Expression of interleukin 1β converting enzyme in 5-FU induced apoptosis in esophageal carcinoma cells

AIM To study the role of interleukin 1β converting enzyme (ICE) in antitumor drug induced apoptosis in tumor cells. METHODS Morphological changes in human esophageal carcinoma Eca 109 cells after treated with 5 fluorouracil (5 FU) were observed under light and electron microscope. Expression of ICE in the tumor cells exposed to 5 FU was examined by the immunocytochemical method. RESULTS The cells treated with 5 FU displayed disappearance of nucleoli, chromatin gathering under nuclear envelope, karyorrhexis, budding and the formation of apoptotic bodies. The expression of ICE was negative in control cells, and 5 FU could induce the ICE expression in Eca 109 cells undergoing apoptosis. The number and the staining intensity of positive cells increased with the extension of action time. CONCLUSION 5 FU may induce apoptosis in human esophageal carcinoma Eca 109 cells; ICE gene may be involved in the regulation of 5 FU induced apoptosis; and ICE protein may mediate apoptosis induced by 5 FU.

Neuronal apoptosis and interleukin 1-beta converting enzyme expression in the frontal cortex and hippocampus of mice with ischemia/reperfusion injury

BACKGROUND： Interleukin 1β-converting enzyme （ICE） gene expression can induce neuronal apoptosis. However, the dynamic changes in ICE gene expression and its effects on neuronal apoptosis under cerebral ischemia/reperfusion conditions remain unclear. OBJECTIVE： To observe neuronal apoptosis and changes in ICE gene expression in the frontal cortex and hippocampus following ischemia/reperfusion injury. DESIGN, TIME AND SETTING： A randomized, controlled animal study was conducted at the Laboratory of Experimental Animal Center, the Second Hospital of Jilin University and Central Laboratory, the Second Hospital of Jilin University, China, from November 2008 to September 2009. MATERIALS： The ICE gene primer sequence （TaKaPa Co., Dalian, China）, FACScan Flow cytometer （Becton Dickinson, Franklin Lakes, N J, USA）, and Perkin Elmer GeneAmp PCR system 2400 （Perkin Elmer, Waltham, MA, USA） were used in this study. METHODS： A total of 45 healthy, adult, male, Kunming mice were randomly assigned to normal control （n = 5）, sham surgery （n = 5）, and model （n = 35） groups. The mice in the model group were equally and randomly subdivided into seven subgroups according to various reperfusion time points （1 hour, 1,3, 7, 14, 28, and 42 days）. Animal models of ischemia/reperfusion injury were established by bilateral carotid artery ligation in the model group. The mice in the sham surgery group only received saline perfusion and surgery for carotid artery exposure. MAIN OUTCOME MEASURES： Neuronal apoptosis in the frontal cortex and hippocampus of mice was measured using flow cytometry. The time course of ICE mRNA levels in the frontal cortex and hippocampus were quantified using reverse transcription-polymerase chain reaction. RESULTS： Neuronal apoptosis in the frontal cortex and hippocampus peaked at 3 days following ischemia/reperfusion injury （P 〈 0.05）. ICE mRNA expression increased in the frontal cortex at 1 day following ischemia/reperfusion injury （P 〈 0.05）, decreased at 3 days, and then peaked at 14 days （P 〈 0.05）. ICE mRNA expression increased in the hippocampus at 3 days following ischemia/reperfusion injury （P 〈 0.05）, peaked at 7 days （P 〈 0.05）, and then decreased gradually to normal levels at 28 days. CONCLUSION： Neuronal apoptosis peaked at 3 days following ischemia/reperfusion injury, and both apoptosis and ICE mRNA levels remained high for 2 weeks after injury. Early apoptosis may result from increased ICE mRNA expression.

