百令胶囊干预肾小球硬化大鼠TGF—β 1 mRNA表达的研究

目的研究百令胶囊对大鼠肾小球硬化的干预作用,并探讨其与转化生长因子（TGF-β1）mRNA表达的关系。方法正常Wistar大鼠36只随机分为正常对照组,模型组和百令胶囊组。模型组和百令胶囊组行5／6肾切除制备进行性肾小球硬化模型,百令胶囊组以百令胶囊混悬液[4g／（kg·d）]灌胃治疗,模型组和正常对照组每天给予等量的自来水灌胃,大鼠自由饮水、进食。末次术后9周各组分别处死大鼠12只,测定血生化指标和尿蛋白排泄量;采用逆转录PCR（RT-PCR）检测大鼠肾组织中TGF-β1mRNA的表达,应用免疫组织化学方法检测。肾组织TGF-β1的表达。结果与模型组相比,百令胶囊组大鼠血浆白蛋白（Alb）增高,胆固醇（choles-terin）和低密度脂蛋白（LDL）降低,尿蛋白排泄（UAE）减少,血肌酐（Cr）、尿素氮（BUN）水平下降;TGF-β1 mRNA的表达明显减少（P〈0．01）,免疫组化检测示TGF-β1的表达减弱（P〈0．05）。结论随着5／6肾切除大鼠肾小球硬化程度加重,TGF-β1的表达增加,提示TGF-β1参与了肾小球硬化的进展;百令胶囊干预后,肾小球硬化大鼠TGF-β1的表达明显下调,可能是百令胶囊延缓。肾小球硬化进展的机制之一。     

汉防己甲素延缓肾小球硬化机理的实验研究

目的 探讨汉防己甲素对肾小球硬化大鼠肾脏转化生长因子（TGF－β1）基因表达的影响。方法 27只雄性Wistar大鼠随机分为正常对照组,肾小球硬化模型组、汉防己甲素组和氨氯地平组,用单侧肾切除加2次注射阿霉素制备肾小球硬化动物模型,用Northern印迹杂交,观察肾皮质TGF－β1mRNA表达。结果 汉防己甲素使肾小球硬化大鼠肾皮质TGF－β1mRNA表达较模型组明显下调,肾小球细胞外基质较模型组明显减少。结论 汉防己甲素通过下调TGF－β1表达而减少肾小球ECM沉积,从而延缓肾小球硬化。     

大黄对肾硬化大鼠肾小球细胞增殖和p27蛋白表达的影响

目的通过大黄对实验性肾硬化大鼠中细胞周期负调控蛋白p27的表达和肾小球细胞增殖的影响，探讨大黄延缓肾硬化进展的作用机制。方法3个月龄雄性Wistar大鼠24只，随机分为对照组、肾硬化组和治疗组，每组8只。肾硬化组大鼠在无菌条件下行左肾摘除，术后第7天给予多柔比星5 mg·kg^-1，尾部静脉注射，再于术后第28天重复注射多柔比星3 mg·kg^-1。治疗组处理同肾硬化组，并于肾摘除当天开始每日灌服大黄抽提液(溶于10%纤维素钠溶液)0.5 mg·kg^-1，对照组大鼠则分别以假手术和静脉注射等体积0.9%氯化钠注射液替代。3组于手术或假手术后第13周处死大鼠。测定各组大鼠血尿素氮(BUN)、血肌酐(SCr)、24 h尿蛋白定量以及肾小球平均截面积和平均体积。分离肾小球，提取肾小球总RNA，用半定量RT-PCR方法检测肾小球内增殖细胞核抗原（PCNA)-mRNA的表达，同时采取免疫组织化学方法检测PCNA和细胞周期负调控蛋白p27的表达。结果治疗组血尿素氮、血肌酐和24 h尿蛋白均较肾硬化组明显降低(P＜0.05或P＜0.01)。组织病理学检查显示治疗组病理损害明显减轻，肾小球截面积和平均体积明显减小。与肾硬化组比较，治疗组肾小球 PCNA mRNA表达明显减弱，增殖指数下降，p27蛋白表达升高(P＜0.01)。结论大黄能上调肾硬化大鼠肾小球细胞周期p27蛋白的表达，减轻肾小球细胞增殖，从而延缓肾硬化的进展。     

