Rho/ROCK信号通路与糖尿病肾病

糖尿病肾病(DN)是糖尿病最严重的微血管并发症之一,并已成为终末期肾脏病(ESRD)的重要原因,成为人类致死、致残的一个重要因素。因此,发现糖尿病肾病的新机制及关于此机制的新药研究,对改善糖尿病肾病预后非常重要。近年来,不断深入的研究提示,Rho/ROCK信号通路可能成为防治糖尿病肾病的药物新靶点。本文就Rho/ROCK信号通路与糖尿病肾病的关系作一综述。     

慢性间断性低氧大鼠认知功能和脑胆碱能神经元的进行性变化

目的：探讨慢性间断性低氧（CIH）大鼠认知功能的进行性变化及其与脑胆碱能神经元变化的关系。方法：成年雄性SD大鼠40只,随机均分为对照组、慢性间断性低氧1,3,5周组。应用Morris水迷宫检测认知功能的变化;利用HE染色在光镜下计数前额叶皮层和海马坏死神经元数;利用免疫组化方法检测前额叶皮层和海马胆碱乙酰转移酶（ChAT）阳性表达。结果：CIH各组大鼠学习记忆能力呈进行性下降趋势;与对照组比较,CIH5w组出现明显学习记忆功能障碍（P〈0.05）。CIH各组前额叶皮层和海马变性坏死神经元数增多,且随低氧时间延长,上述改变呈慢性进行性加重趋势。CIH各组前额叶皮层和海马ChAT阳性表达逐渐下降;与对照组比较,CIH3w组和CIH5w组前额叶皮层和海马ChAT阳性表达明显减少,差异具有显著性（P〈0.05）。结论：慢性间断性低氧大鼠认知功能进行性下降与前额叶皮层和海马神经元病理性损伤、ChAT表达进行性减少有关。     

lncRNA与慢性肾病关系的研究进展

近年来,慢性肾病(chronic kidney disease,CKD)的发病率和死亡率逐年升高,越来越引起人们的重视。其发病机制复杂,病情往往持续性进展,最终发展为终末期肾功能衰竭。lncRNA是长链非编码RNA,显示出高度组织特异性,在最近一些研究中暗示了其在疾病中的重要性,lncRNA可能成为人类疾病未来治疗靶点。本文综述lncRNA在慢性肾病中的研究进展,为科学研究和临床应用提供参考。     

IgA肾病患者肾小球系膜区沉积的IgA1来源的研究进展

IgA肾病(immunoglobulin A nephropathy,IgAN)危害极大,明确系膜区IgA1的来源对于阐明发病机制、探索新的诊疗手段具有重要意义。诸多研究报道显示IgAN患者血液中以及肾小球系膜区沉积的IgA是半乳糖缺失IgA1,目前比较公认IgAN是"免疫复合物引起的肾小球疾病"。有关IgAN患者IgA1的来源,传统观点是粘膜相关的淋巴组织中B淋巴细胞产生过多而"溢出"至血液,或者通过粘膜-骨髓轴引起骨髓浆细胞产生并分泌至循环血液。近年来非B细胞表达免疫球蛋白的报道,以及近期我们在体外原代肾小球系膜细胞中检测到IgA1的表达和分泌的初步研究,提示系膜细胞也可以产生IgA1,并可能参与了IgAN患者系膜区IgA1的沉积及IgAN的发病机制。本文我们将围绕IgAN患者IgA1的来源进行综述。     

单次尾静脉注射法阿霉素大鼠肾病模型的建立

目的建立阿霉素大鼠肾病模型,并观察模型的动态变化。方法 26只Wistar雄性大鼠随机分为模型组(13只)和空白组(13只),模型组单次尾静脉注射阿霉素6.2 mg/kg,空白组注射等容积生理盐水。检测连续10周12 h尿蛋白定量、终末血生化指标,光镜、电镜下观察各组大鼠肾脏病理改变。结果模型组12 h尿蛋白定量造模后1周与空白组差异有显著性(P0.01),第5周达到高峰;血总蛋白、白蛋白均低于空白组,甘油三酯、胆固醇、血尿素氮均高于空白组(均P0.05),血肌酐差异无显著性(P=0.64)。肾脏病理改变:第5周为微小病变型,第10周为局灶性节段性肾小球硬化。结论单次尾静脉注射6.2 mg/kg阿霉素,可以成功建立渐进性的大鼠肾病综合征模型。     

