转化生长因子β1对人肾间质成纤维细胞组织型转谷氨酰胺酶表达的影响

近年来组织型转谷氨酰胺酶（tTG）在组织纤维化的发生和发展中的作用受到了关注。本实验通过观察TGF-β1对体外培养的人。肾间质成纤维细胞tTG表达的影响，探讨TGF-β1与tTG的相互关系及其在。肾问质纤维化中可能的作用。     

组织型转谷氨酰胺酶与肾间质纤维化的研究现状

业已证实，肾问质纤维化（RIF）是各种慢性肾脏疾病进展至终末期肾衰竭的共同病变过程，且其对肾功能的影响较肾小球病变更为重要。对于细胞外基质（ECM）合成和降解失衡导致RIF这一共识问题，既往大量工作着重于降解ECM成分的主要蛋白酶一基质金属蛋白酶的调控研究，近年来，ECM蛋白翻译后被绞联修饰，从而抵抗蛋白酶降解，造成ECM过度沉积这一现象已引起重视。其中组织型转谷氨酰胺酶（tissue transglutarninase，tTG）是一种Ca^2＋依赖的具有转酰胺基作用的酶，它分布广泛，在许多生理和病理条件下均发挥重要作用。tTG分布到细胞外能使很多ECM蛋白成分之间发生绞联，形成稳固结构而抵抗降解；同时在动物模型[2-4J和肾病患者的组织标本中均发现RIF部位tTG蛋白的量和活性明显增加，因此tTG被认为是参与组织纤维化的重要分子。目前有关tTG与RIF的研究尚处于起始阶段，本文就tTG对RIF的影响及其可能机制作一简要综述，以飨同道。     

组织转谷氨酰胺酶特异性抑制剂预防肝纤维化的实验研究

目的 观察组织转谷氨酰胺酶(tTG)特异性抑制剂六甲烯胺对四氯化碳(CCl4)诱导的大鼠肝纤维化形成的影响.方法 雄性SD大鼠100只,按随机数字表法分成正常对照组(20只)、肝纤维化组(40只)、六甲烯胺组(40只).用腹腔内注射CCl4诱导大鼠肝纤维化模型,在注射CCl4的前2 d开始腹腔内注射六甲烯胺(112 mg·kg-1·d-1),直到注射CCl4的第4、8周处死大鼠,摘取肝脏并提取血清,应用RT-PCR检测tTG、α-平滑肌抗体(α-SMA)、Ⅰ型胶原蛋白和基质金属蛋白酶抑制剂-1(TIMP-1)等基因的mRNA表达情况,应用Western blot检测tTG和α-SMA蛋白的表达,肝功能和肝脏组织中羟脯氨酸的含量,同时取大鼠肝脏行组织学检查,并对肝纤维化程度进行半定量计分系统分级评分.采用单因素方差分析检测结果.结果 CCl4注射8周后,肝纤维化组肝损伤明显,ALT、总胆汁酸、Tbil、羟脯氨酸、tTG、α-SMA、Ⅰ型胶原蛋白酶、TIMP-1分别为(1313±157)U/L、(99.9±18.5)μmol/L、(10.9±1.6)μmol/L、(55±12)μg/g、145.6±51.2、130.3±44.6、211.3±75.1、162.4±53.5;而六甲烯胺处理后,肝纤维化程度、肝功能指标均显著降低,ALT、总胆汁酸、Tbil、羟脯氨酸、tTG、α-SMA、Ⅰ型胶原蛋白酶、TIMP-1分别为(378±87)U/L、(61.0±12.7) μmol/L、(9.8±1.7)μmol/L、(70±14)μg/g、48.6±12.3、40.7±12.3、63.9±16.0、59.2±23.4.结论 六甲烯胺能通过抑制tTG途径显著改善CCl4诱导的大鼠肝纤维化程度.     

