雷珠单抗治疗湿性年龄相关性黄斑变性的疗效观察及其对患者血浆miRNA-126、VEGF-A表达的影响

目的探讨玻璃体腔注射雷珠单抗治疗湿性年龄相关性黄斑变性(wAMD)的临床疗效,及其对患者血浆中微小RNA(miRNA)-126、血管内皮生长因子(VEGF)-A的表达水平的影响。方法前瞻性研究。纳入2018年6月—2019年6月蚌埠医学院第一附属医院眼科收治40例(40只眼)wAMD患者为wAMD组,其中男17例、女23例,年龄50~86岁。纳入同期在蚌埠医学院第一附属医院体检中心进行健康体检的中老年人为对照组(40人),其中男18人、女22人,年龄53~81岁。wAMD组患者均接受单次玻璃体腔注射雷珠单抗0.05 mL治疗。采用实时荧光定量PCR(qPCR)法检测对照组和wAMD组治疗前及治疗后1个月血浆miRNA-126相对表达水平,酶联免疫吸附试验检测血浆中VEGF-A水平。(1)比较对照组、wAMD组治疗前miRNA-126及VEGF-A的表达水平。(2)比较wAMD组治疗前、治疗后1个月血浆miRNA-126、VEGF-A表达水平,以及最佳矫正视力(BCVA)、黄斑中央区视网膜厚度(CMT)、脉络膜新生血管(CNV)面积的变化情况。(3)分析wAMD组患者治疗前、治疗后1个月血浆miRNA-126的相对表达量与VEGF-A水平、CMT、CNV面积的相关性。结果(1)wAMD组患者的miRNA-126相对表达量(0.86±0.11)低于对照组(1.04±0.10),VEGF-A水平(329.09±17.58)pg/mL高于对照组(295.74±14.80)pg/mL,差异均有统计学意义(t=8.010、9.179,P值均<0.01)。(2)wAMD组治疗后1个月miRNA-126相对表达量(1.47±0.27)较治疗前(0.86±0.11)增加,VEGF-A水平(315.36±15.44)pg/mL较治疗前(329.09±17.58)pg/mL降低,差异均有统计学意义(t=19.410、8.048,P值均<0.01);BCVA(0.45±0.11)较治疗前(0.83±0.16)增加,CMT与CNV面积分别为(296.53±21.52)μm,(0.58±0.18)mm2,较治疗前的(311.88±37.25)μm、(0.70±0.17)mm2均降低,差异均有统计学意义(t=12.667、3.602、4.906,P值均<0.01)。(3)wAMD组患者治疗前及治疗后1个月血浆miRNA-126相对表达水平与VEGF-A、CMT、CNV面积均呈显著负相关(r治疗前=-0.706、-0.723、-0.552,r治疗后1个月=-0.783、-0.709、-0.620,P值均<0.01)。治疗前拟合VEGF-A、CMT、CNV面积与miRNA-126相对表达量之间的线性回归方程分别为:YVEGF-A=-112XmiRNA-126+426,YCMT=-243XmiRNA-126+522,YCNV=-0.84XmiRNA-126+1.42;治疗后线性回归方程分别为:YVEGF-A=-45.64XmiRNA-126+382,YCMT=-57.53XmiRNA-126+381,YCNV=-0.41XmiRNA-126+1.18。结论雷珠单抗可通过降低VEGF-A的表达,减少CMT及CNV面积,有效改善BCVA。雷珠单抗在取得临床疗效的同时,可能通过增强miRNA-126基因表达活性而延缓wAMD的进展。

Effects of ranibizumab on microRNA-126 and vascular endothelial growth factor-A and its clinical efficacy in patients with wet age-related macular degeneration

Objective To investigate the effects of the intravitreal injection of ranibizumab on the expression levels of microRNA-126 (miRNA-126) and vascular endothelial growth factor A (VEGF-A) and its clinical efficacy in patients with wet age-related macular degeneration (wAMD).Methods A total of 40 cases (40 eyes) of wAMD patients, including 17 males and 23 females, aged 50-86 years, who were admitted to the First Affiliated Hospital of Bengbu Medical College from June 2018 to June 2019 were included in this prospective study as the wAMD group. Meanwhile, the control group consisted of 40 middle-aged and elderly patients, including 18 males and 22 females, aged 53-81 years, who underwent physical examination in the Physical Examination Center of the First Affiliated Hospital of Bengbu Medical College. All the patients in the wAMD group received a single intravitreal injection of ranibizumab (0.05 mL). Real-time fluorescence quantitative polymerase chain reaction method was used to detect the relative expression levels of miRNA-126 in the control and wAMD groups before treatment and 1 month after treatment, and plasma VEGF-A level was measured via enzyme-linked immunosorbent assay. (1) The expression levels of miRNA-126 and VEGF-A before treatment in the control and wAMD groups were compared. (2) Plasma miRNA-126 and VEGF-A expression levels and changes in best corrected visual acuity (BCVA), central macular retinal thickness (CMT), and choroidal angiogenesis (CNV) area in the wAMD group were compared before treatment and 1 month after treatment. (3) The correlations of the relative plasma miRNA-126 expression and VEGF-A levels with the CMT and CNV area of the patients in the wAMD group before treatment and 1 month after treatment were analyzed.Results (1) The relative expression of miRNA-126 in the wAMD group (0.86±0.11) was significantly lower than that in the control group (1.04±0.09) (t=8.010, P<0.01). The VEGF-A level in the wAMD group (329.09±17.58) pg/mL was significantly higher than that in the control group (295.74±14.80)pg/mL (t=9.179, P<0.01). (2) The relative expression of miRNA-126 in the wAMD group was significantly higher 1 month after treatment (1.47±0.27) than before treatment (0.86±0.11) (t=19.410, P<0.01), and the VEGF-A level of the wAMD group 1 month after treatment (315.36±15.44) pg/mL was significantly lower than that before treatment (329.09±17.58) pg/mL (t=8.048, P<0.01). BCVA was significantly increased 1 month after treatment (0.45±0.11) compared with before treatment (0.83±0.16) (t=12.667, P<0.01). The CMT and CNV areas of the wAMD group 1 month after treatment were (296.53±21.52)μm and(0.58±0.18)mm², respectively, which were significantly reduced compared with those prior to treatment(311.88±37.25) μm and(0.70±0.17)mm², respectively], and the differences were statistically significant (t=3.602, 4.906, all P values<0.01). (3) The relative expression level of plasma miRNA-126 was negatively correlated with the VEGF-A, CMT, and CNV of the patients in the wAMD group before treatment and 1 month after treatment (r=-0.706, -0.0.723, -0.552, respectively, before treatment; r=-0.783, -0.709, -0.620, respectively, 1 month after treatment; all P values<0.01). The linear regression equations between the area of VEGF-A, CMT, and CNV and the relative expression of miRNA-126 before treatment were as follows: $\hat{Y}$_VEGF-A=-112X_miRNA-126+426, $\hat{Y}$_CMT=-243X_miRNA-126+522, and $\hat{Y}$_CNV=-0.84X_miRNA-126+1.42. Their linear regression equations after treatment were as follows: $\hat{Y}$_VEGF-A=-45.64X_miRNA-126+382, $\hat{Y}$_CMT=-57.53X_miRNA-126+381, and $\hat{Y}$_CNV=-0.41X_miRNA-126+1.18.Conclusions Ranibizumab can effectively improve BCVA by reducing the expression of VEGF-A and the areas of CMT and CNV. Ranibizumab may slow down the development of wAMD by enhancing the expression activity of the miRNA-126 gene while achieving clinical efficacy.