摘要
目的观察当归多糖对高糖诱导的视网膜神经节细胞(RGC)氧化应激损伤及凋亡的影响,并探讨可能机制。方法取对数期RGC-5细胞,分为对照组(常规培养基培养)、高糖组(含葡萄糖30 mmol/L的培养基培养)及高糖+低、中、高剂量组(分别用含葡萄糖30 mmol/L与当归多糖100、200、400 mg/L的培养基培养)。另取对数期RGC-5细胞,将细胞分为高糖+mi R-NC组、高糖+mi R-148a-3p mimics组、高糖+mi R-148a-3p inhibitor组、高糖+当归多糖+mi R-148a-3p mimics组、高糖+当归多糖+mi R-148a-3p inhibitor组。ELISA法检测细胞超氧化物歧化酶(SOD)活性及丙二醛(MDA)水平;Annexin V/PITC双染法检测细胞凋亡率;Western blot法检测细胞Bcl-2相关X蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)蛋白表达量。结果与对照组比较,高糖组SOD活性、Bcl-2蛋白表达量降低,MDA水平、凋亡率、Bax蛋白表达量升高(P<0.05);与高糖组比较,高糖+低剂量组SOD活性、Bcl-2蛋白表达量升高,MDA水平、凋亡率、Bax蛋白表达量降低(P<0.05);与高糖+低剂量组比较,高糖+中剂量组SOD活性、Bcl-2蛋白表达量升高,MDA水平、凋亡率、Bax蛋白表达量降低(P<0.05);与高糖+中剂量组比较,高糖+高剂量组SOD活性、Bcl-2蛋白表达量升高,MDA水平、凋亡率、Bax蛋白表达量降低(P<0.05)。与高糖+mi R-NC组比较,高糖+mi R-148a-3p mimics组mi R-148a-3p表达量及SOD活性、Bcl-2蛋白表达量升高,MDA水平、凋亡率、Bax蛋白表达量降低(P<0.05),高糖+mi R-148a-3p inhibitor组mi R-148a-3p表达量及SOD活性、Bcl-2蛋白表达量降低,MDA水平、凋亡率、Bax蛋白表达量升高(P<0.05);与高糖+mi R-148a-3p mimics组比较,高糖+当归多糖+mi R-148a-3p mimics组mi R-148a-3p表达量及SOD活性、Bcl-2蛋白表达量升高,MDA水平、凋亡率、Bax蛋白表达量降低(P<0.05);与高糖+mi R-148a-3p inhibitor组比较,高糖+当归多糖+mi R-148a-3p inhibitor组mi R-148a-3p表达量及SOD活性、Bcl-2蛋白表达量升高,MDA水平、凋亡率、Bax蛋白表达量降低(P<0.05)。结论当归多糖可减轻高糖诱导的RGC氧化应激损伤,减少细胞凋亡,且表现为剂量依赖性,其作用机制可能与上调mi R-148a-3p表达有关。
Objective To observe the effect of Angelica polysaccharide on the oxidative stress injury and apoptosis of retinal ganglion cells(RGC)induced by high glucose,and explore its possible mechanism.Methods Log phase RGC-5 cells were taken and divided into a control group(cultured in conventional medium),a high glucose group(cultured in medium containing 30 mmol/L glucose),and high glucose+low-,medium-and high-dose groups(cultured with medium containing 30 mmol/L of glucose and 100,200,and 400 mg/L of Angelica polysaccharide,respectively).Log-phase RGC-5 cells were also taken and divided into a high glucose+miR-NC group,a high glucose+miR-148a-3p mimics group,a high glucose+miR-148a-3p inhibitor group,a high glucose+Angelica polysaccharide+miR-148a-3p mimics group,a high glucose+Angelica polysaccharide+miR-148a-3p inhibitor group.The activity of superoxide dismutase(SOD)and the level of malondialdehyde(MDA)in cells were detected by ELISA.The apoptosis rate of cells was detected by AnnexinV/PITC double staini ng.Western blot was used to detect the expression levels of Bcl-2-related X protein(Bax)and B-lymphocytoma-2(Bcl-2)in cells.Reessuullttss Compared with the control group,the SOD activity and Bcl-2 protein expression were decreased,while the MDA level,apoptosis rate,and Bax protein expression were increased in the high glucose group(P<0.05).Compared with the high-glucose group,the SOD activity and Bcl-2 protein expression were increased,while the MDA level,apoptosis rate,and Bax protein expression were decreased in the high glucose+low-dose group(P<0.05).Compared with the high glucose+low-dose group,the SOD activity and Bcl-2 protein expression were increased,while the MDA level,apoptosis rate,and Bax protein expression were decreased in the high glucose+middle-dose group(P<0.05).Compared with the high glucose+middle-dose group,the SOD activity and Bcl-2 protein expression were increased,while the MDA level,apoptosis rate,and Bax protein expression were decreased in the high glucose+high-dose group(P<0.05).Compared with the high glucose+miR-NC group,the miR-148a-3p expression,SOD activity,and Bcl-2 protein expression were increased,while the MDA level,apoptosis rate,and Bax protein expression were decreased in the high glucose+miR-148a-3p mimics group(P<0.05),and the miR-148a-3p expression,SOD activity,and Bcl-2 protein expression were decreased,while the MDA level,apoptosis rate,and Bax protein expression were increased in the high glucose+miR-148a-3p inhibitor group(P<0.05).Compared with the high glucose+miR-148a-3p mimics group,the miR-148a-3p expression,SOD activity,Bcl-2 protein expression were increased,while the MDA level,apoptosis rate,Bax protein expression were decreased in the high glucose+Angelica polysaccharide+miR-148a-3p mimics group(P<0.05).Compared with the high glucose+miR-148a-3p inhibitor group,the miR-148a-3p expression,SOD activity,Bcl-2protein expression were increased,while the MDA level and apoptosis rate,Bax protein expression were decreased in the high glucose+Angelica polysaccharide+miR-148a-3p inhibitor group(P<0.05).CCoonncclluussiioonn Angelica polysaccharide can alleviate the oxidative stress injury of RGC induced by high glucose and reduce the apoptosis in a dose-dependent manner,whose mechanism may be related to the up-regulation of miR-148a-3p expression.
作者
曹朗
王艳新
贾冠美
CAO Lang;WANG Yanxin;JIA Guanmei(Department of Ophthalmology,Baoding Second Central Hospital,Baoding 072750,China)
出处
《吉林中医药》
2023年第1期73-78,共6页
Jilin Journal of Chinese Medicine
基金
保定市科技计划项目(2141ZF218)。
作者简介
曹朗(1986-),女,主治医师,主要从事眼科疾病研究。
