雷公藤多甙对大鼠肾移植急性排异反应治疗作用的观察

利用大鼠肾移植模型,观察雷公藤多甙(GTW)、雷公藤甲素及小剂量环孢霉素A(CsA)对大鼠肾移植急性排异反应的影响。结果证实,应用GTW或CsA后大鼠移植肾肾长明显减少,肾重明显减轻,肾小球基底膜增厚,肾小管空泡变性、坏死及间质水肿明显减轻;利用微机肾组织切片定量分析示移植术后第四、八天,肾小管面积、数量也明量增加,而间质面积明显减少。雷公藤甲素来证实有上述作用.另外,观察应用GTW治疗者,移植术后第九～第11天存活率明显增加.     

肾移植患者肾组织和尿中巨细胞病毒DNA的动态观察及其临床意义

肾移植患者CMV感染对排异反应的影响认识不一,国内目前尚无资料可考.本文采用原位杂交及多聚酶链反应技术,动态观察了8例肾移植患者移植前供肾、移植后肾活检组织及肾活检当日尿标本中CMV-DNA的检出状况,结合肾组织病理形态学分析,发现①正常肾组织即有少数CMV感染;②肾组织CMV感染率随移植时间延长而增加;③CMV感染与移植肾慢性排异有较为密切的关系,可能是造成慢性排异反应发生的因素之一。     

低剂量免疫抑制剂对肾移植受者淋巴细胞表型的影响

目的探讨肾移植长期低剂量免疫抑制维持用药受者的免疫学特征.方法采用多种单克隆抗体,以流式细胞仪为工具,对存活3年以上,长期低剂量免疫抑制维持用药受者、常规剂量维持用药受者和慢性移植肾失功受者的外周血淋巴细胞表型进行检测,分析细胞表型与临床用药量和移植效果的关系.结果低剂量受者CD95、NK及HLA-DR阳性细胞数目与正常人接近,而在常规剂量组和移植肾失功能组中,CD95明显升高,NK细胞明显减少(P＜0.05).结论CD95、NK及HLA-DR水平有可能用于评价长期移植受者的免疫状态,并作为调整免疫抑制剂用量的参考指标.     

大黄酸对慢性移植肾肾病大鼠转化生长因子β1、结缔组织生长因子和纤维连接蛋白的影响

目的:探讨中药大黄酸对大鼠慢性移植肾肾病(CAN)模型肾功能、组织学、转化生长因子β1(TGF-β1)、结缔组织生长因子(CTGF)和纤维连接蛋白(FN)表达的影响.方法:F344近交系大鼠为供体、Lewis近交系大鼠为受体,左肾原位肾移植,建立30只CAN大鼠模型.将30只移植大鼠分成模型组(16只)和大黄酸治疗组(14只);在移植后1月、2月及4月分别收集大鼠的血、尿标本,检测肾功能及尿蛋白定量.移植后2月及4月分别宰杀大鼠,观察各组大鼠移植肾组织学变化及大黄酸对模型动物TGF-β1、CTGF和FN表达的影响.另取5只Lewis大鼠作对照组,行右肾切除,术后4月末行相应指标检测.结果:与模型组比较(1)大黄酸治疗组大鼠肾功能改善;肾组织学示肾间质纤维化及间质炎症明显改善;(2)大黄酸治疗组大鼠肾组织CTGF表达降低,细胞外基质FN明显降低.结论:大黄酸能改善CAN大鼠肾功能及肾组织慢性病变,降低CTGF表达,减轻肾脏纤维化.     

