Differential effects of phosphoramidon and captopril on NK1 receptor-mediated plasma extravasation in the rat trachea

We sought to confirm the identity of the tachykinin receptor subtype that mediates plasma extravasation in the rat trachea, and assess the respective contributions of neutral endopeptidase (NEP) and angiotensin-converting enzyme (ACE) in regulating this tachykinin-induced response. To achieve these aims, we determined the relative potencies of several natural tachykinins and receptor-selective synthetic agonists, both before and after inhibiting NEP with phosphoramidon and ACE with captopril. We also determined the effects of these peptidase inhibitors, and the NK-1 receptor antagonist L-703,606, on the plasma extravasation produced by capsaicin, which releases tachykinins endogenously from sensory nerve endings. We found that the rank order of potency for producing plasma extravasation in the rat trachea was NK-1 receptor agonist ([Sar⁹, Met(O₂)¹¹] SP)>substance P>neurokinin A> neurokinin B. The NK-2 ([Nle¹⁰]NKA (4–10)) and NK-3 ([MePhe⁷]NKB) receptor agonists were without effect. We observed no change in the relative potencies of these peptides after giving rats phosphoramidon or captopril, which suggests that the different peptide potencies are not simply the consequence of different rates of enzymatic degradation. Nevertheless, the responses to substance P and neurokinin A were clearly potentiated in rats given phosphoramidon, indicating that NEP effectively degrades tachykininsin vivo. No significant potentiation was evident for any peptide in rats given captopril. Similarly, the plasma extravasation produced by capsaicin was potentiated in rats given phosphoramidon, but not in those given captopril. Pretreating rats with L-703,606 abolished the response to capsaicin. We conclude from these observations that NK-1 receptors mediate tachykinin-induced plasma extravasation in the rat trachea, and that NEP regulates this response with little or no contribution from ACE.     

三棱、莪术对肝纤维化大鼠IL-1、IL-6、TNF-α的影响

目的：观察三棱、莪术对免疫性肝纤维化大鼠血清中IL-1、IL-6、TNF-α的影响及其抗肝纤维化的作用机制。方法：用猪血清腹腔注射建立免疫性肝纤维化大鼠模型，以血清中IL-1、IL-6、TNF-α的浓度变化，光、电镜下的肝组织病理学改变为观察指标，观察三棱、莪术对大鼠免疫性肝纤维化的影响。结果：免疫性肝纤维化大鼠血清中IL-1、IL-6、TNF-α水平明显增高；三棱、莪术可下调肝纤维化大鼠模型IL-1、IL-6、TNF-α水平趋向于正常；并能改善肝脏组织病理学变化。结论：三棱、莪术通过减少IL-1、IL-6、TNF-α的合成与释放，发挥抗肝纤维化作用；在抗肝纤维化过程中具有免疫调控作用。     

肝纤维化时肝脏OPN和PAI-1的表达变化

目的:研究骨桥蛋白(OPN)及纤溶酶原激活物抑制物(PAI-1)的表达特征及其在肝纤维化时的变化。方法:采用二甲基亚硝胺制作大鼠肝纤维化模型,大鼠肝脏常规HE和天狼猩红染色,采用SABC法做免疫组织化学染色及Western blotting检测OPN和PAI-1蛋白表达,抽提肝组织总RNA,RT-PCR检测OPN mRNA表达。结果:正常大鼠肝组织OPN和PAI-1表达极弱,肝纤维化大鼠肝脏中OPN表达增强,阳性信号散在或弥漫性分布,主要见于小叶内中央静脉周围、纤维间隔内以及周围巨噬细胞胞浆、枯否氏细胞、汇管区的部分肝细胞、肝窦壁内皮细胞。PAI-1在肝纤维化大鼠肝组织汇管区、肝细胞变性坏死处,肝窦周Disse间隙及毗邻以上部位的肝细胞,组织纤维间隔处及其外周细胞亦见阳性染色。Western blotting检测正常大鼠肝脏OPN的蛋白表达极低,肝纤维化组OPN的蛋白表达较正常组显著增强(P0.01)。与正常组比,肝纤维化组PAI-1表达也显著增强。RT-PCR检测结果显示,正常大鼠肝脏OPN mRNA表达极低,肝纤维化大鼠肝脏OPN mRNA的表达明显增强(P0.05)。研究结果证明,肝纤维化时大鼠肝组织OPN及PAI-1的表达水平显著增高。结论:肝纤维化时大鼠肝组织OPN及PAI-1的表达水平显著增高,OPN可能会促进PAI-1的高表达,从而抑制ECM降解、加速肝纤维化进程。     