Effect of angiotensin Ⅱ type 1 receptor blocker and angiotensin converting enzyme inhibitor on the intraocular growth factors and their receptors in streptozotocin-induced diabetic rats

AIM： To investigate the effect of angiotensin II type 1 receptor blocker （ARB） and angiotensin converting enzyme inhibitor （ACEI） on intraocular growth factors and their receptors in streptozotocin-induced diabetic rats. METHODS： Forty Sprague-Dawley rats were divided into 4 groups： control, diabetes mellitus （DM）, candesartan- treated DM, and enalapril-treated DM （each group, n---10）. After the induction of DM by streptozotocin, candesartan [ARB, 5 mg/（kg · d）] and enalapril [ACEI, 10 mg/（kg · d）] were administered to rats orally for 4Wko Vascular endothelial growth factor （VEGF） and angiotensin II （Ang II） concentrations in the vitreous were measured using enzyme-linked immunosorbent assays, and VEGF receptor 2 and angiotensin II type 1 receptor （ATIR） levels were assessed at week 4 by Western blotting. RESULTS： Vitreous Ang II levels were significantly higher in the DM group and candesartan-treated DM group than in the control （P=0.04 and 0.005, respectively）. Vitreous ATIR increased significantly in DM compared to the other three groups （P〈0.007）. Candesartan-treated DM rats showed higher vitreal ATIR concentration than the enalapril-treated DM group and control （P〈0.001 and P=0.005, respectively）. No difference in vitreous Ang II and ATIR concentration was found between the enalapril- treated DM group and control. VEGF and its receptor were below the minimum detection limit in all 4 groups. CONCLUSION： Increased Ang II and ATIR in the hyperglycemic state indicate activated the intraocular renin-angiotensin system, which is inhibited more effectively by systemic ACEI than systemic ARB.

Plasma Levels of Angiotensin-Converting Enzymes 1 and 2 and <i>AGTR</i>2 (T1247G and A5235G) Gene Polymorphisms Are Associated to Breast Cancer Progression

Background: Breast cancer is the most common type of cancer among women. Diagnosed and treated timely, patients may have good prognostics. In Brazil, in 2012, the estimate of new cases was 52,680 and the number of registered deaths in 2012 was 12,852. The Renin-Angiotensin System (RAS) is known for its role in arterial hypertension and in other cardiovascular diseases. Angiotensin-Converting Enzyme 2 (ACE2) is the key to Ang-(1-7) formation, and counterbalances the ACE1/AngII/AGTR1 axis actions. RAS components have complex interactions with different tissues and their actions are not restricted to the cardiovascular system. Recently, the RAS has been associated with different types of cancers and in particular with gynecological cancers. Objectives: Our aim is to investigate possible associations between allelic distribution of two genetic polymorphisms in the AGTR2 receptor with ACEs 1 and 2 plasma levels among women with breast cancer. Patients and Methods: Patients with breast cancer were genotyped for two polymorphisms of the AGTR2 (T1247G and A5235G). Genotyping assays (TaqMan) were performed with genomic DNA extracted from blood cells. ACEs plasma level measurements were conducted in women from the breast-cancer group (N = 53). ACEs were measured in the plasma of these patients using ELISA kits. Results: SNPs genotype distribution is correlated with ACEs plasma levels. ACEs plasma levels are also correlated with clinical variables and ACE2 high levels are associated with better prognostics. Conclusions: Changes in circulating levels of ECA1/AngII ECA2/ Ang-(1-7) determine the magnitude of the inflammatory response that an individual can trigger and the variation in ACE 1 and 2 plasma level measurements in the blood of breast cancer patients suggests an association with the process of mammary carcinogenesis. Thus, the RAS may be associated with the process of mammary carcinogenesis by both genotypic variations of RAS components and by circulating levels of ACEs.

Serum levels of angiotensin converting enzyme as a biomarker of liver fibrosis

The renin angiotensin system(RAS) is classically conceived as a circulating hormonal system involved in blood pressure control and hydroelectrolyte balance. The discovery that RAS components are locally expressed in a wide range of organs and tissues,including the liver,pointed to a role for this system in the pathogenesis of several conditions including hepatic fibrosis and cirrhosis. It has been widely reported that the classical RAS axis composed by the angiotensin converting enzyme(ACE)-angiotensin(Ang) Ⅱ-Ang type 1(AT1) receptor mediates pro-inflammatory,pro-thrombotic,and pro-fibrotic processes. On the other hand,the alternative axis comprising ACE2-Ang-(1-7)-Mas receptor seems to play a protective role by frequently opposing Ang Ⅱ action. Chronic hepatitis B(CHB) is one of the leading causes of liver fibrosis,accounting for the death of nearly one million people worldwide. Liver fibrosis is a key factor to determine therapeutic interventions for patients with CHB. However,the establishment of non-invasive and accurate methods to detect reversible stages of liver fibrosis is still a challenge. In an elegant study published in the 36 th issue of the World Journal of Gastroenterology,Noguchi et al showed the predictive value of serum ACE levels in detecting not only advanced stages of liver fibrosis but also initial and intermediate fibrotic stages. The serum levels of ACE might represent an accurate,non-invasive,widely available,and easy method to evaluate fibrosis related to CHB. Moreover,therapies involving the inhibition of the classical RAS axis components might be promising in the control of CHB-related liver fibrosis.

