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血清反应因子经组蛋白乙酰化促进大鼠骨髓间充质干细胞向平滑肌细胞分化
Serum response factor promotes differentiation of rat bone marrow mesenchymal stem cells into smooth muscle cells
via histone acetylation- 第三军医大学学报 2021年43卷第4期 页码：303-310
- 作者机构：
1.400038 重庆，陆军军医大学（第三军医大学）第一附属医院妇产科
2.401121 重庆，重庆市人民医院妇产科
- 作者简介：
[ 梁志清，E-mail: zhi.lzliang@gmail.com LIANG Zhiqing, E-mail: zhi.lzliang@gmail.com
- 基金信息：国家自然科学基金面上项目(81170552)
DOI：10.16016/j.1000-5404.202008213
中图分类号： R322.74;R329.21;R329.26收稿：2020-08-27，
修回：2020-09-16，
纸质出版：2021-04-15
- 稿件说明：
移动端阅览
刘晶霞, 梁志清. 血清反应因子经组蛋白乙酰化促进大鼠骨髓间充质干细胞向平滑肌细胞分化[J]. 第三军医大学学报, 2021,43(4):303-310.
Jingxia LIU, Zhiqing LIANG. Serum response factor promotes differentiation of rat bone marrow mesenchymal stem cells into smooth muscle cells
via histone acetylation[J]. Journal of Third Military Medical University, 2021, 43(4): 303-310.刘晶霞, 梁志清. 血清反应因子经组蛋白乙酰化促进大鼠骨髓间充质干细胞向平滑肌细胞分化[J]. 第三军医大学学报, 2021,43(4):303-310. DOI： 10.16016/j.1000-5404.202008213.
Jingxia LIU, Zhiqing LIANG. Serum response factor promotes differentiation of rat bone marrow mesenchymal stem cells into smooth muscle cells
via histone acetylation[J]. Journal of Third Military Medical University, 2021, 43(4): 303-310. DOI： 10.16016/j.1000-5404.202008213.
摘要
目的
2
探讨血清反应因子（serum response factor，SRF）在大鼠骨髓间充质干细胞（mesenchymal stem cells，MSCs）分化为膀胱平滑肌细胞（smooth muscle cells，SMCs）中的作用和机制。
方法
2
贴壁法分离4周龄雌性SD大鼠的骨髓MSCs与膀胱SMCs，通过Transwell小室共培养诱导MSCs分化。收集MSCs，根据共培养不同时间点进行分组，通过实时荧光定量PCR和免疫荧光法检测共培养SRF MSCs中SMCs标志基因表达；通过染色质免疫共沉淀（chromatin immunoprecipitation assay，ChIP）定量PCR技术检测SMCs相关基因启动子区组蛋白乙酰化修饰、组蛋白去乙酰化酶（histone deacetylase，HDAC）和SRF的富集。丁酸钠（sodium butyrate，NaB）预处理MSCs后与SMCs共培养，实验分为正常共培养对照组和NaB预处理干预组，ChIP-qPCR对比两组间组蛋白修饰和SRF富集情况。
结果
2
共培养MSCs中，α-SMA、Calponin和SM-MHC基因表达量增加，在这些基因启动子区H3K9
ace
，H3
ace
和H4
ace
富集量增加（
P
<
0.05），同时MSCs中HDAC1和HDAC2降低（
P
<
0.05）。与对照组相比，NaB干预组SRF、H3K9
ace
和H4
ace
富集量增加（
P
<
0.05）。
结论
2
SRF可能在HDAC1和HDAC2调节下通过结合H3K9
ace
和H4
ace
控制SMCs标志基因表达，从而促进MSCs向SMCs的分化。
Abstract
Objective
2
To explore the role and mechanism of serum response factor (SRF) in differentiation of rat bone marrow mesenchymal stem cells (MSCs) into bladder smooth muscle cells (SMCs).
Methods
2
Rat MSCs and SMCs were isolated respectively from the bone marrow and bladder of 4-week-old female SD rats by adherent method
and then co-cultured together in a Transwell chamber. After the MSCs were harvested at different time points
real-time PCR and immunofluorescense assay were used to detect the expression of SMCs specific marker genes in co-cultured MSCs. Furthermore
the enrichment of histone acetylation modification
histone deacetylase (HDAC) and SRF in the promoter of these genes were tested by chromatin immunoprecipitation assay (ChIP) q-PCR in co-cultured MSCs. Finally
ChIP-qPCR was used to compare the histone acetylation modification and SRF enrichment in sodium butyrate (NaB) pretreatment group with control cells.
Results
2
The expression levels of α-SMA
Calponin and SM-MHC were increased in MSCs co-cultured with SMCs. H3K9ace
H3ace and H4ace in the promoter of these genes were enriched (
P
<
0.05)
and then
HDAC1 and HDAC2 in the induced MSCs were correspondingly decreased significantly (
P
<
0.05). The enrichment of SRF in the NaB pretreatment group shared the same increasing tendency with H3K9ace and H4ace.
Conclusion
2
SRF may bind to H3K9ace and H4ace
through the regulation of HDAC1 and HDAC2 to control the expression of SMCs specific genes
and thus promote the differentiation from MSCs to SMCs.
关键词
Keywords
references
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