转谷氨酰胺酶4在前列腺癌进展及预后中的作用

转谷氨酰胺酶4（TGM4）在前列腺癌组织和正常组织中差异表达，且与前列腺癌细胞的多种生物学特性相关，如前列腺癌细胞间质化，细胞侵袭性和细胞基质粘附等。本研究主要探索TGM4表达水平与前列腺癌病理特征及前列腺癌生化复发之间的关系。取160名接受前列腺癌根治术患者的石蜡标本构建组织芯片，进而利用组织芯片行TGM4免疫组织化学染色；TGM4的表达情况采用染色范围与染色强度相结合的方式来评价。通过查阅病历获得患者的临床及病理信息；随访信息通过查阅病历、调用我院前列腺癌随访数据库和电话随访等多种方式获得。进而，我们比较了TGM4在癌和癌旁组织中的表达差异情况，及TGM4不同表达水平与前列腺癌病理特征和生化复发之间的关系。同癌旁组织相比，TGM4在前列腺癌组织中明显高表达（P〈0．001），并且其表达水平与Gleason评分（P〈0．001）和PSA水平（P=0．005）呈明显正相关。单变量分析提示TGM4高表达是前列腺癌生化复发的高危因素（P=0．042），但多变量分析并不支持TGM4高表达是前列腺癌生化复发的高危因素（P=0．139）。TGM4在前列腺癌中高表达，其表达水平与Gleason评分和PSA水平呈正相关，TGM4高表达可能作为前列腺癌生化复发的潜在预测因子。     

组织转谷氨酰胺酶特异性抑制剂预防肝纤维化的实验研究

Objective To observe the effects of cystamine, a tissue transglutaminase (tTG) inhibitor, on the development of rat liver fibrosis induced by carbon tetrachloride. Methods One hundred male SD rats were randomly divided into control group (n=20), hepatic fibrosis group (n=40) and cystamine group (n=40) . Liver fibrosis model was induced by intraperitoneal injection of carbon tetrachloride. Cystamine (112 mg·kg-1·d-1) was administered by intraperitoneal injection 2 days before injection of carbon tetrachloride. The rats were sacrificed at weeks 4 and 8, and the liver tissues and serum specimens were obtained. The mRNA expression of tTG, smooth muscle-alpha (α-SMA), collagen-Ⅰ and tissue inhibitors of metalloproteinase-1 (TIMP-1) were detected by real time PCR. The protein expression of tTG and α-SMA, liver function and content of hydroxyproline in liver tissues were determined by Western blot. Histological changes of the liver was observed under microscope. The fibrosis conditions of rat liver in each group were evaluated according to the semi-quantita-tive scoring system. All the data were analyzed by one-way ANOVA. Results Eight weeks after the injection of carbon tetrachloride, obvious injury of the liver in liver fibrosis group was observed. The levels of alanine trans-aminase (ALT), total bile acid, total bilirubin, hydroxyproline, tTG, α-SMA, collagen-Ⅰ and TIMP-1 were (1313±157)U/L, (99.9±18.5)μmol/L, (10.9±1.6)μmoL/L, (55±12)μg/g, 145.6±51.2, 130.3±44.6, 211.3±75.1 and 162.4±53.5. After administration of cystamine, the levels of ALT, total bile acid, total bilirubin, hydroxyproline, tTG, α-SMA, collagen-Ⅰ and TIMP-1 were (378±87) U/L, (61.0±12.7) μmol/L, (9.8±1.7) μmol/L, (70±14 ) μg/g, 48.6±12.3, 40.7±12.3, 63.9±16.0, 59.2μ23.4. Conclusion Cystamine can alleviate the carbon tetrachloride-induced rat liver fibrosis by inhibiting the tTG pathway.     

组织转谷氨酰胺酶特异性抑制剂预防肝纤维化的实验研究

Objective To observe the effects of cystamine, a tissue transglutaminase (tTG) inhibitor, on the development of rat liver fibrosis induced by carbon tetrachloride. Methods One hundred male SD rats were randomly divided into control group (n=20), hepatic fibrosis group (n=40) and cystamine group (n=40) . Liver fibrosis model was induced by intraperitoneal injection of carbon tetrachloride. Cystamine (112 mg·kg-1·d-1) was administered by intraperitoneal injection 2 days before injection of carbon tetrachloride. The rats were sacrificed at weeks 4 and 8, and the liver tissues and serum specimens were obtained. The mRNA expression of tTG, smooth muscle-alpha (α-SMA), collagen-Ⅰ and tissue inhibitors of metalloproteinase-1 (TIMP-1) were detected by real time PCR. The protein expression of tTG and α-SMA, liver function and content of hydroxyproline in liver tissues were determined by Western blot. Histological changes of the liver was observed under microscope. The fibrosis conditions of rat liver in each group were evaluated according to the semi-quantita-tive scoring system. All the data were analyzed by one-way ANOVA. Results Eight weeks after the injection of carbon tetrachloride, obvious injury of the liver in liver fibrosis group was observed. The levels of alanine trans-aminase (ALT), total bile acid, total bilirubin, hydroxyproline, tTG, α-SMA, collagen-Ⅰ and TIMP-1 were (1313±157)U/L, (99.9±18.5)μmol/L, (10.9±1.6)μmoL/L, (55±12)μg/g, 145.6±51.2, 130.3±44.6, 211.3±75.1 and 162.4±53.5. After administration of cystamine, the levels of ALT, total bile acid, total bilirubin, hydroxyproline, tTG, α-SMA, collagen-Ⅰ and TIMP-1 were (378±87) U/L, (61.0±12.7) μmol/L, (9.8±1.7) μmol/L, (70±14 ) μg/g, 48.6±12.3, 40.7±12.3, 63.9±16.0, 59.2μ23.4. Conclusion Cystamine can alleviate the carbon tetrachloride-induced rat liver fibrosis by inhibiting the tTG pathway.     

