| 冯梅,江霞,王艳丽,等.富氢水对糖尿病视网膜病变大鼠硫氧还蛋白相互作用蛋白/核苷酸结合寡聚化结构域样受体蛋白 3通路及视网膜血管通透性的影响[J].安徽医药,2022,26(7):1367-1373. |
| 富氢水对糖尿病视网膜病变大鼠硫氧还蛋白相互作用蛋白/核苷酸结合寡聚化结构域样受体蛋白 3通路及视网膜血管通透性的影响 |
| Effects of hydrogen rich water on TXNIP/NLRP3 pathway and retinal vascular permeability in diabetic retinopathy rats |
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| DOI：10.3969/j.issn.1009-6469.2022.07.022 |
| 中文关键词: 氢 糖尿病视网膜病变 富氢水 血-视网膜屏障 硫氧还蛋白相互作用蛋白/核苷酸结合寡聚化结构域样受体蛋白 3 大鼠，Sprague-Dawley |
| 英文关键词: Hydrogen Diabetic retinopathy Hydrogen rich water Blood retinal barrier Thioredoxin interacting protein/nu-cleotide binding oligomerization domain-like receptor protein 3 Rats,Sprague-Dawley |
| 基金项目:武汉市卫生计生委科研项目(WX17Q37) |
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| 中文摘要: |
| 目的探究富氢水对糖尿病视网膜病变大鼠视网膜血管通透性及硫氧还蛋白相互作用蛋白(TXNIP)/核苷酸结合寡聚化结构域样受体蛋白 3(NLRP3)通路的影响。方法 2020年 6—9月，SD大鼠采用随机数字表法选取 15只为空白组，其余大鼠尾静脉注射链脲佐菌素(STZ，60 mg/kg)和高脂饲料喂养构建 DR模型，造模成功的大鼠采用随机数字表法分为模型组、富氢水低、高剂量组(5、10 mL/kg)每组 15只。连续给药 28 d后，记录大鼠体质量，检测空腹血糖(FBG)水平；眼底照相和荧光素血管造影(FFA)观察大鼠右眼新生，血管和荧光素渗漏现象；光学相干断层扫描(OCT)检测视网膜厚度；伊文思蓝-白蛋白复合物渗漏分析视网膜血管通透性；HE染色观察视网膜病理变化；免疫荧光染色观察新生血管形成情况；TUNEL染色检测视网膜细胞凋亡；酶联免疫吸附(ELISA)试剂盒测量视网膜匀浆中血管生成素-1(Ang-1)、血管内皮生长因子(VEGF)、白细胞介素 1β(IL-1β)、白细胞介素 6(IL-6)水平；WB法检测视网膜 TXNIP/NLRP3通路和凋亡相关蛋白的表达。结果与空白组相比，模型组大鼠 FBG［(26.49±2.18)mmol/L比(5.76±1.09)mmol/L］、血-视网膜屏障(BRB)通透性［(32.51±2.05)μg/g比(11.24±1.76)μg/g］、视网膜细胞凋亡［(23.31±2.14)%比(0.07±0.01)%］、Bcl-2相关 X蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)、TXNIP［(0.85±0.09)比(0.40±0.05)］、NLRP3［(0.93±0.10)比(0.48±0.07)］、IL-1β［(0.74±0.05)比(0.41±0.04)］、胱天蛋白酶-1(caspase-1)［(0.78±0.07)比(0.24±0.04)］表达、Bax/Bcl-2比值、视网膜匀浆中 Ang-1、VEGF、IL-1β、IL-6水平显著增加(P<0.05)，体质量、视网膜厚度［(148.31±12.76)μm比(223.57±17.23)μm］显著降低(P<0.05)，视网膜有较多新生血管和血管曲折，血管渗漏严重，视网膜细胞明显水肿，有大量炎性渗出，神经节细胞层(GCL)细胞数量减少；与模型组相比，富氢水低、高剂量组大鼠 FBG［(20.85±3.45) mmol/L、(14.27±2.42)mmol/L比(26.49±2.18)mmol/L］、BRB通透性［(24.67±1.85)μg/g、(17.48±1.63)μg/g比(32.51±2.05)μg/g］、视网膜细胞凋亡［(14.65±1.36)%、(9.85±1.22)%比(23.31±2.14)%］、Bax、Bcl-2、TXNIP［(0.64±0.09)、(0.52±0.08)比(0.85±0.09)］、NLRP3［(0.72±0.09)、(0.61±0.08)比(0.93±0.10)］、IL-1β［(0.62±0.06)、(0.53±0.05)比(0.74±0.05)］、caspase-1［(0.57±0.06)、(0.41±0.05)比(0.78±0.07)］表达、Bax/Bcl-2比值、视网膜匀浆中 Ang-1、VEGF、IL-1β、IL-6水平明显降低(P<0.05)体质量、视网膜厚度［(173.62±14.46)μm、(196.54±15.75)μm比(148.31±12.76)μm］明显增加(P<0.05)视网膜水肿情况减轻， CLG，细胞数量明显增加，损伤得到改善。结论富氢水可抑制新生血管形成，降低 DR大鼠视网膜血管通，透性，减轻视网膜神经元损伤；其作用机制可能与抑制 TXNIP/NLRP3通路的激活有关。 |
| 英文摘要: |
| Objective To investigate the effect of hydrogen rich water on retinal vascular permeability and thioredoxin interactingprotein (TXNIP)/nucleotide binding oligomerization domain-like receptor protein 3 (NLRP3) pathway in diabetic retinopathy rats.Meth. ods Fifteen SD rats purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd. from June to September 2020 wereselected by random number table method as blank group. The rest rats were injected with streptozotocin (STZ, 60 mg/kg) via tail vein andfed with high-fat diet to establish DR model. The successful rats were randomly assigned into model group, low and high-dose hydrogenrich water groups (5, 10 mL/kg) by random number table method, with 15 rats in each group. After 28 days of continuous administration,the body weight of rats was recorded and the level of fasting blood glucose (FBG) was detected; fundus photography and fluorescein angiog-raphy (FFA) were used to observe neovascularization and fluorescein leakage; optical coherence tomography (OCT) was used to measurethe retinal thickness; Evans blue albumin complex leakage was used to analyze retinal vascular permeability; HE staining was used to ob-serve the pathological changes of retina; immunofluorescence staining was used to observe the formation of neovascularization; TUNELstaining was used to detect the apoptosis of retinal cells; Enzyme-linked immunosorbent (ELISA) kit was used to measure the levels of an-giopoietin-1 (Ang-1), vascular endothelial growth factor (VEGF), interleukin-1β (IL-1β) and interleukin-6 (IL-6) in retinal homogenate;WB method was used to detect the expression of TXNIP/NLRP3 pathway and apoptosis related proteins in retina.Results FBG [(26.49± 2.18) mmol/L vs. (5.76±1.09) mmol/L], Blood retinal barrier (BRB) permeability [(32.51±2.05) μg/g vs. (11.24±1.76) μg/g], retinal cell apoptosis [(23.31±2.14)% vs. (0.07±0.01)%], expression of Bax, Bcl-2, TXNIP [(0.85±0.09) vs. (0.40±0.05)], NLRP3 [(0.93±0.10) vs. (0.48±0.07)], IL-1β [(0.74±0.05) vs. |
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