川芎嗪抗腹膜间皮细胞损伤的实验研究

目的：观察含川芎嗪腹透液体外对腹膜间皮细胞活力,细胞增殖的影响。方法：分离,培养大鼠腹膜间皮细胞,用细胞计数法,四唑氨蓝（MTT）法,乳酸脱氢酶（LDH）法检测腹膜间皮细胞活力,细胞增殖能力。结果：培养的腹膜间皮细胞暴露于常规使用的4．25％高糖透析液30min后,间皮细胞数量显降低,MTT法示OD值显低于正常对照组,LDH释放增加,提示间皮细胞活力降低,增殖受到显抑制,加入低剂量川芎嗪后,间皮细胞数,MTTOD值较之单纯高糖透析液组均显增高,LDH显降低,但随着剂量的加大,川芎嗪的上述作用减低乃至消失,结论：适当剂量的川芎嗪能对抗常规腹透液引起的间皮细胞抑制和损伤,从而具有保护间皮细胞的作用。     

细胞周期抑制蛋白p27在慢性腹膜透析大鼠腹膜间皮细胞肥大中的作用

目的探讨含糖腹膜透析(腹透)液作用下慢性腹透大鼠模型腹膜间皮细胞的形态与p27表达的关系.方法 30只SD大鼠随机分为4.25%腹透液组(高糖组)、林格氏液组(林格组)和对照组3组8周后进行细胞印片,并用IBAS 2.0图像分析系统进行形态学分析,免疫组织化学方法检测p27表达水平.结果与对照组和林格组相比,高糖组细胞密度显著减少(P＜0.01),表面积明显增大(P＜0.01)与对照组(22 4±0.99)和林格组(2.52±1.25)相比,高糖组间皮细胞p27的表达显著增多(1O.43±3.78,P＜0.01).腹膜间皮细胞p27的表达上调与高糖诱导的间皮细胞的肥大呈显著的正州关(r=0.998,P＜0.O1).结论 D27的表达上调可能是长期腹膜透析高糖诱导间皮细胞肥大的可能机制之一.     

丙酮酸盐腹膜透析液对中性粒细胞产生超氧自由基的影响

腹膜炎的发生与腹膜透析时腹膜防御功能的降低密切相关,众多资料表明,临床常用的酸性高糖乳酸盐腹膜透析液(lactate-based peritoneal dialysis solution,L-PDS)对宿主防御细胞有明显的毒性作用[1]。近年来研究发现以丙酮酸盐替代乳酸盐作缓冲碱的丙酮酸盐腹膜透析液(pyruvate-based peritoneal dialysis solution,     

腹膜透析液对腹膜间皮细胞凋亡与增殖的影响

目的　探讨PD 2腹膜透析液 (腹透液 )在体内对腹膜间皮细胞凋亡和增殖细胞核抗原(PCNA)表达的影响。方法　在正常二级SD大鼠上建立腹膜透析模型 ,用不同浓度的PD 2腹透液灌注腹腔 ,二次 /天 ,2 0ml/次 ,生理盐水作对照。透析第 8周后杀检 ,取腹膜用LSABkit和TUNELkit作免疫 凋亡双重染色。结果　腹透液显著诱导腹膜间皮细胞凋亡 ,以高渗糖腹透液尤为显著 ,不同浓度腹透液对PCNA影响则不同 ,透析液诱导的凋亡与PCNA表达之间无相关性 ,但与腹透液浓度正相关 ,PCNA表达则与腹透液浓度负相关。结论　腹透液可诱导腹膜间皮细胞凋亡 ,并影响PCNA表达 ,其作用与腹透液浓度不同而异。     