组织转谷氨酰胺酶特异性抑制剂预防肝纤维化的实验研究

Objective To observe the effects of cystamine, a tissue transglutaminase (tTG) inhibitor, on the development of rat liver fibrosis induced by carbon tetrachloride. Methods One hundred male SD rats were randomly divided into control group (n=20), hepatic fibrosis group (n=40) and cystamine group (n=40) . Liver fibrosis model was induced by intraperitoneal injection of carbon tetrachloride. Cystamine (112 mg·kg-1·d-1) was administered by intraperitoneal injection 2 days before injection of carbon tetrachloride. The rats were sacrificed at weeks 4 and 8, and the liver tissues and serum specimens were obtained. The mRNA expression of tTG, smooth muscle-alpha (α-SMA), collagen-Ⅰ and tissue inhibitors of metalloproteinase-1 (TIMP-1) were detected by real time PCR. The protein expression of tTG and α-SMA, liver function and content of hydroxyproline in liver tissues were determined by Western blot. Histological changes of the liver was observed under microscope. The fibrosis conditions of rat liver in each group were evaluated according to the semi-quantita-tive scoring system. All the data were analyzed by one-way ANOVA. Results Eight weeks after the injection of carbon tetrachloride, obvious injury of the liver in liver fibrosis group was observed. The levels of alanine trans-aminase (ALT), total bile acid, total bilirubin, hydroxyproline, tTG, α-SMA, collagen-Ⅰ and TIMP-1 were (1313±157)U/L, (99.9±18.5)μmol/L, (10.9±1.6)μmoL/L, (55±12)μg/g, 145.6±51.2, 130.3±44.6, 211.3±75.1 and 162.4±53.5. After administration of cystamine, the levels of ALT, total bile acid, total bilirubin, hydroxyproline, tTG, α-SMA, collagen-Ⅰ and TIMP-1 were (378±87) U/L, (61.0±12.7) μmol/L, (9.8±1.7) μmol/L, (70±14 ) μg/g, 48.6±12.3, 40.7±12.3, 63.9±16.0, 59.2μ23.4. Conclusion Cystamine can alleviate the carbon tetrachloride-induced rat liver fibrosis by inhibiting the tTG pathway.     