肝纤维化形成过程中Ⅰ型血管紧张素Ⅱ受体表达的研究

目的 :探讨Ⅰ型血管紧张素Ⅱ受体 (AngiotensinⅡType1Receptor,AT1R)在人肝脏组织中的表达情况。方法 :运用免疫组织化学SABC法和间接免疫荧光标记法 ,对 12例正常肝组织及 18例纤维化肝组织进行检测。结果 :AT1R阳性表达主要分布在肝小叶周边及肝窦区星形细胞 (HepaticStellateCell,HSC)的胞浆内。 12例正常肝组织中 8例呈阳性表达 ,18例纤维化肝组织均表达AT1R ,且纤维化组阳性细胞数明显多于正常组 (P <0 0 0 1)。结论 :AT1R在肝星形细胞胞浆中的表达随肝纤维化的发展明显增强 ,并且可能在肝纤维化的发生发展中发挥着一定作用。     

Effects of captopril treatment versus placebo on renal function in type 2 diabetic patients with microalbuminuria: a long-term study

We evaluated the renal effect of long-term antihypertensive treatment (12 months) with the angiotensin-converting enzyme inhibitor captopril compared to placebo in 15 type 2 diabetic patients with microalbuminuria. The patients were randomly allocated to captopril (n = 9) or placebo (n = 6). After 1-year therapy no significant decrease in blood pressure was demonstrated with captopril (139 ± 17/80 ± 9 versus 138±13/76±6 mmHg) or placebo (138 ± 9/75 ± 6 versus 135 ± 14/79 ±10 mmHg). Only in a small hypertensive subgroup (n = 4) treated with captopril did we find a significant reduction in blood pressure (154 ± 2/88 ± 1 versus 142 ± 7/78 ± 5 mmHg,P < 0.05). The urinary albumin excretion rate did not change significantly either in the captopril group (95.6 mg/24 h, 25th percentile 138.4, 75th percentile 25.1; versus 127.8 mg/24 h, 25th percentile 29.3, 75th percentile 222) or in the placebo group (99.2 mg/24 h, 25th percentile 58.5, 75th percentile 125.8; versus 120.9 mg/24 h, 25th percentile 62.1, 75th percentile 179.7). There were also no alterations in renal blood flow or filtration rate. In the hypertensive subgroup treated with captopril a reduction in urinary albumin excretion rate after 3 and 6 months of treatment was observed (captopril 73.4 versus 24 and 41 mg/24 h,P < 0.05), but not after 12 months. Triglyceride and cholesterol levels remained constant before and after treatment while glycosylated hemoglobin decreased significantly after 12 months captopril (7.8 ± 0.9 versus 6.9 ± 0.7 mg%,P < 0.03). We conclude that in patients with type 2 diabetes with microalbuminuria angiotensin-converting enzyme inhibitors may have protective renal effects in so far as a lack of increase in urinary albumin excretion is equivalent to low progression in renal disease.Abbreviations ACE angiotensin-converting enzyme - GFI glomerular filtration rate - HbA glycosylated hemo globin - RPF renal plasma flow - UAER urinary albumin excretion rate Correspondence to: R. Prager     

