研究糖尿病大鼠肾肘蛋白激酶（PK）Cα亚型蛋白表达和mRNA水平均的变化。方法SD大鼠随机分为正常对照和糖尿病组，成模5周后分别采用PAS染色、免疫组化、PT-PCR、Westem blot方法检测单个肾小球面积，肾皮质PKCα的定位和定量以及PKCαmRNA的水平。结果同正常相比，糖尿病组大鼠单个肾小球面积增加，但细胞数并无明显变化；肾皮质PKCα的表达明显增强，并且有自胞浆向胞膜的转位；而肾质中PKCαmRNA的表达也增加了1.67倍。结论早期糖尿病大鼠肾脏中PKCα亚型表达和转录水平上调，提示PKCα可能与糖尿病肾病的发展有关。

研究血管紧张素I型受体（ATI）反义（AS）寡枋苷梭（ODN）对大鼠肾脏局中肾素-血管紧张素系统（RAS）的阻断作用。方法采用单侧肾动脉注射+原位电穿孔的方法，ATI AS-ODN导入到Wistar大鼠的一侧肾脏，观察以FITC荧光标记的ODN在肾脏的转移位置，以ATI放射自显影定量分析的方法观察转移基因的时间-效应曲线。在基因转移3d后以RT-PCR、Western印迹及饮疫组织化学方法检测肾脏ATI的mRNA的蛋白表达与公布，并与正义（Sen)ODM转移、似注射+肾脏原位电穿孔（Sham)作为对照。结果FITC标记的ODN在基因转移10min后主要定位于转移侧肾脏的肾小不，而对侧肾脏FITC荧学阴性。ATI放射自显影定量分析显示在基因转移的3d内，结合有血管紧张素II（Ang II)的ATI较未转移的对侧肾脏减少约70%。AT1 AS-ODA基因转移后3d,转移侧肾脏AT1 mRNA水平较对侧肾脏下降约43%，蛋白水平下降约67%，而与ATI Sen-ODN和Sham组转移侧肾脏比较，AT1 mRNA水平下降分别约47%和51%，蛋白水平下降分别63%和71%。免疫组化检醒发现ATI的阻断以系膜区的表达减少为主。结论本研究所用的单侧肾动肪注射+原位电穿孔的基因转移方法成功地抑制了单侧肾脏的肾小球ATI受体的表达，为研究全身性疾病肾损害中肾脏局部RAS的短期用用机制提供了一个比较理想的整体动作模型。

观察环加氧酶（COX）在梗阴性肾病大鼠肾脏的表达，比较选择性COX-2抑制剂莫比可与消炎痛对COX、纤溶酶原激活物抑制物1（PAI-1）和基质金属蛋白酶组织抑制物-1（TIMP-1）假手术组（C）为对照。4周后应用逆转录，聚合酶链反应（RT-PCR）检测COX-1、COX-2、PAI-1及TIMP-1的mPNA表达，免疫组织化学检测COX-1和COX-2的蛋白水平，并采用放射免疫法测定术侧肾脏残余尿液的血栓浣素B2（TXB2）浓度。结果COX-1的mPNA和蛋白水平在4组间差异无显著性意义。U组大鼠尿TXB2浓度急剧上升，术肾PAI-1和TIMP-的1mPNA表达明显上调，COX-2的基因表达和蛋白南合成大幅增加。莫比可能够显著抑制上述基因的转录（P＜0.05），而I组的数值与U组相仿。M组和I组的尿TXB2浓度明显下降，尤以M组为著。结论COX-2参与了UUO发生，发展的病理过程。选择性COX-2抑制剂能够从抑制COX-2活性、下调COX-2、PAI-1及TIMP-1的基因转录等多方面发挥其积极的治疗作用。

蛋白尿是肾脏病进展的独立危险因素。研究显示，阻断肾素-血管紧张素系统能有效降低血压、减少蛋白尿和延缓慢性肾脏疾病的进展，且已发现若增加增紧张素转换酶抑制剂（ACEI）或血管紧张素II受体拮抗剂（ARB）剂量，虽无明显的降压作用，但可增加降蛋白尿的幅度。本研究观察增加氯沙剂量后地我国慢性肾脏病患乾蛋白尿的疗效。

