肠肌成纤维细胞与IBD相关肠纤维化

 肠纤维化是炎症性肠病(inflammatory bowel disease, IBD)的并发症,肠肌成纤维细胞是肠纤维化的关键细胞。肠肌成纤维细胞及其与炎症细胞的相互作用在IBD相关肠纤维化发生中起重要作用。     

Changes in peritoneal permeability due to chemical fibrosis

A relation between the mass transfer area coefficient (MTAC) and development of chemically induced fibrosis was evaluated. 20ml/kg of 0.08–0.2wt% chlorhexidine gluconate (CHG) was injected into the abdominal space of male Wistar rats (at 8 weeks). After 1–2h dwell, CHG was discarded and then the abdominal space was flushed with 20ml/kg of 0.9% saline solution three times. A peritoneal equilibration test (PET) was carried out every week for 1 month using Lactate Ringer solution containing 100 mg/dl of urea-N, 10 mg/dl of creatinine, and 1.5% of albumin. Eight out of 11 rats survived, 5 out of 8 rats developed adhesions, and 1 developed sclerosing encapsulating peritonitis-like degeneration. Although there was no statistically significant change in MTAC, MTAC increased from 1.18±0.66 ml/min to 1.92±0.6 ml/min in the first 3 weeks (P=0.09), and slightly decreased at the fourth week. It can be concluded that increase of MTAC may be due to increase of peritoneal permeability and/or increase of effective surface area such as angiogenesis.     

腹膜透析患者透出液透明质酸的变化

目的观察腹膜透析(简称腹透)患者透出液透明质酸(HA)的变化.方法对22例尿毒症腹透患者的透出液采用放射免疫法测定HA水平.结果腹透患者透出液HA较血清HA水平明显增高,分别为(665.79±562.81)μg/L和(336.46±235.32)μg/L(P＜0.05),而层黏连蛋白(LN)和Ⅲ型前胶原(PCⅢ)水平无显著差异;腹透患者透析时间大于3年组较小于3年组的透出液HA水平明显减低,分别为(211.16±73.82)μg/L和(410.10±96.35)μg/L(P＜0.005),而LN和PCⅢ水平无显著差异;腹透患者腹膜炎组透出液HA水平较非腹膜炎组显著增高,分别为(845.12±219.68)μg/L和(303.38±117.27)μg/L(P＜0.005);腹膜炎组透出液LN及PCⅢ也明显增高(P＜0.01).结论测定腹透患者透出液HA水平有助于了解腹膜间皮细胞功能及腹腔状态.     

振摇对腹膜表面层和腹膜功能的影响

目的：探讨腹腔振摇对腹膜表面层和腹膜功能的影响。 方法： 将12只雄性SD大鼠随机分成对照组（n=6）和振摇组（n=6）；振摇组大鼠麻醉后经腹部插入静脉留置针，由留置针管注入25 mL含［125I］-白蛋白的4.25%的葡萄糖透析液，在电摇床(100 r/min)上行腹膜动力学试验；对照组大鼠则行常规腹膜动力学试验。腹膜动力学试验完毕后，取对照组和振摇组大鼠的腹膜组织，经含1％四氧化锇（OsO4）的氟化碳(FC75)固定液固定后，行电镜检查，经Bouin液固定后行常规病理检查（HE染色）。 结果： 振摇组大鼠的腹膜通透性显著高于对照组，表现为腹腔液体重吸收率明显增加，经毛细血管的超滤率明显下降，淋巴重吸收率明显增加，净超滤量明显下降，小分子溶质转运明显增高。电镜检查结果显示，振摇组的腹膜表面层明显薄于和疏松于对照组，腹膜组织常规病理学检查两组无明显差异。 结论： 腹腔振摇使腹膜通透性增高，同时破坏了腹膜表面层，提示腹膜表面层在控制腹膜通透性中起重要作用。     

