青藤碱对UUO小鼠肾组织间质纤维化与炎症因子表达的干预作用

目的：通过抗风湿药青藤碱对UUO小鼠肾组织α-平滑肌肌动蛋白（α-smooth muscle actin,α-SMA）、炎症介质细胞间黏附分子（intercellular adhesion molecule1,ICAM-1）、单核细胞趋化因子（monocyte chemoattractant protein1,MCP-1）表达的影响,探讨其防治肾间质纤维化作用及其机制。方法：实验采用小鼠单侧输尿管结扎（UUO）模型,用不同剂量青藤碱进行干预,用血管紧张素受体抑制剂蒙诺作为对照。肾脏病理用HE和Masson染色;肾组织α-SMA表达采用免疫组化;ICAM-1蛋白质表达采用Western-blotting方法检测;MCP-1基因表达采用逆转录-聚合酶链式反应（RT-PCR）方法。结果：UUO模型组小鼠肾间质纤维化程度以及肾组织α-SMA的表达较假手术组显著增高,肾小管间质中炎细胞浸润亦较假手术组显著增强;青藤碱各治疗组肾间质纤维化程度、α-SMA和ICAM-1蛋白、MCP-1基因表达均显著低于模型组;蒙诺治疗亦可显著降低肾间质纤维化程度和肾组织α-SMA的表达,但对于炎细胞浸润和炎症因子抑制作用显著较青藤碱弱。而且未见其具有抑制ICAM-1表达的作用。结论：抗风湿药青藤碱可显著抑制肾间质纤维化和肌成纤维细胞的积聚,可能与其显著抑制肾组织炎症因子的表达及炎细胞浸润有关。     

血管紧张素1a型受体基因敲除小鼠肾脏局部肾素-血管紧张素系统的变化及其对糖尿病肾小球硬化的影响

目的 探讨血管紧张素1a型受体（AT1aR）基因敲除小鼠肾脏局部肾素-血管紧张素系统（RAS）的组分改变对糖尿病肾病（DN）肾小球硬化的影响及其可能机制。 方法 AT1aR基因敲除小鼠和野生型小鼠腹腔注射链脲佐菌素（STZ，300 mg/kg）诱导糖尿病模型12周后，取肾脏组织作冰冻组织切片，用激光捕获微切割技术分离肾小球，提取RNA。用实时定量PCR的方法检测肾小球内AT1aR、血管紧张素1b型受体（AT1bR）、血管紧张素2型受体（AT2R）、血管紧张素原、血管紧张素转化酶（ACE）、肾素、醛固酮合成酶（CYP11B2）的mRNA表达。PAS染色观察肾脏病理变化。免疫组化检测转化生长因子β1（TGF-β1）、1型纤溶酶原激活物抑制物(PAI-1)、单核细胞趋化因子1(MCP-1)和肾素的表达。比较不同基因型小鼠肾小球细胞外基质和各细胞因子的表达变化。 结果 与野生型小鼠相比，AT1aR基因敲除小鼠肾小球内AT1bR、血管紧张素原、肾素、CYP11B2的表达明显上调（P < 0.05），AT2R表达下调，ACE无明显改变；AT1aR基因敲除小鼠肾小球细胞外基质明显增加（P < 0.05），TGF-β1、PAI-1、MCP-1和肾素的表达均明显增加（P < 0.05）。 结论 AT1aR基因敲除并不能使糖尿病小鼠肾脏病变改善。RAS组分的表达改变（AT1bR的上调和AT2 的下调，肾素的上调和CYP11B2的上调）参与糖尿病肾小球病变过程。     