高糖对人腹膜间皮细胞局部内皮素系统表达的影响

目的:研究高浓度葡萄糖对人腹膜间皮细胞(HPMC)局部内皮素系统表达的影响,并应用非特异性内皮素受体拮抗剂即ETA/ETBR拮抗剂PD142893进行干预,探讨高浓度葡萄糖透析液导致腹膜受损的机制.方法:以人腹膜间皮细胞株(HMrSV5)为研究对象,采用放免法检测不同时间(12 h、24 h和48 h)、不同浓度(50、100、150、200、250 mmol/L)的葡萄糖、甘露醇作用下HPMC分泌的ET-1水平.采用半定量逆转录多聚酶链反应(RT-PCR)检测局部内皮素系统ET-1、ETAR、ETBR及ECE的基因表达.采用ELISA检测不同浓度ET-1、葡萄糖作用下IL-1β水平,并在此基础上加入PD142893,观察IL-1β的变化.结果:(1)正常HPMC分泌低水平的ET-1,高浓度的葡萄糖可诱导HPMC分泌 ET-1增加,呈时间和剂量依赖.高浓度的甘露醇作用下,HPMC分泌的ET-1与正常对照组相比未见明显差异.(2)正常HPMC表达完整的ET-1、ETAR、ETBR、ECE mRNA.高糖作用下ET-1、ETAR、ETBR mRNA表达增强,而高糖对ECE mRNA的表达无明显影响.(3)ET-1、高糖刺激HPMC分泌IL-1β,呈剂量依赖.PD142893可明显减少高糖诱导的IL-1β的分泌.结论:正常HPMC存在局部内皮素系统,高浓度葡萄糖诱导其表达增加,高浓度葡萄糖透析液导致腹膜受损可能部分通过内皮素途径,内皮素可能是导致CAPD患者腹膜纤维化的重要因素.     

氨基酸腹膜透析液与传统腹膜透析液对腹膜间皮细胞功能的影响

目的通过体外实验对照观察了含氨基酸腹膜透析液(1.1%Nutrineal)和传统葡萄糖腹膜透析液(1.5%Dianeal)对正常人腹膜间皮细胞(HPMC)的生物相容性及功能的影响。方法用1.1%Nutrineal和1.5%Dianeal进行腹膜透析,4h后引流。观察两种不同透出液及不同时间对体外培养的HPMC功能的影响。采用四甲基偶氮多胍(MTT)和细胞直接计数法测定HPMC增殖程度,以测定乳酸脱氢酶(LDH)水平显示细胞损伤程度。采用ELISA法测定白细胞介素6(IL-6)水平。结果与5%葡萄糖对照组相比,1.5%Dianeal透出液可显著降低HPMC细胞增殖能力、细胞总蛋白合成、IL-6分泌水平和细胞黏附能力(P<0.001、P<0.001、P<0.0001、P<0.0001),而细胞培养液上清中的LDH分泌水平明显提高(P<0.001)。1.1%Nutrineal透出液可降低HPMC细胞增殖能力(P<0.001)和明显增加IL-6的分泌水平(P<0.0001),但对HPMC的LDH分泌水平,细胞黏附能力,蛋白总合成量无明显影响。结论与传统的葡萄糖腹膜透析液相比,氨基酸腹膜透析液能更好地维持HPMC在细胞存活能力、细胞黏附功能及蛋白合成等方面的功能。氨基酸腹膜透析液可诱导HPMC分泌IL-6水平显著升高,其机制有待进一步探讨。     

细菌毒素和肿瘤坏死因子-α对人腹膜间皮细胞白细胞介素8和纤连蛋白分泌的影响

目的：研究细菌毒素和肿瘤坏死因子（TNF）-α对人腹膜间皮细胞分泌白细胞介素（IL）-8和纤连蛋白（FN）的影响。方法：分离人腹膜间皮细胞（HPMC）做体外培养,采用酶联免疫双抗夹心法和逆转录聚合酶链反应,检测细胞产生IL-8和FN水平的情况。采用Lowry方法检测培养液中细胞内总蛋白。结果：脂多糖（10μg/ml)、金黄色葡萄球菌肠毒素（10μg/ml)和TNF-α（1000U/ml)分别刺激HPMC后,培养液内IL-8和FN蛋白质水平显著增高（P＜0.01),并上调IL-8和FN的mRNA表达。结论：细菌毒素和TNF-α可引起人腹膜间皮细胞IL-8和FN蛋白水平增加,并上调HPMC IL-8和FN mRNA表达;FN合成增加可能是导致腹膜纤维化原因之一。     