Effects of Yinchenhao Decoction on Self-regulation of Renin-angiotensin System by Targeting Angiotensin Converting Enzyme 2 in Bile Duct-ligated Rat Liver

Summary： In order to investigate whether Yinchenhao decoction （YCHD） attenuates hepatic fibro- genesis in the bile duct ligation （BDL） model via recovering and restoring the self-regulation and bal- ance of the renin-angiotensin system （RAS）, 33 specific-pathogen-free （SPF） male Sprague-Dawley rats with common BDL and scission were randomly divided into five groups as follows： G1, the sham group （n=4）; G2, BDL 7-day group （n=5）; G3, BDL＋YCHD 430 mg/mL （n=8）; G4, BDL＋losartan 0.65 mg/mL （ARB group, n=8）; G5, model group （BDL without any treatment, n=8）. YCHD and losartan （10 mL.kgl.day-1） were given by gastric gavage for 16 days following BDL in G3 and G4 groups, respec- tively. The effect of YCHD on liver fibrosis and the detailed molecular mechanisms were assessed by liver function including total bilirubin （TBIL）, direct bilirubin （DBIL）, indirect bilirubin （IDBIL）, alanine aminotransferase （ALT）, and aspartate aminotransferase （AST）. Histological changes were ob-. served by transmission electron microscopy （TEM） and Masson trichrome staining. Western blotting was used to detect the protein expression level of the renin-angiotensin system （RAS） components in- cluding angiotensin converting enzyme （ACE）, angiotensin II type 1 receptor （AT1R）, ACE2, angio- tensin II （Ang II） as well as transforming growth factor 131 （TGF131）. The experimental data were ana- lyzed by principle component analytical method of pattern recognition. The results showed that bio- chemically, serum TBIL, DBIL, IDBIL, ALT and AST levels were markedly increased following BDL as compared with the sham group （P〈0.05）. Serum TBIL, IDBIL and DBIL levels in G3 group were dramatically decreased as compared with G5 and G4 groups （P〈0.05）. Serum AST level in G3 was sig- nificantly lowered than in G5 group （P〈0.05）, but there was no significant difference in ALT among G3, G4 and G5 groups （P〉0.05）. Histologically, livers in G3 group showed less hepatocytes necrosis, less bile duct hyperplasia and less collagen formation than in G4 and G5 groups. The protein expression lev- els of ACE2, ACE, Ang II, AT1R and TGF131 in G2, G3 and G4 groups were significantly higher than in sham group （P〈0.05）, and lower than in G5 group （P〈0.05）. However, the differences among G2, G3 and G4 groups were not significant （P〉0.05）. ACE2 protein expression in G3 group was significantly higher than in G2 group （P〈0.05） and there was no significant difference in comparison with G4 group （P〉0.05）. Moreover, the protein expression of TGF131 in G3 group was significantly lower than in G5 and G4 groups （P〈0.05）. Our findings suggest that the antifibrotic effects of YCHD may be associated with the decreased classical RAS pathway components and TGFI31 downexpression so as to recover and rebuild self-regulation of the RAS by elevating the protein expression of ACE2.

Elevated pancreatic enzymes, IgM, soluble interleukin-2 receptor in anti-GADab(+) type 1 diabetes

Type 1 diabetes can be classified into immune-mediated diabetes (type 1A) and idiopathic diabetes, which lacks immunological evidence for beta cell autoimmunity (type 1B). Type 1A diabetes is characterized by the presence of the anti-glutamic acid decarboxylase antibody (anti-GADab). Fulminant type 1 diabetes is classified as type 1B diabetes, and characterized by the absence of anti-GADab, flu-like symptoms, and elevated serum exocrine pancreatic enzymes. We report a type 1 diabetic patient who showed flu-like symptoms, elevated serum exocrine pancreatic enzymes, and an extremely high-titer of anti-GADab, manifesting the characteristics of both type 1A and fulminant type 1 diabetes.