含糖透析液对慢性腹膜透析大鼠腹膜功能及间皮细胞形态的影响

目的:探讨含糖透析液对慢性大鼠腹膜透析模型腹膜功能和腹膜间皮细胞形态的影响及它们之间的关系。方法:40只SD大鼠随机分为4组,除对照组外,余3组每天分别腹腔内注入20mL4.25%透析液(HG)、1.5%透析液(LG)、林格氏液(RG)。8周后进行腹膜功能试验,并行细胞印片进行形态学分析。结果:HG组及LG组腹腔内透出液量和净出超量明显低于对照组和RG组(P<0.01),4h透析液与血浆尿素氮浓度之比(D/P_urea)显著高于对照组和RG组(P<0.05),尿素氮清除率(C_urea)显著低于对照组和RG组(P<0.01)。细胞印片上HG组及LG组间皮细胞的细胞密度显著少于对照组和RG组,表面积显著大于对照组和RG组(P<0.01)。但以上改变在HG组及LG组间无显著差异。使用含糖透析液8周所致的腹膜超滤功能下降和间皮细胞肥大呈负相关(r=-0.896,P<0.05)。结论:长期应用含糖透析液可使腹膜超滤功能下降,这一作用可能与腹膜间皮细胞肥大有关。     

一氧化氮抑制剂对腹膜透析大鼠腹膜形态及表面层的影响

目的:探讨高糖透析液对大鼠腹膜巨噬细胞一氧化氮(NO)的产生、诱导型一氧化氮合酶(iNOS)的表达和腹膜透析通透性的影响以及NO抑制剂的保护作用。方法: 培养生理盐水、1.5％和4.25％葡萄糖透析液透析下的大鼠腹腔内的巨噬细胞, 检测细胞培养上清液NO的浓度和巨噬细胞iNOS的表达, 并观察1.5％及4.25％葡萄糖透析液和NO抑制剂透析下大鼠的液体超滤量和腹膜形态的变化。结果:大鼠腹腔内巨噬细胞NO的产生随透析液中葡萄糖含量增加而增加;腹腔巨噬细胞iNOS的表达, 葡萄糖透析组明显高于生理盐水组;NO抑制剂对光镜下腹膜形态无影响, 但明显减轻4.25%葡萄糖透析液对超滤量和腹膜表面层的减少作用。结论:一氧化氮抑制剂可明显改善高糖透析大鼠的腹膜通透性, 并减轻高糖对腹膜表面层的损伤。     

Protective measures against ultrafiltration failure in peritoneal dialysis patients

Ultrafiltration failure in patients undergoing peritoneal dialysis is a condition with an incidence that increases over time. It is related to increased cardiovascular morbidity and mortality and is a major cause of the abandonment of the treatment technique. Because the number of patients undergoing renal replacement therapy is increasing with society aging and because approximately 10% of this population is treated with peritoneal dialysis, this matter is becoming more common in everyday practice for clinicians involved in the care of patients with chronic renal failure. In this review, we summarize the available measures used to prevent and treat ultrafiltration failure and the current state of research in the field, both in the experimental and clinical settings, focusing on the possible clinical applications of recent findings.     

Microbiology of peritoneal dialysis-related infection and factors of refractory peritoneal dialysis related peritonitis: A ten-year single-center study in Taiwan

BackgroundPeritoneal dialysis (PD)-related infection is a serious complication of patients with PD. Refractory peritonitis may lead to failure of PD, shift to hemodialysis (HD) or death. Besides, microbiologic resistance increased worldwide that might impact the treatment choice for such infections. Investigating the causative pathogens and risk factors of PD-related infections in Taiwan was warranted.MethodsThis is a retrospective study involving patients with PD from 2007 to 2016 in a southern Taiwan hospital. Patient characteristics, microbiological data, outcomes, and factors associated with refractory peritonitis were analyzed.ResultsThere were 190 episodes of PD-related peritonitis in 110 patients from this cohort. Gram-positive organisms were the leading cause of PD-related peritonitis, but gram-negative organisms, esp. Pseudomonas aeruginosa, were predominant for exit site infection and tunnel infection. The incidence of peritonitis was 0.25 episode per patient-year (1 episode per 47.69 months). The refractory rate was 14.2% (27/190). Methicillin resistance was noted in 2 (13.3%) of 15 Staphylococcus aureus isolates. Of 114 isolates, 72.8% (83) were susceptible to either cefazolin or gentamicin. Staphylococcus spp. and Escherichia coli infections were significantly associated with refractory peritonitis. Baseline hyponatremia (<130 mmol/L) was independently associated with refractory peritonitis.ConclusionGram-positive organisms remained major cause of PD-related peritonitis. About three quarters of causative pathogens were susceptible to the recommended empirical treatment for PD-related peritonitis. Baseline hyponatremia (<130 mmol/L) was independently associated with refractory peritonitis. Staphylococcus spp. and E. coli infections had important roles for refractory peritonitis.     