免疫球蛋白样结构域受体2缺失加重缺血再灌注诱导的肾纤维化

目的探究免疫球蛋白样结构域受体2(immunoglobulin-like domain-containing receptor 2, Ildr2)在缺血再灌注诱导肾纤维化中的作用。方法利用CRISPR/Cas9技术构建Ildr2敲除小鼠(KO组), 其对照组为野生型小鼠(WT组)。通过夹闭左侧肾蒂构建单侧肾脏缺血再灌注(unilateral renal ischemia reperfusion, UIR)模型(UIR组), 造模后分为KO-UIR组和WT-UIR组;假手术小鼠(Sham组)不进行缺血处理。采用肌氨酸氧化酶法检测血清肌酐水平;二乙酰肟比色法检测血尿素氮水平;酶联免疫吸附测定法检测尿白蛋白水平, 计算尿白蛋白/肌酐比值;HE、PAS及MASSON染色检测肾组织炎性细胞浸润情况和纤维化程度;实时荧光定量PCR检测Ildr2, 肾损伤相关分子中性粒细胞明胶酶相关载脂蛋白(neutrophil gelatinase-associated lipocalin, NGAL)和肾损伤分子1(kidney injury molecule-1, KIM-1), 纤维化标志分子Ⅰ型胶原α1(...     

IL-17C影响小鼠移植肾存活的机制

目的探讨白细胞介素(IL)-17C在小鼠肾移植中的作用及其机制。方法以Balb/c(H-2Kd小鼠为供体,IL-17C基因敲除(IL-17CKO)小鼠(敲除组)、C57BL/6J(H-2Kb)小鼠(野生组)为受体,建立小鼠生命支持型肾移植模型。术后比较两组小鼠的体质量及存活时间。采用苏木素-伊红(HE)染色及过碘酸-雪夫(PAS)染色对移植肾进行病理学检查。采用逆转录聚合酶链反应(RT-PCR)检测移植肾组织中颗粒酶B、干扰素(IFN)-γ、肿瘤坏死因子(TNF)-α、IL-6及IL-1β的信使核糖核酸(mRNA)表达水平,采用流式细胞术检测移植肾组织中炎症细胞浸润情况。结果移植术后敲除组小鼠存活时间显著短于野生组小鼠(P=0.031),且体质量下降程度更明显,但差异无统计学意义。病理分析发现敲除组小鼠移植肾损伤较野生组小鼠明显加重。敲除组小鼠移植肾组织中颗粒酶B、IFN-γ、TNF-α、IL-6的mRNA表达水平均显著高于野生组(均为P0.01),IL-1βmRNA表达呈降低趋势,但差异无统计学意义(P=0.16)。流式细胞分析发现敲除组小鼠移植肾组织中CD45+CD11b+Ly6G+中性粒细胞和CD45+CD11b+Ly6Chi单核细胞浸润较野生组小鼠明显增加(分别为P0.05,P0.01),而CD45+Ly6Chi F4/80+巨噬细胞浸润无明显变化(P0.05)。结论 IL-17C参与肾移植后炎症反应的调控,可能通过降低促炎细胞因子的表达及炎症细胞的浸润来减轻急性排斥反应,改善移植肾存活情况。     

Nrf2基因敲除小鼠脊髓损伤后脊髓内TNF-α及MMP-9的变化

目的 探讨转录因子NF-E2相关因子2(Nrf2)对小鼠脊髓损伤后脊髓内肿瘤坏死因子-α(TNF-α)及基质金属蛋白酶-9(MMP-9)表达的影响.方法 分别利用野生型Nrf2(+/+)小鼠及Nrf2(-/-)基因敲除小鼠制作脊髓钳夹损伤模型.在小鼠脊髓损伤后48 h用干湿比法测定脊髓水肿程度.在脊髓损伤后24 h,用逆转录聚合酶链式反应(RT-PCR)法测定脊髓组织MMP-9 mRNA含量的变化;酶联免疫吸附试验测定脊髓组织中TNF-α及MMP-9蛋白的含量;明胶酶谱法测定脊髓组织中MMP-9的酶活性.用SPSS 16.0软件进行统计学分析.结果 脊髓损伤后48 h,Nrf2(-/-)基因敲除小鼠及Nrf2(+/+)小鼠的脊髓均出现明显水肿,但Nrf2(-/-)基因敲除小鼠尤为严重(P＜0.05).脊髓损伤后24 h,Nrf2(-/-)基因敲除小鼠及Nrf2(+/+)小鼠的脊髓内TNF-α与MMP-9的表达均增加,但Nrf2(-/-)基因敲除小鼠尤为显著(P＜0.01).同时,Nrf2(-/-)基因敲除小鼠在脊髓损伤后24 h,脊髓内MMP-9明胶酶活性较Nrf2(+/+)小鼠的损伤组明显增强(P＜0.01).结论 Nrf2在小鼠脊髓损伤后可能是通过减少炎性介质TNF-α及MMP-9的表达,降低MMP-9的明胶酶活性来减轻炎症反应,减轻脊髓水肿程度,起到神经保护的作用.     