高糖与炎症因子及黄芪对人腹膜间皮细胞毒性的影响

目的:研究不同浓度的含糖腹透液、多种炎症因子以及二者的共同作用对人腹膜间皮细胞损伤的影响,并探索中药黄芪对此的干预作用.方法:用乳酸脱氢酶(LDH)释放法测定细胞毒性,观察各种因素(不同浓度的含糖腹透液与炎症因子的组合、多种炎症因子及其组合和中药黄芪)对体外培养人腹膜间皮细胞株损伤的影响.结果:4.25%腹透液使细胞破坏明显增加,P＜0.05;4.25%腹透液+TNF-α或(LPS+TNF-α+IL-1β)进一步加剧细胞损伤,P＜0.05和P＜0.01.黄芪明显减少细胞破坏,在1.5%腹透液情况下,P＜0.001;在TNF-α和LPS+TNF-α+IL-1β情况下,P＜0.05.结论:黄芪减少腹膜间皮细胞损伤,在炎症因子或含糖腹透液条件下,黄芪仍有保护间皮细胞的作用.     

葡萄糖透析液与腹膜纤维化

腹膜透析 (PD)目前是治疗终末期肾病的主要措施之一。在PD病人 ,葡萄糖透析液 (浓度一般为 1.5 0 %～4.2 5 % )可引起腹膜纤维化。近年来大量研究已证实 ,PD时腹膜间皮细胞直接浸泡于这种高糖透析液中 ,高糖可抑制间皮细胞生长 ,影响其修复和代谢功能 ,造成细胞外基质的大量沉积 ,这些均与PD相关性腹膜纤维化的发生有关。本文拟就葡萄糖透析液对腹膜间皮层完整性的影响及细胞外基质沉积两方面作一综述。     

丝裂素活化蛋白激酶信号通路在人腹膜纤维化的作用

目的 探讨丝裂素活化蛋白激酶(MAPK)信号通路对转化生长因子(TGF)β1致人腹膜间皮细胞(HPMC)高表达纤连蛋白(FN)、纤溶酶原激活物抑制物(PAI)1的调控作用。方法 采用胰蛋白酶消化法从人腹膜组织中分离间皮细胞,建立稳定的体外培养模型。用四甲基偶氮唑盐比色法(MTT)评估TGF-β1和细胞外信号调节激素(ERK)及p38阻断剂对HPMC的增殖作用。采用酶联免疫双抗夹心法检测HPMC培养液中FN、PAI-1的蛋白质水平。采用逆转录多聚酶链反应(RT-PCR)检测HPMC细胞内FN、PAI-1mRNA的表达。结果 (1)TGF-β1能刺激HPMC增殖(P<0.05),且呈剂量时间依赖性,加ERK阻断剂组或p38阻断剂组较TGF-β1组有明显抑制作用(P<0.01)。(2)TGF-β1能引起HPMC FN、PAI-1蛋白水平显著增高(P<0.01);加ERK阻断剂组或p38阻断剂组能使上述高表达受到抑制(P<0.01)。(3)TGF-β1能使HPMC FN、PAI-1mRNA表达上调,加ERK阻断剂组或p38阻断剂组能使上调水平受到抑制。结论 MAPK对TGF-β1致人腹膜间皮细胞高表达FN和PAI-1起调控作用,为临床防治腹膜纤维化提供新的思路。     

Phosphatidylcholine and peritoneal transport during peritoneal dialysis

Peritoneal effluent of patients on chronic ambulatory peritoneal dialysis (CAPD) contains a surface-active material (SAM) made up of phospholipids and showing phosphatidylcholine on thin-layer chromatography. This substance drastically lowers surface tension, helps to repel water and has a lubricating effect. The presence of stratified phosphatidylcholine on the peritoneum might narrow the stagnant dialysate fluid layer and situations which can alter the quantity or composition of SAM may affect peritoneal transport and also, perhaps, the formation of adherences. This led us to verify, experimentally, the presence of phospholipids in basal conditions, after CAPD and during peritonitis and to check if addition of phosphatidylcholine to dialysis liquid is able to modify water transport in patients with low ultrafiltration and peritonitis. Phospholipids in the dialysis effluent of patients who have been on CAPD for a long time are lower than observed in the first days of peritoneal dialysis. A more drastic, significant decrease in phospholipids was observed in patients with low ultrafiltration and in patients with peritonitis. Mean ultrafiltration significantly increases in patients with low ultrafiltration and in those with peritonitis during dialysis exchanges containing phosphatidylcholine (50 mg/l) indicating that the latter is able to restore normal physiological conditions.     

