金属蛋白酶组织抑制因子-1对心室重构影响的研究进展

心力衰竭是各种原因导致的心脏病的终末阶段,其发生和发展的基本机制是心室重构,而后者主要归结于细胞外基质合成和降解间失衡。金属蛋白酶组织抑制因子-1(tissue matrix metalloproteinase inhibitor-1,TIMP-1)是一种重要的基质金属蛋白酶(matrix metalloproteinases,MMPs)特异性抑制因子,可局部调节MMPs的活性,减少细胞外基质的降解,从而在许多心血管疾病的心室重构过程中发挥了重要作用,本研究就TIMP-1对心室重构影响的研究进展做一综述。     

达格列净对慢性心力衰竭大鼠心肌细胞基质金属蛋白酶及其组织抑制因子-1水平的影响

目的 探讨达格列净对慢性心衰大鼠心肌细胞基质金属蛋白酶-2(MMP-2)、MMP-9及其组织抑制因子-1(TIMP-1)表达的影响。方法 以腹主动脉缩窄(AC)法建立慢性心力衰竭大鼠模型。将50只雄性SD大鼠随机分成假手术组、假手术+达格列净组、模型组、模型+达格列净组。假手术+达格列净组和模型+达格列净组予以每天1次喂养达格列净1 mg·kg^-1,喂养8周。以Masson染色法观察心肌组织胶原纤维;以酸水解法检测心肌组织羟脯氨酸并计算胶原总含量;以实时荧光定量聚合酶链反应检测各组心肌组织MMP-2、MMP-9、TIMP-1表达情况。结果 假手术组、假手术+达格列净组、模型组、模型+达格列净组大鼠心肌组织胶原容积分数分别为(2.01±0.43)%、(1.98±0.80)%、(4.61±0.52)%和(3.48±0.74)%;羟脯氨酸含量分别为(0.27±0.06)、(0.25±0.08)、(0.63±0.05)和(0.47±0.10)μg·mg^-1;胶原总含量分别为(2.05±0.21)、(2.01±0.41)、(4.70±0.32)和(3....     

基质金属蛋白酶-9和金属蛋白酶组织抑制因子-1在妊高症患者体内的表达及临床意义

目的探讨基质金属蛋白酶-9（MMP-9）和金属蛋白酶组织抑制因子-1（TIMP-1）在妊娠期高血压患者胎盘组织中的表达及其临床意义。方法以2011年3月至2013年3月期间,本院诊治的90例妊娠期高血压患者作为观察组,选取同期40例正常妊娠产妇作为对照组,通过免疫组织化学方法 ,检测两组胎盘组织中MMP-9和TIMP-1的表达水平。结果与对照组相比,观察组MMP-9中阳性和强阳性率明显减少,P〈0.05,而TIMP-1阳性率变化不大,差异没有统计学意义。结论胎盘组织中MMP-9表达下降,引发胎盘局部缺血、缺氧,最终导致妊娠期高血压。     

Arresten蛋白通过调控上皮-间质转化抑制宫颈癌细胞的迁移

目的探讨Arresten蛋白对宫颈癌Siha细胞迁移的影响及其可能的作用机制。方法采用四甲基偶氮唑盐(MTT)实验、划痕实验和Boyden小室趋化实验检测Arresten蛋白对宫颈癌Siha细胞增殖及迁移能力的影响;通过免疫细胞化学法检测细胞迁移分子标志基质金属蛋白酶-9(MMP-9)的表达水平;利用实时荧光定量聚合酶链反应(PCR)和蛋白质印迹法检测上皮-间质转化(EMT)标志物E-cadherin与N-cadherin的水平变化。结果经3.2μg/ml Arresten蛋白处理的Siha细胞,其增殖能力没有明显改变(P>0.05)。通过对划痕实验中细胞迁移距离和穿越Boyden小室的细胞数目进行分析,发现经3.2μg/ml Arresten蛋白处理的Siha细胞迁移功能受到抑制(P<0.01),细胞内迁移分子标志基质金属蛋白酶(MMP)-9的表达水平也显著降低(P<0.01)。Arresten蛋白可诱导Siha细胞内上皮标志物E-cadherin表达上调(P<0.01),间充质细胞标志物N-cadherin表达下调(P<0.01)。结论 Arresten蛋白对宫颈癌细胞迁移有抑制作用,可能通过抑制EMT途径发挥作用。。     

