Laboratory of Vaccines and Immunotherapeutics, Institute of Bioscience, Universiti Putra Malaysia, UPM Serdang, Selangor 43400, Malaysia.
BMC Biotechnol. 2012 Sep 19;12:64. doi: 10.1186/1472-6750-12-64.
Gene therapy could provide an effective treatment of diabetes. Previous studies have investigated the potential for several cell and tissue types to produce mature and active insulin. Gut K and L-cells could be potential candidate hosts for gene therapy because of their special features.
In this study, we isolated gut K and L-cells to compare the potential of both cell types to produce insulin when exposed to similar conditions. The isolated pure K and L-cells were transfected with recombinant plasmids encoding insulin and with specific promoters for K or L-cells. Insulin expression was studied in response to glucose or meat hydrolysate. We found that glucose and meat hydrolysate efficiently induced insulin secretion from K and L-cells. However, the effects of meat hydrolysate on insulin secretion were more potent in both cells compared with glucose. Results of enzyme-linked immunosorbent assays showed that L-cells secreted more insulin compared with K-cells regardless of the stimulator, although this difference was not statistically significant.
The responses of K and L-cells to stimulation with glucose or meat hydrolysate were generally comparable. Therefore, both K and L-cells show similar potential to be used as surrogate cells for insulin gene expression in vitro. The potential use of these cells for diabetic gene therapy warrants further investigation.
基因治疗可为糖尿病提供有效的治疗方法。之前的研究已经调查了几种细胞和组织类型产生成熟和有活性胰岛素的潜力。肠 K 和 L 细胞因其特殊特性可能成为基因治疗的潜在候选宿主。
在这项研究中,我们分离了肠 K 和 L 细胞,以比较这两种细胞类型在暴露于相似条件下产生胰岛素的潜力。分离的纯 K 和 L 细胞被转染了编码胰岛素的重组质粒,并带有 K 或 L 细胞的特异性启动子。研究了胰岛素在葡萄糖或肉水解物刺激下的表达。我们发现葡萄糖和肉水解物能有效地诱导 K 和 L 细胞的胰岛素分泌。然而,与葡萄糖相比,肉水解物对胰岛素分泌的作用在两种细胞中更为强烈。酶联免疫吸附试验的结果表明,无论刺激物如何,L 细胞分泌的胰岛素均多于 K 细胞,尽管这种差异无统计学意义。
K 和 L 细胞对葡萄糖或肉水解物刺激的反应通常是可比的。因此,K 和 L 细胞均显示出相似的潜力,可作为体外胰岛素基因表达的替代细胞。这些细胞用于糖尿病基因治疗的潜力值得进一步研究。