血清多配体蛋白聚糖1、血管紧张素转换酶2与川崎病患儿冠状动脉损害和临床疗效的关系

目的研究血清多配体蛋白聚糖1(syndecan-1,SDC-1)、血管紧张素转换酶2(angiotensin-converting enzyme 2,ACE2)与川崎病(Kawasaki disease,KD)患儿冠状动脉损害(coronary artery lesions,CAL)和临床疗效的关系。方法选取2020年6月至2022年3月收治的113例KD急性期患儿作为研究对象并设为KD组,治疗前行超声心动图检查,根据检查结果分为CAL组(n=39例)和无CAL组(n=74例);另选取同期体检的60例健康儿童作为对照组。收集受试者临床资料,采用酶联免疫吸附法检测血清SDC-1、ACE2水平并比较。患儿治疗后进行疗效评估,将治愈与好转患儿纳入治疗有效组(n=89例),无效患儿纳入治疗无效组(n=24例)。采用多因素logistic回归模型分析患儿治疗无效的影响因素,受试者工作特征(ROC)曲线分析血清SDC-1、ACE2等指标及其联合应用模型对治疗无效的预测价值。结果KD组血清SDC-1、ACE2水平显著高于对照组(P<0.05)。CAL组血清SDC-1、ACE2水平高于无CAL组(P<0.05)。治疗有效组与治疗无效组在发热时间、降钙素原(procalcitonin,PCT)、C反应蛋白(C-reactive protein,CRP)、白细胞介素6(interleukin-6,IL-6)、SDC-1、ACE2水平方面比较差异有统计学意义(P<0.05)。多因素logistic回归分析显示:PCT、CRP、IL-6、SDC-1、ACE2为患儿治疗无效的显著影响因素(P<0.05)。ROC曲线分析显示:血清SDC-1、ACE2两指标联合预测患儿治疗无效的曲线下面积(AUC)(95%CI)为0.837(0.726~0.926),预测价值均高于单独检测。血清SDC-1、ACE2及PCT、CRP、IL-6联合预测患儿治疗无效的AUC(95%CI)为0.903(0.861~0.912),预测价值较血清SDC-1、ACE2两指标联合预测有所提升。结论KD患儿血清SDC-1、ACE2水平上升,两者水平越高,其并发CAL风险越高,临床疗效越差,联合检测对临床疗效具有较好的预测价值。

急性心肌梗死PCI术后HF患者外周血NLRP3、Caspase⁃1、IL⁃1β的表达及意义

目的探讨急性心肌梗死(AMI)经皮冠状动脉介入治疗(PCI)术后心力衰竭(HF)患者外周血核苷酸寡聚化结构域样受体热蛋白结构域相关蛋白3(NLRP3)、半胱氨酸天门冬氨酸蛋白水解酶⁃1(Caspase⁃1)、白细胞介素⁃1β(IL⁃1β)的表达及意义。方法选取许昌医院2018年5月至2023年5月收治的82例接受PCI术的AMI患者,根据术后是否发生HF将其分为HF组(33例)与非HF组(49例)。比较两组外周血NLRP3、Caspase⁃1、IL⁃1β的mRNA表达水平以及超声心动图指标[左心室射血分数(LVEF)、左室峰射血率时间(LVT⁃PER)、左室峰射血率(LVPER)]、心肌损伤标志物[肌酸激酶同工酶(CK⁃MB)、肌钙蛋白Ⅰ(cTnⅠ)、肌红蛋白(Mb)]。分析外周血NLRP3、Caspase⁃1、IL⁃1β表达水平与超声心动图指标、心肌损伤标志物的相关性,以及对AMI患者PCI术后HF发生的预测价值。结果HF组外周血NLRP3、Caspase⁃1、IL⁃1β的mRNA水平均高于非HF组,差异有统计学意义(P<0.05)。AMI患者外周血NLRP3、Caspase⁃1、IL⁃1β水平之间均存在显著正相关性(P<0.05)。HF组LVEF、LVT⁃PER均低于非HF组,LVPER、CK⁃MB、cTnⅠ、Mb均高于非HF组,差异有统计学意义(P<0.05)。AMI患者外周血NLRP3、Caspase⁃1、IL⁃1β水平与LVEF、LVT⁃PER存在显著负相关性,与LVPER、CK⁃MB、cTnⅠ、Mb存在显著正相关性(P<0.05)。ROC曲线分析显示,外周血NLRP3、Caspase⁃1、IL⁃1β以及联合对AMI患者PCI术后HF发生均有预测效能(P<0.05),其AUC分别为0.800、0.930、0.978、0.994,其中联合效能最高,敏感性为96.97%,特异性为95.92%。结论PCI术后发生HF的AMI患者外周血NLRP3、Caspase⁃1、IL⁃1β表达增加,三指标可预测术后HF发生,且联合预测效能最高。