多瘤病毒相关性肾病患者肾组织浸润细胞的变化及意义

目的:观察肾移植术后多瘤病毒相关性肾病(BKVAN)患者肾组织浸润细胞的变化,探讨其临床意义。方法:28例BKVAN患者作为研究对象(BKVAN组),选取26例急性排斥反应和19例常规活检患者作为对照组。比较BKVAN组与对照组肾组织中CD4~+、CD8~+、CD68~+、CD20~+细胞,白细胞介素2受体(IL-2R)的阳性表达及肾小管上皮细胞HLA-DR的阳性表达情况;根据血清肌酐(SCr)水平将BKVAN患者分为A1组11例(109.62SCr≤219.23μmol/L)、A2组11例(219.23SCr≤328.85μmol/L)和A3组6例(SCr328.85μmol/L),观察不同SCr水平BKVAN患者移植肾组织中上述指标的变化。结果:BKVAN肾组织中CD4~+、CD8~+、CD68~+、CD20~+细胞、IL-2R的阳性表达较常规活检组均明显增加(P0.01),而肾小管上皮细胞HLA-DR的阳性表达增加不明显;BKVAN组与急性排斥反应组相比,前者肾组织中B淋巴细胞浸润数量增加,但统计学差别不明显,而急性排斥反应组CD8~+细胞和肾小管上皮细胞HLA-DR的阳性表达显著增加(P0.01)。随着SCr水平的升高,BKVAN患者移植肾组织中CD68~+细胞、IL-2R的阳性表达增加,其中A3组与A1组相比CD68~+细胞的浸润具有显著统计学差别(P0.01)。结论:BKV诱发肾组织T淋巴细胞和B淋巴细胞活化,加重移植肾损害;与急性排斥相比BKVAN组受者肾组织中CD20~+细胞的浸润数量更多、肾小管上皮细胞HLA-DR阳性表达和CD8~+细胞明显减少。     

急性排异及其浸润细胞的研究进展

单克隆抗体技术的发展及其广泛应用，使人们对移植肾急性排异时肾组织内细胞浸润的成分有了更进一步的认识。新近研究的焦点集中在肾组织内浸润细胞的类型，如CD4”、CDS”细胞亚群和肾实质性细胞主要组织相容性复合物（MHC）抗原的表达等。内皮细胞和移植肾粘附分子表达的改变也已成为目前研究的重要领域。研究移植肾浸润细胞成分的目的，是寻找浸润细胞表面上某种新标记，特别是寻找与细胞功能相关的标记。本文主要综述肾移植急性排异时，肾组织内不同浸润细胞的免疫学特性及其在临床上的应用价值。是急性排异反应时主要细胞和分子的…     

自然杀伤细胞抑制肝癌肺转移

目的研究自然杀伤(natural killer,NK)细胞对肝癌的抑制作用,为临床应用提供实验依据.方法从人外周血分离培养及鉴定NK细胞.在体外,研究NK细胞抑制肝癌细胞的增殖、迁徙、转移.在体内,检测NK细胞在裸鼠肝脏存活情况.利用人肝癌组织裸鼠肝脏原位移植模型来评估NK细胞在体内对肝癌生长、转移的抑制功能.通过检测NK细胞活化受体、NKB1、穿孔素和颗粒酶的表达情况来评估白介素(interleukin,IL)-2对NK细胞免疫功能的刺激作用.结果采用密度梯度法可以获取较大量的外周血单个核细胞,且能够从中分离到高活力的NK细胞.NK细胞经IL-2激活后活力增高,成簇悬浮繁殖、扩增、生长.在体外,NK细胞可抑制肝癌细胞的增殖、迁移和侵袭.在体内,NK细胞在裸鼠肝脏可长期存活;NK细胞可明显抑制裸鼠肝癌肺转移.然而,NK细胞对肝脏肿瘤生长抑制不明显.IL-2可诱导NK细胞免疫相关分子的表达并提高其肿瘤抑制功能.结论NK细胞的免疫学功能可被IL-2活化从而抑制肝癌的转移.     

辣椒素对大鼠糖尿病肾病的影响及其作用机制

目的 研究辣椒素( Capsaicin)对大鼠糖尿病肾病(DN)的影响.方法 30只清洁级雄性SD大鼠随机分为正常对照组(NC组),糖尿病肾病组(DN组),辣椒素组(Capsaicin组),每日监测大鼠血糖和糖化血红蛋白(HbA1c),8 w后,检测尿微量白蛋白(MAU)的变化,心脏采血后处死动物,观察大鼠肾组织病理变化,检测大鼠肾组织中细胞凋亡率.结果 小剂量Capsaicin腹腔注射能明显降低糖尿病肾病大鼠的血糖和HbA1c,降低尿微量白蛋白,改善肾组织病理损伤,使细胞凋亡率减少.结论 Capsaicin能改善大鼠DN,与降低血糖,降低蛋白尿,减轻肾组织损伤和抑制细胞凋亡有关.     