Encapsulating peritoneal sclerosis in paediatric peritoneal dialysis patients

Encapsulating peritoneal sclerosis (EPS) is a serious complication of chronic peritoneal dialysis (CPD). In contrast to the adult population, there are few studies regarding EPS in paediatric CPD patients, and the majority of reported patients are from Japan. The aim of the present report is to define the incidence of EPS in our paediatric CPD patients and to describe the clinical and laboratory characteristics. A total of 104 paediatric patients were followed from November 1989 to November 2003 and two were diagnosed as EPS (1.9%). The dialysis periods of these patients were 45 and 53 months with 6 and 8 peritonitis episodes, respectively. Clinical signs of EPS developed 7 and 14 days after the removal of the dialysis catheter, and CPD was replaced by haemodialysis because of persistent peritonitis. One patient was well after surgical management but died 6 months later. The second patient who was treated with prednisolone remained well at 16 months. In conclusion, EPS is a rare but important complication of CPD. We recommend that all patients on CPD who develop ultrafiltration failure be evaluated radiologically for the occurrence of EPS. Management should be tailored to the individual patient.     

腹腔喷射通气对家猪腹膜氧合的影响

目的 评价腹腔喷射通气对家猪腹膜氧合的影响.方法 健康家猪24只,雌雄不拘,12～ 16周龄,体重35 ～ 45 kg,采用随机数字表法,将其随机分为3组(n=8):假手术组(S组)、腹腔常频通气组(N组)及腹腔高频通气组(H组).麻醉诱导后经口行气管插管,机械通气,于气道机械通气35 min时进行腹腔喷射通气,驱动压0.5 kg/cm2,纯氧流量1.8 L/min,吸呼比1.0:1.5,N组和H组通气频率分别为16和150次/min,腹腔通气期间每隔30 s采集动脉血样,进行血气分析,当脉搏血氧饱和度≤90％时停止腹腔通气.记录无通气安全时间(腹腔通气开始至PaO2＜ 60 mm Hg的时间).结果 与S组和N组比较,H组PaO2升高,无通气安全时间延长(P＜ 0.05),3组间PaCO2差异无统计学意义(P＞ 0.05).结论 腹腔高频喷射通气可提高家猪腹膜氧合的效果,且可延长无通气安全时间,而腹腔常频喷射通气对家猪腹膜氧合无影响.     

Bidirectional peritoneal transport of albumin in continuous ambulatory peritoneal dialysis

The present study was undertaken in order to assess bidirectionalperitoneal kinetics of albumin after simultaneous i.v. and i.p.injection of radioiodinated albumin tracers (¹²⁵I-RISA and ¹³¹I-RISA)in eight clinically stable uraemic patients undergoing continuousambulatory peritoneal dialysis (CAPD). The plasma volume, intravascularalbumin mass (IVM), and overall extravasation rate of albuminwere not significantly different from that found in healthycontrols. Albumin flux from the plasma into the peritoneal cavitywas 3.71 ± 0.82 (SD) µmol/h, which was only 3%of the overall extravasation rate (137 ± 52 µmol/h).Albumin flux from the peritoneal cavity into the plasma wassubstantially lower (0.22 ± 0.07 µmol/h, P<0.01).The net peritoneal accumulation of the albumin from plasma over4 h was 14 ± 3.2 µmol, which was significantlylower than the intraperitoneal albumin mass at the end of thedialysis (54 ± 19 µmol, P<0.01). This indicatesthat only about 25% of the albumin loss during CAPD occurs directlyfrom the plasma. The initial osmotic net filtration was 508± 302 ml. The volume flow equivalent to the albumin fluxwas 6.3 ± 1.5ml/h into the peritoneal cavity and 7.8± 1.9ml/h back into the plasma. Although minor, as comparedto the osmotic net filtration (508 ml), the albumin flux equivalentvolume (31.2 ml) exceeded the steady state filtration (25.2ml) significantly (P<0.01) during the 4 h investigation. In conclusion, albumin flux into the peritoneal cavity is smallcompared to the overall extravasation rate, but our resultssuggest that CAPD loss of albumin predominantly occurs fromthe subperitoneal interstitial space and only to a minor degreedirectly from the plasma. Albumin flux equivalent volume flowis relatively small and most probably represents peritoneallymph drainage.     

