Molecular analysis of collagens in bladder fibrosis
Molecular analysis of collagens in bladder fibrosis
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DOI：
10.1016/s0022-5347(01)62606-5
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发表时间：
1998-10-01
影响因子：
6.6
通讯作者：
Howard, PS
中科院分区：
文献类型：
--
作者：
Deveaud, CM;Macarak, EJ;Howard, PS
目的：膀胱组织纤维化的特征在于结缔组织在膀胱壁的不同层内的异常沉积，导致低容量、高压膀胱，这可能最终导致肾瘢痕形成和衰竭。这些膀胱在功能上被称为“非顺应性”，并且可能由不同的病因引起：神经源性，包括脊髓发育不良和脊髓损伤，或非神经源性，由于梗阻或放射治疗。为了研究负责这种纤维化的分子机制，我们分析了一个特征良好的儿科患者群体的改变（S）在胶原蛋白I型和III型调节在蛋白质和核酸levels.Materials和方法：免疫组织化学定位的胶原蛋白亚型（I，III和IV）进行了使用类型特异性单克隆抗体。通过羟脯氨酸分析测定总胶原，并且通过竞争性ELISA定量溴化氰提取程序之后胶原蛋白的亚型特异性表达。通过胍/苯酚/氯仿提取总RNA，用放射性标记的人cDNA探针进行狭缝印迹杂交分析，通过光密度法对所得的autoradiographs.Results进行定量：逼尿肌平滑肌束的结缔组织浸润是特异性的III型胶原。蛋白质分析表明：1）总胶原蛋白增加，2）III型：I型胶原蛋白比率的统计学显著增加，和3）非顺应性膀胱组织中III型胶原蛋白的绝对增加。在mRNA水平上，在非神经源性膀胱组织中I型和III型胶原蛋白稳态mRNA的协同增加，而神经源性膀胱组织的特征是III型：I型mRNA转录比的增加。这些数据表明，膀胱纤维化中胶原合成的调节是复杂的，其特征在于转录和转录后机制，这取决于纤维化的病因。
Purpose: Fibrosis of bladder tissue is characterized by an abnormal deposition of connective tissue within different layers of the bladder wall, resulting in a low volume, high pressure vesical which may ultimately contribute to renal scarring and failure. These bladders are functionally referred to as "non-compliant" and may result from different etiologies: neurogenic, which encompasses myelodysplasia and spinal cord injury, or non-neurogenic, owing to obstruction or radiation therapy. To examine the molecular mechanisms responsible for this fibrosis, we have analyzed a well-characterized pediatric patient population for alteration(s) in collagen types I and III regulation at the protein and nucleic acid levels.Materials and Methods: Immunohistochemical localization of collagen subtypes (I, III, and IV) was carried out using type specific monoclonal antibodies. Total collagen was determined by hydroxyproline analysis, and subtype specific expression of collagenous proteins, following cyanogen bromide extraction procedures, was quantified by competitive ELISA. Total RNA was extracted by guanidinium/phenol/chloroform, and slot blot hybridization analyses with radiolabeled human cDNA probes were quantified by densitometry of resulting autoradiograms.Results: Connective tissue infiltration of detrusor smooth muscle bundles was specific for type III collagen. Protein analyses demonstrated: 1) an increase in total collagen, 2) a statistically significant increase in the type III: type I collagen ratio, and 3), an absolute increase in type III collagen protein in non-compliant bladder tissue. At the mRNA level, there was a coordinate increase in both collagen I and III steady-state mRNAs in non-neurogenic bladder tissue, whereas neurogenic bladder tissue was characterized by an increase in the type III: type I mRNA transcript ratio.Conclusions: These data suggest that regulation of collagen synthesis in bladder fibrosis is complex and is characterized by both transcriptional and post-transcriptional mechanisms, depending upon the etiology of the fibrosis.