蛋白酶激活受体2在小鼠肾间质纤维化中的表达及其意义

目的 观察单侧输尿管梗阻(UUO)小鼠模型中蛋白酶激活受体2(PAR-2)在肾小管间质中的表达部位、动态变化及其与肾间质纤维化的关系。方法 制备UUO小鼠模型,采用RT-PCR方法检测UUO术后第1、3、5、7、10、14、21天肾组织中PAR-2mRNA的表达;免疫组化方法检测PAR-2蛋白的表达。分析它与肾间质相对面积、α-平滑肌肌动蛋白(SMA)表达水平的关系。结果 UUO术后第1天肾小管间质可见少量PAR-2mRNA和蛋白的表达,第7-14天表达显著增加,其后维持在较高水平。PAR-2表达量与肾间质相对面积、α-SMA表达量成正相关。PAR-2表达主要定位于肾小管上皮细胞(尤其为近曲小管),也可见于肾毛细血管袢、间质浸润细胞和成纤维细胞。结论 PAR-2在肾小管上皮细胞和肾间质的表达可能参与了肾间质纤维化发生发展的过程。     

脉血康对单侧输尿管梗阻大鼠肾保护作用及机制探讨

目的观察脉血康对单侧输尿管梗阻(unilateral ureteral obstruction,UUO)大鼠肾脏保护作用并探讨其可能作用机制。方法建立UUO大鼠模型,大鼠随机分为假手术组、模型组及脉血康组3组,各组给予相应干预,各组大鼠于术后14 d处死,抽取血清并留取肾组织标本,检测各组大鼠血肌酐(SCr)、尿素氮(BUN)、胆固醇(total cholesterol,TC)、三酰甘油(triglyceride,TG)情况,并观察各组肾脏病理改变情况,采用免疫组化方法观察肾组织中组织型纤溶酶原激活物(tissue type plasminogen activator,t-PA)、尿激酶型纤溶酶原激活物(urokinase type plasminogen activator,u-PA)、纤溶酶原激活物抑制剂1(plasminogen activator inhibitor-1,PAI-1)在各组的表达情况。结果①3组肾脏形态学变化有差异,假手术组双肾大小及体积无改变,颜色红润,模型组及脉血康组术侧肾脏体积明显增大,术侧肾盂肾盏及输尿管结扎段以上全程扩张。模型组肾实质颜色变浅,剖面肾皮质变薄,皮髓质分界不清,脉血康组肾脏颜色稍红润,肾脏皮髓质分界较清。②假手术组肾脏病理未见明显改变,与假手术组比较,模型组及脉血康组肾小管损伤及肾间质纤维化程度显著(P0.01);与模型组比较,脉血康组肾小管损伤及肾间质纤维化程度明显减轻(P0.05)。③与假手术组相比,模型组SCr、BUN、TC、TG水平均升高(P0.05),与模型组相比,脉血康组SCr、BUN、TC、TG水平均下降(P0.05)。④假手术组PAI-1、t-PA、u-PA表达于肾小管上皮细胞及肾小球系膜细胞胞浆,肾间质表达极少;与假手术组比较,模型组及脉血康组PAI-1明显增多(P0.05),而t-PA、u-PA表达明显减少(P0.05),与模型组相比,脉血康组PAI--1表达明显减少,t-PA表达明显增多(P0.05),u-PA无明显差别(P0.05)。结论脉血康可能通过调节脂质代谢下调肾脏PAI-1的表达,上调t-PA的表达从而促进肾脏细胞外基质降解,对UUO大鼠肾间质纤维化及肾功能具有保护作用。     

