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该研究观察了高糖高胰岛素对小鼠胰腺星状细胞(pancreatic stellate cells,PSCs)活化、增殖、细胞外基质(extracellular matrix,ECM)合成和半乳凝素-3(galectin-3,Gal-3)表达的影响。分离PSCs并培养至3~5代后进行实验。PSCs干预分为低糖对照组(5 mmol/L葡萄糖)、高糖组(25 mmol/L葡萄糖)、高胰岛素组(5 mmol/L葡萄糖+100 nmol/L胰岛素)、高糖高胰岛素组(25 mmol/L葡萄糖+100 nmol/L胰岛素)。细胞免疫荧光检测胰岛素受体(insulin receptor,IR)和胰岛素样生长因子-1型受体(insulin like growth factor-1 receptor,IGF-1R)在PSCs的表达;MTT法检测PSCs增殖;RT-PCR和Western blot测定平α-滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、I型胶原(type I collagen,Col I)、纤连蛋白(fi bronectin,Fn)和Gal-3的m RNA和蛋白质水平。结果发现,PSCs细胞表达IR和IGF-1R;与低糖对照组相比,高糖组、高胰岛素组、高糖高胰岛素组均诱导PSCs活化、增殖并促进Col I、Fn生成和Gal-3表达,其中以高糖高胰岛素组最为显著。以上结果说明,2型糖尿病高糖、高胰岛素微环境可能促进PSCs活化、增殖、ECM生成和Gal-3表达,在一定程度上可导致胰腺纤维化。
This study investigated the effects of high glucose and high insulin on the activation and proliferation of mouse pancreatic stellate cells (PSCs), the synthesis of extracellular matrix (ECM) and the expression of galectin-3 (Gal- 3) the impact of expression. PSCs were isolated and cultured to 3 to 5 generations after the experiment. PSCs intervention was divided into low glucose control group (5 mmol / L glucose), high glucose group (25 mmol / L glucose), high insulin group (5 mmol / L glucose +100 nmol / L insulin) mmol / L glucose + 100 nmol / L insulin). The expression of insulin receptor (IR) and insulin-like growth factor-1 receptor (IGF-1R) in PSCs was detected by immunofluorescence staining. The proliferation of PSCs was detected by MTT assay. The expressions of α-smooth muscle actin (α-SMA), type I collagen (Col I), fibronectin (Fn) and Gal-3 m RNA and protein levels. The results showed that PSCs cells expressed IR and IGF-1R. Compared with the low glucose control group, the high glucose group, the high insulin group and the high glucose and high insulin group all induced the activation and proliferation of PSCs and promoted the expression of Col I, Fn and Gal-3 , Among which the group with high glucose and high insulin was the most obvious. These results suggest that high glucose and high insulin microenvironment of type 2 diabetes may promote the activation and proliferation of PSCs, the production of ECM and the expression of Gal-3, to a certain extent, lead to pancreatic fibrosis.