整合素α5β1在脉络膜血管新生中的研究进展

脉络膜血管新生(choroidal neovascularization,CNV)是眼内后极部病理性血管新生的重要表现,是造成视力损伤的主要病因之一。整合素α5β1因在CNV的形成过程中发挥重要作用而受到广泛关注。我们就整合素α5β1的结构、分布、活性调节,及在视网膜色素上皮细胞及脉络膜内皮细胞中的表达及对脉络膜新生血管的调节进行综述,探讨抑制血管新生的有效方法。     

A novel mutation of CYP4V2 gene associated with Bietti crystalline dystrophy complicated by choroidal neovascularization

AIM: To investigate the clinical characteristics and genetic features of a Bietti crystalline dystrophy (BCD) proband in a Chinese family. METHODS: A Chinese female diagnosed with BCD complicated by bilateral choroidal neovascularization (CNV) and her parents underwent complete ophthalmic examinations, including fundus autofluorescence (AF), fundus photography (FP), fundus fluorescein angiography (FFA), visual field testing, full-field electroretinography (ERG), optical coherence tomography (OCT) and optical coherence tomography angiography (OCTA). The sequencing of the CYP4V2 gene was performed to the whole family. RESULTS: Bilateral tiny glittering crystal-like deposits and differing extent of atrophy of the retinal pigment epithelium (RPE) were found in the posterior pole of her fundus. The diffuse hypo-fluorescence shown on AF images and window defects shown on FFA both indicated the atrophy of the RPE and choriocapillaris. OCT showed the thinning of the RPE and choriocapillaris layer, ellipsoid zone (EZ) band defect and CNV in both eyes. OCTA images proofed bilateral type 2 CNV. The visual field test showed central and paracentral scotoma. ERG showed a slightly decreased b-wave in scotopic ERG. Gene sequencing identified three mutations of the CYP4V2 gene, c.802_807del, c.810delT, and c.1388G>A. The mutation c.1388G>A was a novel substitution mutation. CONCLUSION: The novel mutation c.1388G>A may be a possible cause that could induce the clinical phenotype of BCD.     

The effect of intravitreal bevacizumab and transpupillary thermotherapy on choroidal metastases and literature review

Aims:

To represent the effects of transpupillary thermotherapy (TTT) and intravitreal bevacizumab on choroidal metastases and review the literature.

Settings and Design:

A retrospective, interventional, noncomparative case series.

Materials and Methods:

A retrospective, interventional, noncomparative case series of five eyes in three patients with choroidal metastases was conducted. Fundus findings of choroidal metastases were divided into two types: Solitary or diffuse type. The size of the tumor was termed small (<10 mm diameter), medium (10–15 mm diameter) or large (>15 mm diameter). All eyes received one session of TTT followed by 3 weekly intravitreal bevacizumab injections as an adjuvant therapy. The parameters of treatment for TTT were 1.2–3 mm spot size, 150–300 mW, 60 s with the whole lesion covered confluently. The changes in preoperative and postoperative best-corrected visual acuity (BCVA) were recorded. Serial color fundus photography and optical coherent tomography were performed to measure the treatment efficacy.

Results:

All eight choroidal metastases were solitary type. The size of six tumors was small, the size of one tumor was medium, and the size of one tumor was large. All five eyes of the three patients had improvement of BCVA after treatment. Fundus photos revealed tumor shrinkage and the mean shrinkage percentage was 61.27 ± 21.71%. Optical coherence tomography revealed complete resolution of serous retinal detachment. There was no recurrence after 6 months follow-up.

Conclusions:

TTT combined with intravitreal bevacizumab injections brought about beneficial effects in reducing tumor size and improving vision in all five eyes of the three patients. Despite the retrospective nature of our study, the absence of control group and the size limitation that, of course, limit the statistical power, TTT combined with intravitreal bevacizumab seems to be efficient in providing another cost-reducing and time-saving treatment option for patients with choroidal metastases. The antineoplastic properties of bevacizumab make it a viable adjunctive therapy. Studies with more cases and a longer follow-up period are warranted.     

Investigation of the choroidal thickness in patients with hypothyroidism

Purpose:

The effect of hypothyroidism on the choroidal thickness (CT) was investigated in patients with subclinical hypothyroidism and overt hypothyroidism, and biochemically and clinically euthyroid patients receiving levothyroxine treatment. The patients were compared with healthy subjects.

Materials and Methods:

One eye of 71 hypothyroid and 22 healthy subjects between 20 and 40 years of age were included in this study. CT measurements were taken at the fovea and at 2 points that were 1500 μm nasal and temporal to the fovea using spectral-domain optical coherence tomography. Independent sample t-test''s and was used for statistical analysis of the data.

