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β细胞素(betacellulin,BTC)是目前较受关注的胰岛再生因子,但其促胰腺、胰岛再生的机制不清.BTCe是betacellulin的功能片段,促细胞增殖能力与BTC相同.实验通过原核表达方法获得BTCe蛋白,MTT法证实其促3T3-L1细胞增殖能力.将BTC或BTCe作用于原代培养的大鼠胰岛,观察其对胰岛分泌的急性及长期影响作用,实时定量PCR及免疫荧光检测胰岛内关键基因的表达.将质粒pcDNA3.1-BTCe注射入链脲霉素(STZ)诱导的糖尿病大鼠肌肉中,观察对大鼠血糖的影响作用.加入BTC或BTCe可明显提高体外培养大鼠胰岛的GSIS水平,但实时定量PCR及免疫荧光显示胰岛内4种关键基因的表达并无明显变化;pcDNA3.1-BTCe转染糖尿病大鼠15～20天后血糖出现下降,糖耐量明显改善;免疫荧光显示:胰腺内有大量PDX-1+的导管细胞及胰岛素阳性细胞出现.推测BTC及BTCe对体外长期培养的大鼠胰岛具有一定的保护作用,可能通过促进胰腺内PDX-1+的导管细胞及胰岛素阳性细胞的增殖、诱导对STZ诱导的糖尿病大鼠的高血糖具有一定缓解作用.
However, the mechanism of pancreatic islet and pancreatic islet regeneration is unclear.BTCe is a functional fragment of betacellulin, and has the same promoting ability of cell proliferation as BTC.Through the prokaryotic expression method The BTCe protein was obtained by MTT assay, and the proliferation of 3T3-L1 cells was confirmed by MTT assay.Acute and long-term effects of BTCe or BTCe on rat pancreatic islets were observed by real-time quantitative PCR and immunofluorescence assay The expression of key genes within the plasmid pcDNA3.1-BTCe injected streptozotocin (STZ) -induced diabetic rats muscle to observe the effect of blood glucose in rats.Adding BTC or BTCe can significantly improve in vitro cultured rat However, there was no significant change in the expression of four key genes in islets after real-time quantitative PCR and immunofluorescence. The level of blood glucose decreased and the glucose tolerance remarkably improved after 15 to 20 days of pcDNA3.1-BTCe transfection in diabetic rats. Fluorescence showed that a large number of PDX-1 + ductal cells and insulin-positive cells appeared in the pancreas.It is presumed that BTC and BTCe have a protective effect on long-term cultured rat islets in vitro possibly through Proliferation into the pancreatic duct PDX-1 + cells and insulin-positive cells, the induction of STZ-induced diabetic rats with hyperglycemia certain mitigation.