Ang-2在大鼠阿霉素肾病的表达及意义

目的:肾病综合征临床以大量蛋白尿为主要特征,病理都有肾小球脏层细胞(即足细胞)的损伤,越来越多的研究证实,足细胞的损伤,可促进肾小球硬化。本实验旨在探讨大鼠肾小球硬化过程中,内源性血管生成素的异常变化及其病理作用。方法:选择48只健康、平均体重为150~200gWistar大鼠,随机分二组,肾病组和对照组各24只。采用逆转录聚合酶链式反应技术(RT-PCR)检测阿霉素注射后1、2、4及7周肾组织Ang-2 mRNA的表达。结果:(1)实验组第1周未检测到Ang-2mRNA表达,但于第2、4和7周时Ang-2 mRNA表达上调,各时点比较有明显差异(P<0.05)。(2)对照组整个实验期间均未检测到Ang-2 mRNA表达。(3)两组同时点比较有显著性差异(P<0.05)。结论:阿霉素肾病大鼠肾脏在肾小球硬化过程中存在内源性血管生成素的异常变化,Ang-2表达上调,提示Ang-2在实验组的异常表达与足细胞损伤有关。这种改变可能是机体适应代偿机制。     

氟伐他汀对糖尿病肾病鼠血管内皮生长因子表达的影响

目的：探讨氟伐他汀对糖尿病肾病（DN）大鼠肾组织血管内皮生长因子（VEGF）mRNA及蛋白质表达的影响。方法：SD大鼠分为对照组、DN组、氟伐他汀治疗组，链脲菌素腹腔注射诱导DN大鼠模型，诱导成功后氟伐他汀治疗组给予氟伐他汀经胃管灌服。试验12周末检测大鼠体重、血糖、尿肌酐、24h尿白蛋白。采用RT—PCR检测VEGFmRNA及免疫组化检测蛋白质的表达。结果：DN组大鼠体重、血糖、尿白蛋白，VEGFmRNA及蛋白质的表达，肾小球细胞数，细胞外基质（ECM）聚积均高于对照组（P〈0．01）。与DN组相比，氟伐他汀治疗组大鼠体重、血糖、尿白蛋白，肾脏VEGFmRNA及免疫组化检测蛋白质的表达，ECM聚积显著降低（P〈0．01或P〈0．05）。尿VEGF与尿白蛋白呈正相关（r=0．42，P=0．039），与尿肌酐亦呈正相关（r=0．35，P=0．043）．结论：VEGF表达增加可能参与DN的发病机制。氟伐他汀可减轻DN大鼠体重、血糖，减少尿白蛋白的排泄，抑制肾小球系膜增殖及硬化，降低VEGFmRNA及蛋白质的表达。     

苯那普利对肾病幼鼠纤溶蛋白肾组织定位表达的影响

目的　探讨肾病幼年大鼠肾小球硬化及肾间质纤维化病变进展过程中 ,肾组织尿激酶型纤溶酶原激活物 (u PA )、组织型纤溶酶原激活物 (t PA)及其特异性抑制物 (PAI- 1)蛋白定位表达的特点 ,及予血管紧张素转换酶抑制剂 (ACEI)——苯那普利 (benazepril;lotensin)治疗的影响。方法　采用阿霉素诱导的肾病大鼠为动物模型 ,予 ACEI治疗 12周后测大鼠体重、血压、尿蛋白、及血生化各项指标的变化 ,同时用免疫组织化学染色等方法检测各组肾组织 u PA、t PA和 PAI- 1的蛋白表达的变化特点。结果　肾病大鼠肾组织 PAI- 1表达均高于正常对照组 ,而 u PA、t PA均低于正常组 ;经治疗后肾组织 PA I- 1趋于下降 ,而 u PA、t PA表达趋于增高 (P<0 .0 1)。结论　纤溶系统的平衡紊乱是肾病大鼠肾小球硬化和肾间质纤维化进展中的重要病生理变化之一 ,ACEI治疗可改善 PAs/ PAI- 1的异常表达 ,防止细胞外基质的异常沉积 ,阻止肾小球硬化和间质纤维化病变进展。     