Results:

The CT was significantly thicker in hypothyroid patients compared to healthy subjects (P values were 0.013 for subfoveal, 0.015 for temporal and 0.020 for nasal segments). The intraocular pressure (IOP) and body mass index (BMI) were also significantly higher in hypothyroid patients (P values were 0.021 and 0.003, respectively). There was not a statistically significant difference in the BMI and IOP measurements between healthy subjects and euthyroid patients (P > 0.05). However, there was a statistically significant difference in the subfoveal, temporal and nasal CT measurements between healthy subjects and euthyroid patients (P values were 0.006, 0.031 and 0.013, respectively).

Conclusions:

All subgroups of hypothyroid patients had thicker CT compared to healthy subjects. Euthyroid patients receiving levothyroxine treatment had lower IOP, BMI levels, and serum lipid levels than patients with subclinical hypothyroidism and overt hypothyroidism.     

MEK/ERK参与大鼠脉络膜新生血管基质金属蛋白酶-2和基质金属蛋白酶-9的表达调控

目的 探讨基质金属蛋白酶2(matrix metalloproteinase-2,MMP-2)和MMP-9在大鼠脉络膜新生血管(choroidal neovascularization,CNV)内的表达及其可能的调控机制.方法 将23只成年雄性棕色挪威大鼠随机分为2组,一组为玻璃体内注药组,视网膜光凝后即刻玻璃体内注射3μL PD98059;另一组为单纯光凝组,单纯行视网膜光凝.观察时间为光凝后3d、7d和14 d.各时间点处死大鼠后摘除眼球,免疫组织化学法和免疫荧光法观察MMP-2和MMP-9在大鼠CNV内不同时间点的表达特点.眼底荧光血管造影(fundusfluorescence angiography,FFA)和HE法观察玻璃体内注射PD98059对CNV生成的作用,Western blotting检测观察注药对CNV内MMP-2和MMP-9表达的影响.结果 光凝后3d,单纯光凝组光凝区即有MMP-2和MMP-9的阳性表达;光凝后7d和14 d二者表达均逐渐增强.光凝后7d,玻璃体内注药组MMP-2和MMP-9均显著被抑制,分别被抑制约69％和80％.Western blotting检测结果示玻璃体内注射组可显著抑制ERK的磷酸化,光凝后3d及7d的抑制率分别为54％和60％,而对ERK总量的表达无明显作用.FFA和HE显示,玻璃体内注药组光凝后14 d减少光凝局部CNV的荧光素渗漏约51％,抑制CNV厚度达57％.免疫荧光法检测结果示光凝后7d,玻璃体内注药组抑制MMP-2和MMP-9的表达分别为73％和64％.结论 MMP-2和MMP-9参与了大鼠CNV的生成,光凝后3d即有阳性表达,至光凝后14 d表达进一步增强,MEK/ERK通路至少部分参与了MMP-2和MMP-9在CNV内的生成调控.     

玻璃体腔注射雷珠单抗治疗渗出型老年性黄斑变性伴浆液性视网膜色素上皮脱离的疗效观察

目的 观察玻璃体腔注射雷珠单抗(IVR)治疗渗出型老年性黄斑变性(AMD)伴浆液性视网膜色素上皮脱离(PED)的临床效果.方法 临床确诊为渗出型AMD伴浆液性PED并行IVR治疗的23例患者23只眼纳入研究.所有患者采用国际标准对数视力表行最佳矫正视力(BCVA)检查,检测结果转换为最小分辨角对数(logMAR)记录;并采用光相干断层扫描(OCT)检查测量PED高度、PED容积及黄斑中心凹视网膜厚度(CFT).患眼平均logMAR BCVA为0.77±0.39,平均PED高度为(357.2±171.9) μm,平均PED容积为(0.741±1.012) mm3,平均CFT为(317.9±73.8)μm.治疗方法为手术室无菌条件下常规玻璃体腔注射10 mg/ml的雷珠单抗0.05 ml(含雷珠单抗0.5 mg),每一个月注射1次,连续注射3次,此后根据随访情况按需注射.以末次治疗后6个月为疗效判定时间点,对比分析治疗前后患眼BCVA、PED高度、PED容积及CFT变化情况.结果 治疗后患眼平均logMAR BCVA为0.61±0.27,较治疗前明显提高,差异有统计学意义(t=2.601,P＜0.05).23只眼中,视力提高17只眼,视力稳定4只眼,视力下降2只眼.治疗后患眼平均PED高度为(247.7±171.7)μm,平均PED容积为(0.337±0.498) mm3,平均CFT为(302.5±89.3) μm.与治疗前比较,治疗后患眼平均PED高度、平均PED容积均明显减小,差异有统计学意义(t=3.192、2.502,P＜0.05);平均CFT有所降低,但差异无统计学意义(t=0.887,P＞0.05).所有患者在随访期内均未发生眼内炎、葡萄膜炎等眼部不良反应.结论 IVR能安全有效地治疗伴有浆液性PED的渗出型AMD,提高患者视力,降低PED高度,减小PED容积.     

