摘要
背景:研究表明脐带间充质干细胞可以显著改善肝硬化的程度,进而修复肝损伤,但其治疗肝硬化的分子调控机制,尤其是非编码RNA调控的肝内基因变化,目前并没有得到详细的阐释。目的:分析人脐带间充质干细胞移植肝硬化大鼠肝细胞中微小RNA基因表达的变化。方法:采用四氯化碳皮下注射联合乙醇喂服方法建立肝硬化大鼠模型,造模8周后经尾静脉输注人脐带间充质干细胞,每周1次,连续注射4周。最后一次注射治疗1周后收集大鼠肝脏组织进行石蜡切片和提取肝脏RNA进行表达谱基因芯片分析,同时收集血清利用自动生化分析仪测定肝功能指标变化。结果与结论:人脐带间充质干细胞治疗可以显著降低谷丙转氨酶、谷草转氨酶和转肽酶水平,脂肪病变和肝细胞坏死显著减少。微小RNA表达谱芯片杂交分析和PCR验证结果显示rno-mi R-369-5p、rno-mi R-3584-5p和rno-mi R-153*这3种微小RNA基因在造模过程中先下调表达,并在人脐带间充质干细胞治疗后显著上调;而rno-mi R-93、rno-mi R-199a-3p、rno-mi R-195、rno-let-7a和rno-mi R-19a这5种微小RNA基因在造模过程中先上调表达,并在人脐带间充质干细胞治疗后显著下调。以上结果表明人脐带间充质干细胞逆转肝硬化和肝细胞损伤过程中,可能通过上调rno-miR-369-5p、rno-mi R-3584-5p和rno-mi R-153*等miR NA基因表达,下调rno-miR-93、rno-mi R-199a-3p、rno-mi R-195、rno-let-7a和rno-mi R-19a等相关miR NA基因表达发挥治疗作用。
BACKGROUND： Human umbilical cord mesenchymal stem cells（hU C-MSCs） can obviously relieve liver cirrhosis, and thereby repair liver injury. However, the molecular mechanism of h UC-MSCs therapy for liver cirrhosis is limited at present, and especially the non-coding RNA regulation of hepatic gene changes has not been detailed. OBJECTIVE： To investigate the changes of micro RNA after h UC-MSCs therapy in rats with liver cirrhosis. METHODS： Liver cirrhosis models were established in rats using carbon tetrachloride subcutaneous injection plus oral administration of alcohol. At 8 weeks after modeling, h UC-MSCs were injected via the tail vein once aweek for 4 consecutive weeks. At 1 week after the last injection, rat liver tissues were collected for paraffin embedding. Liver RNA was extracted for gene chip analysis. Blood samples were collected and analyzed using an automatic biochemical analyzer to detect the changes of liver function. RESULTS AND CONCLUSION： Alanine aminotransferase, aspartate aminotransferase and gamma-glutamyl transpeptidase were improved significantly after hU C-MSCs therapy. Fat lesions and necrosis of hepatocytes were significantly reduced. MicroR NA expression microarray hybridization analysis and PCR results showed that rno-miR-369-5p, rno-miR-3584-5p and rno-miR-153＊ were down-regulated during modeling and increased after hU C-MSCs therapy. And rno-miR-93, rno-miR-199a-3p, rno-miR-195, rno-let-7a and rno-miR-19 a were firstly up-regulated in the process of modeling and then down-regulated obviously after hU C-MSCs therapy. These results suggest that hU C-MSCs may reverse liver cirrhosis and liver cell damage through up-regulation of rno-miR-369-5p, rno-miR-3584-5p and rno-miR-153＊, and down-regulation of rno-miR-93, rno-miR-199a-3p, rno-miR-195, rno-let-7a and rno-miR-19 a.
出处
《中国组织工程研究》
CAS
北大核心
2015年第23期3674-3680,共7页
Chinese Journal of Tissue Engineering Research
基金
济南市2014科学技术发展计划(201403010)~~