慢性移植性肾病大鼠脾脏中辅助性T细胞和B 细胞免疫状态的研究*

目的:检测慢性移植性肾病(CAN)大鼠脾脏中辅助性T细胞(Th)和B细胞特征性因子表达量的变化,探究这些Th/B细胞免疫状态在CAN病程中的作用。方法:采用Fischer-Lewis左肾原位移植法建立大鼠慢性移植性肾病模型,Lewis-Lewis同种自体移植作为对照组。所有受体大鼠,术后8周处死,取脾脏组织,进行HE染色,拍照后采用双盲法评价脾脏组织病理变化程度及淋巴细胞浸润情况。用Trizol法提取脾脏组织中总RNA,采用实时荧光定量聚合酶链式反应(q RT-PCR)法检测各组脾脏中Th1,Th2,Th17,Treg和B细胞标志性因子的表达情况。结果:与对照组相比,CAN大鼠脾脏出现明显的结构肿胀及淋巴细胞浸润增多,并且Th1细胞特征性因子IFN-γ和T-bet表达量显著增加(P0.001,P0.05);Th2细胞特征性因子GATA-3表达升高(P0.001),但IL-4无变化;IFN-γ/IL-4比例明显上调(P0.001),T-bet/GATA3比例没有显著差异。Th17的特征性因子IL-17未见明显改变,而Treg细胞特征性因子Foxp3表达增加(P0.001),IL-17/Foxp3平衡明显向Treg细胞偏移(P0.05)。B细胞激活相关因子TNFRSF13C和RAG1表达量均显著上调(P0.01,P0.05),而RAG2水平则没有变化。结论:CAN大鼠脾脏中Th1/Th2的活性平衡向Th1偏移,分化平衡未出现显著变化;Th17/Treg的平衡向Treg细胞偏移,B细胞免疫状态也被激活,这些变化在CAN病程的发展中起到了重要作用,并且为临床监测和治疗提供了新的依据。     

姜黄素对5/6肾结扎诱导的慢性肾病小鼠模型肾纤维化的保护作用

目的研究姜黄素对慢性肾病(CKD)肾纤维化的保护作用,并探讨可能作用机制。方法 30只C57BL/6 J小鼠随机分成假手术对照组(NC)、5/6肾结扎模型组(LIG)和姜黄素治疗组(LIG+CUR),每组10只,按照改良的5/6肾结扎方法制作CKD动物模型,姜黄素组小鼠给予含姜黄素饲料(每日100 mg/kg),其余组给予正常饲料,三月后牺牲小鼠,检测纤维化指标α-SMA及参与慢性肾病纤维化的Hippo通路转录激活子Yap。结果肾功能检测结果显示,与假手术组相比,结扎组中的尿素氮(BUN)、血肌酐(Scr)显著增高,给予姜黄素可以有效保护肾功能受损;H&E、Masson和免疫组化结果均显示,结扎组小鼠的肾出现明显肾小管病变和一定程度的纤维化改变,给予姜黄素后纤维化程度显著减轻(P<0.05);而Yap蛋白在造模后mRNA和蛋白水平均出现升高,姜黄素处理后则显著下降(P<0.05)。结论姜黄素可有效改善5/6肾结扎诱导的慢性肾病病症和减轻纤维化病变,机制研究结果初步提示可能与降低Hippo信号通路中的Yap表达有关。     

HIF-1α在大鼠造影剂肾病中的表达及对肾小管损伤的影响

目的:探讨HIF-1α在造影剂肾病大鼠血清中的表达水平及对肾小管损伤的影响。方法:将45只SD大鼠随机分为三组,每组15只。其中空白对照组(A组)大鼠禁食水12 h后,于尾静脉注射氯化钠注射液0.5 m L,共三次,每次间隔15 min。造影剂肾病组(B组)大鼠禁食水12 h后,于尾静脉以10 mg/kg注射吲哚美辛,15 min后以10 mg/kg注射左旋硝基精氨酸甲酯(L-NAME),15 min后再注射碘比醇(3 g I/kg)。阿托伐他汀钙组(C组)大鼠于实验前3 d开始喂食阿托伐他汀钙片,连续喂食3天,剂量为80mg/kg/d,再禁食水12 h后,制作造影剂肾病模型,步骤同造影剂肾病组。观察并比较三组大鼠肾功能指标(BUN、Cr)变化、HIF-1α表达水平及肾小管损伤情况。结果:造影剂肾病组大鼠BUN水平低于阿托伐他汀钙组和空白对照组,而Scr水平高于阿托伐他汀钙组和空白对照组,差异均具有统计学意义(P0.05)。阿托伐他汀钙组大鼠BUN水平低于空白对照组,而Scr水平高于空白对照组,差异具有统计学意义(P0.05)。造影剂肾病组大鼠肾小管损伤高于阿托伐他汀钙组和空白对照组,阿托伐他汀钙组大鼠肾小管损伤高于空白对照组,差异均具有统计学意义(P0.05)。造影剂肾病组大鼠HIF-1α表达高于阿托伐他汀钙组和空白对照组,阿托伐他汀钙组大鼠HIF-1α表达高于空白对照组,差异均具有统计学意义(P0.05)。结论:造影剂肾病发生时存在缺氧情况,阿托伐他汀钙在造影剂肾病中具有保护作用。     