Effects of proteasome inhibitors on rat renal fibrosis in vitro and in vivo

Aim: Transforming growth factor‐β (TGF‐β) is involved in renal tubulointerstitial fibrosis. Recently, the ubiquitin proteasome system was shown to participate in the TGF‐β signalling pathway. The aim of this study was to examine the effects of proteasome inhibitors on TGF‐β‐induced transformation of renal fibroblasts and tubular epithelial cells in vitro and on unilateral ureteral obstruction (UUO) in vivo. Methods: Rat renal fibroblasts NRK‐49F cells and tubular epithelial cells, NRK‐52E, were treated with TGF‐β in the presence or absence of a proteasome inhibitor, MG132 or lactacystin. Rats were subjected to UUO and received MG132 i.p. for 7 days. Results: In cultured renal cells, both MG132 and lactacystin inhibited TGF‐β‐induced α‐smooth muscle actin (α‐SMA) protein expression according to both western blotting and immunofluorescent study results. MG132 also suppressed TGF‐β‐induced mRNA expression of α‐SMA and upregulation of Smad‐response element reporter activity. However, MG132 did not inhibit TGF‐β‐induced phosphorylation and nuclear translocation of Smad2. In contrast, MG132 increased the protein level of Smad co‐repressor SnoN, demonstrating that SnoN is one of the target molecules by which MG132 blocks the TGF‐β signalling pathway. Although the proteasome inhibitor suppressed TGF‐β‐induced transformation of cultured fibroblasts and tubular epithelial cells, MG132 treatment did not ameliorate tubulointerstitial fibrosis in the rat UUO model. Conclusion: Proteasome inhibitors attenuate TGF‐β signalling by blocking Smad signal transduction in vitro, but do not inhibit renal interstitial fibrosis in vivo.     

小鼠单侧输尿管梗阻诱导肾脏纤维化模型中Cadherin6的表达变化

目的研究钙黏蛋白6(cadherin6,CDH6)在肾脏纤维化中表达的变化。方法建立单侧输尿管梗阻(UUO)小鼠模型,术后21天收获肾脏组织,HE染色观察肾间质纤维化程度,RT-q PCR检测肾组织FSP-1、TGF-β1、CDH6 m RNA表达量,免疫组化染色检测肾组织FSP-1、CDH6蛋白表达量;TGF-β1刺激人肾小管上皮细胞HK-2,RT-q PCR及Western Blot分别检测FN1、PAI-1、CDH6 m RNA表达量及CDH6蛋白表达量。结果 HE染色显示UUO组小鼠肾脏出现肾小管萎缩、大量基质沉积,与Sham组相比,UUO组小鼠肾脏组织FSP-1、TGF-β1、CDH6 m RNA表达量及FSP-1、CDH6蛋白表达量均上调;细胞实验结果显示,与空白对照组相比,TGF-β1诱导的HK-2细胞形态向成纤维细胞转变,FN1、PAI-1m RNA表达水平上调,CDH6 m RNA及蛋白表达水平上调。结论 TGF-β1诱导的肾小管上皮细胞CDH6及纤维化相关因子表达上调,伴随上皮细胞向间质表型转化,可能参与单侧输尿管梗阻诱导的小鼠肾脏纤维化的发展。     

加味抵挡汤对单侧输尿管梗阻大鼠肾间质PAI-1表达的影响

目的探讨加味抵挡汤延缓肾间质纤维化的机制。方法将60只大鼠随机分为模型组、加味抵挡汤组、贝那普利组、假手术组、空白组各12只,前3组行单侧输尿管梗阻（UUO）术,术后第14天处死大鼠,留取梗阻侧肾组织行HE染色,观察肾脏病理学变化,并用免疫组化方法测定肾组织纤溶酶原激活剂抑制物-1（PAI-1）的表达。结果加味抵挡汤能改善肾间质损伤和肾间质纤维化程度,降低肾脏组织致纤维化因子PAI-1表达。结论加味抵挡汤具有延缓肾纤维化的作用。     

NAD(P)H‐quinone oxidoreductase 1 silencing aggravates hormone‐induced prostatic hyperplasia in mice

