文献类型：期刊文献

中文题名：黄芪多糖对甲醛染毒人BMSCs染色体损伤的保护作用

英文题名：Protective Effect of Astragulus Polysaccharide on Chromosome Damage in Human BMSCs Exposed to Formaldehyde

作者：舍雅莉[1];张秋菊[1];李亚玲[1];张立[1];张国欣[1];张磊[1];刘永琦[1];李长天[1]

第一作者：舍雅莉

机构：[1]甘肃中医药大学甘肃省高校重大疾病分子医学与中医药防治研究省级重点实验室敦煌医学与转化省部共建教育部重点实验室

第一机构：甘肃中医药大学科研实验中心（甘肃省中医药标准化技术委员会秘书处）

年份：2020

卷号：26

期号：2

起止页码：66

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD2019_2020】；

基金：国家自然科学基金项目(81560667);甘肃省自然科学基金项目(1506RJZA045)

语种：中文

中文关键词：黄芪多糖;甲醛;骨髓间充质干细胞;微核;姐妹染色单体互换

外文关键词：astragalus polysaccharide;formaldehyde;bone marrow mesenchymal stem cell;micronucleus;sister chromatid exchange

摘要：目的:研究黄芪多糖(APS)对甲醛染毒人骨髓间充质干细胞(BMSCs)微核形成、姐妹染色单体互换(SCE)频率升高的保护作用及潜在的机制。方法:体外培养人BMSCs,随机分为空白组,甲醛组,APS 40,100,400 mg·L^-1组。用120μmol·L^-1甲醛染毒人BMSCs,染毒同时,APS 40,100,400 mg·L^-1组分别加入40,100,400 mg·L^-1APS共培养。利用倒置相差显微镜观察细胞形态,微核实验检测微核形成情况,姐妹染色单体互换实验检测SCE发生频率,实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测增殖细胞核抗原(PCNA),着色性干皮病基因B,D,F,G(XPB,XPD,XPF,XPG) mRNA和蛋白表达情况。结果:与空白组比较,甲醛染毒人BMSCs细胞数量明显减少,形态明显改变,APS40,100,400 mg·L^-1作用后,细胞数量和形态均有所恢复。甲醛组微核形成及SCE发生频率较空白组显著升高(P<0.01),PCNA mRNA和蛋白表达明显降低(P<0.05),XPB,XPD,XPF和XPG mRNA和蛋白表达明显升高(P<0.05,P<0.01);与甲醛组比较,APS 40,100,400 mg·L^-1作用后,微核形成及SCE发生频率显著降低(P<0.01),PCNA,XPB,XPD,XPF和XPG mRNA和蛋白表达均明显升高(P<0.05,P<0.01),其中APS 100 mg·L^-1组效果最为明显。结论:APS可以保护甲醛染毒人BMSCs减少微核形成和SCE发生频率,100 mg·L^-1APS保护作用最为明显,其机制可能与上调核苷酸切除修复通路PCNA,XPB,XPD,XPF和XPG基因表达,促进损伤修复有关。
Objective: To study the protective effect of astragalus polysaccharide(APS) on micronucleus and sister chromatid exchange(SCE) in human bone marrow mesenchyml stem cell(BMSCs) exposed to formaldehyde,in order to initially explore the potential mechanism.Method: BMSCs were cultured in vitro,cells were randomly divided into five groups: control group,formaldehyde group,and APS 40,100,400 mg·L^-1 groups.BMSCs were infected with 120 μmol·L^-1 formaldehyde,meanwhile,APS 40,100,400 mg·L^-1 groups were co-cultured with 40,100,400 mg·L^-1 APS.Cell morphology was observed by inverted phase contrast microscope,micronucleus were detected by micronucleus test,SCE was detected by SCE test,and mRNA and protein expressions of proliferating cell nuclear antigen(PCNA),xeroderma pigmentosum B,D,F,G(XPB,XPD,XPF,XPG) were detected by quantitative real-time fluorescence polymerase chain reaction(Real-time PCR) and Western blot.Result: Compared with control group,cell counts decreased,and cell morphology of BMSCs in formaldehyde group significantly changed,they were all recovered gradually in 40,100,400 mg·L^-1 APS groups.Compared with control group,the micronucleus and SCE increased significantly(P<0.01),PCNA mRNA and protein expressions down-regulated significantly(P<0.05),while XPB,XPD,XPF,XPG mRNA and protein expressions up-regulated significantly(P<0.05,P<0.01).Compared with formaldehyde group,BMSCs were treated with APS at 40,100,400 mg·L^-1,micronucleus and SCE decreased significantly(P<0.01),and mRNA and protein expressions of PCNA,XPB,XPD,XPF and XPG up-regulated significantly(P<0.05,P<0.01).Among them,the 100 mg·L^-1 APS group had the most obvious effect.Conclusion: APS can protect formaldehyde-induced BMSCs micronucleus and SCE,especially 100 mg·L^-1 APS has the most obvious effect.The mechanism may be associated with the up-regulation of expressions of PCNA,XPB,XPD,XPF and XPG in the nucleotide exicision repair pathway(NER),which promoted the damage repair.