中华糖尿病杂志
中華糖尿病雜誌
중화당뇨병잡지
CHINES JOURNAL OF DLABETES MELLITUS
2010年
2期
132-136
,共5页
翟清%孙子林%吴同智%查爱云
翟清%孫子林%吳同智%查愛雲
적청%손자림%오동지%사애운
糖尿病%胰岛β细胞%胰腺星状细胞%凋亡
糖尿病%胰島β細胞%胰腺星狀細胞%凋亡
당뇨병%이도β세포%이선성상세포%조망
Diabetes mellitus%Pancreatic beta-cells%Pancreatic stellate cells%Apoptosis
目的 观察胰腺星状细胞对高糖诱导的大鼠胰岛β细胞株Ins-1细胞活力及凋亡的影响,探索胰腺星状细胞在糖尿病胰岛功能衰竭进程中的作用.方法 构建Ins-1及胰腺星状细胞共培养系统,细胞分为Ins-1对照组、Ins-1高糖组(25 mmol/L葡萄糖)、Ins-1高渗组(25 mmol/L甘露醇)、共培养对照组、共培养高糖组(25 mmol/L葡萄糖)、共培养高渗组(25 mmol/L甘露醇).24 h后采用流式细胞法分析Ins-1细胞早期凋亡水平,48 h后分别采用嚷唑蓝(MTT)法和4,6-二氨基-2-苯基吲哚(DAPI)染色法检测细胞活力及凋亡细胞形态.使用单因素方差分析进行数据统计.结果 Ins-1高糖组与Ins-1对照组比较,Ins-1细胞凋亡率显著增加(分别为7.93%±0.41%、3.73%±0.35%,F=55.68,P<0.05);共培养高糖组与共培养对照组比较,Ins-1细胞凋亡率显著增加(分别为11.73%±1.20%、5.03%±0.41%,F=55.68,P<0.05).Ins-1高糖组与Ins-1对照组比较,细胞活力明显下降(分别为2.28±0.13、2.85±0.31,F=97.75,P<0.05);共培养高糖组与共培养对照组比较,细胞活力明显下降(分别为0.62±0.06、1.29±0.19,F=97.75,P<0.05).共培养对照组、共培养高糖组、共培养高渗组与Ins-1对照组、Ins-1高糖组、Ins-1高渗组比较,Ins-1细胞凋亡率显著增加(分别为5.03%±0.41%、3.73%±0.35%;11.73%±1.20%、7.93%±0.41%;7.60%±0.72%、5.60%±0.40%;F=55.68,P<0.05),细胞活力明显下降(分别为1.29±0.19、2.85±0.31;0.62±0.06、2.28±0.13;0.65±0.07、2.35±0.12;F=97.75,P<0.05),并可见凋亡细胞典型形态特征.结论 高糖、胰腺星状细胞可致大鼠胰岛β细胞Ins-1活力下降,凋亡增加;胰腺星状细胞可促进高糖诱导的胰岛β细胞凋亡.
目的 觀察胰腺星狀細胞對高糖誘導的大鼠胰島β細胞株Ins-1細胞活力及凋亡的影響,探索胰腺星狀細胞在糖尿病胰島功能衰竭進程中的作用.方法 構建Ins-1及胰腺星狀細胞共培養繫統,細胞分為Ins-1對照組、Ins-1高糖組(25 mmol/L葡萄糖)、Ins-1高滲組(25 mmol/L甘露醇)、共培養對照組、共培養高糖組(25 mmol/L葡萄糖)、共培養高滲組(25 mmol/L甘露醇).24 h後採用流式細胞法分析Ins-1細胞早期凋亡水平,48 h後分彆採用嚷唑藍(MTT)法和4,6-二氨基-2-苯基吲哚(DAPI)染色法檢測細胞活力及凋亡細胞形態.使用單因素方差分析進行數據統計.結果 Ins-1高糖組與Ins-1對照組比較,Ins-1細胞凋亡率顯著增加(分彆為7.93%±0.41%、3.73%±0.35%,F=55.68,P<0.05);共培養高糖組與共培養對照組比較,Ins-1細胞凋亡率顯著增加(分彆為11.73%±1.20%、5.03%±0.41%,F=55.68,P<0.05).Ins-1高糖組與Ins-1對照組比較,細胞活力明顯下降(分彆為2.28±0.13、2.85±0.31,F=97.75,P<0.05);共培養高糖組與共培養對照組比較,細胞活力明顯下降(分彆為0.62±0.06、1.29±0.19,F=97.75,P<0.05).共培養對照組、共培養高糖組、共培養高滲組與Ins-1對照組、Ins-1高糖組、Ins-1高滲組比較,Ins-1細胞凋亡率顯著增加(分彆為5.03%±0.41%、3.73%±0.35%;11.73%±1.20%、7.93%±0.41%;7.60%±0.72%、5.60%±0.40%;F=55.68,P<0.05),細胞活力明顯下降(分彆為1.29±0.19、2.85±0.31;0.62±0.06、2.28±0.13;0.65±0.07、2.35±0.12;F=97.75,P<0.05),併可見凋亡細胞典型形態特徵.結論 高糖、胰腺星狀細胞可緻大鼠胰島β細胞Ins-1活力下降,凋亡增加;胰腺星狀細胞可促進高糖誘導的胰島β細胞凋亡.
