Rizk Natalie N, Saed Ghassan M, Diamond Michael P
Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Wayne State University, Detroit, Michigan 48201, USA.
Fertil Steril. 2006 Oct;86(4 Suppl):1217-22. doi: 10.1016/j.fertnstert.2006.04.022. Epub 2006 Sep 7.
To determine whether the insulin receptor (IR), insulin-like growth factor-I receptor (IGF-IR), and IGF-I are expressed differentially in fibroblasts isolated from normal peritoneal and adhesion tissue before and after 24-hour treatment with increasing glucose concentrations.
Prospective experimental study.
University medical center.
PATIENT(S): Primary cultures of fibroblasts established from peritoneal and adhesion tissues of the same patients.
INTERVENTION(S): Glucose treatment of the primary cultured fibroblasts for 24 hours with increasing concentrations of glucose (100-850 mg/dL).
MAIN OUTCOME MEASURE(S): Real-time polymerase chain reaction was used to measure messenger RNA (mRNA) levels for IR, IGF-IR, and IGF-I. Enzyme-linked immunosorbent assay was used to determine protein levels.
RESULT(S): At the normal glycemic level (100 mg/dL), adhesion fibroblasts have significantly higher mRNA levels of the IR (7.96 +/- 0.15 vs. 6.97 +/- 0.16; P<.05), IGF-IR (7.72 +/- 0.22 vs. 6.88 +/- 0.06; P<.05), and IGF-I (7.04 +/- 0.10 vs. 5.92 +/- 0.10; P<.05) when compared with normal fibroblasts, respectively. Data are expressed as log(mRNA/microg RNA). Normal fibroblasts respond to increasing glucose concentrations by increasing the expression levels of the IR, IGF-IR, and IGF-I, whereas adhesion fibroblasts respond by decreasing the expression of the IR, IGF-IR, and IGF-I.
CONCLUSION(S): The differential expression of the IR, IGF-IR, and IGF-I in adhesion fibroblasts may contribute to the pathogenesis of fibrosis observed in diabetic patients.
确定在正常腹膜组织和粘连组织分离出的成纤维细胞中,胰岛素受体(IR)、胰岛素样生长因子-I受体(IGF-IR)和IGF-I在葡萄糖浓度递增处理24小时前后是否存在差异表达。
前瞻性实验研究。
大学医学中心。
从同一患者的腹膜和粘连组织建立的成纤维细胞原代培养物。
用浓度递增的葡萄糖(100 - 850mg/dL)对原代培养的成纤维细胞进行24小时处理。
采用实时聚合酶链反应测量IR、IGF-IR和IGF-I的信使核糖核酸(mRNA)水平。采用酶联免疫吸附测定法测定蛋白质水平。
在正常血糖水平(100mg/dL)时,与正常成纤维细胞相比,粘连成纤维细胞的IR(7.96±0.15对6.97±0.16;P<0.05)、IGF-IR(7.72±0.22对6.88±0.06;P<0.05)和IGF-I(7.04±0.10对5.92±0.10;P<0.05)的mRNA水平分别显著更高。数据以log(mRNA/μg RNA)表示。正常成纤维细胞对葡萄糖浓度增加的反应是增加IR、IGF-IR和IGF-I的表达水平,而粘连成纤维细胞的反应是降低IR、IGF-IR和IGF-I的表达。
粘连成纤维细胞中IR、IGF-IR和IGF-I的差异表达可能有助于解释糖尿病患者中观察到的纤维化发病机制。