丹参对肾移植大鼠慢性排斥移植肾病理变化及其TGF-β1表达的影响

目的观察丹参对肾移植大鼠慢性排斥移植肾组织病理变化和转化生长因子-β1（TGF-β1）表达的影响。方法制作SD-W istar大鼠肾移植慢性排斥模型,完整保留受体右肾作为每个移植肾的内对照。选取移植成功受体15只,随机分成治疗组8只与对照组7只。治疗组受体大鼠自术后10 d起每日腹腔注射丹参注射液80 mg/kg至术后12周;对照组给予相同容量的生理盐水腹腔注射。术后12周取受体大鼠移植肾组织行组织病理学检查,并用免疫组化染色法检测移植肾中的TGF-β1。结果移植后12周,两组受体移植肾均存活,体积较正常右肾略小,色泽较苍白,出现不同程度的单个核细胞浸润、肾小球硬化、肾小管萎缩、间质纤维化和小动脉内膜纤维性增厚等慢性排斥组织病理学改变。治疗组大鼠移植肾组织的病理改变Banff评分（6.40±1.04）分,明显低于对照组的（7.87±0.55）分,P〈0.01。TGF-β1主要表达于肾小管和间质细胞。治疗组肾组织的TGF-β1表达强度为6.55±0.95,明显低于对照组的7.74±0.97,P〈0.05。结论丹参能够减轻大鼠肾移植慢性排斥移植肾组织病理学损害,下调移植肾组织TGF-β1的表达。     

低剂量免疫抑制对肾移植受者淋巴细胞表型的影响

目的：探讨肾移植长期低剂量免疫抑制维持用药受者的免疫学特征。方法：采用多种单克隆抗体,以流式细胞仪为工具,对存活3年以上,长期低剂量免疫抑制维持用药受者,常规剂量维持用药受者和慢性移植肾失功受者的外周血淋巴细胞表型进行检测,分析细胞表型与临床用药量和移植效果的关系。结果：低剂量受者CD95,NK及HLA－DR阳性细胞数目与正常人接近,而在常规剂量组和移植肾失功能组中,CD95明显升高,NK细胞明显减少（P＜0．05）。结论：CD95,NK及HLA－DR水平有可能用于评价长期移植受者的免疫状态,并作为调整免疫抑制剂用量的参考指标。     

Acute rejection of kidney transplant is associated with induction of replicative senescence in CD8+ T lymphocytes

Acute renal rejection repeatedly activates immunocompromised CD8 + T cells. Maintained activation of CD8 + T cells can induce a process of replicative senescence. In the present study, we will evaluate in CD8 lymphocytes from patients undergoing acute renal rejection characteristics of replicative senescence such as: a) low expression of CD28 molecule; b) telomere shortening and c) increase production of proinflammatory cytokines. The study was carried out in CD8 + T cells from 14 patients transplanted without clinical evidences of acute renal rejection, 14 patients kidney transplanted with clinical and anatomopathological evidences of acute renal rejection, 8 healthy controls. The results shown that in peripheral blood and renal biopsy of patients with acute renal rejection there is a significant increment of the population of T cells CD28-CD8+, with short telomere length, as compared with healthy controls and patients without acute renal rejection. The presence of senescent cells was associated with high levels of IL-10 and IFN-Y in plasma and urine. In conclusion our study suggest that the CD8 + T cells of patients with acute renal rejection suffer a process of replicative senescence.     

Effects of alkaloid sinomenine on levels of IFN-γ, IL-1β, TNF-α and IL-6 in a rat renal allograft model

Aims: To evaluate the immunosuppressive efficacy of alkaloid sinomenine (SIN) and the synergistic effects in combination with cyclosporin A (CsA) in acute rejection after rat renal allograft. Materials & methods: Animals were treated with saline in group 1, SIN (30 mg/kg/d) in group 2, CsA (2.5 mg/kg/d) in group 3 and SIN (30 mg/kg/d) + CsA(2.5 mg/kg/d) in group 4. Another 12 syngeneic renal transplantation animals were treated with saline as control. Survival time is observed. The levels of serum creatinine (Scr) and blood urea nitrogen (Bun) were detected; the secretion of IFN-γ, IL-1β, TNF-α and IL-6 were detected by ELISA. The kidneys were fixed to perform histological staining. Results: The mean survival time was 8.00 ± 2.10 days in group 1, 10.67 ± 1.21 days in group 2, 11.00 ± 1.41 days in group 3 and 19.67 ± 2.80 days in group 4, while all the recipients survived more than 180 days in the control group. The 24-h urinary volume and urinary time of the other three groups were increased significantly compared with group 1. The levels of Scr and Bun, levels of IFN-γ, IL-1β, TNF-α and IL-6 were significantly higher in group 1 than that in the other three groups; there were significant differences between group 4 and group 2 or 3. Conclusion: Our study showed that SIN had immunosupression effects in rat renal allograft models, it also had a synergistic effect in combination with CsA, which provided a new immunosuppressant for clinical application.     