渗透压制剂对人腹膜间皮细胞透明质酸合成酶的调节作用

目的: 了解不同渗透压制剂对人腹膜间皮细胞(humanperitonealmesothelialcells,HPMCs)透明质酸合成酶(hyaluronansynthase,HAS)的调节作用及对透明质酸(hyaluronan,HA)合成的影响。方法: 分离培养的HPMCs随机分为5组:正常对照组(对照组);90mmol/L葡萄糖组(高糖组);30mmol/L葡萄糖组(低糖组);5%缩合葡萄糖(缩合葡萄糖组)及90mmol/L甘露醇组(甘露醇组)。给予上述刺激后继续培养24h,以RT-PCR法检测HAS-2mRNA和HAS-3mRNA的表达;收集细胞培养上清液,以放射免疫法检测HA浓度;以微粒排除法观察细胞胞衣样结构的面积。结果: 高糖组、低糖组、缩合葡萄糖组、甘露醇组HAS-2mRNA的表达均显著高于对照组(P均<0.05);高糖组HAS-3mRNA的表达亦明显增加(P<0.05),低糖组、缩合葡萄糖组、甘露醇组HAS-3mRNA表达与对照组相比差异无显著(P>0.05)。各实验组细胞胞衣样结构面积与细胞体面积的比值无显著差异(P均>0.05)。对照组、高糖组、低糖组、缩合葡萄糖组、甘露醇组细胞培养上清液HA的浓度均显著高于对照组(P均<0.05);高糖组HA浓度显著高于低糖组、缩合葡萄糖组和甘露醇组(P<0.05)。结论: 与高浓度葡萄糖相比,缩合葡萄糖虽增强HPMCsHAS-2mRNA表达,但对与炎症反应相关的HAS-3mRNA表达无影响,提示缩合葡萄糖可能在对腹膜组织生物相容性方面优于高浓度葡萄糖。     

醛固酮在腹膜透析大鼠腹膜纤维化中的作用

目的:研究醛固酮在腹膜透析大鼠腹膜纤维化中的作用,以及醛固酮受体拮抗剂是否能够减轻腹膜纤维化。方法:通过腹腔注射脂多糖(第1、3、5、7天,0.6 mg/kg)+透析液(每天腹腔注射4.25%含糖透析液100 m L/kg)建立伴有腹膜纤维化的腹膜透析大鼠模型。同时给予醛固酮受体拮抗剂(螺内酯,100 mg·kg-1·d-1)灌胃。4周后取大鼠腹膜行病理和免疫组化检测,同时采用real-time PCR和Western blot法检测醛固酮合成酶(CYP11B2)、11β-羟基类固醇脱氢酶2型(11β-HSD2)、盐皮质激素受体(MCR)和炎症因子的表达情况。结果:成功建立伴有腹膜纤维化的腹膜透析大鼠模型,发现腹膜间皮细胞受损,形态改变,胶原纤维沉积,腹膜增厚,腹膜和腹透液中巨噬细胞浸润增多。与正常大鼠相比,腹膜上MCR、11β-HSD2和CYP11B2表达增高,醛固酮含量增多。同时腹膜上NF-κB/MCP-1表达增加。而给予螺内酯治疗后腹膜纤维化减轻,NF-κB/MCP-1表达减少。结论:局部产生的醛固酮可能通过NF-κB/MCP-1途径参与腹膜纤维化过程。醛固酮受体拮抗剂螺内酯能够减轻腹膜透析大鼠的腹膜纤维化程度。     