목적 관찰이선성상세포대고당유도적대서이도β세포주Ins-1세포활력급조망적영향,탐색이선성상세포재당뇨병이도공능쇠갈진정중적작용.방법 구건Ins-1급이선성상세포공배양계통,세포분위Ins-1대조조、Ins-1고당조(25 mmol/L포도당)、Ins-1고삼조(25 mmol/L감로순)、공배양대조조、공배양고당조(25 mmol/L포도당)、공배양고삼조(25 mmol/L감로순).24 h후채용류식세포법분석Ins-1세포조기조망수평,48 h후분별채용양서람(MTT)법화4,6-이안기-2-분기신타(DAPI)염색법검측세포활력급조망세포형태.사용단인소방차분석진행수거통계.결과 Ins-1고당조여Ins-1대조조비교,Ins-1세포조망솔현저증가(분별위7.93%±0.41%、3.73%±0.35%,F=55.68,P<0.05);공배양고당조여공배양대조조비교,Ins-1세포조망솔현저증가(분별위11.73%±1.20%、5.03%±0.41%,F=55.68,P<0.05).Ins-1고당조여Ins-1대조조비교,세포활력명현하강(분별위2.28±0.13、2.85±0.31,F=97.75,P<0.05);공배양고당조여공배양대조조비교,세포활력명현하강(분별위0.62±0.06、1.29±0.19,F=97.75,P<0.05).공배양대조조、공배양고당조、공배양고삼조여Ins-1대조조、Ins-1고당조、Ins-1고삼조비교,Ins-1세포조망솔현저증가(분별위5.03%±0.41%、3.73%±0.35%;11.73%±1.20%、7.93%±0.41%;7.60%±0.72%、5.60%±0.40%;F=55.68,P<0.05),세포활력명현하강(분별위1.29±0.19、2.85±0.31;0.62±0.06、2.28±0.13;0.65±0.07、2.35±0.12;F=97.75,P<0.05),병가견조망세포전형형태특정.결론 고당、이선성상세포가치대서이도β세포Ins-1활력하강,조망증가;이선성상세포가촉진고당유도적이도β세포조망.
Objective To explore the influence of pancreatic stellate cells (PSCs) on the apoptosis of pancreatic islet cell line Ins-1 induced by high concentration of glucose. Methods A co-culture system was established, and Ins-1 apoptosis was evaluated by flow cytometry with Annexin IV/PI labeling after 24-hour treatment with and without 25 mmol/L glucose both in Ins-1 group and Ins-1 + PSCs group, respectively. Ins-1 vitality and nuclei morphological changes were observed with MTY and DAPI-staining respectively after 48-hour intervention with and without 25 mmol/L glucose. Resets Both in the Ins-1 groups and the co-cultured groups, apoptosis rates of high glucose groups were higher than those of the controls (7.93%±0.41%, 3.73%±0. 35%; 11.73% ± 1.20%, 5.03%±0.41% ;F = 55.68, P <0.05), and optical densities (ODs) of the high glucose groups were lower than those of the controls (2.28±0.13, 2.85±0.31; 0.62±0.06, 1.29±0.19; F=97.75, P<0.05). Apoptesis rates of the controls, high glucose group and hyperosmotic group in co-cultured groups were significantly higher than those of corresponding group in Ins-1 single groups (5.03%±0. 41%, 3. 73% ± 0. 35% ; 11.73% ±1.20%, 7.93%±0.41%; 7.60%±0.72%, 5.60%±0.40%; F=55.68, P<0.05), and ODs were significantly lower ( 1.29±0. 19, 2. 85±0. 31 ; 0. 62 ± 0. 06, 2. 28 ± 0. 13 ; 0. 65 ± 0. 07, 2. 35 ± 0. 12;F = 97.75, P < 0. 05 ), and the typical nuclei morphological changes of apoptosis cells were observed. Conclusion Hyperglycemia contributes to the deterioration of beta-cell line Ins-1, and increases its apoptosis rate, possibly intensified in the context of PSC involvement.
