CAJ | 학술논문

目的：建立人骨髓间充质干细胞( hMSCs)与视网膜色素上皮细胞( RPE)低氧高糖培养的细胞共培养模型,研究在与hMSCs共培养条件下低氧及高糖时RPE细胞增殖、凋亡和移行等生物学特征的影响,对糖尿病刺激脉络膜新生血管( CNV)的机制进行初步探讨。 　　方法：使用Transwell间接共培养模型。将共培养模型按照培养氧浓度及糖浓度培养基类型分为4组：常氧常糖组(21% O2加5.56mmol/L葡萄糖培养液,A组)、常氧高糖组(21% O2加30mmol/L葡萄糖培养液,B组)、低氧常糖组(5% O2加5.56mmol/L葡萄糖培养液,C组)和低氧高糖组(5% O2加30mmol/L 葡萄糖培养液,D 组)。通过CCK-8检测间接共培养模型在12,24,48 h时RPE细胞的增殖能力、Transwell 检测间接共培养模型在24 h 时 RPE细胞的移行情况、流式细胞仪检测间接共培养模型中24 h时RPE细胞的凋亡情况。 　　结果：与对照组相比,12,24和48 h时, B组(1.61±0.41,1.80±0.34,1.91±0.35)、C 组(1.34±0.46,1.94±0.40,2.14±0.41)及D组(1.98±0.47,2.26±0.42,2.55±0.40)中的细胞增殖能力均较对照组(0.92±0.45,1.27±0.32,1.59±0.41)增强(P<0.05),并且低氧高糖联合作用组增殖能力最强。在24h时,B 组(149.5±9.19)、C 组(140±9.90)、D组(170.5±7.78)较对照组(114.5±7.78)的RPE 　　细胞移行能力增强(P<0.05)。而在24h时各组间的凋亡情况无明显差异(P>0.05)。 　　结论：与hMSCs共培养条件下,低氧及高糖环境促进RPE细胞的增殖和移行,而对其凋亡并无影响。
목적：건립인골수간충질간세포( hMSCs)여시망막색소상피세포( RPE)저양고당배양적세포공배양모형,연구재여hMSCs공배양조건하저양급고당시RPE세포증식、조망화이행등생물학특정적영향,대당뇨병자격맥락막신생혈관( CNV)적궤제진행초보탐토。 　　방법：사용Transwell간접공배양모형。장공배양모형안조배양양농도급당농도배양기류형분위4조：상양상당조(21% O2가5.56mmol/L포도당배양액,A조)、상양고당조(21% O2가30mmol/L포도당배양액,B조)、저양상당조(5% O2가5.56mmol/L포도당배양액,C조)화저양고당조(5% O2가30mmol/L 포도당배양액,D 조)。통과CCK-8검측간접공배양모형재12,24,48 h시RPE세포적증식능력、Transwell 검측간접공배양모형재24 h 시 RPE세포적이행정황、류식세포의검측간접공배양모형중24 h시RPE세포적조망정황。 　　결과：여대조조상비,12,24화48 h시, B조(1.61±0.41,1.80±0.34,1.91±0.35)、C 조(1.34±0.46,1.94±0.40,2.14±0.41)급D조(1.98±0.47,2.26±0.42,2.55±0.40)중적세포증식능력균교대조조(0.92±0.45,1.27±0.32,1.59±0.41)증강(P<0.05),병차저양고당연합작용조증식능력최강。재24h시,B 조(149.5±9.19)、C 조(140±9.90)、D조(170.5±7.78)교대조조(114.5±7.78)적RPE 　　세포이행능력증강(P<0.05)。이재24h시각조간적조망정황무명현차이(P>0.05)。 　　결론：여hMSCs공배양조건하,저양급고당배경촉진RPE세포적증식화이행,이대기조망병무영향。
AIM: To evaluate cell proliferation, apoptosis and migration of human retinal pigment epithelial cells ( RPE) when co - cultured with human marrow mesenchymal stem cells ( hMSCs ) in condition of hypoxia and hyperglycemia so as to explore possible mechanisms of diabetes aggravating choroidal neovascularization ( CNV) preliminarily. METHODS:Both hMSCs and RPE cells were co-cultured in a transwell system. The experiment was divided into four groups: 21% O2 with 5. 56mmol/L glucose ( control group, A ), 21% O2 with 30mmol/L glucose ( hyperglycemia and normoxia group, B ) , 5% O2 with 5.56mmol/L glucose ( normoglycemia and hypoxia group, C ) and 5% O2 with 30mmol/L glucose ( hyperglycemia and hypoxia group, D) . Cell Counting Kit-8 (CCK-8) was used to detect the proliferation of RPE cells in each group at 12, 24 and 48h respectively. Flow cytometry was performed to observe apoptosis of RPE cells at 24h. Additionally, we assessed migration capabilities of RPE via transwell assay under the condition of hyperglycemia and hypoxia by co-culturing of hMSCs.RESULTS:In this co-culturing system, at 12, 24 and 48h, group B (1. 61±0. 41, 1. 80±0. 34;1. 91±0. 35), C (1.34±0. 46, 1. 94±0. 40, 2. 14±0. 41) and D (1. 98±0. 47, 2.26±0.42, 2. 55±0. 40) showed significantly higher proliferation rate than group A (0. 92±0. 45, 1. 27±0. 32, 1.59±0. 41, P<0. 05). The migration capabilities of RPE in group B (149. 5±9. 19), C (140±9. 90) and D (170. 5±7. 78) increased dramatically compared with group A ( 114. 5±7.78, P<0.05) at 24h, whereas there was no significant difference of apoptosis ratio among four groups (P>0. 05). CONCLUSION:By coexistence with hMSCs, the synergy of hyperglycemia and hypoxia can improve migration and proliferation of RPE cells, and have no effect on apoptosis of RPE cells within short period.