高热量高蛋白饮食诱导GK大鼠糖尿病肾病模型的建立

目的运用高热量高蛋白饮食诱导GK大鼠2型糖尿病肾病模型的建立,并探讨其可能的作用机制。方法 28周龄GK大鼠24只,随机分成对照组、模型组,每组各12只,模型组给予高热量高蛋白饮食,对照组给予正常饮食,共8周。于第0、4、8周观察24 h尿微量白蛋白、24 h尿蛋白、尿肌酐、尿微量白蛋白/尿肌酐比值水平;于第0、8周观察空腹血糖和血清肌酐、尿素氮、总胆固醇、甘油三脂、一氧化氮水平;实验结束时取双肾称重并计算肾肥大指数,取肾组织观察病理形态学变化,检测肾组织钠钾ATP酶活性。结果与对照组比,模型组大鼠24 h尿微量白蛋白、24 h尿蛋白、尿微量白蛋白/尿肌酐比值、空腹血糖、总胆固醇、甘油三脂、一氧化氮、肾肥大指数水平和肾组织钠钾ATP酶活性显著提高,模型组肾小球体积增大,系膜基质增生,基底膜增厚明显。结论运用高热量高蛋白饮食诱导GK大鼠可成功建立2型糖尿病肾病模型。血糖血脂的上升是糖尿病肾病形成的重要因素,同时钠钾ATP酶活性增强进一步损伤肾小管功能,一氧化氮升高促使肾小球高灌注、高滤过,也是加速GK大鼠肾病形成的原因。     

慢性充血性心力衰竭大鼠模型的建立

目的:探讨建立慢性充血性心力衰竭大鼠模型的方法。方法:采用腹主动脉缩窄法制备CHF大鼠模型。于右肾动脉分支处上方缩窄腹主动脉至0.6mm,同时设立假手术组及正常对照组。12周后行一般情况观察,超声心动图及血液动力学检测心脏结构和功能、计算心脏肥厚指数,HE及Masson染色检测心肌病理改变,综合评价CHF大鼠心功能。结果:与假手术及正常对照组比较,CHF模型组术后12周均出现进食减少、精神萎靡、少动、被毛无光;IVS、LVPW、LVM、LVEDD、LVESD、SV显著增加,EF、FS显著降低;HR显著增加;VSP、+dP/dtmax显著降低,T-dP/dtmax显著延长;LVEDP显著升高,-dP/dtmax绝对值显著降低;心脏湿重及心脏肥厚指数均增加;病理结果表明CHF模型组心肌细胞肥大、排列紊乱、心肌胶原纤维增多。结论:应用腹主动脉缩窄术可成功制备CHF大鼠模型。     

Facility for chronic exposure of rats to ELF magnetic fields

The facility consists of a 12 × 11.5 × 2.4 m high room containing six sets of exposure apparatus and the other equipment necessary to maintain a pathogen-free system. The apparatus sets produced 5 mT (rms), 0.5 mT, or a sham exposure. The apparatus was arranged in the room to minimize the fringing field of the 5 mT set at the sham position. Each set was 3.85 × 1.80 × 0.66 m in outside dimension, containing 24 cages in the magnetically homogeneous region. The apparatus was designed using Harvey's figure-eight-configuration and generated a horizontal sinusoidal alternating field. In order to save electric power, the coil of the apparatus constituted a 50 Hz LC resonance circuit with a condensor bank to which electric power was supplied to compensate losses. Magnetic flux density was kept constant by controlling the coil current. Although mild steel was used in the skeleton of the building, the fringing flux at the sham was as low as 0.1 to 1 μT. Stainless steel was used for ventilating ducts, racks for the cages, cage covers, feeder baskets, and watering nozzles. The homogeneity of the field was measured to be ± 10% in the animal residence area, and food and water consumption was found to be unaffected by the field. At 5 mT, the coil current was 370 A, and the hollow coil was cooled by a stream of 20°C water to prevent both heat and dew on the coil surface. Vibration and acoustic noise was prevented by fiber reinforced plastic framework of the coil. High harmonic distortion was not observed at the output terminal of the coil driver. The facility has operated without trouble for 2 years. © 1993 Wiley-Liss, Inc.     

