AIM:To evaluate if topical use of αB-crystallin minipeptides supports corneal healing following flap surgery.METHODS:Cultured corneal cells were treated with fluorescent taggedαB-crystallin mini-peptides to assess i...AIM:To evaluate if topical use of αB-crystallin minipeptides supports corneal healing following flap surgery.METHODS:Cultured corneal cells were treated with fluorescent taggedαB-crystallin mini-peptides to assess its internalization.Cultured corneal cells pre-treated with or without the mini-peptides were exposed to H_(2)O_(2) 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 withαB-crystallin mini-peptides or protein.Cultured trigeminal neurones were pre-treated either with αB-crystallin mini-peptides or crystallin protein and exposed to H_(2)O_(2) and presence of beading in the dendrites and axons was assessed.Corneal flap surgery was conducted on rabbit cornea and treated topically either withαB-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.RESULTS:Internalization ofαB-crystallin mini-peptides was ascertained by the detection of fluorescence within the corneal cells.The MTT assay revealed that treatment withαB-crystallin mini-peptide reduced cell death induced by H_(2)O_(2) treatment.The mini-peptides did not influence the elongation of trigeminal neurites,but significantly(P<0.05)reduced beading in the neurites.In rabbit eye,the treated corneas showed reduced hyper-reflective zones(P<0.05)and suppression in the expression of inflammatory cytokines.Histopathological examination also revealed reduction of inflammatory response in treated corneas.CONCLUSION:TheαB-crystallin mini-peptides restrict the damage to corneal cells and neurons and aids in corneal healing.展开更多
AIM:To evaluate the effect of epigallocatechin gallate(EGCG) in preventing lens opacity and the aggregation of lens αB-crystallin in model rats of diabetes mellitus(DM).METHODS:This experimental study included Wistar...AIM:To evaluate the effect of epigallocatechin gallate(EGCG) in preventing lens opacity and the aggregation of lens αB-crystallin in model rats of diabetes mellitus(DM).METHODS:This experimental study included Wistar rats for DM as in vivo models and divided into 5 groups.The treatment groups were administered EGCG by orally for 20d and were then assessed for their degree of lens opacity with binocular microscope and lens αB-crystallin expression from Western blot analyze.RESULTS:Pearson correlation test and regression analysis on EGCG exposure and final random blood sugar(RBS) obtained a significance level of P<0.05.EGCG exposure can significantly lower RBS with an R~2 of 0.5634(56.34%).The same analysis on EGCG exposure and the degree of lens opacity obtained a significance level of P<0.05 and increased exposure to EGCG can significantly lower the degree of lens opacity with an R~2 of 0.8577(85.77%).Correlation analysis between EGCG and the expression of lens αB-crystallin can be concluded that the higher the EGCG exposure administered,the higher the native lens αB-crystallin expression and the lower the aggregate lens αB-crystallin expression.There was also significant effect in which every 1 mg/kg body weight dose of EGCG can increase the native lens αB-crystallin expression by 0.0063 and decrease the aggregate lens αB-crystallin expression by 0.0076.CONCLUSION:The administration of EGCG at a dose of 300,600,and 1200 mg shows a significant effect on preventing lens opacity and aggregation of αB-crystallin in diabetic rat models and this research could be a biomolecular prevention of cataract.展开更多
目的β-淀粉样蛋白(Aβ)的异常聚集,导致细胞外老年斑和细胞内神经原纤维缠结的形成,从而引发的细胞毒性诱导神经元凋亡,导致渐进性认知功能障碍的临床表现,是阿尔茨海默病(AD)常见发病机制之一。因此,我们研究PNU282987激活SD新生乳鼠...目的β-淀粉样蛋白(Aβ)的异常聚集,导致细胞外老年斑和细胞内神经原纤维缠结的形成,从而引发的细胞毒性诱导神经元凋亡,导致渐进性认知功能障碍的临床表现,是阿尔茨海默病(AD)常见发病机制之一。因此,我们研究PNU282987激活SD新生乳鼠大脑皮质星形胶质细胞α7尼古丁乙酰胆碱能受体(n ACh R)后通过PI3K/Akt/HSF1信号通路介导内源性αB-crystallin(Cryab)对Aβ聚集的影响。方法分离出生24 h内SD乳鼠皮质层的星形胶质细胞进行培养、纯化、并传代,通过细胞免疫荧光的方法对星形胶质细胞进行鉴定;体外制备Aβ1-42寡聚体并进行鉴定;将细胞分为空白对照组、MLA处理组、PNU282987处理组、PNU282987+MLA处理组、LY294002处理组、PNU282987+LY294002处理组、Aβ处理组、PNU282987+Aβ处理组、PNU282987+LY294002+Aβ处理组及HSF1敲低组。收集细胞总蛋白,通过Western蛋白印迹法检测p-Akt,Cryab,HSF1和Aβ的蛋白表达水平。结果细胞免疫荧光鉴定显示培养的原代星形胶质细胞占总细胞98%以上;Western蛋白印迹法对细胞中Cryab,p-Akt,HSF1和Aβ表达水平的结果显示:与空白对照组相比较,PNU282987处理后,Cryab,p-Akt,HSF1蛋白都表现出上升的趋势(P<0.05),而单独MLA组或LY294002组无明显差异;在HSF1敲低组,PNU282987上调Cryab的作用受到明显的抑制;正常对照组未检测到Aβ的表达,与Aβ组相比,PNU282987组的Aβ表达降低(P<0.05);与PNU282987+Aβ组相比,PNU282987+LY294002+Aβ组的Aβ表达升高(P<0.01)。结论在原代星形胶质细胞模型上,PNU282987可通过激活星形胶质细胞的α7 n ACh R介导HSF1上调内源性Cryab从而抑制Aβ集聚,PI3K/Akt信号通路很可能是这个过程中的重要参与因素,为探讨Cryab和HSF1可能是治疗AD的一个潜在靶点提供了一定的基础研究。展开更多