组织转谷氨酰胺酶特异性抑制剂预防肝纤维化的实验研究

Objective To observe the effects of cystamine, a tissue transglutaminase (tTG) inhibitor, on the development of rat liver fibrosis induced by carbon tetrachloride. Methods One hundred male SD rats were randomly divided into control group (n=20), hepatic fibrosis group (n=40) and cystamine group (n=40) . Liver fibrosis model was induced by intraperitoneal injection of carbon tetrachloride. Cystamine (112 mg·kg-1·d-1) was administered by intraperitoneal injection 2 days before injection of carbon tetrachloride. The rats were sacrificed at weeks 4 and 8, and the liver tissues and serum specimens were obtained. The mRNA expression of tTG, smooth muscle-alpha (α-SMA), collagen-Ⅰ and tissue inhibitors of metalloproteinase-1 (TIMP-1) were detected by real time PCR. The protein expression of tTG and α-SMA, liver function and content of hydroxyproline in liver tissues were determined by Western blot. Histological changes of the liver was observed under microscope. The fibrosis conditions of rat liver in each group were evaluated according to the semi-quantita-tive scoring system. All the data were analyzed by one-way ANOVA. Results Eight weeks after the injection of carbon tetrachloride, obvious injury of the liver in liver fibrosis group was observed. The levels of alanine trans-aminase (ALT), total bile acid, total bilirubin, hydroxyproline, tTG, α-SMA, collagen-Ⅰ and TIMP-1 were (1313±157)U/L, (99.9±18.5)μmol/L, (10.9±1.6)μmoL/L, (55±12)μg/g, 145.6±51.2, 130.3±44.6, 211.3±75.1 and 162.4±53.5. After administration of cystamine, the levels of ALT, total bile acid, total bilirubin, hydroxyproline, tTG, α-SMA, collagen-Ⅰ and TIMP-1 were (378±87) U/L, (61.0±12.7) μmol/L, (9.8±1.7) μmol/L, (70±14 ) μg/g, 48.6±12.3, 40.7±12.3, 63.9±16.0, 59.2μ23.4. Conclusion Cystamine can alleviate the carbon tetrachloride-induced rat liver fibrosis by inhibiting the tTG pathway.     

组织转谷氨酰胺酶特异性抑制剂预防肝纤维化的实验研究

Objective To observe the effects of cystamine, a tissue transglutaminase (tTG) inhibitor, on the development of rat liver fibrosis induced by carbon tetrachloride. Methods One hundred male SD rats were randomly divided into control group (n=20), hepatic fibrosis group (n=40) and cystamine group (n=40) . Liver fibrosis model was induced by intraperitoneal injection of carbon tetrachloride. Cystamine (112 mg·kg-1·d-1) was administered by intraperitoneal injection 2 days before injection of carbon tetrachloride. The rats were sacrificed at weeks 4 and 8, and the liver tissues and serum specimens were obtained. The mRNA expression of tTG, smooth muscle-alpha (α-SMA), collagen-Ⅰ and tissue inhibitors of metalloproteinase-1 (TIMP-1) were detected by real time PCR. The protein expression of tTG and α-SMA, liver function and content of hydroxyproline in liver tissues were determined by Western blot. Histological changes of the liver was observed under microscope. The fibrosis conditions of rat liver in each group were evaluated according to the semi-quantita-tive scoring system. All the data were analyzed by one-way ANOVA. Results Eight weeks after the injection of carbon tetrachloride, obvious injury of the liver in liver fibrosis group was observed. The levels of alanine trans-aminase (ALT), total bile acid, total bilirubin, hydroxyproline, tTG, α-SMA, collagen-Ⅰ and TIMP-1 were (1313±157)U/L, (99.9±18.5)μmol/L, (10.9±1.6)μmoL/L, (55±12)μg/g, 145.6±51.2, 130.3±44.6, 211.3±75.1 and 162.4±53.5. After administration of cystamine, the levels of ALT, total bile acid, total bilirubin, hydroxyproline, tTG, α-SMA, collagen-Ⅰ and TIMP-1 were (378±87) U/L, (61.0±12.7) μmol/L, (9.8±1.7) μmol/L, (70±14 ) μg/g, 48.6±12.3, 40.7±12.3, 63.9±16.0, 59.2μ23.4. Conclusion Cystamine can alleviate the carbon tetrachloride-induced rat liver fibrosis by inhibiting the tTG pathway.     