血清IL-41及ACE2水平诊断小儿川崎病的临床价值

目的探索川崎病(KD)患儿血清白介素41(IL-41)、血管紧张素转化酶2(ACE2)表达水平及临床诊断价值。方法选取2018年12月—2022年12月合肥市妇幼保健院普儿科诊治KD患儿80例为KD组,根据超声心动图结果,分为冠状动脉损伤(CAL)亚组(n=26)和非CAL亚组(n=54),以同期急性上呼吸道感染伴发热患儿为对照2组(n=40),同期行择期手术的腹股沟斜疝患儿为对照1组(n=40)。采用酶联免疫吸附法检测血清IL-41、ACE2水平;采用Pearson相关分析血清IL-41、ACE2与临床资料的相关性;多因素Logistic回归分析影响KD患儿发生CAL的影响因素;绘制受试者工作特征曲线分析血清IL-41、ACE2预测KD患儿发生CAL的价值。结果血清IL-41、ACE2水平比较,KD组>对照2组>对照1组,差异均有统计学意义(F/P=519.731/<0.001,1115.501/<0.001);KD组患儿血清IL-41、ACE2水平与发热时间、C反应蛋白及降钙素原呈正相关(IL-41:r/P=0.562/<0.001,0.589/<0.001,0.613/<0.001;ACE2:r/P=0.622/<0.001,0.609/<0.001,0.574/<0.001)。CAL亚组发热时间、C反应蛋白、降钙素原、IL-41、ACE2均高于非CAL亚组,差异有统计学意义(t/P=3.459/0.001,11.187/<0.001,11.377/<0.001,12.299/<0.001,25.882/<0.001)。血清IL-41、ACE2、发热时间、C反应蛋白、降钙素原升高是影响KD患儿CAL发生的独立危险因素[OR(95%CI)=1.598(1.271~2.010),1.573(1.241~1.994),1.384(1.142~1.667),1.496(1.171~1.912),1.513(1.159~1.975)]。血清IL-41、ACE2及两项联合预测KD患儿发生CAL的AUC分别0.812、0.815、0.878,两项联合的AUC大于血清IL-41、ACE2单一检测(Z/P=5.116/<0.001、4.217/0.009)。结论KD患儿血清IL-41、ACE2升高,与发热时间、C反应蛋白及降钙素原有关,两者联合对KD患儿CAL发生具有较高的预测价值。

ACE2-Ang(1-7)-MasR轴对心血管系统保护作用的研究进展

肾素-血管紧张素系统(RAS)是人体重要的体液调节系统之一,在维持机体血压稳定和水电解质平衡中发挥了重要作用。近年来,发现了RAS系统的新支路,ACE2和Ang(1-7)等是心血管系统的关键保护因子,ACE2-Ang(1-7)-MasR轴成为了心血管疾病领域的研究热点。非经典的RAS系统ACE2-Ang(1-7)-MasR轴能够拮抗经典的ACE-AngⅡ-AT1R轴,二者共同维系机体的平衡,该轴在高血压、冠心病、心律失常和心力衰竭等心血管疾病治疗中可能成为新的治疗靶点。

