摘要
目的 研究人瘢痕疙瘩组织中沉默信息调节因子2同系物1(SIRT1)的表达及其与氧化应激指标的相关性.方法 回顾性选取2023年1月至2024年1月入沧州市人民医院的80例瘢痕疙瘩患者作为研究对象,参考不同疾病进展阶段分为早期组(0～6个月,n=31)、中期组(6～12个月,n=26)、晚期组(12个月以上,n=23).参考生长期将瘢痕疙瘩组织划分为3个部位,即老化部位、增生部位及浸润部位.采用实时荧光定量PCR(qPCR)法测定瘢痕疙瘩组织与正常组织内的SIRT1 mRNA及SIRT1蛋白表达水平;采用蛋白质免疫印迹法测定p-p65蛋白、p65蛋白、SIRT1蛋白表达水平,同时采用免疫化学法测定氧化应激指标[晚期氧化蛋白产物(AOPP)、丙二醛、超氧化物歧化酶(SOD)]水平.结果 早、中、晚期组组内比较,瘢痕疙瘩组织内SIRT1 mRNA及SIRT1蛋白水平均低于正常组织,差异均有统计学意义(P＜0.05);早、中、晚期组组间比较,瘢痕疙瘩组织与正常组织内SIRT1 mRNA及SIRT1蛋白水平差异均无统计学意义(P＞0.05).早、中、晚期组组内比较,瘢痕疙瘩组织内浸润部位SIRT1 mRNA及SIRT1蛋白水平低于增生、老化部位,且增生部位低于老化部位,差异均有统计学意义(P＜0.05);早、中、晚期组组间比较,瘢痕疙瘩组织内老化、增生及浸润部位SIRT1 mRNA及SIRT1蛋白水平比较,差异均无统计学意义(P＞0.05).瘢痕疙瘩组织与正常组织p65蛋白水平比较,差异无统计学意义(P＞0.05);瘢痕疙瘩组织p-p65、p-p65/p65与AOPP、丙二醛、SOD水平均高于正常组织,差异均有统计学意义(P＜0.05).SIRT1 mRNA及SIRT1蛋白表达在人瘢痕疙瘩组织内与AOPP、丙二醛及SOD均呈负相关(P＜0.05).结论 人瘢痕疙瘩组织内SIRT1表达水平低于正常组织,与瘢痕疙瘩患者的疾病进展无相关性,但与不同表达部位联系紧密,且与氧化应激指标呈负相关.
Abstract
Objective To study the expression of silence-regulating factor-2 homologue 1(SIRT1)in human keloid tissue and its cor-relation with oxidative stress index.Methods A total of 80 patients with keloid who were admitted to Cangzhou Peoples Hospital from January 2023 to January 2024 were retrospectively selected as the research objects.According to different stages of disease progression,they were divided into the early group(0-6 months,n=31),the middle group(6-12 months,n=26)and the late group(more than 12 months,n=23).The keloid tissue was divided into three parts according to the reference growth period,namely,the aging part,the hyperplasia part and the infil-tration part.The expression levels of SIRT1 mRNA and SIRT 1 protein in keloid tissue and normal tissues were determined by real-time PCR(qPCR);p-p65 protein,p65 protein,SIRT 1 protein were determined by immunoblot,and oxidative stress indicators[late oxidized protein product(AOPP),malondialdehyde,superoxide dismutase(SOD)]by immunochemistry.Results In the early,middle and late groups,the SIRT1 mRNA and SIRT1 protein levels in keloid tissue were lower than those in normal tissue,less significant,the differences were statistically significant(P＜0.05);in the early,middle and late groups,there were no statistically significant differences in SIRT1 mRNA and SIRT1 pro-tein levels between keloid tissue and normal tissue(P＞0.05).In the early,middle and late groups,the SIRT1 mRNA and SIRT1 protein levels in the infiltrating sites were lower than the hyperplasia and aging sites,and the hyperplasia sites were lower than the aging sites,the differences were statistically significant(P＜0.05);in the early,middle and late groups,there were no statistically significant difference in the SIRT1 mR-NA and SIRT1 protein levels in the aging,hyperplasia and infiltration sites(P＞0.05).There was no statistically significant difference in p65 protein levels between keloid and normal tissues(P＞0.05);the levels of p-p65,p-p65,AOPP,malondialdehyde,and SOD were higher than normal tissues,the differences were statistically significant(P＜0.05).SIRT1 mRNA And SIRT1 protein expression was negatively associ-ated with AOPP,malondialdehyde and SOD(P＜0.05).Conclusion SIRT1 expression levels in human keloid tissue are lower than those in normal tissue and show no correlation with disease progression in keloid patients,but are strongly linked with different expression sites and nega-tively correlated with oxidative stress indicators.
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