CAJ | 학술논문

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-miR-369-5p、rno-miR-3584-5p和rno-miR-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-miR-93、rno-miR-199a-3p、rno-miR-195、rno-let-7a和rno-miR-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-miR-3584-5p和rno-miR-153*等miRNA基因表达，下调rno-miR-93、rno-miR-199a-3p、rno-miR-195、rno-let-7a和rno-miR-19a等相关miRNA基因表达发挥治疗作用。
배경：연구표명제대간충질간세포가이현저개선간경화적정도，진이수복간손상，단기치료간경화적분자조공궤제，우기시비편마RNA조공적간내기인변화，목전병몰유득도상세적천석。목적：분석인제대간충질간세포이식간경화대서간세포중미소RNA기인표체적변화。방법：채용사록화탄피하주사연합을순위복방법건립간경화대서모형，조모8주후경미정맥수주인제대간충질간세포，매주1차，련속주사4주。최후일차주사치료1주후수집대서간장조직진행석사절편화제취간장RNA진행표체보기인심편분석，동시수집혈청이용자동생화분석의측정간공능지표변화。결과여결론：인제대간충질간세포치료가이현저강저곡병전안매、곡초전안매화전태매수평，지방병변화간세포배사현저감소。미소RNA표체보심편잡교분석화PCR험증결과현시rno-miR-369-5p、rno-miR-3584-5p화rno-miR-153*저3충미소RNA기인재조모과정중선하조표체，병재인제대간충질간세포치료후현저상조；이rno-miR-93、rno-miR-199a-3p、rno-miR-195、rno-let-7a화rno-miR-19a저5충미소RNA기인재조모과정중선상조표체，병재인제대간충질간세포치료후현저하조。이상결과표명인제대간충질간세포역전간경화화간세포손상과정중，가능통과상조rno-miR-369-5p、rno-miR-3584-5p화rno-miR-153*등miRNA기인표체，하조rno-miR-93、rno-miR-199a-3p、rno-miR-195、rno-let-7a화rno-miR-19a등상관miRNA기인표체발휘치료작용。
BACKGROUND:Human umbilical cord mesenchymal stem cels (hUC-MSCs) can obviously relieve liver cirrhosis, and thereby repair liver injury. However, the molecular mechanism of hUC-MSCs therapy for liver cirrhosis is limited at present, and especialy the non-coding RNA regulation of hepatic gene changes has not been detailed. OBJECTIVE:To investigate the changes of microRNA after hUC-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, hUC-MSCs were injectedvia the tail vein once a week for 4 consecutive weeks. At 1 week after the last injection, rat liver tissues were colected for paraffin embedding. Liver RNA was extracted for gene chip analysis. Blood samples were colected 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 hUC-MSCs therapy. Fat lesions and necrosis of hepatocytes were significantly reduced. MicroRNA 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 hUC-MSCs therapy. And rno-miR-93, rno-miR-199a-3p, rno-miR-195, rno-let-7a and rno-miR-19a were firstly up-regulated in the process of modeling and then down-regulated obviously after hUC-MSCs therapy. These results suggest that hUC-MSCs may reverse liver cirrhosis and liver cel 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-19a.