摘要目的 检测原发性开角型青光眼(POAG)患者外周血补体C3水平和C3外显子基因多态性,探讨补体与POAG的相关性.方法 采用前瞻性病例对照研究设计,收集2014年12月至2015年12月于复旦大学附属眼耳鼻喉科医院确诊的45例POAG患者,同时收集与病例组年龄、性别相匹配的45名健康体检者作为对照组.采用免疫比浊法测定血清C3浓度.通过提取外周血有核细胞基因组DNA,经PCR扩增补体C3外显子后直接测序作基因序列分析.采用SPSS 20.0软件进行统计学分析,独立样本t检验分析不同组间C3水平的差异性,卡方检验分析两组之间C3外显子基因多态性分布的差异性,单因素方差分析比较C3外显子不同基因型间血清C3水平的差异性.结果 POAG组血清C3浓度相比较于对照组显著降低[(104.81 ±29.15) mg/dl vs(121.06±18.39) mg/dl,t=-3.162,P=0.002],差异有统计学意义.C3外显子基因测序共检测到7个单核苷酸多态性(SNP)(rs2230201、rs2230204、rs2230205、rs428453、rs423490、rs7951、rs539822147),未发现突变.SNPs的频率及分布在POAG组和对照组之间差异均无统计学意义,且未发现SNPs不同基因型间血清C3水平的显著差异.结论 POAG患者血清中补体C3水平显著降低,提示补体C3可能参与青光眼的病理机制.C3外显子基因检测并无异常发现,可基本排除基因多态性或突变对血清C3水平的影响.

abstractsObjective In order to explore the association between complement C3 and primary open angle glaucoma (POAG),the serum complement C3 level was detected and the polymorphism of the C3 exon gene was analyzed in patients with POAG.Methods A prospective case-control study was designed.A total of 45 patients with POAG visiting Eye &ENT hospital of Fudan University were collected from December 2014 to December 2015,and 45 age-/gender-matched healthy subjects from yearly health screening were collected as normal controls.Serum C3 concentration was detected by immunoturbidimetric assay.Meanwhile,genomic DNA was extracted from peripheral blood leukocytes,and sequencing for C3 exons was followed with PCR to analyze the gene polymorphism.Statistical analysis was performed by use of SPSS 20.0 software.Student's t test was used to evaluate the difference of serum C3 level between two groups.Chisquare test was used to analyze the difference in the distribution of C3 exon gene polymorphism between two groups.The one-way ANOVA was utilized to analyze the difference of serum C3 level among different genotypes of C3 exon.Results The serum level of C3 in POAG (104.81 ± 29.15)mg/dl was significantly lower (t =-3.162,P =0.002) compared to controls (121.06 ± 18.39) mg/dl.There were a total of seven single nucleotide polymorphisms (SNPs) (rs2230201,rs2230204,rs2230205,rs428453,rs423490rs7951,rs539822147) within the C3 exon gene region,but no mutation was detected.There was no difference in the frequency and distribution of these SNPs between POAG and controls,as well as,there was also no significant difference in serum C3 level among these SNPs genotypes.Conclusions Decreased serum C3 level in POAG patients indicated that complement C3 might be involved in the pathomechanism of glaucoma.No significant abnormality of C3 exon gene was detected,so the gene polymorphism having an impact on serum C3 level could be excluded as a reason probably.