组织转谷氨酰胺酶特异性抑制剂预防肝纤维化的实验研究

Objective To observe the effects of cystamine, a tissue transglutaminase (tTG) inhibitor, on the development of rat liver fibrosis induced by carbon tetrachloride. Methods One hundred male SD rats were randomly divided into control group (n=20), hepatic fibrosis group (n=40) and cystamine group (n=40) . Liver fibrosis model was induced by intraperitoneal injection of carbon tetrachloride. Cystamine (112 mg·kg-1·d-1) was administered by intraperitoneal injection 2 days before injection of carbon tetrachloride. The rats were sacrificed at weeks 4 and 8, and the liver tissues and serum specimens were obtained. The mRNA expression of tTG, smooth muscle-alpha (α-SMA), collagen-Ⅰ and tissue inhibitors of metalloproteinase-1 (TIMP-1) were detected by real time PCR. The protein expression of tTG and α-SMA, liver function and content of hydroxyproline in liver tissues were determined by Western blot. Histological changes of the liver was observed under microscope. The fibrosis conditions of rat liver in each group were evaluated according to the semi-quantita-tive scoring system. All the data were analyzed by one-way ANOVA. Results Eight weeks after the injection of carbon tetrachloride, obvious injury of the liver in liver fibrosis group was observed. The levels of alanine trans-aminase (ALT), total bile acid, total bilirubin, hydroxyproline, tTG, α-SMA, collagen-Ⅰ and TIMP-1 were (1313±157)U/L, (99.9±18.5)μmol/L, (10.9±1.6)μmoL/L, (55±12)μg/g, 145.6±51.2, 130.3±44.6, 211.3±75.1 and 162.4±53.5. After administration of cystamine, the levels of ALT, total bile acid, total bilirubin, hydroxyproline, tTG, α-SMA, collagen-Ⅰ and TIMP-1 were (378±87) U/L, (61.0±12.7) μmol/L, (9.8±1.7) μmol/L, (70±14 ) μg/g, 48.6±12.3, 40.7±12.3, 63.9±16.0, 59.2μ23.4. Conclusion Cystamine can alleviate the carbon tetrachloride-induced rat liver fibrosis by inhibiting the tTG pathway.     

氯沙坦对糖尿病大鼠肾组织TLR4表达的影响

目的观察氯沙坦对糖尿病大鼠肾组织TOLL样受体4（TLR4）表达的影响。方法采用链脲佐菌素（STZ,65mg·kg^-1）腹腔注射建立糖尿病大鼠模型,将其分为对照组、模型组、治疗组。治疗组在造模成功后1周给予氯沙坦20mg·kg^-1灌胃。12周后测3组24h尿蛋白、血肌酐、肾重／体重、血CRP及TNF-α水平;观察肾脏病理形态学变化;应用免疫组化法检测肾组织TLR4、核因子-κB（NF-κB）的蛋白表达。结果模型组24h尿蛋白、血肌酐、尿素氮、肾重／体重明显升高,肾小球肥大、细胞增生,PAS阳性物质沉积增多。血CRP、TNF-α含量及肾组织TLR4、NF-κB的蛋白表达上调。经氯沙坦治疗后,血CRP、TNF-α含量下降,TLR4、NF-κB的表达下调。结论氯沙坦对糖尿病肾病具有保护作用,其作用机制可能是通过下调NF-κB、TLR4的表达,降低血浆CRP、TNF-α含量来实现的。     

厄贝沙坦对糖尿病大鼠肾脏骨桥蛋白表达及肾纤维化的影响

目的 探讨不同剂量厄贝沙坦对糖尿病大鼠肾组织骨桥蛋白( OPN)表达及小管间质纤维化的影响.方法 将63只8周龄雄性Wistar大鼠按随机数字表法分为正常对照组(Ctrl组,n=7)、糖尿病组(DM组,n=14)、30 mg/kg肼屈嗪干预组(DM+ Hyd组,n=12)、25mg·kg-1· d-1厄贝沙坦干预组(DM+Irb25组,n=10)、50 mg·kg-1·d-1厄贝沙坦干预组(DM+Irb50组,n=9)和200 mg · kg-1·d-1厄贝沙坦干预组(DM+Irb200组,n=11).糖尿病模型造模成功后4周,灌胃法给予相应剂量的肼屈嗪和厄贝沙坦.第12周末,观察各组大鼠24h尿白蛋白排泄率(UAER)、内生肌酐清除率(Ccr);PAS及Masson染色观察各组大鼠肾脏病理形态和胶原纤维沉积;ELISA法测定各组大鼠肾组织AngⅡ含量;实时定量PCR检测大鼠肾组织转化生长因子(TGF)β1、OPN mRNA的表达.结果 药物干预各组大鼠UAER、Ccr较DM组显著减少(均P＜ 0.05).DM组大鼠肾小球肥大,系膜基质增生,肾小球及小管间质有大量胶原纤维沉积;药物干预后,肾小球及肾小管上述病变减轻.DM组大鼠肾组织AngⅡ水平及TGF-β1、OPN mRNA表达均显著升高(均P＜0.05),给予肼屈嗪及厄贝沙坦干预后,AngⅡ、TGF-β1、OPN mRNA表达显著降低(均P＜0.05),且随着厄贝沙坦剂量的递增而递减.相关分析结果显示,DM组大鼠肾组织AngⅡ水平与OPN mRNA的表达呈正相关(r=0.74,P＜0.01).结论 厄贝沙坦通过减少肾脏局部AngⅡ水平,进而减少肾脏TGF-β1、OPN mRNA的表达,最终减轻小管间质纤维化,发挥肾脏保护作用,且这种保护作用具有剂量依赖性.     