尿毒清颗粒对糖尿病大鼠肾脏炎症损伤的影响

目的探讨尿毒清颗粒对糖尿病大鼠肾脏炎症损伤的保护作用。方法选用链脲佐菌素（STZ）诱导的糖尿病大鼠模型。实验分3组：正常对照组（N组）、糖尿病未干预组（D组）、尿毒清颗粒治疗组（Q组,2．6g·kg-1·d-1灌胃）,于第4、8周末检测血肌酐（SCr）、尿肌酐（Ucr）,计算肌酐清除率（Ccr）,检测血清超敏C反应蛋白（hs—CRP）、肿瘤坏死因子α（TNF-α）、白细胞介素6（IL-6）,光镜观察肾组织病理改变,免疫组织化学检测肾组织核因子xB（NF-kB）、单核细胞趋化蛋白1（MCP-1）表达,检测肾皮质MCP-1mRNA表达。结果尿毒清颗粒可以提高糖尿病大鼠Ccr（P〈0．01）,改善肾脏病理损伤,减少血清hs-CRP、肾组织NF-kB、MCP-1表达（P〈0．1）5,P〈0．01）,对血清TNF-α、IL-6影响不显著。结论炎症反应参与糖尿病大鼠肾脏损伤,尿毒清颗粒能降低血清及肾脏炎症介质表达,对糖尿病大鼠肾脏炎症损伤起保护作用。     

虫草菌丝对5/6肾切除大鼠肾组织纤维化保护作用的研究

目的研究虫草菌丝对于改善5/6肾切除慢性肾衰大鼠肾组织纤维化的保护作用。方法按经典5/6(Ablation/Infarction,A/I)肾切除法制作慢性肾衰大鼠模型,随机分为模型组、虫草组,另设假手术组。给予相应干预,60 d后检测大鼠血红蛋白、肾功能等生化指标,测定尾动脉血压,病理切片HE染色观察肾脏组织情况,Western blot法检测肾组织肿瘤坏死因子α(tumor necrosis factors-α,TNF-α)蛋白表达,实时定量PCR检测残余肾组织肾皮质和髓质中TNF-αmRNA表达。结果虫草组的血肌酐、血尿素氮较治疗前明显降低(P0.01),内生肌酐清除率明显升高(P0.01)。虫草组降低血肌酐、血尿素氮水平,升高内生肌酐清除率水平均优于模型组(P0.01)。与模型组比较,虫草组的血红蛋白值明显提高(P0.01),收缩压明显下降(P0.01),舒张压有所下降(P0.05)。肾组织病理显示,虫草组病理变化明显减轻,优于模型组。与假手术组比较,在模型组和虫草组皮质和髓质组织中TNF-α的蛋白表达、TNF-αmRNA表达均明显升高(P0.01);与模型组比较,虫草组的皮质和髓质组织中TNF-α的蛋白表达TNF-αmRNA均明显减少(P0.01)。结论虫草菌丝能够改善慢性肾衰大鼠肾功能,减轻贫血,改善肾组织病理变化。这一作用机制可能与虫草菌丝抑制TNF-α生成、减轻慢性肾衰大鼠组织的微炎症状态有关。     