NAD(P)H‐quinone oxidoreductase 1 (NQO1) is a highly inducible flavoprotein known to involve in various cellular defence mechanisms. In this study, we explored whether NQO1 deletion affects hormone‐induced prostatic hyperplasia. Testosterone propionate (3 mg/kg, IP) was injected into wild‐type (WT) and NOQ1 knockout C57BL/6 mice (NQO1^?/?) for 14 consecutive days, and the samples were collected for biological and histochemical studies. The testosterone‐treated NQO1^?/? showed about 140% higher prostate weight than the testosterone‐treated WT, with enhanced connective tissue and hyperplastic glands formations. However, increased dihydrotestosterone level after testosterone treatment was not significantly different between the WT and NQO1^?/?. In contrast, the enhanced nuclear expression of proliferating cell nuclear antigen in NQO1^?/? prostate confirmed aggravated prostatic hyperplasia in NQO1^?/?. Moreover, the expression of heat shock protein (HSP) 90‐α was markedly increased in the NQO1^?/?, and this was supported by increased testosterone‐induced nuclear androgen receptor expression in NQO1‐silenced LNCaP cells. Testosterone‐induced prostate‐specific antigen expression was not reversed in NOQ1‐silenced cells after finasteride treatment. Although the exact role of NQO1 in prostatic hyperplasia remains unclear, the hyperplasia exacerbation due to NQO1 deletion might be independent of type 2 5α‐reductase and might be related to enhanced androgen receptor affinity due to enhanced HSP90‐α expression.     

Effect of combined angiotensin‐converting enzyme and aldosterone inhibition on plasma plasminogen activator inhibitor type 1 levels in chronic hypertensive patients

Aim: A possible link between the renin–angiotensin–aldosterone system (RAAS) and fibrinolysis has recently been suggested. Systemic infusion of angiotensin II results in an increase in plasminogen activator inhibitor type 1 (PAI‐1) levels and angiotensin‐converting enzyme inhibitors (ACEI) have been shown to decrease PAI‐1 levels. Moreover, recent data indicated that plasma aldosterone levels were positively correlated with plasma PAI‐1 levels. This study was designed to compare the effects of an ACEI with an ACEI in combination with an aldosterone antagonist on PAI‐1 levels in chronic hypertensive patients. Methods: Patients were randomized into two groups and were treated with either low salt diet plus fosinopril (group 1, n = 43) or low salt diet plus fosinopril plus spironolactone (group 2, n = 42). Plasma PAI‐1, tissue plasminogen activator (tPA) and plasma renin activity (PRA) levels were measured before and after 24 week treatment in both groups. Results: The mean basal PRA levels were similar in both groups. After antihypertensive therapy, the mean PRA increased significantly in both groups (P < 0.005). The mean plasma PAI‐1 levels were reduced in both treatment groups (P < 0.005). However, the reduction in group 2 was more pronounced (P < 0.05). Although after the treatment mean plasma levels of PAI‐1 significantly reduced in both groups, the reduction of PAI‐1 levels was more pronounced in group 2. Conclusion: Although the plasma levels of PAI‐1 significantly reduced after treatment in both groups, the reduction of PAI‐1 levels was more pronounced in group 2. These data indicated that administration of aldosterone antagonists in combination with ACEI had additional benefit on fibrinolysis in chronic hypertensive patients.     

Additive effects of hyperbaric oxygen and platelet‐derived growth factor‐BB in chondrocyte transplantation via up‐regulation expression of platelet‐derived growth factor‐β receptor

The present study investigated the effects of hyperbaric oxygen (HBO) and platelet‐derived growth factor‐BB (PDGF‐BB) in chondrocyte transplantation. In vitro, chondrocytes were treated with HBO, PDGF‐BB, and HBO combined with PDGF‐BB (H+P). Cell growth was analyzed using cell counting, MTT assay, and FACS analysis. mRNA expression of the PDGF‐α receptor (PDGFR‐α) and β receptor (PDGFR‐β) was detected by RT‐PCR. Protein expression of PDGFR‐β was detected by Western blotting. In vivo, chondrocytes and PDGF‐BB were suspended in alginate as a transplantation system. Cartilage defects were grafted with this system and with or without HBO treatment. Released PDGF‐BB concentration was quantified by ELISA. After 8 weeks, animals were sacrificed and the repaired tissues were examined. In vitro data suggested that each treatment increased cell growth via the up‐regulated mRNA expression of PDGFR‐α and increased cell accumulation in the S‐phase. The H+P treatment was more additive in cell growth and in mRNA and protein expression of PDGFR‐β than HBO or PDGF‐BB. In vivo results suggested that PDGF‐BB delivery lasted for more than 5 weeks. Scoring results showed that each treatment significantly increased the cartilage repair. Safranin‐O and type II collagen staining confirmed the hyaline‐like cartilage regeneration in the repaired tissues. In situ up‐regulation of PDGFR‐β expression partially explains the additive effect of H+P treatment in cartilage repair. Accordingly, H+P offers a potential treatment method for cartilage repair. © 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27:1439–1446, 2009     

