摘要
目的探讨叶酸对糖尿病小鼠视网膜的保护作用及其抗氧化和抗炎机制。方法采用随机数字表法将32只16周龄SPF级雄性db/db小鼠随机分为模型组和叶酸组,另选择16周龄SPF级雄性C57BL/KsJ小鼠16只作为对照,叶酸组小鼠用(2 ml)叶酸灌胃每日1次,剂量为71μg/kg,连续60 d;对照组和模型组小鼠以相同的方法给予相同体积的生理盐水,定期观察并记录各组小鼠运动、精神状态、体质量、空腹血糖(FPG)水平变化。灌胃结束后摘取小鼠眼球并分离视网膜;采用苏木精-伊红染色法评估各组小鼠视网膜组织病理学变化;收集小鼠球后血标本,采用ELISA法测定小鼠血清中同型半胱氨酸(Hcy)水平;采用实时荧光定量PCR法检测各组小鼠视网膜中肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)mRNA水平;采用Western blot法检测小鼠视网膜中B淋巴细胞瘤2蛋白(bcl-2)、bcl-2相关X蛋白(bax)、3-硝基酪氨酸(3-NT)和4-羟基壬氨酸(4-HNE)水平变化;采用生化试剂盒检测小鼠视网膜中超氧化物歧化酶(SOD)活性、8-羟基脱氧鸟苷(8-OHdG)和丙二醛(MDA)含量;采用免疫荧光染色法测定视网膜中NADPH氧化酶4(NOX4)含量。结果实验期间对照组小鼠FPG正常,体质量缓慢平稳增加;模型组小鼠FPG>16.7 mmol/L,体质量快速增加,叶酸组小鼠FPG逐渐下降,体质量增加较模型组平缓。实验后2个月,模型组小鼠血清Hcy浓度为(27.18±3.18)μmol/L,高于对照组的(8.28±2.18)μmol/L和叶酸组的(13.73±2.54)μmol/L,差异均有统计学意义(均P<0.05);对照组小鼠视网膜结构完整,模型组小鼠视网膜厚度变薄,毛细血管生成增加,可见炎症细胞浸润,叶酸组小鼠视网膜厚度较模型组增加,新生血管和炎症细胞减少。模型组视网膜中TNF-α和IL-6 mRNA相对表达量均高于对照组,叶酸组视网膜组织中TNF-αmRNA、IL-6 mRNA相对表达量低于模型组,差异均有统计学意义(均P<0.05)。叶酸组小鼠视网膜中bax�
Objective To explore the protective effects of folic acid on retinas and its anti-oxidative stress mechanism in diabetic mice.Methods Thirty-two 16-week-old SPF degree male db/db mice were randomized into model group and folic acid group,and 16 matched C57BL/KsJ mice were used as controls.Folic acid was used to the mice by oral gavage once per day with the dose of 71μg/kg(2 ml)for 60 days in the folic acid group,and the same volume of normal saline solution was used in the model group and control group in the same way.The activities,mental state,body weight,and fasting plasma glucose(FPG)of the mice were recorded during experiment.At the end of the intervention,the mice were sacrificed and the retinas and blood sample were obtained.The histopathology of the retinas was examined with hematoxylin-eosin staining;serum homocysteine(Hcy)was detected by ELISA assay;the relative expressions of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)mRNA were detected in the retinas by real-time fluorescence quantitative PCR;the relative expressions of B lymphoma 2 protein(bcl-2),bcl-2 related X protein(bax),3-nitrotyrosine(3-NT)and 4-Hydroxynonine(4-HNE)proteins were assayed by Western blot assay;superoxide dismutase(SOD),8-hydroxydeoxyguanosine(8-OHdG)and malondialdehyde(MDA)levels in the retinas were detected by biochemical kits,and immunofluorescence assay was used to detect the expression of NADPH oxidation 4(NOX4)in the retinas.The use and care of the experimental animals adhered to the ARVO Statement by the American Association for Vision and Ophthalmology Research and this study protocol was approved by Ethic Committee of Qinghai University(QHDX 2018-35).Results Over the experimental period,The FPG was normal and body weight was gradually increased in the mice of control group.The FPG>16.7 mmol/L and the mice appeared obese.In the folic acid group,both body mass and FPG of the mice were gradually reduced.At the end of drug administration,serum Hcy concentration of the mice was(27.18±3.18)μmol/L in the model gro
作者
李英兰
吴敏
薛小琴
李长兴
雷有芳
代冬芳
Li Yinglan;Wu Min;Xue Xiaoqin;Li Changxing;Lei Youfang;Dai Dongfang(Department of Ophthalmology,Qinghai Provincial People's Hospital,Xining 810007,China;School of Medicine,Qinghai University,Xining 810016,China)
出处
《中华实验眼科杂志》
CAS
CSCD
北大核心
2020年第1期23-30,共8页
Chinese Journal Of Experimental Ophthalmology
基金
青海省科技厅项目(2018-ZJ-730)。
关键词
糖尿病/并发症
视网膜
保护作用
叶酸
氧化应激
动物模型
小鼠
Diabetes/complication
Retina
Protective effect
Folic acid
Oxidative stress
Disease model
Mice