慢性肾盂肾炎肾间质纤维化与雌激素及其受体相关性的实验研究

目的：探讨雌激素及其受体与慢性肾盂肾炎肾间质纤维化的有相关性。方法：应用雌性大鼠卵巢切除（去势）后和/或大肠杆菌由膀胱经不完全梗阻输尿管逆行感染制作慢性肾盂肾炎肾间质纤维化动物模型,观察正常或去势后、或合并慢性肾盂肾炎肾间质纤维化、及用氟哌酸或/和倍美力治疗后的大鼠雌激素受体在肾脏组织中的表达,及α-SMA及TGF-β在肾脏组织中表达的改变,探讨雌激素、雌激素受体与慢性肾盂肾炎、肾间质纤维化的关系。结果：去势后慢性肾盂肾炎组大鼠尿蛋白定量有所升高。去势、慢性肾盂肾炎均造血清雌激素水平降低,氟哌酸、倍美力、氟哌酸加倍美力治疗后血清雌激素水平均有不同程度的升高。肾脏ER-β主要定位于肾小管上皮细胞、集合管上皮细胞的胞浆。单纯去势组、去势后慢性肾盂肾炎大鼠肾脏ER-β较正常组升高。与正常组比较,慢性肾盂肾炎组、去势后慢性肾盂肾炎组大鼠肾脏表达间质纤维化标志蛋白α-SMA及TGF-β1明显增高,肾间质纤维化明显,氟哌酸加倍美力组可降低α-SMA的表达,缓解肾间质纤维化。血清E2与肾脏ER-β呈负相关,与肾间质纤维化指标α-SMA、TGF-β1负相关,肾脏ER-β与TGF-β1有一定的正相关,而与α-SMA无明显关系。结论：雌激素及其受体与慢性肾盂肾炎肾间质纤维化具有相关性。     

清肝益肾祛风复方抗高血压肾损伤作用及机制研究

目的:探究清肝益肾祛风方对自发性高血压大鼠的降压、抗高血压肾损伤作用及其可能的作用机制,以期为临床治疗高血压肾损伤提供科学依据。方法:采用4周龄雄性SHR大鼠作为高血压模式动物,适应性饲养一周后随机分为SHR模型组、坎地沙坦治疗组,清肝益肾祛风方(QYQ)治疗组。正常对照组选用同周龄雄性WKY大鼠。从6周龄开始,QYQ治疗组和坎地沙坦治疗组每天分别给予2 ml的清肝益肾祛风方水煎液和坎地沙坦悬液灌胃。SHR组和WKY组采用同等体积的纯水做为对照,连续灌胃至20周龄。采用小动物无创血压计每2周测量一次血压;连续给药至20周龄后,10%水合氯醛腹1腔注射麻醉、腹主动脉取血后处死动物。留取双侧肾脏、采用离心机分离血清。左肾脱水固定包埋后做石蜡切片,Masson’s trichrome染色观察左侧肾脏组织病理变化;采用全自动生化检测仪检测大鼠血清中尿素、肌酐、尿酸水平; Realtime PCR方法检测右侧肾脏组织氧化应激标志物相关基因的mRNA表达水平、炎症因子基因的mRNA表达水平及胶原相关基因的mRNA表达水平。结果:与WKY组相比,SHR组大鼠血压从6周龄开始显著升高(P0.05),从12周龄开始达到稳定;与SHR组相比较,清肝益肾祛风方与坎地沙坦均能显著降低SHR大鼠血压(P0.05); Masson’s trichrome染色结果:WKY组肾脏组织未见病理损伤,SHR组大鼠肾脏组织出现较大面积纤维化。与SHR组相比,QYQ治疗组与坎地沙坦治疗组肾脏组织纤维化面积均显著降低(P0.05);生化检测结果提示:与WKY组相比,SHR组血清中尿酸、尿素含量显著升高(P0.05)。与SHR相比较,QYQ治疗组血清尿酸含量显著降低(P0.05)。Real-time PCR结果:与WKY组相比,SHR组肾脏组织氧化应激相关基因mRNA表达水平显著升高(P0.05),IL-23,TNF-α及TGF-β1等炎症因子相关基因的mRNA表达水平显著升高(P0.05),IL-10基因的mRNA表达水平显著降低(P0.05),胶原相关基因的mRNA表达水平显著上升(P0.05)。与SHR组相比,QYQ组肾脏组织氧化应激相关基因mRNA表达水平显著降低(P0.05),IL-23,TNF-α等炎症因子相关基因的mRNA表达水平显著降低(P0.05),IL-10的mRNA表达水平显著升高(P0.05),Col5a2及α-SMA基因的mRNA表达水平显著降低(P0.05)。结论:清肝益肾祛风方能显著降低SHR大鼠血压、显著减轻SHR大鼠肾脏组织纤维化,具有降压、抗高血压肾损伤的作用,其作用机制可能与减轻肾脏组织氧化应激反应、炎症反应及减少肾组织胶原纤维合成相关。     