组织型转谷氨酰胺酶在局灶节段性肾小球硬化大鼠肾脏中的表达和意义

目的探讨组织型转谷氨酰胺酶(tTG)在局灶节段性肾小球硬化(FSGS)模型大鼠肾脏中的表达及其意义。方法FSGS大鼠模型采用左颈静脉插管单次注射嘌呤霉素氨基核苷(PAN)方法建立,对照组大鼠注射等量生理盐水,20周后处死各组大鼠。肾组织切片用PAS染色,光镜下观察病理变化;用免疫组织化学、底物掺入免疫荧光法、Western印迹法分别观察肾内tTG蛋白分布、原位活性和蛋白水平变化;用免疫组化半定量方法分析细胞外基质纤连蛋白(FN)表达。结果模型组肾组织病理呈典型局灶节段肾小球硬化病变,伴大量蛋白尿。对照组肾组织tTG蛋白表达较弱,分布于肾小球内;模型组肾组织tTG蛋白分布于肾小球硬化部位。Western印迹结果显示模型组肾组织tTG蛋白水平比对照组增加2.61倍(P<0.05)。对照组肾组织tTG原位活性微弱;模型组肾小球tTG活性明显增强。模型组FN主要分布在肾小球,表达水平明显高于对照组(3.73±0.57比2.50±1.00,P<0.05):并且LTG蛋白水平与FN水平呈显著正相关(r=0.73,P<0.05)。结论tTG可能通过交联细胞外基质蛋白,抵抗降解,参与FSGS发生发展。     

厄贝沙坦对糖尿病肾病大鼠病理改变及足细胞凋亡的影响

目的 观察厄贝沙坦对糖尿病肾病大鼠病理改变及足细胞凋亡情况的影响,探讨厄贝沙坦对糖尿病肾损伤的保护作用机制.方法 腹腔注射链脲佐菌素建立糖尿病肾病大鼠模型.将36只雄性Wistar清洁级健康大鼠按随机数字表法分为正常对照组(N组)、糖尿病肾病组(DN组)和厄贝沙坦干预组(E组),于用药第12周取肾组织行苏木素-伊红染色及PAS染色,光镜下观察各组肾脏病理改变情况,采用TUNEL法计数凋亡足细胞数目.结果 12周时,DM组、E组大鼠24 h尿蛋白定量与N组比较差异有统计学意义(P＜0.01);E组大鼠24 h尿蛋白定量与DM组比较差异有统计学意义(P＜0.01);光镜下各组肾脏病理改变显示,N组大鼠肾小球形态未见异常变化,采用TUNEL法观察足细胞形态结构正常,未见凋亡足细胞;DN组大鼠肾脏体积较N组增大,系膜区增宽,可见不同程度的系膜细胞增生;采用TUNEL法显示足细胞体积肿胀增大,凋亡足细胞数目明显增多(P＜0.01);E组大鼠肾小球毛细血管基底膜增厚明显改善,凋亡足细胞数目较DM组明显减少(P＜0.01).结论 厄贝沙坦对糖尿病肾病的保护作用机制可能是通过改善肾小球滤过膜结构、抑制足细胞凋亡机制来减少糖尿病肾病大鼠大量蛋白尿,改善糖尿病肾病的预后.

Abstract：

Objective To investigate the influnce of irbesartan on the structure of glomerular filtration memberance and apoptosis of podocytes in diabetic nephropathy (DN) rats.Methods Give intraperitoneal injection of streptozotocin(STZ) to establish models of DN rats. 36 male Wistar clean rats were divided the into 3 groups,Normal Comparison group (N group), diabetic nephropathy group (DN group) and treatment group with Irbesartan (50 mg/kg. d E group) . After 12 weeks we observe 24 h urinary Protein count and the pathological changes of the kidney by extracting renal tissue to do HE staining and PAS staining; to count the apoptosis of podocytes of rats using TUNEL method. Results the level of 24 hour Protein in group E and group DM increased compared with those in the group N.,while the level of 24 hour Protein in group E decreased compared with that in the group DN after 12 weeks (P ＜ 0.01). For N group rats: glomerulus shapes are normal by optical microscopes, the structures of the podocytes are normal, using TUNEL method and there is no apoptotic podocytes. For DM group rates, the volume of kidney is bigger than N group under optical microscope, mesangial region becomes wider and there is different degrees of mesangial hyperplasia. Apoptotic podocytes increased than that of N group( P ＜ 0.01). As for the E group, basement membrane of glomerulus capillary improved significantly, the number of apoptotic podocytes decreased noticeably compared with those of the DM group (P ＜ 0.01). Conclusion The protection mechanism of the irbesartan to the diabetic nephropathy (DN) is probably through the improvement of the structure of glomerular filtration memberance, refraining the mechanism of the apoptosis of podocytes.     

