胰腺星状细胞对高糖诱导的胰岛β细胞凋亡的影响

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.

Effect of pancreatic stellate cell on apoptosis of beta-cell induced by high concentration of glucose

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.