淫羊藿苷对急性肾损伤向慢性肾脏病转化小鼠PPAR信号通路影响的研究

目的:探讨淫羊藿苷是否具有缓解缺血再灌注诱导急性肾损伤向慢性肾脏病转化的作用以及对PPAR信号通路的影响。方法:选用SPF级10周龄雄性C57BL/6小鼠随机分为3组,对照组(Control)、模型组(IRI)、模型+250 mg/kg淫羊藿苷组(IRI+I),每组6只,4周后小鼠处死,行病理染色,RT-PCR检测肾小管损伤和肾纤维化指标以及PPAR信号通路相关基因的变化。结果:Masson染色显示,IRI模型组肾间质纤维化严重,淫羊藿苷组与模型组相比肾间质纤维化程度明显改善(P<0.05),同时其肾损伤评分较模型组也有不同程度的改善(P<0.05);实时定量PCR显示,IRI组与Control组相比肾小管损伤、肾纤维化及炎症等基因Havcr-1、 FN、Col3a1、TGF-β₁、TNF-α表达量均增加(P<0.05),IRI+I组与IRI组相比肾小管损伤、肾纤维化及炎症等基因Havcr-1、FN、Col3a1、TGF-β₁、TNF-α表达量显著减少(P<0.05);与对照组相比,模型组Pparα mRNA、Cpt1...     

Effect of SOST gene deletion on the progression of renal interstitial fibrosis in obstructive kidney injury

Abstract

Objectives: The role of SOST/sclerostin in mediating tissue fibrogenic response to injury/inflammation remains largely unknown. Thus, we conducted this study to determine whether SOST/sclerostin plays a role in renal interstitial fibrosis (RIF) for the first time. Methods: Unilateral ureteral obstruction (UUO) was performed to create obstructive kidney injury model. Twelvemale SOST knockout (SOST KO) mice and 12 age-matched wild-type (WT) mice were divided into three groups: sham surgery, UUO 3?d and UUO 7?d. The mice were sacrificed at each time point and kidney tissues were collected. Histopathological changes were evaluated by hematoxylin and eosin and Masson staining, while α-smooth muscle actin (α-SMA), type I collagen (Col-I) and fibronectin (FN) expression levels were detected by RT-PCR and western-blot. Results: In sham control group, neither WT nor SOST KO exhibited fibrotic change. On 3 days after UUO, total renal histopathological score and fibrotic area were aggravated and α-SMA, Col-I and FN expressions were upregulated, but no difference was observed between WT and SOST KO. On 7 days after UUO, compared with WT, SOST KO mice showed higher total renal histopathological score and fibrotic area percentage, as well as a higher level of fibrogenic marker mRNA/protein expression (except for α-SMA mRNA and FN mRNA). Conclusion: It is supposed that SOST gene is involved in the regulation of RIF progression. In obstructive kidney injury, SOST gene deletion would probably enhance renal fibrogenic response and promote the progression of RIF. But more evidences are needed to further identify the role of SOST/sclerostin in mediating RIF progression.     