乙酰肝素酶在糖尿病肾病大鼠蛋白尿发生中的作用

目的 观察糖尿病肾病大鼠肾组织乙酰肝素酶（HPA）的表达，探讨其在糖尿病肾病大鼠蛋白尿发生中的作用。 方法 SD健康大鼠被随机分为健康对照组（n = 6）、糖尿病6周组(DM6w, n = 10)和糖尿病12周组(DM12w, n = 10)，采用一次性腹腔注射链脲菌素（STZ）的方法建立糖尿病大鼠模型。分别于造模后6周和12周末测定各组大鼠相对肾质量、血糖、尿素氮、血肌酐、24 h尿量及尿蛋白量(24 h)等指标，并观察肾脏病理改变。RT-PCR和免疫组织化学法检测各组大鼠肾组织HPA mRNA和蛋白表达变化，并分析其与蛋白尿发生的相关性。 结果 （1）DM6w和DM12w组大鼠的相对肾质量、血糖、尿素氮、24 h尿量及尿蛋白量(24 h)与健康对照组相比明显升高, 差异均有统计学意义(P < 0.05或P < 0.01)。（2）DM6w和DM12w组大鼠HPA mRNA和蛋白表达比健康对照组均显著增高(P < 0.01)。（3）大鼠肾组织HPA mRNA和蛋白表达与尿蛋白量(24 h)之间均呈正相关 (r = 0.783，P < 0.01；r = 0.793，P < 0.01)。 结论 HPA在糖尿病肾病中的表达升高可能参与了糖尿病肾病蛋白尿的发生。     

大黄酸对2型糖尿病肾病大鼠疗效观察

目的：观察大黄酸防治2型糖尿病肾病的作用。方法：以高糖高脂饮食联合低剂量链脲佐菌素（STZ）注射方法诱导出2型糖尿病肾病大鼠模型后，分模型组、预防组和治疗组，后2组分别在STZ注射后1周和1个月后给予大黄酸100mg/kg剂量灌胃，每日1次，连续6个月，在不同时间进行形态学和有关生化指标的观察。结果：大黄酸预防给药和治疗后的观察结果显示，大黄酸明显降低糖尿病大鼠的24h尿蛋白排泄，改善肾重指数。肾脏组织病理学显示，大黄酸处理后的糖尿病大鼠肾小球和丝球体的面积明显减小，襻腔扩张减轻，系膜增生和细胞外基质减少。大鼠肾小球纤连蛋白沉积明显减弱。大黄酸也明显降低糖尿病大鼠的血脂水平。以上作用随着给药时间的延长，效果越来越明显。胰岛素抑制试验表明，在大黄酸预防给药和治疗6个月后，糖尿病大鼠的血浆稳态葡萄糖水平（SSPG）显著降低。以上结果比较，以预防组效果为佳。结论：大黄酸通过降低糖尿病大鼠的尿蛋白排泄、减轻肾脏肥大、改善胰岛素敏感性、降低血脂水平，有效地防治2型糖尿病肾病。     

Effects of pirfenidone on renal fibrosis in mice with diabetic nephropathy and its mechanisms