血管紧张素转换酶2-血管紧张素(1-7)-Mas受体轴抗炎机制研究进展

肾素-血管紧张素系统(RAS)与炎症反应关系密切。血管紧张素Ⅱ(AngⅡ)是RAS的主要参与者,可激活与组织损伤、炎症相关的信号通路,而血管紧张素转换酶2(ACE2)-血管紧张素(Ang)(1-7)-Mas受体轴可发挥与AngⅡ相反的作用,抑制炎症反应。本文拟对ACE2-Ang(1-7)-Mas受体轴的基本特性及其在心血管疾病、肾病、肺损伤、神经性疾病中的抗炎机制的研究进展作一综述。

溃疡性结肠炎患者血清Rac1、ACE2水平与肠道功能损害的相关性

目的探讨溃疡性结肠炎(ulcerative colitis,UC)患者血清Ras相关的C3肉毒素底物1(Rac1)、血管紧张素转化酶2(ACE2)水平与肠道功能损害的相关性。方法选取2020年3月至2022年3月驻马店中心医院就诊的UC患者60例为病例组,依据Mayo评分将病例组分为活动期组(36例)、缓解期组(24例),同时以同期健康体检者40例为对照组。分别静脉抽取所有研究对象空腹血液,ELISA法检测血清ACE2水平;q RT-PCR检测血清中Rac1 m RNA水平;同时检测肠道屏障指标(血清二胺氧化酶、细菌内毒素、D-乳酸)水平;采用Pearson法分析UC患者血清中ACE2、Rac1 m RNA水平的相关性以及两者分别与血清二胺氧化酶、细菌内毒素、D-乳酸的相关性。结果与对照组相比,病例组血清中ACE2、Rac1 m RNA水平显著下降,二胺氧化酶、细菌内毒素、D-乳酸水平显著增加(P<0.05);与缓解期组相比,活动期组ACE2、Rac1 m RNA水平显著下降(P<0.05);Pearson法结果显示,UC患者血清中ACE2与Rac1水平呈明显正相关(r=0.390,P<0.05),且分别与二胺氧化酶、细菌内毒素、D-乳酸呈负相关(P<0.05)。结论UC患者血清中ACE2与Rac1水平呈正相关,且分别与肠道屏障指标呈负相关,检测两者水平对于评定肠道功能损害可能具有一定临床价值。

Changes of chymase,angiotensin converting enzyme and angiotensin Ⅱ type 1 receptor expressions in the hamster heart during the development of heart failure

Background Little is known about the role of dual angiotensin Ⅱ forming pathways during heart failure. In the present study, the changes of chymase and angiotensin converting enzyme （ACE） expressions in the failing hearts of hamsters were analysed. Methods Heart failure was induced by ligation of left anterior descending branch of the coronary artery. Chymase, ACE and angiotensin Ⅱ type 1 receptor （AT1R） mRNA levels were analysed by reverse transcription polymerase chain reaction （RT-PCR）. The activities of chymase and ACE were determined by radioimmunoassay （RIA）. Myocardial collagen fibre analysis was performed under optical microscope. Results Left ventricular systolic pressure （LVSP） and maximum left ventricular developed pressure increase rate （ dp/dtmax, mmHg/s） gradually moved lower at 2, 3,4 and 8 weeks after operation. On the other hand, left ventricular end-diastolic pressure （LVEDP） increased gradually after operation. Compared with the control group （3.55±0. 06, 4.79±0.70）, the heart weight/body weight ratio in operation group had increased significantly at 4 weeks and 8 weeks （4. 28±0. 43, 6. 17±0.73） （P 〈0. 01 ）. Collagen staining showed that the quantity of myocardial collagen fibre increased significantly in the operation group. RT-PCR showed that the chymase mRNA level in the operation group was consistently greater than that in the control group. ATIR mRNA level was also increased significantly at 3 weeks and 4 weeks, both being 1.3 times that of the control group （ P〈0.01 ）, whereas ACE mRNA level was not changed. Higher activity of chymase was detected in operation group, being 4, 8, 13 and 19 times that of the control group at 2, 3, 4 and 8 weeks （P〈0.01 ）, respectively. ACE activity was also significantly higher at the same time, being 7, 10, 10 and 3.5 times that of the control（P〈0.01）. Angiotensin Ⅱ （Ang Ⅱ） level in operation group increased significantly, being 2.5, 2.7, 3.5 and 2 times that of the control group at 2, 3, 4 and 8 weeks, respectively （P 〈 0. 01 ）. Conclusions A dual Ang Ⅱ forming pathway from both ACE and chymase in the hamster hearts plays an important role during the development of heart failure. At the decompensatory stage, the reduction of AngⅡ level may be associated with the decrease of ACE activity.