巨噬细胞Bruton酪氨酸激酶基因特异性敲除减轻糖尿病小鼠肾脏损害的作用及机制

目的探讨巨噬细胞Bruton酪氨酸激酶(Btk)基因特异性敲除对糖尿病小鼠肾脏损害的作用及机制。方法选用巨噬细胞Btk基因特异性敲除(Btk-/-)小鼠和C57BL/6N小鼠链脲菌素(STZ,50 mg/kg)造模成功后随机分为正常组、糖尿病组、Btk-/-组和Btk-/-糖尿病组。12周后测定小鼠一般指标,观察肾组织病理学改变,免疫荧光检测肾组织巨噬细胞标志物CD68表达,免疫组化检测足细胞标志物WT1和Nephrin的表达,Western印迹检测细胞外基质纤维连接蛋白(FN)、IV型胶原(IV-Col)及转化生长因子β1(TGF-β1),巨噬细胞激活标志物诱导型一氧化氮合酶(iNOS)、磷酸化(p)-Btk,炎性因子白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)和丝裂原活化蛋白激酶(MAPK)信号通路p-p38MAPK、p-JNK、p-ERK以及核因κB(NF-κB)通路p-p65、p-IκB蛋白水平变化;实时荧光定量PCR检测炎性因子IL-1β、TNF-α、单核细胞趋化蛋白1(MCP-1)mRNA表达。结果与糖尿病组相比,Btk-/-糖尿病组尿白蛋白明显减少,肾组织损伤明显减轻,肾脏巨噬细胞CD68表达明显减少,足细胞标志物WT1及Nephrin表达明显增加,细胞外基质FN、IV-Col及TGF-β1表达明显减少,炎性因子IL-1β、TNF-α及MCP-1表达明显降低,p-JNK、p-ERK、p-p38MAPK及p-p65、p-IκB表达明显下调(均P<0.05)。结论巨噬细胞Btk特异性敲除可能通过MAPK、NF-κB通路降低炎性因子的表达从而对糖尿病小鼠肾脏起到保护作用。     

AMPK attenuates inflammation to reduce fibrosis induced by acute ischemia reperfusion injury in mice

Objective To observe the effect of adenosine monophosphate activated protein kinase (AMPK) on attenuating inflammation in fibrosis induced by acute ischemia reperfusion injury (IRI) in mice. Methods Forty eight male C57BL/6 mice were randomly divided into four groups: sham operation group (sham group), IRI group, AMPK inhibitor+IRI group (AMPK/IRI group) and normal saline+IRI group (NS/IRI group), 12 mice each group. The mice with renal IRI were occluded for 30 min through clipping bilateral renal pedicle, then released renal perfusion. Mice in sham group were performed the separation of renal pedicle without clipping. Mice in AMPK/IRI group and NS/IRI group were respectively intraperitoneal injected AMPK inhibitor and normal saline before IRI. At the 2 d after operation, 6 randomly-selected mice from each group were blooded by extraction eyeball to detect BUN and Scr. The renal histopathological changes were observed through HE staining. The mRNA expression of IL-1β, IL-6 and TNF-α was detected by real time PCR, and the level of AMPK phosphorylation was detected by Western blotting. At the 14 d after operation, Collagen 1 (COL1), α-SMA and fibronectin (FN) were detected by immunofluorescence and Western blotting in 6 remained mice from each group. The degree of kidney fibrosis was observed through sirus red staining. Results Compared with those in sham group, tubular interstitial damage was aggravated (P＜0.05), BUN and Scr were increased (P＜0.05), the mRNA expression of IL-1β, IL-6 and TNF-α was increased at the 2 d after operation (all P＜0.05), and the level of AMPK phosphorylation was activated in IRI group and NS/IRI group (all P＜0.05); the degree of kidney fibrosis and the expression of COL1, α-SMA and FN were increased obviously at the 14 d (all P＜0.05). Compared with those in IRI group, in AMPK/IRI group tubular interstitial damage was aggravated (P＜0.05), BUN and Scr were increased (all P＜0.05), the mRNA expression of IL-1β, IL-6 and TNF-α was increased at the 2 d (all P＜0.05), and the level of AMPK phosphorylation was decreased (P＜0.05). Moreover, the degree of kidney fibrosis and the expression of COL1, α-SMA and FN were increased obviously at the 14 d in AMPK/IRI group (all P＜0.05). Conclusions AMPK can ameliorate the acute renal ischemia reperfusion injury induce fibrosis in mice, and the mechanism may be related to the decrease of inflammatory reaction.     