基于HSC的肝纤维化发病及治疗研究进展

肝纤维化主要是由肝星状细胞（HSC）的活化、细胞外基质（ECM）的合成与降解失衡导致ECM的过度增生和沉积所致。HSC的激活和增生是肝纤维化发生、发展的中心环节,抑制HSC的活化是防治肝纤维化形成的关键。抗肝纤维化的治疗措施包括抑制HSC增殖或诱导HSC凋亡,抑制胶原蛋白的产生或促进胶原蛋白的降解、细胞因子的调控以及间质干细胞的灌注等,因而早期肝纤维化的防治研究具有重要意义。     

Autoradiography of angiotensin-converting enzyme in fixed and unfixed rat brain using the specific enzyme inhibitor [I]351A or a polyclonal antibody and [I]staphylococcal protein A

Angiotensin-converting enzyme (ACE, kininase II, EC 3.4.15.1) was visualized in unfixed rat brain sections by autoradiography after incubation with a polyclonal goat anti-rabbit lung converting-enzyme antibody and [¹²⁵I]protein A. Paraformaldehyde fixation interfered with the recognition of ACE by its antibody in brain nuclei but not in the choroid plexus. Conversely, incubation of brain sections with the specific ACE inhibitor [¹²⁵I]351A allowed ACE visualization in either unfixed and paraformaldehyde-fixed brain sections, since [¹²⁵I]351A binding was not affected by our fixation conditions. Our results indicate that ACE could be visualized in unfixed brain sections directly by incubation with the specific inhibitor [¹²⁵I]351A or indirectly by radioimmunohistochemistry and autoradiography. Different autoradiographic methods could be used to visualize and quantify ACE in unfixed or fixed tissues.     

依那普利和倍他乐克对长期血透病人左心肥大的影响

目的和方法：比较依那普利和倍他乐克对血透病人左心功能及心肌重量的影响。血两 被分为３组,比较１２个月后血压、左心功能及左心肥大的改变。结果：组１（依那普利）和组２（倍他乐克）在治疗后血压有同等程度不下降,而心脏检查示：治疗后,组１舒张末期左这人径９ＬＶＥＤＤ）、左室心肌重量（ＬＶＭ）、左 室心肌重量指数（ＬＶＭＩ）、室间隔厚度（ＩＶＳ）、左室后壁厚度（ＬＶＰＷＤ）、和二尖瓣舒张晚期最大充盈速度（Ａ     

The predictive value of CD38 positive hepatic stellate cell count for assessing disease activity and fibrosis in patients with chronic hepatitis

The activation of hepatic stellate cells (HSCs) is a critical event in hepatic fibrosis. The objectives of this study were to find out if cluster of differentiation 38 (CD38) can be demonstrated immunohistochemically on HSCs in liver biopsies from patients with chronic liver disease and if CD38 immunopositive HSC count is correlated with METAVIR inflammatory and fibrosis scores. Immunohistochemical labelling for CD38 was performed on 100 liver biopsies from patients with chronic liver disease. The CD38 immunopositive HSCs were identified and counted. The CD38 immunopositive HSC count was found to be associated with both the METAVIR score and the fibrosis scores. The CD38 immunopositive HSC count was able to discriminate between no fibrosis and stages 2, 3 or 4 fibrosis, but could not discriminate between no fibrosis and stage 1 fibrosis. Using receiver operating characteristic (ROC) curves, a cut-off point of 10 HSCs per 10 high power field (hpf), or 25 per 100 hepatocytes, is 80% sensitive and 70% specific for predicting fibrosis. The specificity rose to 100% in patients with hepatitis C viral (HCV) infection. We conclude that CD38 positive HSCs can be demonstrated immunohistochemically and that the count is highly predictive of moderate to severe hepatic fibrosis.     