广东虫草对腺嘌呤诱导的慢性肾衰竭大鼠的治疗作用

广东虫草Cordyceps guangdongensis是广东省微生物研究所近年发现并成功驯化的独有新品种,前期研究发现其具有抗氧化、抗疲劳、延缓衰老等药理作用,但其对慢性肾衰竭的治疗作用还未见报道。实验采用饲喂腺嘌呤诱导大鼠慢性肾衰竭（CRF）模型,观察比较了阴性对照组、阳性对照组、广东虫草子实体低、中、高剂量组CRF模型大鼠及正常健康大鼠对照组的血尿素氮、肌酐、24h尿量、尿蛋白量,以及肾组织病理变化。结果表明广东虫草子实体组能显著降低CRF大鼠血尿素氮和肌酐,能促进机体生成白蛋白和总蛋白,改善肾功能衰竭大鼠的临床症状及肾脏水肿、变大和病变程度。由此证明广东虫草子实体对大鼠的慢性肾衰竭有明显的治疗作用。     

Expression of Fc alpha/mu receptor by human mesangial cells: a candidate receptor for immune complex deposition in IgA nephropathy.

IgA nephropathy is characterized by the deposition of IgA immune complexes in the glomerular mesangium, but the mechanisms responsible for this are not well understood. Human mesangial cells (HMCs) can bind IgA but do not express known IgA receptors. We show here that primary HMCs express mRNA for a novel receptor, the Fc alpha/mu receptor (Fcalpha/muR), and that receptor expression is upregulated by IL-1. We also detected mRNA for a novel receptor variant in HMCs that may encode a soluble form of the receptor. Fcalpha/muR was expressed in a heterologous system which showed that the receptor was approximately 58 kDa in weight and was only minimally N-glycosylated. As predicted from the characteristics of the murine homologue, the expressed human Fcalpha/muR was able to bind IgA and IgM, but not IgG. These results suggest that Fcalpha/muR may be the receptor responsible for mesangial IgA deposition in IgA nephropathy.     

Augmenter of liver regeneration ameliorates renal fibrosis in rats with obstructive nephropathy

Renal fibrosis is a hallmark in CKD (chronic kidney disease) and is strongly correlated to the deterioration of renal function that is characterized by tubulointerstitial fibrosis, tubular atrophy, glomerulosclerosis and disruption of the normal architecture of the kidney. ALR (augmenter of liver regeneration) is a growth factor with biological functions similar to those of HGF (hepatocyte growth factor). In this study, our results indicate that endogenous ALR is involved in the pathological progression of renal fibrosis in UUO (unilateral ureteral obstruction) rat model. Moreover, we find that administration of rhALR (recombinant human ALR) significantly alleviates renal interstitial fibrosis and reduces renal-fibrosis-related proteins in UUO rats. Further investigation reveals that rhALR suppresses the up-regulated expression of TGF-β1 (transforming growth factor β1) induced by UUO operation in the obstructed kidney, and inhibits Smad2 and Smad3 phosphorylation activated by the UUO-induced injury in the animal model. Therefore we suggest that ALR is involved in the progression of renal fibrosis and administration of rhALR protects the kidney against renal fibrosis by inhibition of TGF-β/Smad activity.     

