CAJ | 학술논문

目的：：探讨单细胞海藻(MPPT)提取物对人脐带间充质干细胞(hUC-MSCs)的抗衰老作用。方法：取剖宫产新生儿脐带,分离、传代培养 hUC-MSCs,显微镜下观察细胞形态,收集第3、10、15代细胞,qRT-PCR 分析不同代细胞中 p16、p21、p53、PCNA、Sirt2 mRNA 的表达情况。 CCK-8法检测不同质量浓度 MPPT 提取物对第15代 hUC-MSCs 增殖的影响；流式细胞术检测 MPPT 提取物对 hUC-MSCs 细胞周期和凋亡的影响；β-半乳糖苷酶染色法检测MPPT 提取物对 hUC-MSCs 衰老的影响；qRT-PCR 检测 MPPT 提取物对 hUC-MSCs 中 p16、p21、p53、PCNA、Sirt2 mR-NA 表达的影响。结果：第3代细胞呈梭形贴壁细胞,第15代细胞呈不规则状态,易脱壁。随着传代次数的增加, hUC-MSCs 中 PCNA、Sirt2 mRNA 表达减少(F =76.944、25.649,P <0.001),而 p16、p21和 p53 mRNA 表达增加(F =46.714、77.645、845.676,P <0.001)；与第15代细胞相比,1 g/ L 的 MPPT 提取物可促进第15代 hUC-MSCs 增殖,促进细胞周期进入 S 期,抑制细胞凋亡(P 均<0.05)；MPPT 提取物处理组衰老细胞率降低(F =773.557,P <0.001),细胞中 PCNA、Sirt2 mRNA 表达增加,p16、p21和 p53 mRNA 表达降低(P 均<0.05)。结论：MPPT 提取物可能通过调节 hUC-MSCs 中 p16、p21、p53、PCNA 和 Sirt2等衰老相关基因的表达发挥抗衰老作用。
목적：：탐토단세포해조(MPPT)제취물대인제대간충질간세포(hUC-MSCs)적항쇠로작용。방법：취부궁산신생인제대,분리、전대배양 hUC-MSCs,현미경하관찰세포형태,수집제3、10、15대세포,qRT-PCR 분석불동대세포중 p16、p21、p53、PCNA、Sirt2 mRNA 적표체정황。 CCK-8법검측불동질량농도 MPPT 제취물대제15대 hUC-MSCs 증식적영향；류식세포술검측 MPPT 제취물대 hUC-MSCs 세포주기화조망적영향；β-반유당감매염색법검측MPPT 제취물대 hUC-MSCs 쇠로적영향；qRT-PCR 검측 MPPT 제취물대 hUC-MSCs 중 p16、p21、p53、PCNA、Sirt2 mR-NA 표체적영향。결과：제3대세포정사형첩벽세포,제15대세포정불규칙상태,역탈벽。수착전대차수적증가, hUC-MSCs 중 PCNA、Sirt2 mRNA 표체감소(F =76.944、25.649,P <0.001),이 p16、p21화 p53 mRNA 표체증가(F =46.714、77.645、845.676,P <0.001)；여제15대세포상비,1 g/ L 적 MPPT 제취물가촉진제15대 hUC-MSCs 증식,촉진세포주기진입 S 기,억제세포조망(P 균<0.05)；MPPT 제취물처리조쇠로세포솔강저(F =773.557,P <0.001),세포중 PCNA、Sirt2 mRNA 표체증가,p16、p21화 p53 mRNA 표체강저(P 균<0.05)。결론：MPPT 제취물가능통과조절 hUC-MSCs 중 p16、p21、p53、PCNA 화 Sirt2등쇠로상관기인적표체발휘항쇠로작용。
Aim: To explore the anti-aging effects of marine phytoplankton(MPPT) on human umbilical cord derived mesenchymal stem cells(hUC-MSCs). Methods: hUC-MSCs were isolated from umbilical cords of mature healthy newbo-rns delivered by caesarean section, and then cultured and passaged, while the morphology was observed under microscope. The passage 3, 10, and 15 cells were collected and the mRNA expressions of p16, p21, p53, PCNA, and Sirt2 in different passages of hUC-MSCs were detected by qRT-PCR. CCK-8 assay was performed to detect the effect of MPPT on cell prolif-eration and flow cytometry was used to detect cell cycle and apoptosis in the passage 15 cells. β-galactosidase staining was used to detect the effect of MPPT on cell senescence, and the effect of MPPT on the mRNA expressions of p16, p21, p53, PCNA, and Sirt2 of hUC-MSCs were detected by qRT-PCR. Results: The cell morphology of passage 3 was fusiform and easily adherent, however, the passage 15 hUC-MSCs were flat and poorly adherent. The mRNA expressions of PCNA and Sirt2 reduced gradually along with passaging(F = 76. 944,25. 649,P < 0. 001), while those of p16, p21, and p53 in-creased(F = 46. 714,77. 645,845. 676, P < 0. 001). Compared with the passage 15 cells, 1 g/ L MPPT could promote the proliferation, induce cells into S phase, inhibit apoptosis of the passage 15 hUC-MSCs(P < 0. 05), decrease the positive rate of β-galactosidase positive staining cells(F = 773. 557,P < 0. 001), and increase the mRNA expressions of PCNA and Sirt2,while decrease the mRNA expressions of p16, p21, and p53(P < 0. 05). Conclusion: MPPT has anti-aging effects on hUC-MSCs, and the mechanism is partly through regulating the expressions of p16, p21, p53, PCNA, and Sirt2.