| IntroductionPeritoneal Dialysis ( PD) is an essential part of one - body renal replacement therapies, and during Peritoneal Dialysis, water and soluble substances are exchanged through peritoneum, a semipermeable membrane. Peritoneal Dialysis - related peritonitis is the one of severe complications, and once it cann' t be controlled well, it can lead to dialysis failure.Peritoneal Dialysis -related peritonitis causes ultrafiltration failure(UF) in peritoneal funtion. UF may be devided into four types ( I - IV type). During the earlier period, PD - related peritonitis manifests I type membrane failure, which is characterized by rapid absorption of glucose ( Glc) and earlier disap-pearation of osmotic pressure difference. II type membrane failure means less product of multiplication of peritoneal permeability and peritoneal area, and fl[ type membrane failure means higher contraultrafiltration ability, but average level of capiliary ultrafiltration ability. Recently IV type membrane failure is raised, whose cause is abnormal aquaporin. As far, UF is still the research focus.Hexokinase (HK) is the first rate -limit enzyme in the glucose metabolis-tic pathway. Our previous research showed that there was high - activity HK in peritoneal mesothelial cells ( PMCs) and HK played an important role in I type ultrafiltration failure. Then does HK have the same effect in PD - related peritonitis? In this research the regulation of HKs and net Glc Utilization in PMCs by lipopolysaccharide (LPS) will be discussed, in order to illumilate the mecha-nism of UF during PD - related peritonitis.Tumor necrosis factor (TNF) is also named by cachectin, which includes two subtypes: TNF and TNF. LPS is their strong stimulus. Recent researches focused on TNF and proved that TNF participated in many physical and pathological periods, and the inflammation factors from the drain of peritoneal dialysis , for example TNF, IL - 1, IL - 6, all joined in these periods. Many reports showed that PMCs were able to secrete some factors, for example IL - 1, IL - 6 and so on, but whether PMCs can express TNF during peritonitis hasn' t been reported yet. This experiment will try to explain the damages to peritoneum caused by PD - related peritonitis and the function of PMCs during the process.Materials1. Experimental cells: peritoneal mesothelial cells were isolated by primary culture from male rat of the Sprague Dawley; L929 cell strain (rat fibroblasts strain ).2. Reagents relating to cell culture and passage.3. Lipopolysaccharide.4. Reagents relating to measurement of hexokinases activity and glucose concentration.5. Reagents relating to measurement of TNF activity.Methods1. Enzymatic disaggregation was used for primary culture and passage.2. Standard G6PDH - coupled assay was used for measurement of HKs activity to observe the regulation effects of LPS on HKs activity.3. Colorimetric assay was used for measurement of net Glc utilization.4. Utilize cytotoxicity effect of TNF on L929 cells to measure TNF activity.Results1. Different - concentration LPS induced HKs activity with high Glc.(1 ) LPS concentration and glucose concentration had a cross - effect on HKs activity ( F=423.55, P<0.0001).(2) When LPS stimulated PMCs, Glc induced HKs activity in a dose - dependent manner ( F = 24.13 ,P < 0.0001).(3) LPS induced HKs activity in a dose - dependent manner (F =7. 15, P <0.0001).2. LPS induced HKs activity with high Glc for different periods.(1) LPS - effect time and glucose concentration both influenced HKs activity, and there is a cross -effect between them on HKs activity (F =54. 88, P < 0. 0001).(2) When LPS stimulated PMCs for different periods, Glc induced HKs activity in a dose - dependent manner( F = 31. 63 , P < 0. 0001).(3) LPS induced HKs activity in a time - dependent manner( F = 5. 03, P <0.01).3. LPS accelerated the net Glc utilization of PMCs.(1) Compared to the control group, the net Glc utilization of PMCs increased significa...
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