中药淫羊藿苷逆转肝癌HepG2.2.15细胞恶性表型及诱导分化研究

目的:探讨淫羊藿苷(ICA)诱导HepG2.2.15细胞分化凋亡的作用机制.方法:MTT法检测细胞增殖;流式细胞术(FACS)检测细胞周期分布;RT-PCR方法检测P27基因、FLIP基因mRNA表达水平的变化;AnnexinⅤ-FITC/PI双染法检测ICA处理后HepG2.2.15细胞凋亡的变化;电化学发光法和速率散射比浊法分别检测ICA处理后HepG2.2.15细胞上清液中甲胎蛋白(AFP)和转铁蛋白(Tf)水平变化.结果:ICA作用HepG2.2.15细胞增殖呈抑制作用,具有时间依赖性.ICA处理后,HepG2.2.15细胞周期各时相分布与对照组相比发生变化,G0/G1期升高,S期减小,与对照组相比有显著性差异(56.26±1.56%vs49.68±1.34%,19.95±1.24%vs28.02±1.03%;P<0.01).ICA分别上调HepG2.2.15细胞P27和下调FLIP基因mRNA的表达水平(0.78vs0.27,0.54vs0.90);HepG2.2.15细胞凋亡率增加,AFP下降,Tf水平升高,与对照组相比均有显著性意义(7.09%vs0.59%,156±46mg/Lvs285±58mg/L,152.1±26mg/Lvs67.1±24mg/L;P<0.05).结论:ICA可能通过升高G0/G1期,减少S期抑制HepG2.2.15细胞的增殖;上调P27mRNA和Tf水平,下调FLIPmRNA的表达和AFP合成的水平来诱导细胞分化.

ACE-1基因多态性与缺血性脑卒中的遗传易感性研究

目的 研究血管紧张素 - 1转换酶 ( ACE- 1)基因多态性与缺血性脑卒中 ( IS)的关系。方法 运用多聚酶链反应 ( PCR)技术检测 143例 IS及 15 4例对照组患者 ACE- 1基因多态性。结果 IS组 ACE- 1基因 DD基因型和 D等位基因与对照组比较差异无统计学意义 ;重型组 ID基因型与对照组相比有统计学意义 ,D等位基因与临床IS相关。结论 ACE- 1基因 ID基因型可能是 IS危险因子 ,ID基因型与重型 IS有关。

ACEI联合ARB对维持性血液透析患者CRP、IL-1和IL-6的影响

目的探讨联合应用血管紧张素转换酶抑制剂(ACEI)和血管紧张素Ⅱ受体拮抗剂(ARB)对维持性血液透析(MHD)患者C-反应蛋白(CRP)、白细胞介素-1(IL-1)和白细胞介素-6(IL-60)的影响。方法选取2012年2月至2014年12月期间在解放军第323医院肾病血透中心治疗的无明显感染症状的MHD患者50例,口服盐酸贝那普利10 mg/d和厄贝沙坦150 mg/d,于用药前和用药1、3、6个月测量血压、血红蛋白、血浆白蛋白、Kt/V值、残余尿量,检测炎症指标CRP、IL-1、IL-6水平,统计分析各项指标差异。结果 42例患者入组获得分析数据。服药6个月时血红蛋白、血浆白蛋白均呈现升高趋势,与用药前比较差异均有统计学意义(P<0.05);服药3、6个月时,CRP显著降低,与用药前比较差异均有统计学意义(P<0.05);IL-1和IL-6水平在1、3、6个月均表现出降低趋势,与用药前比较差异均有统计学意义(P<0.05);服药期间Kt/V值、残余尿量无显著变化(P>0.05)。结论联合应用ACEI和ARB类药物可降低血液透析患者的CRP、IL-1和IL-6水平,可能对改善患者微炎症状态带来益处。

