骨髓间充质干细胞通过调节LPS-TLR4通路抑制肝星形细胞活化的作用机制

谢婵; 彭亮; 叶一农; 张耿林; 谢仕斌; 高志良

骨髓间充质干细胞通过调节LPS-TLR4通路抑制肝星形细胞活化的作用机制

中山大学附属第三医院

基金项目:

中华医学会百洋肝纤维化基金资助项目(2010-004)；国家自然基金资助项目(30971356)；

详细信息

中图分类号: R575.2
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出版历程
- 出版日期: 2011-05-20

Bone marrow mesenchymal stem cells regulated hepatic stellate cells activation through LPS-TLR4 pathway

Research funding:

摘要

摘要: 目的研究人骨髓间充质干细胞（BMSC）是否可通过干预LPS-TLR4通路,调控肝星状细胞（HSC）LX2活化,阐明BMSC在肝纤维化形成中的作用。方法体外共培养BMSC、LX2,分为BMSC+LX2（LPS刺激）组、TLR4阻断剂+LX2（LPS刺激）组、LX2（LPS刺激）组、BMSC（LPS刺激）组、LX2（无LPS刺激）组,培养6、12、36、48 h;收集上清,ELISA检测IL-8及TGFβ表达,RT-PCR检测LX2的TLR4、Myd88及NF-κB的表达,Western Blot检测TGFβ、SMA、ColⅠ、MyD88、TLR4表达。免疫荧光检测NF-κB细胞内表达情况。结果 LPS可以刺激LX2活化,活化的LX2分泌IL-8及TGFβ增多,TGFβ、SMA、ColⅠ、MyD88、TLR4表达增加,NF-κB p65主要为核内表达。与BMSC共培养后,LPS导致的LX2活化减弱,LX2的TGF-β、SMA、ColⅠ、MyD88、TLR4的表达下降,NF-κB p65胞浆表达为主。结论 BMSC可以通过干预LPS-TLR4通路抑制LX2的活化。
- 骨髓细胞 /
- 间质干细胞 /
- 肝细胞,星形细胞 /
- LPS-TLR4通路
Abstract: Objective To investigate whether bone marrow mesenchymal stem cells (BMSC) affect the hepatic stellate cells (LX2) through inhibiting LPS-TLR4 pathway, and to clarify the mechanism of LPS causing liver fibrosis by activating LX2.Methods Human BMSC and LX2 were isolated and cultured in vitro separately or co-cultured in transwell under different treatments in four groups, which were 2×105 BMSC and 2×105 LX2 (with 100μg/ml LPS stimulation) co-culture group, 2×105 BMSC and 2×105 LX2 (no LPS stimulation) co-culture group, 2×105 LX2 (with 100μg/ml LPS stimulation) group, 2×105 BMSC (with 100μg/ml LPS stimulation) group and 2×105 LX2 (no LPS stimulation) group.Supernatant of 6, 12, 36 and 48 hours after treatment was collected and detected for IL-8 and TGFβ expression by ELISA.The TLR4, Myd88 and NF-κB gene expression of LX2 were detected by RT-PCR.The SMA, TGFβ, TLR4, Myd88 and ColⅠ expression were detected by Western Blotting.NF-κB p65 expression in cell was observed by immunofluorescence.Results LPS can stimulate LX2 activation.Activated LX2 secreted more IL-8 and TGFβ than static LX2 respectively, P<0.05.Gene expression of α-SMA, TGFβ and ColⅠ were increased in activated LX2.NF-κB p65 was mainly expressed in nucleus of activated LX2.After co-culture with BMSC, LX2 activation by LPS was inhibited significantly as the SMA, TGFβ, TLR4, Myd88 and ColⅠ were reduced.NF-κB p65 was mainly expressed in cytoplasm.Conclusion BMSC can inhibit LX2 activation through LPS-TLR4 pathway.
- bone marrow cell /
- mesenchymal stem cells /
- hepatocytes /
- astrocytes

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参考文献(12)

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