敦煌疗风虚瘦弱方对大鼠慢性心衰的治疗作用及其机制

目的：观察敦煌疗风虚瘦弱方（LXD）对慢性心衰大鼠的治疗作用及机制。方法：48只Wistar雄性大鼠随机分为6组（n=8）：正常组、模型组、卡托普利组和LXD高、中、低剂量组。除正常组外,其余各组大鼠尾静脉注射阿霉素（2.5 mg/kg）,每周1次,连续6周复制慢性心衰模型,末次注射阿霉素60 min后,卡托普利组灌胃卡托普利,LXD高、中、低剂量组分别以灌胃LXD水煎液80、40、20 g/kg （生药/体重）,正常组和模型组灌胃等容量生理盐水,每天1次,连续6周,同时每周腹腔注射15 mg/kg异丙肾上腺素1次。记录各组大鼠的呼吸、皮毛颜色、活动、体重、力竭游泳时间、心率（HR）、平均动脉压（MAP）、左室收缩压（LVSP）、左室舒张末压（LVEDP）、左室内压最大上升速率（+dp/dtmax）,左室内压最大下降速率（-dp/dtmax）,检测血清白介素-6（IL-6）及肿瘤坏死因子-α（TNF-α）水平,评价抗氧化活性和心室质量指数,光镜下观察心肌组织形态变化。结果：与正常组比较,模型大鼠体重增长减慢,力竭游泳时间明显缩短,心脏功能减弱,出现细胞因子激活、自由基损伤、心肌酶释放增多,HE染色显示心肌组织形态出现损伤,心肌纤维断裂等。与模型组比较,各剂量LXD对模型大鼠有不同程度的治疗作用,给药第42天,LXD各剂量组体重明显增长;LXD高、中剂量组力竭游泳时间明显延长（P<0.05）,MAP、血清IL-6、TNF-α、MDA、LVMI和RVMI显著降低（P<0.05,P<0.01）,LVSP、+dp/dtmax及-dp/dtmax均显著升高（P<0.05,P<0.01）,LXD高剂量组LVEDP显著降低（P<0.01）,LXD中、低剂量组CK和AST显著降低（P<0.05,P<0.01）;LXD各剂量组血清SOD升高。结论：敦煌疗风虚瘦弱方对阿霉素诱导的慢性心衰模型大鼠具有一定的治疗作用,作用机制与改善血流动力学,降低心肌组织损伤有关。     

Downregulation of miR‑214-3p attenuates mesangial hypercellularity by targeting PTEN‑mediated JNK/c-Jun signaling in IgA nephropathy

Mesangial cell (MC) proliferation and matrix expansion are basic pathological characteristics of IgA nephropathy (IgAN). However, the stepwise mechanism of MC proliferation and the exact set of related signaling molecules remain largely unclear. In this study, we found a significant upregulation of miR-214-3p in the renal cortex of IgAN mice by miRNA sequencing. In situ hybridization analysis showed that miR-214-3p expression was obviously elevated in MCs in the renal cortex in IgAN. Functionally, knockdown of miR-214-3p alleviated mesangial hypercellularity and renal lesions in IgAN mice. In vitro, the inhibition of miR-214-3p suppressed MC proliferation and arrested G1-S cell cycle pSrogression in IgAN. Mechanistically, a luciferase reporter assay verified PTEN as a direct target of miR-214-3p. Downregulation of miR-214-3p increased PTEN expression and reduced p-JNK and p-c-Jun levels, thereby inhibiting MC proliferation and ameliorating renal lesions in IgAN. Moreover, these changes could be attenuated by co-transfection with PTEN siRNA. Collectively, these results illustrated that miR-214-3p accelerated MC proliferation in IgAN by directly targeting PTEN to modulate JNK/c-Jun signaling. Therefore, miR-214-3p may represent a novel therapeutic target for IgAN.     

丁苯酞对慢性酒精中毒大鼠海马、杏仁核H2S/CBS通路及学习记忆能力的影响*

目的: 探讨丁苯酞(NBP)对慢性酒精中毒大鼠学习与记忆相关的能力、海马及杏仁核内硫化氢(H₂S)含量、胱硫醚-β-合成酶(CBS)表达和线粒体ATP酶活性的影响。方法: 随机将90只SD雄性大鼠均分为3组:空白对照组(NC)、模型组(M)和治疗组。除对照组外,其余两组均将含6%(v/v)酒精的水溶液作为唯一饮水来源。喂养14d后,治疗组按5mg/Kg比例腹腔注射NBP,每日一次,连续14d,其余两组注射等剂量生理盐水;对照组随后使用Morris水迷宫法,通过观察和记录动物入水后搜索藏在水下平台所需时间、采用策略和它们的游泳轨迹,从而可分析和推断动物的学习、记忆等方面的能力,并检测海马和杏仁核组织中H₂S浓度、CBS表达及线粒体ATP酶活性。结果: 与正常对照组大鼠相比,模型组大鼠第2-4日潜伏期、游泳距离均增加,海马及杏仁核内H₂S含量、CBS平均光密度值均升高,海马以及杏仁核组织中线粒体ATP酶活性均降低,且均有极显著性差异(P＜0.01)。与模型组大鼠相比较,NBP治疗组大鼠学习记忆成绩第2-4日潜伏期、游泳距离减小,海马及杏仁核H₂S含量、CBS平均光密度值均降低,海马以及杏仁核组织中线粒体ATP酶活性均提高,且均有极显著性差异(P＜0.01)。结论: NBP能够减轻酒精对大鼠的学习与记忆能力的影响,可能与NBP影响海马、杏仁核内H₂S浓度和CBS的表达以及线粒体ATP酶活性有关。