CTSB和bFGF在高氧诱导小鼠视网膜新生血管形成中的表达

Expression of CTSB and b FGF in hyperoxia-induced retinal neovascularization in mice

目的研究组织蛋白酶B(CTSB)和碱性成纤维细胞生长因子(b FGF)在高氧诱导小鼠视网膜新生血管形成中的表达及其相关性。方法7日龄C57BL/6J小鼠56只被随机分为对照组、高氧诱导组、DMSO组和CA-074Me组。对照组小鼠在普通环境下饲养;高氧诱导组、DMSO组和CA-074Me组在高氧环境中饲养5 d建立氧诱导视网膜病变模型;DMSO组和CA-074Me组小鼠玻璃体腔分别注射1μl DMSO和1μl CA-074Me,各1次。各组处理后的小鼠转至标准环境饲养5 d后,采用q PCR和Western blot法分别检测各组小鼠视网膜组织中CTSB和b FGF mRNA和蛋白相对表达水平。结果正常对照组、高氧诱导组、DMSO组和CA-074Me组间小鼠视网膜中b FGF和CTSB mRNA相对表达量总体比较,差异有统计学意义(b FGF:F=226.89,P=0.000;CTSB:F=25.23,P<0.05)。四组小鼠视网膜中b FGF和CTSB蛋白相对表达量总体比较,差异有统计学意义(b FGF:F=121.84,P<0.05;CTSB:F=30.69,P=0.000)。与正常对照组相比较,高氧诱导组和DMSO组中CTSB与b FGF的mRNA和蛋白表达量显著增高;与高氧诱导组相比较,CA-074Me组中CTSB与b FGF的mRNA和蛋白表达量则降低(均P<0.05)。正常对照组和CA-074Me组比较,高氧诱导组和DMSO组比较,CTSB与b FGF的mRNA和蛋白表达量差异均无统计学意义(P>0.05)。结论CA-074 Me抑制高氧诱导小鼠视网膜组织中CTSB的表达,同时下调b FGF表达。这一机制可能是,CA-074 Me通过抑制CTSB抑制b FGF的表达;也可能是CA-074 Me直接作用于b FGF,其具体机制需要在后期的实验中进一步验证。

Objective To investigate the expression of cathepsin B(CTSB)and basic fibroblast growth factor(b FGF)and their correlation in hyperoxia-induced retinal neovascularization in mice.Methods Fifty-six C57 BL/6 J mice were randomly divided into control group,hyperoxia group,DMSO group and CA-074 Me group.The mice were fed in the normal environment in control group,while the mice in hyperoxia group,DMSO group and CA-074 Me group were fed in a high-oxygen environment for 5 d to establish an oxygen-induced retinopathy model.Moreover,the mice in DMSO group and CA-074 Me group were injected with 1μl DMSO and 1μl CA-074 Me(one time)in the vitreous cavity,respectively.The mice in each group were transferred to the standard environment for 5 d after treatment,and then the relative mRNA and protein expression levels of CTSB and b FGF in the retinal tissues were detected by q PCR and Western blot.Results The relative expression of b FGF and CTSB mRNA in the retina was statistically different among control group,hyperoxia group,DMSO group and CA-074 Me group(b FGF:F=226.89,P=0.000;CTSB:F=25.23,P<0.05).The relative protein expression levels of b FGF and CTSB were statistically significantamong four groups(b FGF:F=121.84,P<0.05;CTSB:F=30.69,P=0.000).Compared with normal control group,the mRNA and protein levels of CTSB and b FGF were significantly increased in hyperoxia group and DMSO group(P<0.05).Compared with hyperoxia group,the mRNA and protein levels of CTSB and b FGF were decreased in CA-074 Me group(P<0.05).There was no significant difference in mRNA and protein levels of CTSB and b FGF between normal control group and CA-074 Me group,and between hyperoxia group and DMSO group(P>0.05).Conclusion CA-074 Me may inhibit the expression of CTSB in hyperoxia-induced retinal neovascularization in mice,and b FGF expression is down-regulated.The underlying mechanism may be that CA-074 Me inhibits the expression of b FGF by inhibiting CTSB,or CA-074 Me acts directly on b FGF,and its specific mechanism needs further verification in later experiments.