The Effect of Overexpression of Proteasome PSMB5 on Antioxidative Ability of Human Lens Epithelial Cells
Tieying Zhang1 , Zhaohui Yuan2 , Bingsheng Lou2
1 Department of Ophthalmology, Affiliated Hexian Memorial Hospital, Southern Medical University, Hexian Memorial Medical Hospital of Panyu District, Guangzhou 511400, China 2 Zhongshan Ophthalmic Center, Sun Yat-Sen University, State Key Laboratory of Ophthalomology, Guangzhou 510060, China
Abstract:Objective: To discuss the antioxidative ability of proteasome subunit β5 (PSMB5) over expression in human lens epithelial cells. Methods: This is an experimental study. Recombinant plasmid pcDNA3.1- PSMB5 was constructed and transfected into a human lens epithelium strain SRA01/04 to form a stable transfection (experimental group), and empty pcDNA3.1 was also transfected into lens epithelial cells at the same time as a control. Two groups of cells were treated with 40 μmol/L H2O2. The nuclear morphology of the two group cells was observed by Hoechst 33342 stain, and cell apoptosis of the two group cells was dectected by flow cytometry. Data was analyzed by a paired t test. Results: Under a 40 μmol/L H2O2 treated environment, Staining by Hoechst 33342 indicated that the nuclei in the control group were significantly shrunken and condensed compared with those of the experimental group. The apoptotic percentages detected by flow cytometry in the experimental group and control group were (9.3±0.9)% and (15.7±1.9)%,and both had increased compared to pre-treatment with H2O2, which were (5.9±0.8)% and (7.1±1.1)%, respectively. The apoptotic percentage in the control group increased to a higher level than in the experimental group (t=3.742, P=0.008). Conclusions: Overexpression of proteasome PSMB5 can effectively improve the antioxidative ability of human lens epithelial cells.
张铁英1 袁钊辉2 娄秉盛2. 蛋白酶体β5过表达对人晶状体上皮细胞抗氧化能力的影响[J]. 中华眼视光学与视觉科学杂志, 2020, 22(4): 305-309.
Tieying Zhang1,Zhaohui Yuan2,Bingsheng Lou2. The Effect of Overexpression of Proteasome PSMB5 on Antioxidative Ability of Human Lens Epithelial Cells. Chinese Journal of Optometry Ophthalmology and Visual science, 2020, 22(4): 305-309. DOI: 10.3760/cma.j.cn115909-20190829-00236
Sharma K, Santhoshkumar P. Lens aging: Effects of crystallins. Biochim Biophys Acta, 2009, 1790(10): 1095-1108. DOI: 10.1016/j.bbagen.2009.05.008.
[2]
Taylor A. Mechanistically linking age-related diseases and dietary carbohydrate via autophagy and the ubiquitin proteolytic systems. Autophagy, 2012, 8(9): 1404-1406. DOI: 10.4161/ auto.21150.
[3]
Maupin-Furlow J. Proteasomes and protein conjugation across domains of life. Nat Rev Microbio, 2011, 10(2): 100-111. DOI: 10.1038/nrmicr02696.
[4]
Matyskiela M, Martin A. Design principles of a universal protein degradation machine. J Mol Biol, 2013, 425(2): 199-213. DOI: 10.1016/j.jmb.2012.11.001.
[5]
Budenholzer L, Cheng CL, Li Y, et al. Proteasome structure and assembly. J Mol Bio, 2017, 429(11): 3500-3524. DOI: 10.1016/ j.jmb.2017.05.027.
[6]
Wang X, Yen J, Kaiser P, et al. Regulation of the 26S proteasome complex during oxidative stress. Sci Signal, 2010, 3(12): 1-17. DOI: 10.1126/scisignal.2001232.
Kruk J, Kubasik-Kladna K, Aboul-Enein HY. The role oxidative stress in the pathogenesis of eye diseases: Current status and a dual role of physical activity. Mini-Rev Med Chem, 2016, 16(3): 241-257. DOI: 10.2174/1389557516666151120114605.
[9]
Radwan M, Wood RJ, Sui X, et al. When proteostasis goes bad: Protein aggregation in the cell. IUBMB Life, 2017, 69(2): 49- 54. DOI: 10.1002/iub.1597.
[10]
Shang F, Taylor A. Role of the ubiquitin-proteasome in protein quality control and signaling: Implicatn in the pathogenesis of eye diseases. Prog Mol Biol Transl Sci, 2012, 109(6): 347-396. DOI: 10.1016/B978-0-12-397863-9.00010-9.
[11]
Lachke SA, Ho JWK, Kryukov GV, et al. iSyTE: Integrated systems tool for eye gene discovery. Invest Ophthalmol Vis Sci, 2012, 53(3): 1617-1627. DOI: 10.1167/iovs.11-8839.
[12]
Baraibar MA, Friguet B. Changes of the proteasomal system during the aging process. Prog Mol Biol Transl Sci, 2012, 109(6): 249-275. DOI: 10.1016/B978-0-12-397863-9.00007-9.
[13]
Zetterberg M, Petersen A, Sjostrand J, et al. Proteasome activity in human lens nuclei and correlation with age, gender and severity of cataract. Curr Eye Res, 2003, 27(1): 45-53. DOI: 10.1076/ceyr.27.2.45.15457.
[14]
Andersson M, Sjostrand J, Kadsson JO. Differential inhibition of three peptidase activities of the proteasome in human lens epithelium by heat and oxidation. Exp Eye Res,1999, 69(1): 129-138. DOI: 10.1006/exer.1999.0688.
[15]
Tomaru U, Takahashi S, Ishizu A, et al. Decreased proteasomal activity causes age-related phenotypes and promotes the development of metabolic abnormalities. Am J Patho, 2012, 180(3): 963-972. DOI: 10.1016/j.ajpath.2011.11.012.
[16]
Zhang Y, Jiang W, Xie Z, et al. Vitamin E and risk ofage-related cataract: A meta-analysis. Public Health Nutr, 2015, 18(15): 2804-2814. DOI: 10.1017/S1368980014003115.
[17]
Tweeddale HJ, Hawkins CL, Janmie JF, et al. Cross-linking of lens crystallin proteins induced by tryptophan metabolites and Metal ions: Implications for cataract Development. Free Radic Res, 2016, 50(10): 1116-1130. DOI: 10.1080/10715762.2016. 1210802.
[18]
Elmazar HM, Elmadbouh I, Mandour SS, et al. Association between cataract progression and ischemia-modified albumin in relation to oxidant-antioxidant profiles in the serum, aqueous humor, and lens. J Cataract Refr Surg, 2018, 44(2): 134-139. DOI: 10.1016/j.jcrs.2017.10.051.
[19]
Nowotny K, Jung T, Grune T, et al. Accumulation of modified proteins and aggregate formation in aging. Exp Gerontol, 2014, 57(9): 122-131. DOI: 10.1016/j.exger.2014.05.016.