目的探讨过氧化物酶体增殖物活化受体（PPAR）γ对高糖诱导肾小球系膜细胞（GMc）外基质（ECM）积聚的抑制作用及其机制。方法以表达野生型小鼠PPAγl的质粒pIREs2-EGFP-mPPARγl/WT转染系膜细胞（wT），然后予30mmol/L的高糖（HG）刺激48h，以EusA法检测细胞上清液中TGFβ1、纤连蛋白（FN）的浓度。GMc中c-fos、c-jun、葡萄糖转运蛋白l（GLuT-1）mRNA的表达检测采用半定量RT-PcR法，并以weslem印迹检测胞浆中核因子抑制物（I-KB）、核因子（NF-KB）及胞核中NF-KB、磷酸化细胞外调节激酶（p-ERK）的蛋白表达水平。同时以2umol/L的PPARγ激活剂吡咯列酮（Pi0）、转染表达功能缺陷的突变型（dominant negative，DN）PPARγ1的质粒pIREs2-EGFP．mPPARγl/DN（DN）或空白质粒pIREs2-EGFP作为对照。PPARγ的活性以其与PPARγ反应元件（PPRE）的结合能力表示。结果HG培养时系膜细胞的PPARγ活性较正常糖浓度（5mmol/L）培养时明显上升（P<0.01），HG+wT组的PPAγ活性显著高于HG+DN、HG-pIREs2-EGFP和HG+Pio组。与HG+DN、HG+pIREs2．EGFP相比，wT转染所致的PPARγl高表达能显著抑制高糖诱导GMc的TGFB-1、FN生成增多，减轻c-fos和c-jun的mRNA表达的上调，改善p-ERK蛋白水平的增加、胞浆I-kB表达的降低以及NF-KB由胞浆向胞核转移的增加（P均<O.05）。wT对HG时高表达的GLuT-1mRNA无显著影响。Pio除对P-ERK蛋白水平、NF-KB胞核／浆比例具有显著性改善作用外，对上述其他指标无明显作用。结论PPARγl过表达能抑制由高糖诱导的GMc EcM的增多，其机制可能是通过抑制了ERK／激活蛋白（AP）1及NF-KB等信号分子的传递。提示在糖尿病状态下，增高PPARγ活性可能具有直接的抗纤维化效应。

探讨异性环氧合酶-2（COX-2）抑制剂罗非昔布（rofecoxib)延缓肾大部切除大鼠进生性肾损害的机制。方法将大鼠随机分为假手术组、肾大部分切除（SNX）、罗非昔布组、吲哚美组、氯沙坦组、罗非昔而与氯沙坦合用组，第组6只。6周后后检测大鼠血压、悄蛋白及组化的方法检测纤维连接蛋白（FN）表达，逆转录-聚全印迹法检测肾皮质COX-2、COX-1、纤溶酶原激活物抑制剂-1（PAI-1）及血管紧张素1型受体（AT1）的表达。结果与假手术组相比，XNX组大鼠尿白及肾皮质COX-2的表达与尿TXB2排汇明显增多，COX-1表达无明显变化：系统血压及AngII浓度明显增高，肾皮质TβRI及TβRII的mRNA及PAI-1和ATI的表达显著上调，肾小球硬化指数及小管间质损伤指数增高。罗非昔布能明显减速轻蛋白尿（P＜0.05）、肾皮质TβRI、TβRII的mRNA及PAI-1的表在；较SNX组分别下调36.44%、45.02%和31.16%，与氯沙坦作用相当。吲哚美组大鼠尿蛋白排汇及肾小球三硬华指数较肾大部切孤芳自赏组降低，便肾小管间质损伤指数增高。罗非昔布和吲哚美组大鼠血压幸免无明显降低。罗非昔布和氯沙坦合组尿蛋白较单用组更低，但差异无统计学意义。结论特异性COX-2抑制剂罗非昔布通过部分抑制COX-2的活性，调整肾内活跃的肾素血管紧张素系统，下调TβRI、TβRII、mRNA及PAI-1的表达，从布减轻蛋折尿，延缓肾小球硬化及小管间质纤维化，其确切的机制尚待进一步研究。

测定狼疮性肾炎（LN）患乾的热休史蛋白70（HSP70）、泛素、血管张素II1型受体（AT1R）、血管紧张素II2型受体（AT2R）、血管紧张素转酶（ACE）等，探讨LN患者肾素血管紧张素系统（RAS）是否被激活及其与伴侣蛋白可能存在的关系。方法27例LN患者（LN组）根据系统性红斑狼疮（SLE）活动指数（SLE-DAI）进一步分为活动亚组（13例，SLE-DAI≥15）和非活动亚级（14例，SLE-DAI≥15）；对照组为9例匿性肾炎患者（肾组织活检结果为轻微病变）。对所有肾穿刺标本行HSP70、泛素、AT1R、AT2R、ACE测定。采用TUNEL法测定肾内凋亡细胞的表达。对SLE活动度、病理变化，RAS活性与伴侣蛋白的表达进行相关分析。结果LN组的HSP70表达显著低于对照组，而泛素、AT1R、AT2R、ACE和凋谢细胞的表达明显升高（P值均＜0.05＝。活动亚组与非活动亚组的HSP70及泛素表达的差异无显性（P＜0.05＝，海参动亚组的ACE、AT2R和凋谢细胞的表达显著高于非活动亚组（P值均＜0.05＝。小管间质中，HSP70表达与病慢性指数相关（γ=0.132,P=0.028）；肾小球与肾小管中，泛素表达与AT1R或AT2R均相关（γ的值分别0.574、0.616。P值均＜0.05＝结论伴侣蛋白和RAS可能参与了LN病理变化的发病机制。