大鼠肝纤维化模型肝组织及血清中基质金属蛋白酶抑制因子-1的动态变化

目的观察二甲基亚硝胺(DMN)构建大鼠肝纤维化模型过程中肝组织及血清中TIMP-1及其相关指标的动态变化。方法1%DMN按10mg/Kg腹腔注射方法建立大鼠肝纤维化模型,第1w连续注药3d,第2、3、4、5、6w各连续2d。分别于第1、2、4、6、8w随机取血及大鼠肝组织。ELISA方法测血清基质金属蛋白酶抑制因子-1(TIMP-1)、基质金属蛋白酶-1(MMP-1)、透明质酸(HA)、层粘连蛋白(LN)、Ⅰ型前胶原(Ⅰ-CP)、Ⅲ型前胶原(Ⅲ-CP)含量。肝组织HE、Masson染色观察肝病理变化、胶原纤维含量,原位杂交观察TIMP-1mRNA表达;免疫组化观察TIMP-1蛋白表达量。结果注药后1w,血清TIMP-1即开始升高,4w后维持高值(131.1±13.16)ng/ml,肝组织内TIMP-1mRNA及蛋白同步增高;1w时,MMP-1含量增高,2w处于高峰(79.3±8.45)ng/ml,随后缓慢降低,8w时含量最低(46.6±6.31)ng/ml,然而仍高出正常对照组(32.1±3.66)ng/ml;1w时,HA含量升幅达对照组2倍,后持续增高,至6w时处高峰(350.8±40.26)ng/ml;LN于1w、2w、4w均小幅递增,6w达高峰(132.8±15.1)ng/ml;CP-I含量注药后1w始较对照组升高,6w处高峰(27.2±3.60)ng/ml;CP-Ⅲ含量用药后小幅升高,6w处于高峰(9.06±1.22)ng/ml。肝胶原纤维百分含量4w小幅递增,4w后,肝胶原纤维显著持续增高,8w时最高。大鼠肝组织病理变化显示:1w可见肝细胞变性及小点状坏死灶,4w时,纤维间隔伴小叶结构紊乱,符合肝纤维化3级,6w小部分3级,大部4级;8w,肝硬化表现。结论DMN构建大鼠肝纤维化模型肝组织及血清相关指标呈动态递进发展,为进一步选择合适时机干预肝纤维化进程提供依据、奠定基础。     