瘦素及I和III型胶原蛋白在肝纤维化大鼠肝组织中的变化

 摘要： 目的 研究瘦素（Leptin）及I、 III型胶原蛋白及基因在肝纤维化模型组织中的动态表达水平。方法 四氯化碳（CCl4）皮下注射法制备肝纤维化模型,分别以Western blot及RT-PCR法检测Leptin及I、 III型胶原蛋白及基因在肝纤维化组织中的动态表达。结果 Leptin以及I、 III型胶原蛋白及基因在正常对照组肝脏中均有微量表达,CCl4注射2周后,三者的表达均开始增强,随着纤维化发展呈梯度增加。其mRNA表达水平在模型组明显高于正常组 (P<0.05);在肝纤维化过程中,Leptin与I型胶原(r=0.595,P=0.017)及Leptin与III型胶原(r=0.478,P=0.011) 的动态改变呈显著正相关。结论 Leptin的表达随着纤维化的程度加重而逐步增强,在肝纤维化过程中,Leptin可能参与了细胞外基质成分（ECM）的合成与降解。     

卡托普利对高氧新生儿肺损伤保护作用的实验研究

目的探讨ACE抑制剂卡托普利(CPT) 对高氧致新生儿慢性肺疾病(CLD)的影响.方法新生Wistar大鼠随机分为3组:治疗组即高氧 卡托普利(HO CPT),单纯高氧组(HO),正常对照组(NC).治疗组和高氧组每日分别胃管内灌注CPT(50mg/kg/d) 和生理盐水进行干预;对照组胃管内灌注生理盐水代替.各组动物均于胃管内灌注后7、14、21d分别处死16只,其中8只取肺组织做HE染色;另外8只行支气管肺泡灌洗(BAL),收集BALF检测细胞数及总蛋白含量;利用显微镜图像分析系统测定各组肺泡间隔宽度.结果 CPT能显著减轻肺泡炎和肺纤维化的程度,肺泡间隔宽度、BALF中细胞数和总蛋白水平可间接反应肺组织的损伤程度.结论 CPT能减轻高氧诱导的大鼠肺泡炎和肺纤维化.     

暴发性肝衰竭血脑屏障通透性改变对肝性脑病发生发展的影响

采用硫代乙酰胺所致大鼠暴发性肝衰模型，以脑组织浸泡液中Ｅｖａｎｓ蓝含量为血脑屏障通透性指标。结果表明，在肝性脑病初期血脑屏障通透性就已增加，至脑病晚期则明显增加，并伴脑水肿发生，脑组织浸泡液中Ｅｖａｎｓ蓝含量与血浆内毒素呈正相关。以上结果提示，暴发性肝衰竭时，肝性脑病发生发展与内毒素所致血脑屏障通透性增加密切相关。     

ACEI/ARB对AF患者血浆BNP水平的影响及临床意义

目的探讨血管紧张素转换酶抑制剂（ACEI）/血管紧张素受体拮抗剂（ARB）对心房颤动（AF）患者血浆脑钠肽（BNP）水平的影响及其临床意义。方法选取心功能l～4级AF患者126例,采用干性免疫法测定患者AF发作时血浆BNP浓度,将患者分为使用ACEI/ARB组（62例）和未使用ACEI/ARB组（64例）,比较分析两组血浆BNP浓度的差异。结果使用ACEI/ARB组患者AF发作时血浆BNP水平为（242.31±182.15）pg/ml,明显低于未使用ACEI/ARB组的（453.84±365.57）pg/ml,差异有统计学意义（P〈0.05）。结论 ACEI/ARB对AF患者血浆BNP水平有一定的影响,提示ACEI/ARB可能干扰临床医生根据BNP水平变化对AF、心衰等情况作出评估。     

P物质受体拮抗剂对缺氧大鼠模型肝功能及肝细胞凋亡的影响

将30只雄性SD大鼠(体重240~300 g)随机分为3组,分别为对照组(不进行缺氧处理)、低氧组及NK-1R拮抗剂组,建立大鼠缺氧模型,NK-1R拮抗剂组腹腔注射P物质受体(NK-1R)拮抗剂(S3144)进行干预。观察各组大鼠肝组织病理学变化,检测肝功能指标及Tunnel法测肝凋亡细胞百分比,探讨P物质受体(NK-1R)拮抗剂对缺氧大鼠模型肝损伤的影响及其机制。结果显示:(1)缺氧模型的大鼠肝细胞肿胀,排列紊乱,气球样变性较多,并有点状坏死灶,损伤较NK-1R拮抗剂组严重。(2)NK-1R拮抗剂组GGT[(6.750 0±1.669 1)U/]明显低于低氧组[(9.000 0±1.414 2)U/L)](P<0.05);ALT、AST、ALP各组之间比较无明显差异(P>0.05)。(3)NK-1R拮抗剂组肝细胞凋亡百分比[(26.626 3±10.934 8)%]明显低于低氧组[(44.581 4±17.324 5)%](P<0.05)。研究表明:P物质可能与缺氧性肝损伤有关,通过P物质受体(NK-1R)拮抗剂可能减轻缺氧性肝损伤。     