罗格列酮减轻阿霉素肾病大鼠肾小球硬化的实验观察

目的观察罗格列酮干预治疗对单侧肾切除加重复阿霉素注射诱导的肾小球硬化大鼠肾脏的保护作用。方法建立单侧肾切除加重复阿霉素注射的肾小球硬化大鼠模型,分为罗格列酮治疗组和肾病组,设假手术组为对照组。检测各组大鼠第0、4、8、12周尿蛋白,并观察第12周肾组织病理改变,计算肾小球硬化指数。结果罗格列酮治疗组比肾病组尿蛋白排泄量明显减少(P<0.01),肾小球硬化程度明显减轻。结论罗格列酮对阿霉素肾病大鼠肾脏病变具有保护作用。     

苯钠普利对肾小球硬化大鼠肾组织MCP-1的影响

目的观察苯钠普利对肾小球硬化大鼠肾组织单核细胞趋化蛋白-1(monocyte chemotaltilprotein-1,MCP-1)表达的影响,探讨血管紧张素转化酶抑制剂(angiotensin converting enzyme inhibitor,ACEI)对肾脏的保护作用。方法雄性SD大鼠36只随机分为:正常对照组、肾小球硬化组、苯钠普利治疗组。切除肾小球硬化组和苯钠普利治疗组大鼠左肾,1周后按5mg/kg于尾静脉推注阿霉素,正常对照组行假手术及尾静脉推注等量生理盐水,注射阿霉素后次日始苯钠普利治疗组予苯钠普利6mg/(kg.d)每天清晨灌胃1次,正常对照组和肾小球硬化组仅灌以2ml自来水。灌胃12周后处死大鼠,切取右肾用于肾脏病理分析,免疫组化法测定肾组织MCP-1。结果肾脏病理分析,肾小球硬化组多数肾小球呈节段性硬化,许多肾小球及肾小管受累;苯钠普利治疗组肾脏病变轻于肾小球硬化组,且肾小球硬化指数(GSI)明显低于肾小球硬化组(P<0.01);②免疫组化结果显示正常对照组肾小管中有少量MCP-1的表达,在肾小球内几乎不表达,在肾小球硬化组和苯钠普利治疗组肾小球及肾小管内表达皆强于正常对照组(P<0.01)。苯钠普利治疗组MCP-1表达较肾小球硬化组弱(P<0.01)。结论在单侧肾切除 阿霉素注射诱导的肾小球硬化大鼠模型中,苯钠普利通过抑制血管紧张素转换酶(ACE),减少了血管紧张素II(AngII)的生成,降低了MCP-1在肾脏中的表达,抑制或减轻了巨噬细胞浸润,进而延缓了肾小球硬化的进展。     

罗格列酮对阿霉素致肾小球硬化症大鼠的尿蛋白排泌量及肾小球蛋白(synaptopodin)表达的影响

目的 观察罗格列酮(胰岛素增敏药)对阿霉素致肾病大鼠的尿蛋白排泌量及肾小球蛋白表达的影响.方法 用罗格列酮治疗阿霉素诱导的局灶节段性肾小球硬化(FSGS)大鼠模型;用双缩脲法动态检测大鼠24 h尿蛋白量;分别用荧光实时定量PCR和免疫组织化学法检测大鼠肾组织synaptopodin mRNA和蛋白表达量.结果 罗格列酮能显著减缓FSGS大鼠尿蛋白排泄量的增加(P＜0.05);恢复大鼠肾组织蛋白表达量(P＜0.05),而不影响其mRNA表达量(P＞0.05);FSGS大鼠肾组织蛋白表达量与尿蛋白排泄量呈负相关(P＜0.01).结论 罗格列酮能减轻FSGS模型尿蛋白排泌量,其机制可能部分与维持足细胞足突肌动蛋白微丝骨架系统的稳定有关.     