Effect of Chenodeoxycholic Acid on Fibrosis, Inflammation and Oxidative Stress in Kidney in High-Fructose-Fed Wistar Rats

Background: Recent studies indicate farnesoid X receptor (FXR) plays an important role in regulating lipid metabolism in kidney disease. The purpose of the present study is to investigate the effect of chenodeoxycholic acid (CDCA), a FXR agonist, on fibrosis, inflammation and oxidative stress in kidney in rats fed on high fructose. Methods: Twenty-four healthy male Wistar rats were randomly divided into three groups (n=8): normal control group, high fructose group and chenodeoxycholic acid group. Rats were sacrificed by the end of 16 weeks after feeding. Blood urea nitrogen, serum creatinine, fast glucose, lipid concentration were observed, spot urine samples were obtained to measure the albumin and creatinine levels. Triglyceride of renal cortices was detected. The mRNA level and protein contents of the fibrosis-inducing growth factor transforming growth factor β1 (TGF-β1) and plasminogen activator inhibitor (PAI-I), inflammatory cytokines tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6), oxidative stress index NADPH oxidase 2 (Nox2) and p22phox in kidney were examined. The pathological changes of kidney were examined by PAS staining and immunohistochemical staining. Electron microscope sections were made to measure glomerular basement membrane (GBM) width. Results: Renal injuries including mesangial expansion, GBM thickness and podocyte foot process effacement were found in fructose-fed Wistar rats, FXR agonist CDCA modulates renal lipid metabolism, decreases proteinuria and improves renal fibrosis, inflammation and oxidation stress. High-fructose-feeding may cause lipid nephrotoxicity through down-regulated farnesoid X receptor and increases expression of profibrotic growth factors, proinflammatory cytokines, and oxidative stress in Wistar rats. Conclusion: FXR activation by chenodeoxycholic acid can prevent the injury in kidney induced by high fructose feeding.     

Genetic or pharmacologic blockade of EGFR inhibits renal fibrosis

Although enhanced activation of the EGF receptor (EGFR) associates with the development and progression of renal fibrosis, the mechanisms linking these observations are not completely understood. Here, after unilateral ureteral obstruction (UUO), wild-type mice exhibited sustained EGFR phosphorylation in the kidney and developed renal fibrosis that was more severe than the renal fibrosis observed in waved-2 mice, which have reduced EGFR tyrosine kinase activity. Waved-2 mice also showed fewer renal tubular cells arrested at G2/M, reduced expression of α-smooth muscle actin (α-SMA), downregulation of multiple genes encoding profibrogenic cytokines, including TGF-β1, and dephosphorylation of Smad3, STAT3, and ERK1/2. Administration of the specific EGFR inhibitor gefitinib recapitulated this phenotype in wild-type mice after UUO. Furthermore, inactivation of either EGFR or STAT3 reduced UUO-induced expression of lipocalin-2, a molecule associated with the pathogenesis of CKD. In cultured renal interstitial fibroblasts, inhibition of EGFR also abrogated TGF-β1- or serum-induced phosphorylation of EGFR, STAT3, ERK1/2, and Smad3 as well as expression of α-SMA and extracelluar matrix proteins. Taken together, these data suggest that EGFR may mediate renal fibrogenesis by promoting transition of renal epithelial cells to a profibrotic phenotype, increased production of inflammatory factors, and activation of renal interstitial fibroblasts. Inhibition of EGFR may have therapeutic potential for fibrotic kidney disease.     

