Effects of probucol on high glucose-induced specificity protein 1/Keap1/Nrf2/glutamate-cysteine ligase catalytic in the cultured human müller cells
10.3760/cma.j.issn.1005-1015.2019.02.015
- VernacularTitle:普罗布考对高糖培养人视网膜Müller细胞特化蛋白1/细胞骨架相关蛋白/核因子E2相关因子2/半胱氨酸连接酶催化亚基表达的影响
- Author:
Chenxiang LI
1
;
Shibei AI
;
Zhongping CHEN
;
Xuxia ZHOU
Author Information
1. 中南大学爱尔眼科学院
- Keywords:
Probucol/therapeutic use;
Spl transcription factor;
NF-E2-related factor 2;
Glutamate-cysteine ligase;
Müller cell;
Kelchlike ECH assoiated proteinl
- From:
Chinese Journal of Ocular Fundus Diseases
2019;35(2):187-191
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the expression ofprobucol on high glucose-induced specificity protein 1 (SP 1),kelchlike ECH associated protein 1 (Keap 1),NF-E2-related factor 2 (Nrf2) and glutamatecysteine ligase catalytic (GCLC) in the cultured human müller cells and preliminary study the antioxidation of the probucol on müller cells.Methods Primary cultured human müller cells were randomly divided into four groups:normoglycaemia group (5.5 mmol/L glucose),normoglycaemia with probucol group (5.5 mmol/L glucose+100 μmol/L probucol),hyperglycemia group (25.0 mmol/L glucose),hyperglycemia with probucol group (25.0 mmol/L glucose + 100 μmol/L probucol).Immunofluorescence staining was used to assess distribution of SP1,Keapl,Nrf2,GCLC in human Müller cells.SP1,Keapl,Nrf2 and GCLC messenger RNA (mRNA) expression was evaluated by quantitative real-time RT-PCR (qRT-PCR).Independent sample t test was used to compare the data between the two groups.Results All müller cells expressed glutamine synthetase (＞ 95％),which confirmed the cultured cells in vitro were the purification of generations of müller cells.The expressions of SP 1,Keap 1,Nrf2,and GCLC protein were positive in human müller cells.qRT-PCR indicated that SP1 (t=28.30,P＜0.000),Keap1 (t=5.369,P=0.006),and Nrf2 (t=10.59,P=0.001) mRNA in the hyperglycemia group increased obviously compared with the normoglycaemia group;GCLC (t=4.633,P=0.010)mRNA in the hyperglycemia group decreased significantly compared with the normoglycaemia group.However,SP1 (t=12.60,P=0.000) and Keapl (t=4.076,P=0.015) in the hyperglycemia with probucol group decreased significantly compared with the hyperglycemia group;Nrf2 (t=12.90,P=0.000) and GCLC (t=l 5.96,P＜0.000)mRNA in the hyperglycemia with probucol group increased obviously compared with with the hyperglycemia group.Conclusion Probucol plays an antioxidant role by inhibiting the expression of SP 1,Keap 1 and upregulating the expression of Nrf2,GCLC in müller cells induced by high glucose.