应激状态下胃粘膜组织ECE-1 mRNA表达变化及其意义

为探讨内皮素转化酶(ECE) 1mRNA表达在应激性溃疡(SU)发病中的意义 ,以大鼠冷束缚应激性 (CRS)胃溃疡为模型 ,采用放免、逆转录聚合酶链反应 (RT PCR)及斑点杂交 (Dotblot)等方法 ,动态检测应激前后血浆胃粘膜组织内皮素 1(ET 1)含量、ECE 1mRNA表达水平、胃粘膜血流量(GMBF)及溃疡指数(UI)等指标的变化情况。结果显示 :与正常对照组相比 ,应激组大鼠血浆胃粘膜组织ET 1水平明显升高 (P <0 0 5 ) ,GMBF明显下降 (P <0 0 1)及UI明显增加 (P <0 0 1) ;胃粘膜组织ET 1水平与UI呈显著正相关 (r=0 98,P <0 0 1) ,而与GMBF呈显著负相关 (r=- 0 89,P<0 0 5 )。RT PCR及Dotblot显示各应激组胃粘膜组织中ECE 1mRNA表达水平均较正常对照组明显升高 (P <0 0 1) ,且分别与胃粘膜ET 1水平及UI呈显著正相关 (r=0 93 ,0 95 ,P <0 0 1) ,与GMBF水平呈显著负相关(r=- 0 91,P <0 0 5 )。提示在冷束缚应激诱发大鼠SU形成过程中 ,胃粘膜组织可显著增加ECE 1mRNA的表达水平与ET 1水平 ,从而参与SU的病理生理过程 。

氨对大鼠胃上皮细胞凋亡和白介素1β-转化酶mRNA表达的影响

探讨幽门螺杆菌（Ｈｐ）感染诱发胃上皮细胞凋亡的机制。方法： ３０只Ｗｉｓｔａｒ大鼠随机分成实验组和正常对照组，实验组大鼠饮用氨水３个月，造成胃粘膜损伤模型。采用ＴＵＮＥＬ方法检测鼠胃上皮细胞凋亡情况；采用ＲＴ－ＰＣＲ方法检测胃粘膜白介素１β－转化酶（ＩＣＥ）ｍＲＮＡ的表达。结果：与正常对照组大鼠相比，实验组大鼠胃粘膜肉眼观察见充血、水肿，病理检查示炎症和轻度萎缩改变。实验组胃粘膜上皮细胞凋亡指数（６．９±１．３）显著高于对照组（１．２±０．５， Ｐ＜０．０１），实验组ＩＣＥ ｍＲＮＡ经ＲＴ－ＰＣＲ后的电泳条带用自动图象仪处理测得相对值为０．８３±０．２０，而正常对照组为０．４７±０．１０，差异有显著性（Ｐ＜０．０１）。结论：氨所致的胃粘膜损伤可能与其诱发ＩＣＥ ｍＲＮＡ表达，引起胃粘膜上皮细胞凋亡有关。

高血压肾损害患者IL-6-174基因多态性及其与贝那普利治疗反应的相关性研究

目的分析白细胞介素-6(IL-6)-174基因多态性与高血压肾损害患者之间,及其与贝那普利治疗反应的相关性。方法选取284例高血压病患者,根据24 h尿蛋白排泄率将患者分为高血压组152例(24 h尿蛋白排泄率<20μg/min)和高血压肾损害组132例(24 h尿蛋白排泄率≥20μg/min);同时选取160例健康体检者作为正常血压组。所有入选对象留取血清标本,检测IL-6水平(酶联免疫检测法)及IL-6-174基因多态性(PCR-RFLP法)。以贝那普利对高血压肾损害患者进行干预,比较IL-6-174不同基因型对贝那普利的治疗反应。结果高血压患者的IL-6-174以CC基因型为多见,其次为GG、GC,G/C等位基因频率分别为47%、53%。高血压肾损害患者的IL-6-174以GG基因型最为多见,其次为GC和CC,G/C等位基因频率分别为68%、32%,基因型及G/C等位基因频率比较差异有统计学意义(P<0.05)。IL-6-174的CC基因型贝那普利治疗前后尿蛋白排泄率和IL-6的改变幅度最大,其次是GC和GG基因型,3组间比较差异有统计学意义(P<0.05)。结论 IL-6-174的G等位基因频率与宁夏高血压肾损害相关,高血压肾损害患者的IL-6-174的基因型以GG最为多见。IL-6-174的CC基因型对贝那普利治疗的反应最佳,尿蛋白排泄率下降最快。