Maity Namrata, Konar Aditya, Hazra Sarbani
Department of Veterinary Surgery & Radiology, West Bengal University of Animal and Fishery Sciences, Kolkata 700037, India.
CSIR-IICB, Kolkata 700032, India.
Int J Ophthalmol. 2024 Oct 18;17(10):1772-1779. doi: 10.18240/ijo.2024.10.02. eCollection 2024.
To evaluate if topical use of αB-crystallin mini-peptides supports corneal healing following flap surgery.
Cultured corneal cells were treated with fluorescent tagged αB-crystallin mini-peptides to assess its internalization. Cultured corneal cells pre-treated with or without the mini-peptides were exposed to HO and cell viability was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Elongation of neurites of cultured trigeminal neurones was examined following treatment either with αB-crystallin mini-peptides or protein. Cultured trigeminal neurones were pre-treated either with αB-crystallin mini-peptides or crystallin protein and exposed to HO and presence of beading in the dendrites and axons was assessed. Corneal flap surgery was conducted on rabbit cornea and treated topically either with αB-crystallin peptide (0.5 mg/mL thrice daily for 14d) or phosphate-buffered saline (PBS). Corneal healing was evaluated under slit-lamp biomicroscope, mRNA expression of inflammatory cytokines were assessed and the corneas were evaluated by histopathology.
Internalization of αB-crystallin mini-peptides was ascertained by the detection of fluorescence within the corneal cells. The MTT assay revealed that treatment with αB-crystallin mini-peptide reduced cell death induced by HO treatment. The mini-peptides did not influence the elongation of trigeminal neurites, but significantly (<0.05) reduced beading in the neurites. In rabbit eye, the treated corneas showed reduced hyper-reflective zones (<0.05) and suppression in the expression of inflammatory cytokines. Histopathological examination also revealed reduction of inflammatory response in treated corneas.
The αB-crystallin mini-peptides restrict the damage to corneal cells and neurons and aids in corneal healing.
评估局部应用αB-晶状体蛋白小肽是否有助于瓣下手术后角膜愈合。
用荧光标记的αB-晶状体蛋白小肽处理培养的角膜细胞,以评估其内化情况。对用或未用小肽预处理的培养角膜细胞进行过氧化氢(HO)处理,并通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法检测细胞活力。在用αB-晶状体蛋白小肽或蛋白质处理后,检测培养的三叉神经元神经突的伸长情况。用αB-晶状体蛋白小肽或晶状体蛋白对培养的三叉神经元进行预处理,然后进行HO处理,评估树突和轴突中串珠状结构的存在情况。对兔角膜进行角膜瓣手术,局部用αB-晶状体蛋白肽(0.5mg/mL,每日3次,共14天)或磷酸盐缓冲盐水(PBS)处理。在裂隙灯生物显微镜下评估角膜愈合情况,检测炎性细胞因子的mRNA表达,并通过组织病理学评估角膜。
通过检测角膜细胞内的荧光确定了αB-晶状体蛋白小肽的内化。MTT分析显示,αB-晶状体蛋白小肽处理可减少HO处理诱导的细胞死亡。小肽不影响三叉神经突的伸长,但显著(<0.05)减少了神经突中的串珠状结构。在兔眼中,处理后的角膜高反射区减少(<0.05),炎性细胞因子的表达受到抑制。组织病理学检查还显示处理后的角膜炎症反应减轻。
αB-晶状体蛋白小肽可限制对角膜细胞和神经元的损伤,并有助于角膜愈合。