Antibacterial peritoneal defence in automated peritoneal dialysis: Advantages of tidal over continuous cyclic peritoneal dialysis?

In tidal peritoneal dialysis (TPD) only a part of the infuseddialysate is drained with each exchange, leaving a residualvolume on top of which fresh fluid is cycled. As the persistentpresence of a buffered intraperitoneal reserve volume mightfavour peritoneal macrophage (PMO) function, PMO obtained fromeight patients during a 3-h continuous cyclic peritoneal dialysis(CCPD) or TPD session were studied in a randomized cross-overtrial. PMO were studied for uptake of E. coli (complement-dependent)and S. epidermidis (antibody-dependent), as well as for theirkilling capacity and peak chemiluminescence response. In addition,dialysate was sampled during both treatment sessions and studiedfor pH, osmolality, and effect on the viability of donor phagocytesand mesothelial cells. TPD-derived PMO were significantly better able to phagocytoseE. coli than CCPD-PMO (48 ±8 versus 33±6% uptake,P<0.05), whereas the other tested functional capacities revealedno significant difference between TPD- and CCPD-PMO. DuringTPD dialysate pH ranged from 6 to 7 as compared to a pH rangefrom 5 to 7 in CCPD. The presence of a residual dialysate volumeresulted in less wash-out of cells and opsonins early in thetreatment, and to some extent blunted the noxious effects offresh dialysis solutions. Overall, however, tidal PD appearedto have no advantage over CCPD regarding preservation of peritonealdefences.     

High peritoneal residual volume decreases the efficiency of peritoneal dialysis

BACKGROUND: Wide variation in peritoneal residual volume (PRV) is a common clinical observation. High PRV has been used in both continuous ambulatory peritoneal dialysis (CAPD) and automated peritoneal dialysis to minimize the time of a dry peritoneal cavity and to achieve better dialysis. However, the impact of PRV on peritoneal transport is not well established. In this study, we investigated the effect of PRV on peritoneal transport characteristics. METHODS: Peritoneal effluents were collected in 32 male Sprague-Dawley rats after a five-hour dwell with 1.36% glucose solution. Forty-eight hours later, a four hour dwell using 25 ml of 3.86% glucose solution and frequent dialysate and blood sampling was done in each rat with 125I-albumin as a volume marker. Before the infusion of the 3.86% glucose solution, 0 (control), 3, 6, or 12 ml (8 rats in each group) of autologous effluent (serving as PRV) was infused to the peritoneal cavity. RESULTS: After subtracting the PRV, the net ultrafiltration was significantly lower in the PRV groups as compared with the control group: 13.4 +/- 0.5, 12.0 +/- 1.0, 11.7 +/- 1.7, and 8.9 +/- 0.4 ml for 0, 3, 6, and 12 ml PRV groups, respectively (P < 0.001). The lower net ultrafiltration associated with higher PRV was due to (a) a significantly lower transcapillary ultrafiltration rate (Qu) caused by a lower osmotic gradient, and (b) a significantly higher peritoneal fluid absorption rate (KE) caused by an increased intraperitoneal hydrostatic pressure. No significant differences were found in the diffusive mass transport coefficient for small solutes (glucose, urea, sodium, and potassium) and total protein, although the dialysate over plasma concentration ratios values were higher in the high-PRV groups. The sodium removal was significantly lower in the PRV groups as compared with the control group (P < 0.01). CONCLUSION: Our results suggest that a high PRV may decrease net ultrafiltration through decreasing the Qu, which is caused by a decreased dialysate osmolality, and increasing the KE caused by an increased intraperitoneal hydrostatic pressure. The high volume of PRV also decreased the solute diffusion gradient and decreased peritoneal small solute clearances, particularly for sodium. Therefore, a high PRV may compromise the efficiency of dialysis with a glucose solution.     

Establishment of a novel mouse peritoneal dialysis-associated peritoneal injury model

Clinical and Experimental Nephrology - Peritoneal fibrosis induced by various factors during peritoneal dialysis (PD) can eventually lead to ultrafiltration failure and termination of PD treatment....