STAT3抑制剂S3I－201对实验性肾小管间质纤维化的保护作用

目的探讨STAT3抑制剂S3I－201对小鼠实验性肾小管间质纤维化的保护作用。 方法采用单侧输尿管梗阻手术的方法建立肾小管间质纤维化模型。将实验小鼠随机分为药物假手术组(Sham＋S3I－201),安慰剂假手术组(Sham＋Vehicle),药物造模组(UUO＋S3I－201),安慰剂造模组(UUO＋Vehicle)4组,通过腹腔注射S3I－201溶液(药物)或0.05%DMSO PBS(安慰剂)给药,每天给药一次。造模第7天时留取肾脏标本,用Masson染色和颜色面积测算法评估胶原蛋白沉积的情况。用qRT－PCR法检测肾组织内趋化因子配体16(CXCL16),白介素－1β(IL－1β),细胞间黏附分子1(ICAM－1),转化生长因子－β(TGF－β),肿瘤坏死因子(TNF－α)的mRNA表达,用免疫组化法染色和免疫印迹法检测PDGFRβ蛋白在梗阻肾脏内的表达。 结果UUO＋Vehicle小鼠的肾间质胶原蛋白沉积显著高于Sham＋Vehicle组(P<0.05)。UUO＋Vehicle小鼠肾组织CXCL16,IL－1β,ICAM－1,TGF－β,TNF－α的mRNA表达显著高于Sham＋Vehicle组(P<0.05),UUO＋Vehicle小鼠肾组织血小板来源生长因子受体β(PDGFRβ)蛋白表达显著高于Sham＋Vehicle组(P<0.05)。经过S3I－201治疗7 d后,UUO＋S3I－201小鼠的上述各项指标均显著低于UUO＋Vehicle(P<0.05)。 结论S3I－201通过抑制多种细胞因子的mRNA表达,以及降低PDGFRβ蛋白的表达,减轻实验性肾小管间质纤维化小鼠的肾间质炎症反应,从而发挥肾脏保护作用。     

Expression of peroxisome proliferator-activated receptor (PPAR) in human prostate cancer

BACKGROUND: Recent studies have demonstrated that peroxisome proliferator activator-receptors (PPAR)-gamma is expressed in some cancer cells such as breast, lung, and gastric cancer, and its ligand induces growth arrest of these cancer cells through apoptosis. However, the expression and localization of PPARs in prostate have not been examined. In this study, PPARs expression was investigated in human prostate cancer (PC), prostatic intraepithelial neoplasia (PIN), benign prostatic hyperplasia (BPH), and normal prostate (NP) tissues. METHODS: Tumor specimens were obtained from 156 patients with PC, 15 with PIN, 20 with BPH, and 12 patients with NP tissues. The expressions were investigated by RT-PCR and immunohistochemical methods. RESULTS: Immunoreactive PPAR-alpha and -beta were significantly apparent in PC tissues. Marked expressions of PPAR-alpha and -beta were also detected in PIN, BPH, and NP groups. However, very weak or no expression of immunoreactive PPAR-gamma was found in BPH and NP cases. In contrast, we found significant expression of immunoreactive PPAR-gamma in cancer cells in PC group and in PIN group. CONCLUSIONS: Our results demonstrated that PPAR-gamma is induced in PC, and suggest that PPAR-gamma ligands may mediate its own potent antiproliferative effect against PC cells through differentiation.