人血白蛋白对近端肾小管上皮细胞ACE2表达的影响

目的： 研究人血白蛋白对人近端肾小管上皮细胞（HK-2）血管紧张素转换酶2（ACE2）及血管紧张素转换酶（ACE）表达的影响，探讨白蛋白对肾脏损害的机制。方法: 不同浓度人血白蛋白（0、1、5、10、15 g/L）分别加入到培养的人近端肾小管上皮细胞培养48 h，用RT-PCR法和Western blotting法分别检测HK-2细胞ACE2和ACE mRNA和蛋白表达。结果: 正常培养状态下HK-2细胞（空白对照组）存在ACE2 mRNA和蛋白表达，加入不同浓度（5-15 g/L）的人血白蛋白剂量依赖抑制HK-2细胞ACE2 mRNA和蛋白表达（P<0.01）。正常培养状态下HK-2细胞ACE mRNA和蛋白表达水平较低，随着加入白蛋白浓度的增加其表达水平逐渐升高，10 g/L白蛋白显著促进了HK-2细胞ACE mRNA和蛋白表达（P<0.05）。结论: 在体外培养的HK-2细胞存在ACE及ACE2 mRNA和蛋白表达，人血白蛋白可抑制其ACE2表达而促进ACE表达,这可能是白蛋白促肾小管间质损伤及肾小球硬化、间质纤维化的机制之一。     

槲皮素对四氯化碳诱导的大鼠肝纤维化抑制作用和肝保护作用

目的:探讨槲皮素对四氯化碳(CCl4)诱导大鼠肝纤维化的保护作用,并阐明槲皮素的抗肝纤维化机制.方法:Wistar大鼠随机分为5组:对照组、模型组、低剂量组(EXP-L)、中剂量组(EXP-M)、高剂量组(EXP-H).HE染色检测肝组织病理损伤情况;试剂盒检测谷丙转氨酶(ALT)和谷草转氨酶(AST)活力;羟脯氨酸(...     

ACEI对大鼠缺氧性肺动脉高压血管内皮保护作用的研究

目的: 本研究旨在观察西拉普利对大鼠缺氧性肺动脉高压血管内皮的保护作用机制。方法: 观察大鼠缺氧性肺动脉高压的血液动力学的变化及循环内皮细胞、血管紧张素转化酶、脂质过氧化物的变化。结果: 缺氧性肺动脉高压时脂质过氧化物产生增加, 血管紧张素转换酶活性降低, 循环内皮细胞数量增加(P<0.01); 西拉普利治疗后脂质过氧化物降低, 循环内皮细胞数量减少(P<0.01)。结论: 西拉普利对血管内皮细胞有保护作用。     

JNK信号转导通路对白细胞介素-1β促肝星状细胞增殖的调控作用

目的：探讨JNK信号转导通路在白细胞介素- 1β（IL-1β）介导的促肝星状细胞（HSCs）增殖中的作用。方法:应用 Western印迹法检测JNK的活化程度。应用活细胞计数试剂盒-CCK-8检测HSCs增殖并观察JNK 特异性阻断剂SP600125对IL-1β促HSCs增殖的影响。结果：IL-1β有明 显促大鼠HSCs增殖作用，而经JNK特异性阻断剂 SP600125预处理后，IL-1β促HSCs增殖作用 受到抑制（1.560±0.110 vs 1.427±0.113，P<0.05）。IL-1β以时间依赖 方式激活JNK。IL-1β作用HSCs后0、5、15、30、60和120 min，JNK 活性分别为0.982±0 .299、1.501±0.720、2.133±0.882、3.360±0.452、2.181±0.789、1.385±0 .368。结论:IL-1β可刺激HSCs增殖，细胞内JNK信号转导通路参与了 IL-1β促HSCs增殖作用。     

