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Experimental Study of Aquaporin 1, 3 and Basic Fibroblast Growth Factor in Human Peritoneal Mesothelial Cells for Peritoneal Ultrafiltration

Author: LiangXianHui
Tutor: LiuZhangSuo
School: Zhengzhou University
Course: Department of Internal Medicine Nephrology
Keywords: Peritoneal ultrafiltration Human peritoneal mesothelial cells Aquaporin Basic fibroblast growth factor
CLC: R459.5
Type: Master's thesis
Year: 2007
Downloads: 119
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Abstract


Background and Purpose peritoneal ultrafiltration failure (ultrafiltration failure, UFF) is a major complication of long-term peritoneal dialysis (peritonealdialysis, PD), and become one of the main reasons was forced to withdraw from dialysis patients with PD. Water channel proteins (aquaporins, AQPs) is a group-mediated cross-membrane water transport membrane protein, recent research found that peritoneal mesothelial cells of AQP1 expression of AQP3 in and cross with peritoneal cell transport of water and pathological circumstances UFF is closely links. Histopathological detection of PD patients with peritoneal peritoneal vascular formation increased peritoneal small solutes high transit speculated that increased neovascularization can lead to lead to osmotic pressure gradient disappears the clinical performance UFF. Known angiogenesis by the variety of angiogenic factor regulation, such as vascular endothelial growth factor (vascular endothelial cell growth factor, VEGF), basic fibroblast growth factor (basic fibroblast growth factor, bFGF), transforming growth factor-β1 (transforming growth factorβ1, TGFβ1). Which bFGF promote mitosis and neovascularization peptide factor plays an important role in the development of tissue fibrosis. In recent years, scholars have found that bFGF mRNA expression can be detected in the PD patients with dialysis effluent mesothelial cells may prompt secretion of bFGF potential to, and participation in the peritoneal angiogenesis process. Non-physiological compatibility and high dialysate concentrations of glucose and glucose degradation products (GDP) is widely recognized as one of the cause the UFF of the main reasons for, in addition to chronic renal failure patients with PD uremia environment is also considered trigger UFF The main cause. About high sugar and uremia environment changes in peritoneal mesothelial cells AQPs in peritoneal ultrafiltration currently little research, The bFGF research still is blank. The purpose of this experiment is to observe AQP1, AQP3 and bFGF expression changes in different environments (high sugar, hypertonic uremic environment), find their expression variation in PD, and analysis of AQP1, AQP3 and bFGF between related to a preliminary understanding of the UFF occurrence provide experimental evidence for the existence of a common regulatory mechanism for the prevention and treatment of clinical UFF. Methods Cultured human peritoneal mesothelial cells (human peritoneal mesothelial cells, HPMCs). Take the growth state of good cells, until their integration into films mimic in vivo PD environment, were given glucose uremic serum stimulation, the cells were divided into normal group, high glucose group, uremia group, high-sugar uremia combined stimulation group, while establishment of mannitol group of AQP3 in and bFGF to judge hypertonic environment cell of AQP1. Cell immunohistochemistry and reverse transcriptase polymerase chain reaction (reverse transcript polymerse chainreaction, RT-PCR) method for determination of the different factors that stimulate HPMCs cultured in of AQP1, AQP3, of bFGF protein and mRNA expression levels change, and comparison and correlation analysis. Results 1. The normal HPMCs AQP1, AQP3 and bFGF protein expression of the mRNA levels measured. 2. Compared with the normal group, high glucose group, mannitol group of AQP1 of AQP3 in protein expression enhanced the strong positive expression was seen in the uremic group and high sugar uremia group. AQP1, AQP3 mRNA expression semi-quantitative analysis showed that: compared with the normal group, high glucose group, uremia group, the high sugar uremia group of AQP1 AQP3mRNA increased expression of comparison between groups was statistically significant (P <0.05). The group of high sugar and mannitol group cells AQP1 mRNA (P = 0.15), AQP3 mRNA (P = 0.45) to express the difference was not statistically significant. 3. Compared with the normal group, high glucose group, of uremia groups and high sugar uremia group bFGF protein expression enhanced, while the the mannitol group of bFGF protein expression similar to the normal group. bFGF mRNA expression in the semi-quantitative analysis showed that: compared with the normal group, high glucose group, uremia group, high sugar uremia group bFGF mRNA expression increased (P <0.001), while the the mannitol group of mRNA expression increased, but the difference was not statistically significance (P = 0.32); High sugar group (r = 0.887, P = 0.045), uremia group (r = 0.910, P = 0.032) and high sugar uremia group (r = 0.899, P = 0.038) AQP1 mRNA expression of bFGF positive correlation trend of AQP3 and bFGF mRNA expression was not correlated (P> 0.05). Summary 1. High sugar uremic serum raised HPMCs AQP1 and AQP3 expression, affect peritoneal ultrafiltration; 2. Induced by high glucose of AQP1 of AQP3 in expression through increased osmotic pressure to achieve; 3. Sugar, the uremic serum allows HPMCs bFGF to express increase, suggesting that can cause an increase in peritoneal blood vessels, leading to the UFF occurrence;

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