Wilms瘤1基因在肾小管上皮细胞转分化中的表达及作用

目的 探讨Wilms 肿瘤1基因(WT1)在肾小管上皮细胞(TEC)向肌成纤维细胞转分化中的可能作用&#65377;方法 将体外原代培养TEC分别置含IL-1α(10 ng/ml)&#65380; IL-1α+抗WT1中和抗体(10 μg/ml)的DMEM/F12培养基中培养, 观察TEC的形态变化及免疫学特征&#65377;采用RT-PCR检测各组TEC中WT1及α-SMA的表达&#65377;结果 经IL-1α掺入培养5 d后,体外培养的TEC形态特征趋于成纤维细胞化,细胞拉长&#65380;梭形变,失去原有的呈铺路石样的生长方式&#65377;电镜下细胞极性丧失,表面微绒毛消失&#65377;正常情况下,成年TEC不表达WT1&#65377;经IL-1α掺入培养1 d后,TEC重新表达WT1,同时伴有α-SMA的表达;3 d后WT1表达消失,呈一过性特征,而α-SMA的表达则随时间的延长而逐渐增强&#65377;在培养中掺入抗WT1抗体以中和WTl基因产物后,尽管仍给予IL-1α刺激,TEC大都保持原有特征不变,α-SMA及WT1 mRNA仅呈微弱表达&#65377;结论 高浓度IL-1α可导致TEC向肌成纤维细胞的转分化&#65377;在TEC的转分化过程中,WT1基因的重新&#65380;一过性表达可能起着重要作用&#65377;成年TEC重新获得WT1表达,可能是其发生转分化的内在启动机制&#65377;体外中和WT1的基因产物可明显抑制TEC的转分化进程,并可能籍此影响肾脏的纤维化发生&#65377;     

Erbin在肾间质纤维化中的表达和作用

目的 探讨Erbin在肾脏间质纤维化中表达量的变化及上调Erbin对转化生长因子β1（TGF-β1）诱导大鼠近端肾小管上皮细胞（NRK52E）转分化的影响。 方法 体内实验采用SD大鼠5/6肾切除法建立肾纤维化动物模型,收集并检测各组血清中Scr、BUN水平;Masson染色观察肾间质纤维化程度;免疫组化及Western印迹检测Erbin的分布与表达。 体外实验采用TGF-β1（10 μg/L）刺激NRK52E细胞72 h建立上皮细胞-间充质转分化（EMT）细胞模型;免疫荧光及Western印迹法检测E钙黏蛋白（E-cadherin）和α平滑肌肌动蛋白（α-SMA）的表达变化;RT-PCR及Western 印迹法检测Erbin的表达变化。用脂质体2000将质粒 Prk5-myc-Erbin瞬时转染至NRK52E细胞,Western印迹法观察上调Erbin表达后对上述各种指标的影响。 结果 （1）假手术组大鼠肾功能正常[Scr（33.96±7.28） μmol/L、BUN（8.11±2.55） mmol/L],Masson染色未见肾间质纤维化,Erbin在肾小管表达较少;模型组大鼠Scr [（140.52±61.11） μmol/L]、BUN[（34.23±7.66） mmol/L] 均显著高于假手术组（均P < 0.05）,肾间质可见明显纤维化,Erbin 在肾小管表达也明显增加,是假手术组的2.9倍（P < 0.01）。（2）正常NRK52E 细胞表达E-cadherin,少量表达Erbin和α-SMA。TGF-β1刺激后,NRK52E细胞E-cadherin表达显著减少,Erbin和α-SMA则表达增加（均P < 0.05）;而转染质粒Prk5-myc-Erbin可逆转TGF-β1诱导的NRK52E细胞E-cadherin表达下调,并可抑制α-SMA表达上调（均P < 0.05）。 结论 Erbin在肾间质纤维化中表达增加,上调Erbin表达可抑制TGF-β1诱导NRK52E发生EMT, 提示Erbin在肾脏纤维化中可发挥保护作用。