黄葵胶囊对肾小球硬化防治作用的实验研究

目的研究黄葵胶囊对5/6肾切除慢性肾衰大鼠模型肾小球硬化的治疗作用与可能机制。方法将5/6肾切除SD大鼠随机分为对照组和治疗组,另设假手术组作为对照。术后4周治疗组给予黄葵胶囊灌胃干预,治疗1个月后检测大鼠血清BUN、Scr和血红蛋白水平,图像分析肾组织α-平滑肌肌动蛋白-SMA(α-SMA)的表达量。结果黄葵胶囊可以降低BUN、Scr,减少α-SMA在肾小球的表达。结论黄葵胶囊减轻肾小球硬化的机理在于抑制肌成纤维细胞增殖和抑制ECM合成。     

Changes in angiopoietin expression in glomeruli involved in glomerulosclerosis in rats with daunorubicin-induced nephrosis

AIM: To study the potential pathological role of endogenous angiopoietins in daunorubicin-induced progressive glomerulosclerosis in rats. METHODS: Seventy male Wistar rats were allocated randomly into a daunorubicin group (DRB; n=40) or a control group (n=30). The rats in the DRB group were injected with DRB (15 mg/kg), in their tails. Subsequently, at intervals of 1, 2, 4, 6, 8, and 12 weeks, 5 male Wistar rats in each group were chosen randomly for 24 h urinary protein quantitative measurements (24 h UPQM), and determination of plasma tumor necrosis factor alpha (TNF-alpha), angiopoietin-1 (Ang1), and angiopoietin-2 (Ang2) levels. Kidney sections were examined by electron microscopy, Periodic Acid Schiff (PAS) staining, immunohistochemical staining and in situ hybridization histochemistry. RESULTS: As glomerulosclerosis progressed in the DRB group, expression of Ang1 mRNA and protein in glomeruli decreased and expression of TNF-alpha protein, Ang2 mRNA and protein in glomeruli increased. Expression of Ang1 mRNA and protein in glomeruli were negatively correlated with 24 h UPQM, Fn protein expression, and mean area of extracellular matrix (MAECM). In comparison, expression of Ang2 mRNA and protein in glomeruli were positively correlated with 24 h UPQM, Fn protein expression and MAECM; furthermore, there was a positive correlation between plasma Ang2 and 24 h UPQM. Plasma TNF-alpha and expression of TNF-alpha in glomeruli were positively correlated with expression of Ang2 mRNA and protein in glomeruli. There was a negative correlation between Ang1 protein expression and Ang2 protein expression in glomeruli. CONCLUSION: During DRB-induced glomerulosclerosis, podocyte injury led to a shift in the balance of Ang1 and Ang2 in glomeruli. Increased TNF-alpha in plasma and glomeruli may upregulate Ang2 expression in glomeruli. Elevated Ang2 in both plasma and glomeruli may mediate protein permeability through the glomerular filtration barrier. Moreover, local expression of Ang2 may facilitate the progress of glomerulosclerosis by upregulating a component expression of extracellular matrix.     

先兆子痫肾小球病病理观察与临床研究

目的 研究先兆子痫肾小球病病理改变及与临床的关系。方法对8例先兆子痫肾小球病患者的肾活检标本进行光镜、免疫荧光及电镜检查,观察病理特征并进行临床病理分析。结果病理改变：光镜以肾小球内皮细胞增生为特征,电镜表现更为显著,内皮细胞增生肿胀,上皮细胞足突大部融合,可表现节段性硬化。临床与病理联系：产后时间与内皮细胞增生肿胀程度呈负相关（r=-0．945,P〈0．01）。而尿蛋白量与上皮足突融合程度正相关（r=0．718,P〈0．05）。表现为节段硬化者,尿蛋白量较多,恢复较慢。结论内皮细胞损伤为本病病理特征,肾活检有助于诊断及病情轻重判断。     

Activation of p44/42 in human natural killer cells decreases cell-surface protein expression: Relationship to tributyltin-induced alterations of protein expression