熊果酸对香烟诱导肺气肿大鼠干预作用及其对肺组织MMP-9和TIMP-1表达的影响

目的 通过观察熊果酸对香烟诱导大鼠肺气肿的干预作用及其对肺组织MMP-9和TIMP-1表达的影响,探讨熊果酸减轻肺气肿的作用机制.方法 建立香烟诱导肺气肿大鼠模型,将40只SPF级雄性Wistar大鼠按随机数字表法分为健康对照组、模型组、熊果酸10mg/kg(UA10)组、熊果酸20 mg/kg(UA20)组以及熊果酸40 mg/kg(UA40)组,每组8只.取每只大鼠的右下叶肺组织,石蜡包埋切片,常规HE染色.对HE染色标本进行肺气肿评分即每视野的平均肺泡数和平均肺泡直径的测定.进行TIMP-1、MMP-9免疫组化染色,主要应用链霉素亲生物素蛋白-过氧化酶复合技术.对石蜡包埋的肺组织标本行MMP-9、TIMP-1相对含量的测定.结果 与对照组比较,模型组大鼠肺泡腔扩大、部分肺泡间隔断裂、肺泡腔融合、肺气肿形成.熊果酸各组大鼠可见局部轻度肺气肿形成,主要表现为肺泡间隔略增宽,肺泡腔呈局限性轻度扩大;模型组大鼠肺泡平均内衬间隔(120.17±10.86)μm明显高于对照组(47.76±3.55)pm、平均肺泡数(14.25±10.23)/mm2明显低于对照组(338.08±26.08)/mm2、MMP-9相对阳性面积(18.16±4.62)％明显高于对照组(5.38±0.93)％、TIMP-1 相对阳性面积(5.36±2.15)明显低于对照组(12.80±1.31)％,MMP-9/TIMP-1比值(3.38±0.79)明显高于对照组(0.42±0.16),均具有统计学差异(P＜0.01);与模型组比较,熊果酸10 mg/kg,20 mg/kg,40 mg/kg组平均内衬间隔分别为(104.48±10.85)μm、(80.82±7.68)μm、(53.90±9.92)μm均显著减少,平均肺泡数分别为(165.12±16.65)/mm2、(206.98±25.55)/mm2、(279.94±29.53)/mm2均显著增加,MMP-9蛋白表达含量分别为(14.31±1.90)％、(10.52±1.88)％、(6.90±0.43)％均显著减少,TIMP-1 蛋白表达含量分别为(7.07±1.3)％、(9.86±1.80)％、(11.56±1.46)％均升高,MMP-9/TIMP-1 比值分别为(2.02±0.54)、(1.06±0.37)、(0.52±0.23)均降低,均具有统计学差异(P＜0.01).结论 熊果酸有减轻香烟所致大鼠肺气肿作用,且熊果酸可能通过调节蛋白酶/抗蛋白酶失衡减轻肺气肿.     

2型糖尿病大鼠肺组织基质金属蛋白酶－2、9及其抑制因子－1的表达变化

目的通过检测2型糖尿病大鼠肺组织中基质金属蛋白酶-2（MMP-2）、基质金属蛋白酶－9（MMP-9）及基质金属蛋白酶抑制因子－1（TIMP—1）的表达变化,探讨MMP－2、MMP－9及TIMP－1在糖尿病肺损伤中的作用。方法通过高糖、高脂饮食加腹腔注射小剂量链脲佐菌素（streptozotocin,STZ）的方法,建立2型糖尿病大鼠模型,采用光镜动态观察12周、24周时对照组、糖尿病组的大鼠肺组织的形态学改变,用MassoN三色染色观察肺组织胶原沉积情况,应用免疫组织化学方法检测2组大鼠肺组织MMP－2、MMP－9及TIMP－1的动态表达变化情况。结果糖尿病大鼠肺组织胶原含量明显增多,糖尿病大鼠肺组织MMP－2、MMP－9、TIMP－1表达增多,同时MMP－2／TIMP－1、MMP-9／TIMP－1比例也增加。随病情进展,上述变化更加明显。结论糖尿病大鼠出现了肺间质纤维化,MMP-2、9及TIMP－1表达增多及其比例紊乱参与了糖尿病肺组织病变的发生发展,可能是糖尿病肺纤维化发生的机制之一。     

儿童心肌炎基质金属蛋白酶组织抑制因子-1检测与临床意义

目的 评价血液中基质金属蛋白酶组织抑制因子(TIMP)-1与儿童心肌炎的关系,验证临床检测TIMP-1的意义.方法 将年龄为1～14岁临床诊断为心肌炎患者9例与相匹配的对照组9例取血标本,测定门冬氨酸氨基转移酶(AST)、乳酸脱氢酶(LDH)、肌酸激酶(CK)、α-羟丁酸脱氢酶(HBDH)、肌酸激酶同工酶(CKMB),心肌肌钙蛋白Ⅰ(cTnI)、C反应蛋白及TIMP-1,并行超声心动图等检查.结果 超声心动图示心肌炎患儿射血分数显著降低、左心室扩大.儿童心肌炎患者TIMP-1水平较对照组显著降低.心肌炎患儿与对照组心肌酶、心肌肌钙蛋白Ⅰ、C反应蛋白及检测差异均无统计学意义.结论 对儿童心肌炎患者来说,血液中的TIMP-1水平可能能够显示其疾病的严重程度,且TIMP-1降低可能是心肌炎进展为扩张性心肌病的机制之一.     

