Protective Effect of Astragulus Polysaccharide on Chromosome Damage in Human BMSCs Exposed to Formaldehyde

Ya-li SHE; Qiu-ju ZHANG; Ya-ling LI; Li ZHANG; Guo-xin ZHANG; Lei ZHANG; Yong-qi LIU; Chang-tian LI

doi:10.13422/j.cnki.syfjx.20192205

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Protective Effect of Astragulus Polysaccharide on Chromosome Damage in Human BMSCs Exposed to Formaldehyde

Pharmacology | 更新时间：2021-04-16

- Protective Effect of Astragulus Polysaccharide on Chromosome Damage in Human BMSCs Exposed to Formaldehyde
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 2, Pages: 66-71(2020)
- 作者机构：
  
  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
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20192205
  CLC： R2-0; R22; R285.5
- Received：05 May 2019，
  
  Published Online：07 August 2019，
  
  Published：20 January 2020
- 稿件说明：
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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：

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.

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：

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(

0.01)，PCNA mRNA和蛋白表达明显降低(

0.05)，XPB，XPD，XPF和XPG mRNA和蛋白表达明显升高(

0.05，

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(

0.01)，PCNA，XPB，XPD，XPF和XPG mRNA和蛋白表达均明显升高(

0.05，

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

-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。

Abstract

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

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 (

0.01)

PCNA mRNA and protein expressions down-regulated significantly (

0.05)

while XPB

XPD

XPF

XPG mRNA and protein expressions up-regulated significantly (

0.05

0.01). Compared with formaldehyde group

BMSCs were treated with APS at 40

100

400 mg·L

-1

micronucleus and SCE decreased significantly (

0.01)

and mRNA and protein expressions of PCNA

XPB

XPD

XPF and XPG up-regulated significantly (

0.05

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.

关键词

Keywords

references

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