabetic retinopathy group ( NDR group, n = 52) and diabetic retinopathy group ( DR group, n = 56) by angiography. 52 healthy persons were enrolled in the study as normal control group (NC group). CD14+ cells were isolated from PBMC using density-gradient centrifugation and immunomagnetic beads. The protein level of PEDF in the tPBMC and PBMC (exCD14+) was determined by western blot. The content of white blood cell count was measured.【Results】 The counts of monocytes were similar in NDR and DR groups (P > 0.05) but lower in NC group (P < 0.05). The counts of lymphocytes in DR groups were lower in NDR groups (P<0.05). The level of PEDF in PBMC excluding CD14 positive cells was significantly lower than that of total PBMC (P < 0.05). 【Conclusion】 Monocytes are the main cells to secret PEDF in PBMC.
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