基于网络药理学及实验验证探讨丹皮酚抑制血管内皮细胞间质转化的作用机制

目的 结合网络药理学与体外实验验证探讨丹皮酚抑制血管内皮细胞间质转化（EndMT）的关键靶点及作用机制。方法 通过PharmMapper、HERB、SymMap、ChEMBL、SwissTargetPredicition数据库获取丹皮酚的作用靶点;从GeneCards数据库中检索与EndMT相关的靶标,将两者进行交集分析与可视化;将交集靶点基因上传至STRING数据库联用Cytoscape3.8.2软件构建蛋白相互作用（PPI）网络并筛选核心靶点基因,并进行基因本体论（GO）功能富集和京都基因与基因组百科全书（KEGG）信号通路富集分析。采用Autodock Vina软件对丹皮酚与核心靶点基因进行分子对接验证。体外实验将HUVECs细胞分为空白组、模型[转化生长因子β1(TGF-β1)]组、丹皮酚低、中、高剂量组（30、60、120μmol/L）,通过蛋白质印迹法（Western blotting）筛选TGF-β1诱导EndMT最佳浓度并通过酶联免疫吸附测定（ELISA）检测基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)的蛋白表达量。结果 共筛选出丹皮酚作用靶点414个...     

基质金属蛋白酶抑制因子－1在肝纤维化过程中的表达及动态变化

为探讨基质金属蛋白酶抑制因子－1（TIMP－1）在肝纤维发生发展过程中的表达及动态变化,用皮下注射四氯化碳方法复制大鼠化学性肝纤维化模型,用免疫组织化学方法在4、8、12周分别检测肝组织中TIMP－1的表达,结果表明,随造模时间延长,肝纤维化面积逐渐增加,TIMP－1表达量进行性升高,明显高于对照组（P＜0．05）。结论：TIMP－1参与了肝纤维化的进展,可能起促进作用。     

Inhibition of PI3K and calcineurin suppresses chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2)-dependent responses of Th2 lymphocytes to prostaglandin D(2)

Interaction of prostaglandin D2 (PGD2) with chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2) triggers chemotaxis and pro-inflammatory cytokine production by Th2 lymphocytes. We have investigated the role of inhibitors of various cell-signalling pathways on the responses of human CRTH2+ CD4+ Th2 cells to PGD2. Phosphatidylinositol 3-kinase (PI3K) and Ca2+/calcineurin/nuclear factor of activated T cells (NFAT) pathways were activated by PGD2 in Th2 cells in a CRTH2-dependent manner. Inhibition of the PI3K pathway with LY294002 significantly reduced both PGD2-induced cell migration and cytokine (interleukin-4, interleukin-5 and interleukin-13) production. The inhibitory effect of LY294002 on cell migration is likely to be related to cytoskeleton reorganization as it showed a similar potency on PGD2-induced actin polymerization. The calcineurin inhibitors, tacrolimus (FK506) and cyclosporin A, had no effect on cell migration but completely blocked both cytokine production and the nuclear translocation of NFATc1 suggesting that Ca2+/calcineurin/NFAT is involved in CRTH2-dependent cytokine production but not chemotaxis. The promotion of NFAT nuclear location by PI3K activation may be mediated by negative regulation of glycogen synthase kinase-3beta (GSK3beta), since the PGD2-stimulated increase in phospho-GSK3beta was down-regulated by LY294002, and inhibition of GSK3beta by SB216763 enhanced PGD2-induced Th2 cytokine production and reversed the inhibitory effect of LY294002. These data suggest that PI3K and Ca2+/calcineurin/NFAT signalling pathways are critically involved in pro-inflammatory responses of Th2 cells to PGD2.     