腹透液的生物相容性对2', 5'-寡腺苷酸合成酶活性的影响

目的：观察使用商业腹透液时2', 5'-寡腺苷酸合成酶活性的动态变化和腹膜病理改变,探讨长期腹膜透析腹透液生物相容性对机体免疫功能影响和腹膜间皮的病理改变。方法：①使用3种常用腹透液建立腹膜透析动物模型;②用纸层析法动态测定实施和停止腹膜透析时,2', 5'-寡腺苷酸合成酶活性的消长;③在光镜下观察腹膜间皮病理变化。结果：腹膜透析时,各腹膜透析组和生理盐水组2', 5'-寡腺苷酸合成酶活性均升高;停止腹膜透析,生理盐水组2', 5'-寡腺苷酸合成酶活性恢复正常,各腹膜透析组则持续降低;在腹膜透析时,各腹膜透析组均发生程度不等的间皮细胞脱落、腹膜增厚和急、慢性炎症细胞浸润;而Baxter组无急性炎症发生。随着停止腹膜透析时间延长,腹膜透析组病理改变逐渐修复。结论：长期腹膜透析时,腹膜透析液对机体免疫功能均有长时间抑制,并造成不同程度的腹膜病理改变。腹透液的非生物相容性是导致机体免疫功能降低,腹膜损伤的重要因素。     

The size-dependent efficacy and biocompatibility of hyperbranched polyglycerol in peritoneal dialysis

Glucose is a common osmotic agent for peritoneal dialysis (PD), but has many adverse side effects for patients with end-stage renal disease. Recently, hyperbranched polyglycerol (HPG) has been tested as an alternative osmotic agent for PD. This study was designed to further examine the efficacy and biocompatibility of HPG over a range of different molecular weights. HPGs of varying molecular weights (0.5 kDa, 1 kDa, 3 kDa) were evaluated in a preclinical rodent model of PD. HPG PD solutions were standardized for osmolality and compared directly to conventional glucose-based Physioneal™ PD solution (PYS). The efficacy of HPG solutions was measured by their ultrafiltration (UF) capacity, solute removal, and free water transport; biocompatibility was determined in vivo by the histological analysis of the peritoneal membrane and the cell count of detached peritoneal mesothelial cells (PMCs) and neutrophils, and in vitro cytotoxicity to cultured human PMCs. All the different sized HPGs induced higher UF and sodium removal over a sustained period of time (up to 8 h) compared to PYS. Urea removal was significantly higher for 1–3 kDa than PYS, and was similar for 0.5 kDa. Our analyses indicated that the peritoneal membrane exhibited more tolerance to the HPG solutions compared to PYS, evidenced by less submesothelial injury and neutrophil infiltration in vivo, and less cell death in cultured human peritoneal mesothelial cells. Free water transport analysis of HPG indicated that these molecules function as colloids and induce osmosis mainly through capillary small pores. We attribute the differences in the biocompatibility and osmotic activity of different sized HPGs to the differences in the polymer bound water measured by differential scanning calorimetry. These preclinical data indicate that compared to PYS, low MW HPGs (0.5–3 kDa) produces superior fluid and waste removal with better biocompatibility profile, suggesting that they are promising osmotic agents for PD.     

透明质酸在大鼠慢性腹膜透析模型中的作用

目的:了解透明质酸在大鼠慢性腹膜透析中的作用。方法:20只雄性SD大鼠随机分为3组。高糖组(n=8)和透明质酸组(n=6)每天分别腹腔内注射25mL的4.25%葡萄糖腹膜透析液或含0.01%透明质酸(分子量为1600kD)的4.25%葡萄糖腹膜透析液,共7d;正常对照组(n=6),不进行腹腔注射。第8d进行4h腹膜功能实验并于不同时点留取血和腹透液样品进行分析。结果:透明质酸组的腹腔内液容积(IVP)显著高于高糖组(P＜0.01)。高糖组腹腔液体吸收率(K_E)显著高于正常对照组和透明质酸组;高糖组和透明质酸组的淋巴液体吸收率(K_EB)差异无显著性,均显著高于对照组。高糖组的组织间质吸收率(K_ET)显著高于对照组和透明质酸组,后两者区别不明显。结论:在大鼠慢性腹膜透析模型中多次重复使用透明质酸,可以防止由于长期使用以葡萄糖为渗透压制剂的腹膜透析液所引起的腹腔吸收率增加,增加超滤量,保护腹膜功能。     