Accelerated fibrosis and collagen deposition develop in the renal interstilium of angiotensin type 2 receptor null mutant mice during ureteral obstruction. We examined the role of angiotensin in renal remodeling that is specifically channeled through the angiotensin type 2 receptor (2 receptor). Previously, we observed that in mouse embryonic kidneys the AT1 mRNA is predominantly expressed in the mesenchynle. We therefore chose a model of unilateral ureteral obstrtryction, character-ized by activation of the renin-angiotensin system, while fibrosis develops prominently within the renal interstitium. Male wild-type mice (Ago-2/Y) and mice null mutanl for the AT2 gene (Agtr2-/N) were subujected to a complete unilateral ureteral ligation for 5 or 14 days. Obstructed idneys of Agtr2-/Y mice showed more severe interstitial fibrosis than Ihose of Agtr2-/Y mice. confirmed by increased collagen hy point-counting on Masson trichrome stained sections, and increased oH (I) collagen mRNA expression by Northern blot. Lmmunohistoehernistry staining for PCNA (a marker of cell proliferation), 14: 80 (a marker of acmphages), vimendn (a marker of flbroblasls), and aSMA (a marker of myofibroblasts) revealed that, while the two groups were comparable in the degree of cell proliferation and macrophagc infiltration, fibroblasts/myofibrofilasts were present in a greater abundance in obslructed kidneys of Agtr2-/Y mice than in Agtr2+/Y al both 5 and 14 days after obstruction. Moreuver. cclls undergoing apopiosis were significantly tess in Agtr2-/Y tilth in Agtr2+/Y. Thus, the AT: receptor significantly impacls the remodeling process within renal inlcrstitium, potentially by reguhuing the population) of colblgen-producing cells. While inost of the known effects of angiotensin 11 (Aug 11) are mediated by its type 1 receplor (AT1 [1], the function of type 2 receptor (AT2) has not been fully elucidated. Some investigators suggest that AT2, is involved in control of cell prolileration and differentiation [2. 3] Several recent studies indicate thai AT,. Is involved in apoplosis [4. 5]. Although AT, expression is down- regulated after birth, il is speculated that AT2 plays a role in tissue remodeling or repair of vascular and cutaneous tissues under some pathophysiological conditions [6-9]. It has been widely accepted that, in a variety of renal diseases,

A previous study showed that exogenous angiotensin II (AngII) induces proliferation of glomerular cells through systemic actions of AngII. In the present study, the authors examined the mode of actions of endogenous AngII in injured kidneys that were made deficient in AT1 by using in vivo transfection of antisense oligodeoxynucleotide (AS-ODN). Thy-1 nephritis was induced in rats by injection of mAb 1-22-3. Four days later, glomerular transfection was performed by unilateral whole-kidney electroporation after AT1 AS-ODN delivery through the left renal artery (n=7). The expression of renal AT1 was assessed by autoradiography. The effect of the AS-ODN transfection was assessed 3 d later and compared with transfection with control ODN (n=6), systemically administered pharmacologic AT1 antagonist losartan (n=5) as well as untreated Thy-1 animals (n=5). Fluorescencelabeled AS-ODN was found transfected in almost all glomeruli and localized primarily to the mesangium. Compared with the contralateral untransfected kidney in both normal and Thy-1 rats, AS-ODN suppressed cortical AT1 expression by some 70%. The AS-ODN transfected kidneys of Thy-1 rats had significantly lower glomerular mesangial cell proliferation (7.38

为阐明水孔蛋白（AQP-2）是否与其它AQPs一样具有分布的广泛性。方法用秀行设计的引物以逆转录聚合酶反应法（PT-PCR）检测大鼠体内AQPs mPNA的分布及其在林水状态下的变化。结果AQPs mPNA不仅在肾脏有高表达，在脑、主动肪、肺、肝、脯、小肠、结肠、背上腺、聚丸、卵巢亦有弱表达，禁水46小时后，上术表达均较正常时有明显增强。结论AQP2广泛分布于肾外组织。