大鼠原位肝移植后血清、肝组织NO含量和TNF变化的意义

探讨一氧化氮（NO）在BN至LEW鼠原位肝移植急性排斥反应中的诊断作用。我们应用BN至LEW鼠肝移植急性排斥反应及同基因肝移植动物模型，检测肝移植术后血清NO和肿瘤坏死因子的变化。大鼠原位肝移植急性排斥反应时血浆中NO明显升高，其变化早于TNF，FK506可显著抑制NO的合成，氨基胍可减轻排斥反应的程度。同基因组NO不升高。NO可作为大鼠肝移植急性排斥反应的诊断指标。抑制大鼠肝移植发生急性排斥反应NO的升高，可延长移植物的存活时间。     

CXCR3阻断剂Genistein对大鼠胰腺移植后排斥反应的影响

目的 探讨Genistein对大鼠移植胰腺急性排斥反应的影响以及作用机制.方法 以Wistar和SD大鼠为供体,SD大鼠为受体,建立胰腺移植模型.分成同种系移植组、异种系移植组和Genistein治疗组(在异种系移植后每天腹腔注射Genistein 10 mg/kg体重,共7 d),每组lO只.术后7 d切除移植物行病理学观察及CXCR3免疫组化染色,抽血检测血清IFN-γ、IL-2浓度.结果 同种系移植组移植物无急性排斥反应,无CXCR3表达.血清IFN-γ、IL-2浓度分别为(2.76±0.66)pg/ml和(11.83±1.86)pg/ml;异种系移植组的移植胰腺发生强烈的镜下排斥反应,CXCR3强阳性表达,血清IFN-γ、IL-2浓度分别为(32.64±2.52)PS/ml和(29.59±1.71)PS/ml,较同种系移植组显著升高(P<0.05);Genistein治疗组的移植胰腺排斥反应减轻,CXCR3弱阳性表达,血清IFN-γ、IL-2浓度分别为(15.94±1.95)Pg/ml和(22.62±1.76)ps/ml,较异种系移植组显著降低(P<0.05),但高于同种系移植组(P<0.05).结论 Genistein通过阻断CXCR3的表达,可有效减轻胰腺移植后急性排斥反应.     

Adenovirus mediated CTLA4Ig gene inhibits infiltration of immune cells and cell apoptosis in rats after liver transplantation

AIM: To investigate the role of adenovirus-mediated CTLA4Ig gene therapy in inhibiting the infiltration of macrophages and CD8+T cells and cell apoptosis after liver transplantation. METHODS: The rat orthotopic liver transplantation model was applied. The rats were divided into three groups: group I: rejection control (SD-to-Wistar); group II: acute rejection treated with intramuscular injection of CsA 3.0 mg/(kg·d) for 12 d (SD-to-Wistar+CsA); groupIII: injection of 1×109 PFU adenovirus-mediated CTLA4Ig gene liquor in dorsal vein of penis 7 d before liver transplantation (SD-to-Wistar+CTLA4Ig). Immunohistochemistry and transferase-mediated dUTP nick-end labeling (TUNEL) were used to analyze the expression of CTLA4Ig gene in liver, infiltration of macrophages and CD8+T cells, cell apoptosis in grafts at different time-points after liver transplantation. Histopathological examination was done. RESULTS: CTLA4Ig gene expression was positive in liver on d 7 after administering adenovirus-mediated CTLA4Ig gene via vein, and remained positive until day 60 after liver transplantation. Infiltration of macrophages and CD8+T cells in CTLA4Ig-treated group was less than in rejection control group and CsA-treated group. The apoptotic index of rejection group on d 3, 5, and 7 were significantly higher than that of CTLA4Ig-treated group. A good correlation was found between severity of rejection reaction and infiltration of immune activator cells or cell apoptotic index in grafts. CONCLUSION: CTLA4Ig gene is constantly expressed in liver and plays an important role in inducing immune tolerance.     