Tributyltin (TBT) activates the mitogen activated protein kinase (MAPK), p44/42 in human natural killer (NK) cells. TBT also reduces NK cytotoxic function and decreases the expression of several NK-cell proteins. To understand the role that p44/42 activation plays in TBT-induced loss of NK cell function, this study investigated how selective activation of p44/42 by phorbol 12-myristate 13-acetate (PMA) affects NK cells. Previously it was shown that PMA caused losses of lytic function similar to those seen with TBT exposures. This study examined activation of p44/42 in the regulation of NK-cell protein expression and how this regulation may explain the protein expression changes seen with TBT exposures. NK cells exposed to PMA were examined for levels of cell-surface proteins, granzyme mRNA, and perforin mRNA expression. The expression of CD11a, CD16, CD18, and CD56 were reduced, perforin mRNA levels were unchanged, and granzyme mRNA levels were increased. To verify that activation of p44/42 was responsible for the alterations seen in CD11a, CD16, CD18, and CD56 with PMA, NK cells were treated with the p44/42 pathway inhibitor (PD98059) prior to PMA exposures. In the presence of PD98059, PMA caused no decreases in the expression of the cell-surface proteins. Results of these studies indicate that the activation of p44/42 may lead to the loss of NK cell cytotoxic function by decreasing the expression of CD11a, CD16, CD18, and CD56. Further, activation of p44/42 appears to be at least in part responsible for the TBT-induced decreases in expression of CD16, CD18, and CD56.     

Activation of p44/42 in human natural killer cells decreases cell-surface protein expression: Relationship to tributyltin-induced alterations of protein expression

Tributyltin (TBT) activates the mitogen activated protein kinase (MAPK), p44/42 in human natural killer (NK) cells. TBT also reduces NK cytotoxic function and decreases the expression of several NK-cell proteins. To understand the role that p44/42 activation plays in TBT-induced loss of NK cell function, this study investigated how selective activation of p44/42 by phorbol 12-myristate 13-acetate (PMA) affects NK cells. Previously it was shown that PMA caused losses of lytic function similar to those seen with TBT exposures. This study examined activation of p44/42 in the regulation of NK-cell protein expression and how this regulation may explain the protein expression changes seen with TBT exposures. NK cells exposed to PMA were examined for levels of cell-surface proteins, granzyme mRNA, and perforin mRNA expression. The expression of CD11a, CD16, CD18, and CD56 were reduced, perforin mRNA levels were unchanged, and granzyme mRNA levels were increased. To verify that activation of p44/42 was responsible for the alterations seen in CD11a, CD16, CD18, and CD56 with PMA, NK cells were treated with the p44/42 pathway inhibitor (PD98059) prior to PMA exposures. In the presence of PD98059, PMA caused no decreases in the expression of the cell-surface proteins. Results of these studies indicate that the activation of p44/42 may lead to the loss of NK cell cytotoxic function by decreasing the expression of CD11a, CD16, CD18, and CD56. Further, activation of p44/42 appears to be at least in part responsible for the TBT-induced decreases in expression of CD16, CD18, and CD56.     

CD44v6和hTERT基因在胃癌患者中的表达及临床意义

目的检测胃癌患者组织中CD44v6、hTERT基因的表达情况及与临床病理因素的关系。方法采用免疫组织化学染色的方法检测胃黏膜肠化生、异型增生及胃癌组织中CD44v6、hTERT蛋白表达情况,并结合患者的临床病理资料进行分析。结果胃黏膜肠化生、异型增生及胃癌组织中CD44v6、hTERT蛋白表达阳性率显著高于正常胃黏膜,差异有统计学意义(P<0.05)。CD44v6、hTERT的表达与胃癌的分化程度、肿瘤大小无关(P>0.05),而与肿瘤的浸润深度、淋巴结转移及TNM分期密切相关(P<0.05)。结论 CD44v6、hTERT过表达与胃癌发生发展及浸润转移有关。     