大肠杆菌脂多糖对腹膜间皮细胞己糖激酶活性的影响

目的:观察大肠杆菌脂多糖对原代培养的大鼠腹膜间皮细胞己糖激酶活性的影响;探讨其活性与葡萄糖净利用的关系,初步阐释腹膜透析相关性腹膜炎引起腹膜透析失超滤的机制。方法:利用胰蛋白酶消化法行大鼠腹膜间皮细胞的原代培养及传代，第2代细胞经鉴定及同步化后进入实验；用含不同浓度的大肠杆菌脂多糖（0、0.1、1.0、10、50、100 mg/L）刺激细胞 3 h，另用 10 mg/L 脂多糖分别刺激1、3、6、12、24 h，利用6-磷酸葡萄糖脱氢酶偶联比色法观察腹膜间皮细胞己糖激酶活性的变化；利用比色法观察细胞葡萄糖净利用量。结果:大肠杆菌脂多糖对大鼠腹膜间皮细胞己糖激酶活性的影响呈剂量和时间依赖性并伴有细胞葡萄糖净利用的增加。结论:大肠杆菌感染性腹膜炎时，其脂多糖通过影响腹膜间皮细胞己糖激酶的活性参与腹膜透析失超滤的过程。     

腹膜透析治疗老年慢性肾功能衰竭的临床分析

目的 分析持续性腹膜透析(PD)治疗老年慢性肾功能衰竭(CRF)患者的临床效果,探讨PD在老年CRF患者中的应用经验.方法 回顾性分析89例老年CRF患者接受PD(简称老年PD组)治疗后的临床表现和生化指标、主要并发症、存活率、死亡原因等,并与53例非老年PD组相比较.结果 老年PD组的原发疾病以糖尿病(37.08%)、原发性高血压(30.34%)为主,而非老年PD组则以肾小球肾炎(54.72%)为主.两组患者的1年与3年存活率差异无统计学意义,腹膜炎仍是两组患者退出PD的主要原因.老年组的病死率明显高于非老年组,感染与心脑血管病是两组患者的主要并发症和死亡原因.低钾血症在老年PD患者中非常普遍.结论 腹膜透析在老年CRF患者的治疗中有效,但长期存活仍受腹膜炎、心脑血管病并发症的影响,老年PD患者的低钾血症应引起足够的重视.     

腹膜透析治疗老年慢性肾功能衰竭的临床分析

目的分析持续性腹膜透析(PD)治疗老年慢性肾功能衰竭(CRF)患者的临床效果,探讨PD在老年CRF患者中的应用经验。方法回顾性分析89例老年CRF患者接受PD(简称老年PD组)治疗后的临床表现和生化指标、主要并发症、存活率、死亡原因等,并与53例非老年PD组相比较。结果老年PD组的原发疾病以糖尿病(37.08%)、原发性高血压(30.34%)为主,而非老年PD组则以肾小球肾炎(54.72%)为主。两组患者的1年与3年存活率差异无统计学意义,腹膜炎仍是两组患者退出PD的主要原因。老年组的病死率明显高于非老年组,感染与心脑血管病是两组患者的主要并发症和死亡原因。低钾血症在老年PD患者中非常普遍。结论腹膜透析在老年CRF患者的治疗中有效,但长期存活仍受腹膜炎、心脑血管病并发症的影响,老年PD患者的低钾血症应引起足够的重视。     

