CAJ | 학술논문

背景：研究miR-378对缺血缺氧条件下骨髓间充质干细胞生长增殖、细胞凋亡等生物学行为的影响，为进一步改善骨髓间充质干细胞在梗死心肌中的生存提供新方法。 　　目的：观察miR-378转染后的骨髓间充质干细胞耐受缺血缺氧及促进血管形成的能力。 　　方法：体外培养的骨髓间充质干细胞分为未转染组和转染组，未转染组为未转染miR-378的骨髓间充质干细胞，转染组则采用化学合成的miR-378模拟物转染SD大鼠骨髓间充质干细胞；将两组骨髓间充质干细胞置于缺血缺氧(无血清，体积分数为1%O2、5%CO2，94%N2)环境中培养24 h后，应用锥虫蓝染色计数法、MTS法检测两组细胞在不同时间点的生长及增殖情况，TUNEL法检测细胞凋亡情况；将两组缺血缺氧后的培养基上清液分别刺激人脐静脉内皮细胞，观察血管形成情况。 　　结果与结论：缺血缺氧干预后48 h和72 h，转染组的骨髓间充质干细胞活细胞数明显高于未转染组(均为P <0.01)；转染组的骨髓间充质干细胞表现出较高的增殖能力，缺血缺氧干预后24 h及48 h细胞增殖升高较未转染组明显，差异有显著性意义(P <0.01，P <0.05)；缺血缺氧后转染组骨髓间充质干细胞的凋亡比例明显下降(P <0.05)。两组细胞均可促进血管腔样结构形成，但转染组血管管腔样结构较未转染组显著增多(P <0.01)。研究结果提示miR-378可促进缺血缺氧后骨髓间充质干细胞的生长增殖并抑制其在缺血缺氧条件下的细胞凋亡，同时可提高其促血管生成的能力。
배경：연구miR-378대결혈결양조건하골수간충질간세포생장증식、세포조망등생물학행위적영향，위진일보개선골수간충질간세포재경사심기중적생존제공신방법。 　　목적：관찰miR-378전염후적골수간충질간세포내수결혈결양급촉진혈관형성적능력。 　　방법：체외배양적골수간충질간세포분위미전염조화전염조，미전염조위미전염miR-378적골수간충질간세포，전염조칙채용화학합성적miR-378모의물전염SD대서골수간충질간세포；장량조골수간충질간세포치우결혈결양(무혈청，체적분수위1%O2、5%CO2，94%N2)배경중배양24 h후，응용추충람염색계수법、MTS법검측량조세포재불동시간점적생장급증식정황，TUNEL법검측세포조망정황；장량조결혈결양후적배양기상청액분별자격인제정맥내피세포，관찰혈관형성정황。 　　결과여결론：결혈결양간예후48 h화72 h，전염조적골수간충질간세포활세포수명현고우미전염조(균위P <0.01)；전염조적골수간충질간세포표현출교고적증식능력，결혈결양간예후24 h급48 h세포증식승고교미전염조명현，차이유현저성의의(P <0.01，P <0.05)；결혈결양후전염조골수간충질간세포적조망비례명현하강(P <0.05)。량조세포균가촉진혈관강양결구형성，단전염조혈관관강양결구교미전염조현저증다(P <0.01)。연구결과제시miR-378가촉진결혈결양후골수간충질간세포적생장증식병억제기재결혈결양조건하적세포조망，동시가제고기촉혈관생성적능력。
BACKGROUND:To investigate the influence of miR-378 on bone marrow mesenchymal stem cels proliferation and apoptosis under hypoxic-ischemic condition wil provide a new method for further improving the survival of bone marrow mesenchymal stem cels in the infarcted myocardium. OBJECTIVE:To observe the viability of bone marrow mesenchymal stem cels and their ability of angiogenesis under hypoxic-ischemic condition after miR-378 transfection. METHODS:The bone marrow mesenchymal stem cels of Sprague-Dawley rat were culturedin vitro and divided into the non-transfection group and the transfection group. Mesenchymal stem cels were transfected with miR-378 mimic in the transfection group while the non-transfection group was taken as the control group. After transfection, the two groups were incubated under hypoxic-ischemic condition (serum-free medium,1% O2, 5% CO2, 94% N2) at 37℃ for 24 hours. Viability and proliferation of bone marrow mesenchymal stem cels were evaluated by the trypan-blue exclusion assay and MTS assay respectively. The cellapoptosis was detected by TUNEL assay. The culture supernatant of the two groups was colected separately and used as the conditioned medium after their exposure to hypoxic-ischemic environment. Endothelial cels were then co-cultured with the conditioned medium for the vasculature formation assay. RESULTS AND CONCLUSION: After the experience of hypoxia-ischemia for 24 hours and 72 hours, there was a larger number of the living cels in the transfection group in contrast to the non-transfection group (bothP < 0.01). Compared with the non-transfection group, miR-378-MSCs group presented a stronger ability of proliferation, and their proliferation rates were obviously higher at 24 and 48 hours (P < 0.01 andP < 0.05, respectively). The percentage of apoptotic cels in the transfection group was significantly declined under the hypoxic-ischemic condition (P < 0.05). The vasculature formation assay indicated that the lumen-like structures were found in both of the two groups, however, the number of lumen-like structures was remarkably increased in the transfection group (P < 0.01). miR-378 could promote the proliferation of bone marrow mesenchymal stem cels and suppress their apoptosis under hypoxic-ischemic condition. It could also enhance their ability of vasculogenesis.