基金Supported by the DST Nano-mission,Govt of India,Grant No DST No.SR/NM/NS-1067/2016Facilities were provided by the West Bengal University of Animal&Fishery Sciences and CSIR-IICB for conducting this research。
文摘AIM:To evaluate if topical use of αB-crystallin minipeptides supports corneal healing following flap surgery.METHODS:Cultured corneal cells were treated with fluorescent taggedαB-crystallin mini-peptides to assess its internalization.Cultured corneal cells pre-treated with or without the mini-peptides were exposed to H_(2)O_(2) 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 withαB-crystallin mini-peptides or protein.Cultured trigeminal neurones were pre-treated either with αB-crystallin mini-peptides or crystallin protein and exposed to H_(2)O_(2) and presence of beading in the dendrites and axons was assessed.Corneal flap surgery was conducted on rabbit cornea and treated topically either withαB-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.RESULTS:Internalization ofαB-crystallin mini-peptides was ascertained by the detection of fluorescence within the corneal cells.The MTT assay revealed that treatment withαB-crystallin mini-peptide reduced cell death induced by H_(2)O_(2) treatment.The mini-peptides did not influence the elongation of trigeminal neurites,but significantly(P<0.05)reduced beading in the neurites.In rabbit eye,the treated corneas showed reduced hyper-reflective zones(P<0.05)and suppression in the expression of inflammatory cytokines.Histopathological examination also revealed reduction of inflammatory response in treated corneas.CONCLUSION:TheαB-crystallin mini-peptides restrict the damage to corneal cells and neurons and aids in corneal healing.
文摘AIM:To evaluate the effect of epigallocatechin gallate(EGCG) in preventing lens opacity and the aggregation of lens αB-crystallin in model rats of diabetes mellitus(DM).METHODS:This experimental study included Wistar rats for DM as in vivo models and divided into 5 groups.The treatment groups were administered EGCG by orally for 20d and were then assessed for their degree of lens opacity with binocular microscope and lens αB-crystallin expression from Western blot analyze.RESULTS:Pearson correlation test and regression analysis on EGCG exposure and final random blood sugar(RBS) obtained a significance level of P<0.05.EGCG exposure can significantly lower RBS with an R~2 of 0.5634(56.34%).The same analysis on EGCG exposure and the degree of lens opacity obtained a significance level of P<0.05 and increased exposure to EGCG can significantly lower the degree of lens opacity with an R~2 of 0.8577(85.77%).Correlation analysis between EGCG and the expression of lens αB-crystallin can be concluded that the higher the EGCG exposure administered,the higher the native lens αB-crystallin expression and the lower the aggregate lens αB-crystallin expression.There was also significant effect in which every 1 mg/kg body weight dose of EGCG can increase the native lens αB-crystallin expression by 0.0063 and decrease the aggregate lens αB-crystallin expression by 0.0076.CONCLUSION:The administration of EGCG at a dose of 300,600,and 1200 mg shows a significant effect on preventing lens opacity and aggregation of αB-crystallin in diabetic rat models and this research could be a biomolecular prevention of cataract.
文摘目的β-淀粉样蛋白(Aβ)的异常聚集,导致细胞外老年斑和细胞内神经原纤维缠结的形成,从而引发的细胞毒性诱导神经元凋亡,导致渐进性认知功能障碍的临床表现,是阿尔茨海默病(AD)常见发病机制之一。因此,我们研究PNU282987激活SD新生乳鼠大脑皮质星形胶质细胞α7尼古丁乙酰胆碱能受体(n ACh R)后通过PI3K/Akt/HSF1信号通路介导内源性αB-crystallin(Cryab)对Aβ聚集的影响。方法分离出生24 h内SD乳鼠皮质层的星形胶质细胞进行培养、纯化、并传代,通过细胞免疫荧光的方法对星形胶质细胞进行鉴定;体外制备Aβ1-42寡聚体并进行鉴定;将细胞分为空白对照组、MLA处理组、PNU282987处理组、PNU282987+MLA处理组、LY294002处理组、PNU282987+LY294002处理组、Aβ处理组、PNU282987+Aβ处理组、PNU282987+LY294002+Aβ处理组及HSF1敲低组。收集细胞总蛋白,通过Western蛋白印迹法检测p-Akt,Cryab,HSF1和Aβ的蛋白表达水平。结果细胞免疫荧光鉴定显示培养的原代星形胶质细胞占总细胞98%以上;Western蛋白印迹法对细胞中Cryab,p-Akt,HSF1和Aβ表达水平的结果显示:与空白对照组相比较,PNU282987处理后,Cryab,p-Akt,HSF1蛋白都表现出上升的趋势(P<0.05),而单独MLA组或LY294002组无明显差异;在HSF1敲低组,PNU282987上调Cryab的作用受到明显的抑制;正常对照组未检测到Aβ的表达,与Aβ组相比,PNU282987组的Aβ表达降低(P<0.05);与PNU282987+Aβ组相比,PNU282987+LY294002+Aβ组的Aβ表达升高(P<0.01)。结论在原代星形胶质细胞模型上,PNU282987可通过激活星形胶质细胞的α7 n ACh R介导HSF1上调内源性Cryab从而抑制Aβ集聚,PI3K/Akt信号通路很可能是这个过程中的重要参与因素,为探讨Cryab和HSF1可能是治疗AD的一个潜在靶点提供了一定的基础研究。