腹透液增强腹膜间皮细胞CD40表达及其意义

目的:研究腹膜透析液对大鼠腹膜间皮细胞CD40表达的影响及CD40活化后与细胞间粘附分子-1(ICAM-1)分泌的相关性, 以揭示腹膜透析时腹腔局部炎症的发生机制。方法: 分离、培养大鼠腹膜间皮细胞, 分别用IFN-γ、4.25%腹透液、4.25%腹透液+IFN-γ作为刺激因子, 通过逆转录-多聚酶链反应(RT-PCR)及流式细胞仪(FACS)检测间皮细胞CD40表达;并通过CD40单克隆抗体(CD40mAb)活化高表达的间皮细胞CD40, 用FACS检测间皮细胞ICAM-1的表达。结果:腹膜间皮细胞结构性表达低水平CD40mRNA及蛋白。用4.25%腹透液刺激后, 间皮细胞CD40mRNA及蛋白的表达显著高于对照组。用4.25%腹透液+IFN-γ刺激后, 间皮细胞表面CD40mRNA及蛋白的表达进一步增加, 且明显高于单纯腹透液刺激组。活化CD40受体可显著增强间皮细胞ICAM-1的表达。结论:腹膜间皮细胞可表达CD40, 4.25%透析液及IFN-γ均能显著增加腹膜间皮细胞CD40的表达。间皮细胞CD40的上调, 可能是腹透过程中腹腔局部炎症启动、放大的重要机制之一。     

预防性大网膜部分切除对腹膜透析导管功能障碍发生的影响

目的：观察预防性切除部分大网膜对腹膜透析导管功能障碍的发生以及术后腹腔出血、腹透液渗漏、腹膜透析相关性腹膜炎和透析效率的影响。方法：研究分为大网膜部分切除组和对照组。大网膜部分切除组患者于腹膜透析置管时,切除部分大网膜;对照组予常规开放式手术置管。比较2组术后导管功能障碍及其它并发症的发生率以及置管前及透析 3-6月后血肌酐(Scr)、尿素（urea）和二氧化碳总量(TCO2)差异。结果：大网膜部分切除组导管功能障碍发生率明显低于对照组（P<0.05）,2组血性透出液、腹透液渗漏、腹膜炎发生率无明显差异。置管前,观察组Scr（P<0.05）、urea（P<0.05）明显高于对照组,2组之间TCO2无明显差异;透析后2组Scr、urea、TCO2无明显差异;2组透析后Scr、urea、TCO2较腹膜透析前均明显改善。结论：预防性切除部分大网膜可明显降低导管功能障碍发生率,但不降低腹膜透析效率,也不增加术后腹腔出血、腹透液渗漏和腹膜透析相关性腹膜炎发生率。     

一种改良的尿毒症腹膜透析大鼠模型

 目的：研制一种改良的尿毒症大鼠腹膜透析模型。方法：采用二次手术法切除大鼠5/6肾脏建立尿毒症模型,造模后4周取尾静脉血测肌酐值,以达到正常血肌酐值2~3倍者为尿毒症大鼠模型;使用改良的硅胶腹透管置管制作腹膜透析大鼠模型,手术切口于背部,隧道出口于颈后两耳中点下方2~3 cm处。24只SD大鼠随机分为4组：A组(n=6),假手术组;B组(n=6),尿毒症组,不插管也不透析;C组(n=6),尿毒症插管组,插管但不透析;D组(n=6),尿毒症高糖透析组,4.25%葡萄糖透析液透析2周。D组大鼠停止透析24 h后各组行腹膜功能检查,测超滤量;透出液行白细胞计数;腹膜组织行HE染色,测量腹膜间皮至肌层厚度。结果：B、C、D组血肌酐值为A组的2~3倍,尿毒症大鼠模型制作成功;D组超滤量较A、B、C组显著减少（P<0.05）;B、C、D组透出液白细胞计数略高于A组（P<0.05）,但都未发现感染;HE染色显示D组腹膜间皮至肌层厚度较A、B和C组显著增加（P<0.05）,高糖透析液刺激腹膜组织呈现明显的慢性损伤病理特征。结论：我们通过改良的导管插入法建立了一个理想的尿毒症大鼠腹膜透析模型,该模型为研究腹膜透析功能保护奠定了实验基础。