CD133在不同病理级别脑胶质瘤中的表达及临床意义

目的检测CD133在脑胶质瘤中的表达,探讨其对脑胶质瘤诊断及预后的作用。方法应用辣根过氧化物酶化学法检测38例脑胶质瘤（脑胶质瘤组）和5例正常脑组织（正常对照组）中CD133的表达。结果 CD133在脑胶质瘤组中的阳性表达率为71.1%（27/38）,在正常对照组中的表达为0.0（0/5）,二者比较,差异有统计学意义（P〈0.05）;CD133阳性细胞例数在不同病理级别的脑胶质瘤组织中的表达存在相关性,即胶质瘤级别越高,CD133阳性细胞例数越多,差异有统计学意义（r=0.012,P〈0.01）;不同病理级别的脑胶质瘤中CD133阳性细胞率与患者的复发率存在相关性,即不同病理级别的脑胶质瘤中,CD133阳性细胞率越高,患者复发率越高,差异有统计学意义（r=0.358,P〈0.05）。结论在脑胶质瘤中CD133的表达情况可以用于指导肿瘤的临床诊断和预后评估。     

Estrogen deficiency results in enhanced expression of Smoothened of the Hedgehog signaling in the thymus and affects thymocyte development

Aromatase is an essential enzyme for estrogen synthesis. We investigated the role of estrogen in thymocyte development using aromatase-deficient (ArKO) mice. Like its role as a regulator of bone metabolism through regulating osteoprotegerin (OPG) production, estrogen is involved in the processes of thymocyte development although aromatase mRNA was not detectable in the thymus. Thymic regression and reduced cellularity were evident in ArKO mice. The major difficulties in thymocyte development of ArKO mice were observed during the CD44+ CD25- stage at the cortico-medullary junction and during the CD44- CD25- stage at the subcapsular region where the estrogen receptor was expressed in the stromal cells. The proportion of thymocytes during the CD44+ CD25- stage was reduced. The progression of CD44- CD25+ cells to the CD44- CD25- stage was accelerated in ArKO mice possibly due to insufficient osteoprotegerin production in estrogen-deficiency. However, the expression of Smoothened of the Hedgehog signaling was enhanced in CD4- CD8- double negative cells. This enhancement may result in impaired progression of CD44- CD25- cells to the CD4+ CD8+ double positive stage and impaired proliferation of CD4+ CD8+ double positive cells since Smoothened (Smo) is known to arrest cells as non-proliferating cells. This could be the reason why the proportion of CD3+ TCRbeta(high) cells during the late phase of thymocyte maturation was reduced in ArKO mice. From these observations, we propose that estrogen supports thymocyte development and maturation at many stages through many regulatory pathways including the sonic hedgehog- and the osteoprotegerin ligand (OPGL)-mediated signaling.     

Th17细胞在类风湿关节炎发病机制中的作用研究进展

类风湿关节炎(rheumatoid arthritis,RA)是临床常见炎症性自身免疫疾病,以早期进行性关节滑膜炎症为主要临床特征,晚期则多以关节软骨破坏及骨侵蚀为主要病理特征。RA病因复杂,病理机制至今未明,发病率高,5年期致残率高。Th17细胞作为CD4+T细胞的亚群之一,其分泌的IL-17、IL-21等促炎性细胞因子,在RA关节滑膜炎症和关节软骨破坏及骨侵蚀等多个病理环节均发挥重要作用。基于此,本文对近年来Th17细胞参与RA关节滑膜炎症和关节软骨破坏及骨侵蚀病程的相关研究文献进行综述和讨论,以期为RA发病机制研究提供新思路,为以Th17细胞为作用靶点的创新药物开发提供参考。     

CD44v6 mRNA及其蛋白表达与胃癌预后的关系

目的：探讨CD44v6 mRNA及其蛋白表达与胃癌预后的关系。方法：应用催化信号放大系统原位杂交和免疫组织化学技术，对17例早期胃癌、21例中期胃癌和57例晚期胃癌组织进行CD44v6 mRNA及其蛋白检测。结果：在胃癌中，CD44v6 mRNA及其蛋白的表达阳性率分别为85．3％和82．1％。CD44v6 mRNA及其蛋白表达阳性率在晚期胃癌明显高于早、中期胃癌（P＜0．05）。CD44v6 mRNA表达及其蛋白表达均与胃癌浆膜浸润、淋巴结转移和患者预后呈正相关（P＜0．05）。结论：CD44v6 mRNA及其蛋白表达可反映胃癌分化不良或晚期阶段，可作为预测预后的客观指标。