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引用:张凤久,张丽敏,王宏杰,李海波,王海明,彭向东,杨建玲.发光二极管照射对大鼠高糖视网膜血管内皮细胞的光生物调节作用及其机制.国际眼科杂志 2020;20(8):1339-1342,doi:10.3980/j.issn.1672-5123.2020.8.08
发光二极管照射对大鼠高糖视网膜血管内皮细胞的光生物调节作用及其机制
Photobiological regulation of high glucose retinal vascular endothelial cells in rats by light emitting diode irradiation and its mechanisms
投稿时间:2019-11-19  修订日期:2020-07-08
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DOI:10.3980/j.issn.1672-5123.2020.8.08
关键词:  发光二极管  大鼠  血管内皮细胞  光生物  机制
Key Words:  light emitting diodes  rats  vascular endothelial cells  photobiology  mechanism
基金项目:国家自然科学基金资助项目(No.61465001); 内蒙古自治区高等学校科学研究项目(No.NJZY18214)
Fund Project:National Natural Science Foundation of China(No.61465001); Scientific Research Foundation of Inner Mongolia Autonomous Region Higher Education Institutions(No.NJZY18214)
                    
作者单位
张凤久 中国内蒙古自治区赤峰市,赤峰学院附属医院眼科
张丽敏 中国内蒙古自治区赤峰市,赤峰学院附属医院眼科
王宏杰 中国内蒙古自治区赤峰市,赤峰学院附属医院眼科
李海波 中国内蒙古自治区赤峰市,赤峰学院附属医院眼科
王海明 中国内蒙古自治区赤峰市,赤峰学院附属医院眼科
彭向东 中国内蒙古自治区赤峰市,赤峰学院附属医院眼科
杨建玲 中国内蒙古自治区赤峰市,赤峰学院附属医院眼科
                    
AuthorInstitution
Feng-Jiu Zhang Department of Ophthalmology, Affiliated Hospital of Chifeng College,Chifeng , Inner Mongolia Autonomous Region, China
Li-Min Zhang Department of Ophthalmology, Affiliated Hospital of Chifeng College,Chifeng , Inner Mongolia Autonomous Region, China
Hong-Jie Wang Department of Ophthalmology, Affiliated Hospital of Chifeng College,Chifeng , Inner Mongolia Autonomous Region, China
Hai-Bo Li Department of Ophthalmology, Affiliated Hospital of Chifeng College,Chifeng , Inner Mongolia Autonomous Region, China
Hai-Ming Wang Department of Ophthalmology, Affiliated Hospital of Chifeng College,Chifeng , Inner Mongolia Autonomous Region, China
Xiang-Dong Peng Department of Ophthalmology, Affiliated Hospital of Chifeng College,Chifeng , Inner Mongolia Autonomous Region, China
Jian-Ling Yang Department of Ophthalmology, Affiliated Hospital of Chifeng College,Chifeng , Inner Mongolia Autonomous Region, China
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目的:观察发光二极管照射对大鼠高糖视网膜血管内皮细胞中的光生物调节作用及其机制。

     方法:大鼠视网膜血管内皮细胞随机分为三组:正常对照组、高糖模型组、高糖模型发光二极管照射组,高糖模型发光二极管组的细胞在造模48h后开始采用发光二极管对培养箱中的细胞进行照射。MTT细胞凋亡实验检测各组细胞凋亡率; 激光共聚焦显微镜观察各组视网膜血管内皮细胞胞内钙离子变化; Western blot 法检测各组磷酸化丝氨酸-苏氨酸激酶(P-AKT)蛋白表达。

     结果:正常对照组、高糖模型组、高糖模型发光二极管照射组凋亡率分别为7.54%±2.67%,31.69%±5.74%,21.65%±3.52%(P<0.05)。正常对照组细胞质微弱Ca2+荧光染色呈现出绿色的荧光,其荧光像素值为192.65±50.54; 高糖模型组中,细胞质呈现较强烈的绿色荧光,其荧光像素值为710.69±100.38; 发光二极管照射组中,绿色荧光像素值为430.47±80.67,明显高于正常对照组,但明显低于高糖模型组。三组间细胞中内Ca2+荧光像素值有差异(P<0.05)。这三组细胞P-AKT蛋白量分别为10.26±2.47、2.35±0.16、7.46±1.64(P<0.05)。

     结论:高糖环境抑制苏氨酸激酶通路活性,对大鼠视网膜血管内皮细胞钙稳态产生影响,促使细胞凋亡,低强度的发光二极管照射可激活苏氨酸激酶通路,降低高糖引起的细胞凋亡率。

Abstract:
      AIM: To observe the application of light emitting diode(LED)irradiation in rat retinal vascular endothelial cells with high glucose and its effect and mechanism on photobioregulation.

     METHODS:Rat retinal vascular endothelial cells were randomly divided into three groups: normal control group, high glucose model group, high glucose model light emitting diode irradiation group, and cells in the high glucose model light emitting diode group began to use light emitting diodes in the incubator 48h after modeling. The cells are irradiated. MTT cell apoptosis experiment was used to detect the apoptosis rate of each group; laser confocal microscope was used to observe the changes of intracellular calcium in retinal vascular endothelial cells; Western blot was used to detect the phosphorylated serine-threonine kinase(P-AKT)protein in each group expression.

     RESULTS: The apoptosis rates of normal control group, high glucose model group, and high glucose model light emitting diode irradiation group were 7.54%±2.67%, 31.69%±5.74%, and 21.65%±3.52%, respectively(P<0.05). In the normal control group, the cytoplasm with weak Ca2+ fluorescence showed green fluorescence with a pixel value of 192.65±50.54. In the high-sugar model group, the cytoplasm showed a stronger green fluorescence with a fluorescent pixel value of 710.69±100.38. The green fluorescent pixel value was 430.47±80.67, which was significantly higher than the normal control group, but significantly lower than the high-sugar model group. The intra-Ca2+ fluorescence pixel values in the three groups were statistically significant(P<0.05). The amount of phosphorylated serine-threonine kinase(P-AKT)protein in these three groups of cells was 10.26±2.47, 2.35±0.16, 7.46±1.64, respectively(P<0.05).

     CONCLUSION: High-glucose environment inhibits the activity of threonine kinase pathway, which has an effect on calcium homeostasis of rat retinal vascular endothelial cells and promotes apoptosis. Low-intensity led irradiation can activate threonine kinase pathway and reduce the apoptosis rate caused by high glucose, which is of great application value.

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