Objective To investigate effects of pirfenidone (PFD) on diabetic nephropathy model in db/db mice and to explore its possible mechanisms. Methods (1) Wild-type mice were as the normal control group, and db/db mice were divided into model group and PFD group, with 6 mice in each group. In the PFD group mice were administered continuously by 250 mg?kg-1?d-1 PFD for 18 weeks, and mice in the other two groups were administered by 0.5% sodium carboxymethyl cellulose. Blood glucose and 24 h urinary albumin were measured. The pathological changes of renal tissue were evaluated by PAS staining, PASM staining, Masson staining and Sirius red staining. The expression of collagen type Ⅳ in kidney tissues was detected by immunohistochemistry. (2) Mouse mesangial cells (SV40 MES-13 cells) were cultured as research objects. They were divided into control group, hyperosmolar group, high glucose (HG) group, and 50, 100, 200, 400, 800, 1600 mg/L PFD+HG group. BrdU cell proliferation test was used to evaluate cell proliferation rate. Cells were divided into control group, hyperosmolar group, HG group and PFD+HG group. The mRNA expressions of α-smooth muscle actin (α-SMA), collagen type Ⅰ, collagen type Ⅳ, transforming growth factor-β1 (TGF-β1), interleukin (IL)-1β, IL-6 and monocyte chemotactic protein-1 (MCP-1) were detected by real-time PCR. Results (1) Compared with normal control group, the model mice had higher weight, blood glucose and 24 h urinary albumin, accompanied with glomerular hypertrophy, mesangial area expansion, tubulointerstitial fibrosis and deposition of collagen type Ⅳ (all P＜0.05). Compared with those in model group, in PFD group 24 h urinary albumin decreased, glomerular hypertrophy, mesangial area expansion and tubulointerstitial fibrosis alleviated, and the protein expression of collagen type Ⅳ inhibited (all P＜0.05). (2) Compared with those in HG group, MES-13 cell proliferation rates of 100, 200, 400, 800, 1600 mg/L PFD+HG groups decreased (all P＜0.05), and the mRNA expressions of α-SMA, collagen type Ⅰ, collagen type Ⅳ, TGF-β1, IL-1β, IL-6 and MCP-1 reduced in 400 mg/L PFD+HG group (all P＜0.05). Conclusions PFD can inhibit high glucose-induced proliferation and activation of glomerular mesangial cells, decrease the expression of TGF-β1 and proinflammatory factors, as well as reduce the synthesis of collagen, which improve renal fibrosis of db/db mice.     

2型糖尿病肾病相关生化指标检测及临床意义

目的分析糖尿病肾病与各相关因素的关系以指导临床及早期防治。方法对56例2型糖尿病患者及健康对照组相关指标进行统计分析。结果 CAU组、MAU组及NAU组HbA1c、UA均高于NC组（P〈0.05）;4组HCY、ADPN呈递增趋势;hs-CRP水平随病变程度而增加。结论多种因素影响DN的发生,积极地控制各种危险因素对预防和延缓DN的发生与发展有重要的意义。     

糖尿病肾病与糖尿病视网膜病变的相关性研究进展

糖尿病肾病(DN)和糖尿病视网膜病变(DR)均是糖尿病的微血管病变，是目前成人终末期肾病(ESRD)和致盲的重要原因，两者在发生、发展过程中具有一定平行性，又存在不平行性。DN和DR可预测彼此的发生、发展，但目前对于两者之间的关系尚未明确。因此，本文就DN与DR之间的相关性的研究进展作一综述，为临床诊断治疗提供帮助。     

IL-18、IL-6、IL-10在糖尿病大鼠肾脏的表达

目的检测IL-18、IL-6、IL-10在糖尿病大鼠肾脏的表达,探讨其在糖尿病肾脏病变过程中的作用。方法将40只雄性Wistar大鼠随机分为正常对照4周组（NC1组）、8周组（NC2组）、糖尿病4周组（DMI组）、8周组（DM2组）,每组10只。DM组给予一次性腹腔内注射60mg／kg链脲佐菌素建立糖尿病大鼠模型。检测各组大鼠体重、肾重、尿微量白蛋白排泄率（UAER）。用免疫组化方法检测肾组织IL-18、IL-6、IL-10的表达,利用计算机图像分析系统进行定量分析。结果DM组大鼠的肾重指数（KWI）、UAER较NC组显著增高;DM2组较DM1组明显增高;IL-18、IL-6在NC组肾组织中仅少量表达,而在DM1组表达明显增多,在DM2组表达增高则更为显著。IL-10在NC组丰富表达,而在DM1组表达明显减弱,在DM2组几乎无表达。肾脏局部IL-18、IL-6与IL-10的表达呈显著负相关。结论糖尿病大鼠肾脏局部IL-18、IL-6表达明显升高、IL-10水平显著降低,IL-18、IL-6与IL-10之间平衡的紊乱在糖尿病肾脏损害病程中发挥重要的作用。