
浏览全部资源
扫码关注微信
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,兰州 730000
[第一作者] 舍雅莉,博士,副教授,硕士生导师,从事中医药防治肿瘤研究,E-mail: sheyali@sina.com
*李长天,硕士,教授,硕士生导师,从事中医药防治肿瘤研究,E-mail: 715357951@qq.com
收稿日期:2019-05-05,
网络出版日期:2019-08-07,
纸质出版日期:2020-01-20
移动端阅览
舍雅莉, 张秋菊, 李亚玲, 等. 黄芪多糖对甲醛染毒人BMSCs染色体损伤的保护作用[J]. 中国实验方剂学杂志, 2020,26(2):66-71.
Ya-li SHE, Qiu-ju ZHANG, Ya-ling LI, et al. Protective Effect of Astragulus Polysaccharide on Chromosome Damage in Human BMSCs Exposed to Formaldehyde[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(2): 66-71.
舍雅莉, 张秋菊, 李亚玲, 等. 黄芪多糖对甲醛染毒人BMSCs染色体损伤的保护作用[J]. 中国实验方剂学杂志, 2020,26(2):66-71. DOI: 10.13422/j.cnki.syfjx.20192205.
Ya-li SHE, Qiu-ju ZHANG, Ya-ling LI, et al. Protective Effect of Astragulus Polysaccharide on Chromosome Damage in Human BMSCs Exposed to Formaldehyde[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(2): 66-71. DOI: 10.13422/j.cnki.syfjx.20192205.
目的:
2
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:
2
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120 μmol·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
-1
APS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:
2
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,APS 40,100,400 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:
2
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
-1
APS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:
2
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:
2
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:
2
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:
2
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.
K A Moore , I R Lemischka . Stem cells and their niches [J]. Science , 2006 , 311 ( 5769 ): 1880 - 1885 .
S Méndez-Ferrer , T V Michurina , F Ferraro , et al . Mesenchymal and haematopoietic stem cells form a unique bone marrow niche [J]. Nature , 2010 , 466 ( 7308 ): 829 - 834 .
X LIANG , J ZHANG , W SONG , et al . Formaldehyde exposure in indoor air from public places and its associated health risks in kunshan city, China [J]. Asia Pac J Public Health , 2018 , 30 ( 6 ): 551 - 560 .
Y L SHE , Y LI , Y Q LIU , et al . Formaldehyde induces toxic effects and regulates the expression of damage response genes in BMSCs [J]. Acta Biochim Biophys Sin , 2013 , 45 ( 12 ): 1011 - 1120 .
刘星 , 李啸红 , 陈阳 , 等 . 续断水提液对小鼠急性毒性和遗传毒性的影响 [J]. 中国实验方剂学杂志 , 2016 , 22 ( 5 ): 160 - 165 .
李钦 , 胡继宏 , 高博 , 等 . 黄芪多糖在免疫调节方面的最新研究进展 [J]. 中国实验方剂学杂志 , 2017 , 23 ( 2 ): 199 - 206 .
阎力君 , 洪涛 , 雒江菡 , 等 . 黄芪多糖对结肠癌SW620细胞增殖及凋亡作用的影响 [J]. 中国实验方剂学杂志 , 2017 , 23 ( 22 ): 97 - 101 .
S Costa , P Coelho , C Costa , et al . Genotoxic damage in pathology anatomy laboratory workers exposed to formaldehyde [J]. Toxicology , 2008 , 252 ( 1/3 ): 40 - 48 .
L ZHANG , X TANG , N Rothman , et al . Occupational exposure to formaldehyde,hematotoxicity,and leukemia-specific chromosome changes in cultured myeloid progenitor cells [J]. Cancer Epidemiol Biomarkers Prev , 2010 , 19 ( 1 ): 80 - 88 .
舍雅莉 , 刘永琦 , 李屹 , 等 . 甲醛对鼠BMSCs细胞系的遗传毒性影响 [J]. 基础医学与临床 , 2014 , 34 ( 4 ): 454 - 458 .
R K Singh , M Krishna . DNA damage induced nucleotide excision repair in saccharomyces cerevisiae [J]. Mol Cell Biochem , 2006 , 290 ( 1/2 ): 103 - 112 .
J CHEN , W Bozza , Z ZHUANG . Ubiquitination of PCNA and its essential role in eukaryotic translesion synthesis [J]. Cell Biochem Biophys , 2011 , 60 ( 1/2 ): 47 - 60 .
D Butkiewicz , A Drosik , R Suwinski , et al . Influence of DNA repair gene polymorphisms on prognosis in inoperable non-small cell lung cancer patients treated with radiotherapy and platinum-based chemotherapy [J]. Int J Cancer , 2012 , 131 ( 5 ): 1 - 9 .
向生光 , 胡维新 . 增殖细胞核抗原PCNA在DNA修复中的作用 [J]. 生命的化学 , 2009 , 29 ( 4 ): 472 - 475 .
刘珍财 , 赵海霞 , 马娜 , 等 . 基于PI3K/Akt通路观察五子衍宗方对老龄大鼠精原干细胞增殖的影响 [J]. 中国实验方剂学杂志 , 2018 , 24 ( 23 ): 119 - 125 .
S Sharma , C E Canman . REV1 and DNA polymerase zeta in DNA interstrand crosslink repair [J]. Environ Mol Mutagen , 2012 , 53 ( 9 ): 725 - 740 .
Z Kelman . PCNA: structure,functions and interactions [J]. Oncogene , 1997 , 14 ( 6 ): 629 - 640 .
0
浏览量
19
下载量
1
CSCD
关联资源
相关文章
相关作者
相关机构
京公网安备11010802024621