The effects of angiotensin-converting enzyme inhibitors on peritoneal protein loss and solute transport in peritoneal dialysis patients

OBJECTIVE:

The objective of this study was to examine the effects of angiotensin-converting enzyme inhibitors on peritoneal membrane transport, peritoneal protein loss, and proteinuria in peritoneal dialysis patients.

METHODS:

Fifty-four peritoneal dialysis patients were included in the study. The patients were divided into two groups. Group 1 (n = 34) was treated with angiotensin-converting enzyme inhibitors. Group 2 (n = 20) did not receive any antihypertensive drugs during the entire follow-up. Eleven patients were excluded from the study thereafter. Thus, a total of 30 patients in Group 1 and 13 patients in Group 2 completed the study. We observed the patients for six months. Group 1 patients received maximal doses of angiotensin-converting enzyme inhibitors for six months. Parameters at the beginning of study and at the end of six months were evaluated. ClinicalTrial.gov: {"type":"clinical-trial","attrs":{"text":"NCT01575652","term_id":"NCT01575652"}}NCT01575652.

RESULTS:

At the end of six months, total peritoneal protein loss in 24-hour dialysate effluent was significantly decreased in Group 1, whereas it was increased in Group 2. Compared to the baseline level, peritoneal albumin loss in 24-hour dialysate effluent and 4-hour D/P creatinine were significantly increased in Group 2 but were not significantly changed in Group 1. A covariance analysis between the groups revealed a significant difference only in the decreased amount of total protein loss in 24-hour dialysate. Proteinuria was decreased significantly in Group 1.

CONCLUSION:

This study suggests that angiotensin-converting enzyme inhibitors reduce peritoneal protein loss and small-solute transport and effectively protect peritoneal membrane transport in peritoneal dialysis patients.     

黄芪对肺纤维化大鼠血清细胞因子及肺超微结构的影响

目的：研究黄芪水提物、黄芪皂苷对博莱霉素（BLM）所致肺纤维化大鼠血Th1/Th2型细胞因子平衡、TNF-α表达的调节作用及对肺上皮细胞超微结构的影响,探讨黄芪阻抑肺纤维化的效应机制。方法：Wistar大鼠随机分为空白组、BLM模型组、地塞米松组、黄芪水提物组、黄芪皂苷组；气管内注入BLM复制大鼠肺纤维化模型,造模后第2天开始药物干预,14天采用酶联免疫吸附法测定血IFN-γ、IL-4、TNF-α的含量,14天、28天观察肺上皮细胞超微结构变化情况。结果：模型组大鼠血IFN-γ含量降低,IL-4、TNF-α的含量升高（均P〈0.05）；与模型组比较,黄芪水提物组、黄芪皂苷组、地塞米松组使大鼠血IFN-γ含量明显升高（P〈0.05）,IL-4、TNF-α含量明显降低（P〈0.05）；黄芪水提物组、黄芪皂苷组与地塞米松组比较,上述指标均无统计学差异（P〉0.05）,超微结构观察,模型组肺泡Ⅱ型上皮细胞数量减少,细胞微绒毛稀少,核不规则,核染色质凝集粗块状,板层小体减少空泡样变,线粒体明显肿胀,肺泡间隔成纤维细胞增生明显,胞质内胶原纤维增多；各治疗组均可改善肺泡Ⅱ型上皮细胞超微结构的异常改变。结论：黄芪水提物、黄芪皂苷对肺纤维化大鼠肺泡Ⅱ型上皮细胞超微结构具有保护作用,调节Th1/Th2型细胞因子的平衡及TNF-α含量可能是其阻抑肺纤维化发生的机理之一。