PDT联合ranibizumab玻璃体腔注射治疗年龄相关性黄斑变性合并CNV

目的：观察 PDT 联合玻璃体腔注射 ranibizumab (雷珠单抗)治疗老年性黄斑变性脉络膜新生血管( choroidal neovascularization,CNV)的疗效。
　　方法：将符合纳入标准,经吲哚青绿脉络膜血管造影(indocyanine green angiography, ICGA)、光学相干断层扫描( optical coherence tomography, OCT)检查确诊为黄斑区脉络膜新生血管( CNV)患者27例27眼,经PDT治疗后3~7 d内行 ranibizumab 玻璃体腔注射。观察治疗后1,3,6 mo、末次随访时行最佳矫正视力、FFA、ICGA、OCT 检查及有无并发症发生情况。
　　结果：最佳矫正视力提高17眼(63%),最佳矫正视力稳定6眼(22%),最佳矫正视力下降4眼(15%)。27例27眼治疗前平均渗漏面积为1005.69±105.47μm,治疗后1,3mo后平均875.54±103.27,423.37±79.68μm,与治疗前比较差异有统计学意义(P<0.01),视网膜黄斑中央厚度27例27眼治疗前平均厚度为485.58±122.59μm,治疗后1,3mo后平均398.84±105.32,297.74±89.18μm,与治疗前比较差异有统计学意义(P<0.01)。
　　结论：PDT 封闭 CNV 后,联合玻璃体内腔内注射ranibizumab,有效阻断新生血管复发,减少PDT再次治疗次数和并发症,可提高治疗效果。     

糖尿病大鼠玻璃体腔注射缺氧诱导因子-1α siRNA对血管内皮生长因子蛋白表达的影响

目的：观察缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)小干扰RNA(siRNA)对糖尿病大鼠视网膜组织中血管内皮生长因子(vascular endothelial growth factor,VEGF)蛋白表达的抑制作用,及其用于糖尿病新生血管性疾病治疗的可行性。

方法：以pSilencer2.1-U6neo为质粒载体,构建HIF-1α.siRNA 重组质粒。选取雄性Sprague-Dawley(SD)大鼠54只,随机分为正常对照组(15只)和实验组(39只)。实验组采用尾静脉注射链尿佐菌素(STZ)的方法建立糖尿病大鼠模型,造模成功后将实验组随机分为糖尿病组(15只)、基因治疗组(12只)和空载体组(12只)。正常对照组及糖尿病组大鼠均不做转染; 基因治疗组和空载体组分别转染HIF-1α.siRNA 重组质粒和pSilencer空载体质粒。行苏木精-伊红染色(hematoxylin-eosin staining,HE染色)观察视网膜组织形态,并采用免疫组织化学法检测VEGF蛋白的表达。分别于干扰后24、48、72h,1wk时计算VEGF蛋白的抑制效率。采用单因素方差分析和LSD-t检验进行统计学分析。

结果：HIF-1α.siRNA 重组质粒经酶切、测序鉴定,确定为目的序列。HE染色显示：正常对照组大鼠视网膜各层细胞排列整齐,细胞形态基本正常。糖尿病大鼠视网膜各层细胞排列紊乱,内界膜不完整,新生血管芽、新生血管簇呈垂直状突破内界膜生长。免疫组织化学染色显示：VEGF阳性表达为细胞浆出现棕黄色颗粒,主要位于神经节细胞层。正常对照组VEGF蛋白呈弱阳性表达,而DR对照组和空载体组表达明显增强,基因治疗组较DR组和空载体组表达明显减少,差异有统计学意义(P<0.05)。VEGF蛋白抑制率24、48、72h和1wk时分别为：27.4%、40.6%、47.5%、64.5%。

结论：HIF-1α.siRNA重组质粒能够抑制糖尿病大鼠视网膜中VEGF蛋白的表达,可能成为一种治疗糖尿病性新生血管疾病的新方法。     

CCR7信号通路与视网膜新生血管

视网膜新生血管性疾病是致盲的主要原因。趋化因子受体7(C-C chemokine receptor type 7, CCR7)可通过细胞外信号调节激酶(extracellular signal regulate kinase,ERK)通路促进血管内皮生长因子(vascular endothelial growth factor, VEGF)的表达,导致血管渗漏、血管内皮细胞增生以及新生血管形成等改变。趋化因子受体7的检测可指导视网膜新生血管性疾病的诊治。