无创正压通气对急性加重期 COPD 患者痰液及血液炎性因子水平的影响

目的：观察无创正压通气（ NPPV）对慢性阻塞性肺疾病急性加重期（ AECOPD ）患者痰液及血液中炎性因子水平的影响,探讨NPPV治疗和机制。方法将30例AECOPD患者随机分为NPPV组和对照组各15例,两组均予常规AECOPD治疗药物,在此基础上NPPV组辅助NPPV治疗。分别于治疗前和治疗72 h后评价两组医学研究会呼吸困难量表（ MRC）评分、动脉血气指标,收集痰及空腹静脉血标本测定IL-6、IL-8、肿瘤坏死因子（ TNF）-α含量。结果两组治疗后呼吸困难均缓解、动脉血气指标均改善,尤以NPPV组为著（P均＜0．05）;两组治疗后痰液及血液中IL-6、IL-8、TNF-α水平均较治疗前下降,尤以NPPV组为著（ P均＜0．05）。结论 NPPV治疗后AE-COPD患者痰液及血液中炎性因子水平下调,此可能为NPPV治疗AECOPD的机制之一。     

The role of serum C-reactive protein in acute ischemicreperfusion injury of kidney

ObjectiveBlocking early C-reactive protein-mediated inflammatory reaction may have therapeutic implications in improving the prognosis of acute renal failure with severe ischemic-reperfusion injury. Therefore, the role of serum C-reactive protein in acute renal ischemic-reperfusion injury was investigated.MethodsFourteen New Zealand albino rabbits were selected and divided into a treated and a control group at random. An acute renal ischemia-reperfusion injury model was induced by clamping the right renal artery for 45 minutes with simultaneous contralateral nephrectomy, followed by right renal reperfusion. The treated group was injected with dexamethasone (1 mg/kg) 2 minutes before renal reperfusion. Serum C-reactive protein, blood urea nitrogen, creatinine, and urine volume were recorded at designed time phases in both groups. Data were expressed as mean ± standard deviation and analyzed using the Student's t test.ResultsIn the control group, there was a steady increase of serum C-reactive protein that reached its peak at 6-hour reperfusion, and a positive correlation between C-reactive protein and blood urea nitrogen and creatinine (r = 0.62 and 0.53, respectively); there was a negative correlation between C-reactive protein and urine volume (r = −0.52). Compared with the control group, C-reactive protein values in the treated group remained mainly in the baseline levels after reperfusion, with C-reactive protein peaking at 4-hour reperfusion (p<0.01), whereas urine volume increased significantly (p<0.01).ConclusionThis study indicates that C-reactive protein is involved in the pathogenesis of acute renal ischemic-reperfusion injury; blocking early C-reactive protein-mediated inflammatory reaction may have therapeutic implications in improving the prognosis of acute renal failure with severe ischemic-reperfusion injury.     

IL-10对实验性肝纤维化大鼠转化生长因子β-表达的影响

目的:探讨TGFβ_1在实验性肝纤维化过程中的表达状况及IL-10对肝纤维化大鼠转化生长因子β_1(TGFβ_1)表达的影响。方法:建立大鼠肝纤维化模型并行IL-10干预实验。从正常对照组(C组)和CCl_4诱导肝纤维化模型组(M组)及IL-10干预肝纤维化组(T组)中取肝脏组织,采用S-P免疫组织化学方法检测分析不同组大鼠在肝纤维化进程的不同阶段肝组织中TGFβ_1表达的情况。结果:成功建立大鼠肝纤维化模型;随着肝纤维化程度的加重,TGFβ_1在肝组织中阳性表达明显增强;经Ridit分析,C组与M组间TGFβ_1阳性表达水平有显著性差异(P<0.01);T组TGFβ_1阳性表达较M组明显减弱,经Ridit分析,组间差异有显著性(P<0.01)。结论:TGFβ_1的阳性表达随着肝纤维化程度的进展升高,外源性IL-10对CCl_4诱导的肝纤维化中TGFβ_1的表达具有明显拮抗作用。