Curcumin (diferuloylmethane) inhibits constitutive NF-kappaB activation, induces G1/S arrest, suppresses proliferation, and induces apoptosis in mantle cell lymphoma

Human mantle cell lymphoma (MCL), an aggressive B cell non-Hodgkin's lymphoma, is characterized by the overexpression of cyclin D1 which plays an essential role in the survival and proliferation of MCL. Because of MCL's resistance to current chemotherapy, novel approaches are needed. Since MCL cells are known to overexpress NF-kappaB regulated gene products (including cyclin D1), we used curcumin, a pharmacologically safe agent, to target NF-kappaB in a variety of MCL cell lines. All four MCL cell lines examined had overexpression of cyclin D1, constitutive active NF-kappaB and IkappaB kinase and phosphorylated forms of IkappaBalpha and p65. This correlated with expression of TNF, IkappaBalpha, Bcl-2, Bcl-xl, COX-2 and IL-6, all regulated by NF-kappaB. On treatment of cells with curcumin, however, downregulated constitutive active NF-kappaB and inhibited the consitutively active IkappaBalpha kinase (IKK), and phosphorylation of IkappaBalpha and p65. Curcumin also inhibited constitutive activation of Akt, needed for IKK activation. Consequently, the expression of all NF-kappaB-regulated gene products, were downregulated by the polyphenol leading to the suppression of proliferation, cell cycle arrest at the G1/S phase of the cell cycle and induction of apoptosis as indicated by caspase activation, PARP cleavage, and annexin V staining. That NF-kappaB activation is directly linked to the proliferation of cells, is also indicated by the observation that peptide derived from the IKK/NEMO-binding domain and p65 suppressed the constitutive active NF-kappaB complex and inhibited the proliferation of MCL cells. Constitutive NF-kappaB activation was found to be due to TNF, as anti-TNF antibodies inhibited both NF-kappaB activation and proliferation of cells. Overall, our results indicate that curcumin inhibits the constitutive NF-kappaB and IKK leading to suppression of expression of NF-kappaB-regulated gene products that results in the suppression of proliferation, cell cycle arrest, and induction of apoptosis in MCL.     

Induction of interleukin 8 release from the HMC-1 mast cell line: synergy between stem cell factor and activators of the adenosine A(2b) receptor

The HMC-1 mast cell line has both adenosine A(3) and A(2b) receptors on its surface, but only agonists of the A(2b) receptor are effective at releasing interleukin 8. Object of this study was to look for co-factors for adenosine A(2b) receptor activation. There was a powerful and statistically significant synergy for release of IL-8, both at the mRNA level (measured after 4 hr) and protein level (measured after 24 hr), between adenosine A(2b) receptor agonists and stem cell factor (SCF). Suitable concentrations for showing synergy were 100 ng/mL SCF and 3 microM 5'-N-ethylcarboxamidoadenosine (NECA). At these concentrations, the IL-8 released into the culture medium after SCF and NECA together was typically 3-5-fold greater in amount than the sum of the amounts of IL-8 released after exposure to the same concentrations of NECA and SCF separately. Since mast cells may be exposed to both adenosine and stem cell factor in the diseased lung, the synergy observed in this model system may have implications for asthma.     

Nrf2 protects against 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced oxidative injury and steatohepatitis

Previous studies demonstrate that Nrf2, a master regulator of antioxidative responses, is essential in mediating induction of many antioxidative enzymes by acute activation of the AhR. However, the role of Nrf2 in protecting against oxidative stress and DNA damage induced by sustained activation of the AhR remains unknown and was investigated herein. Tissue and blood samples were collected from wild-type (WT) and Nrf2-null mice 21 days after administration of a low-toxic dose (10 μg/kg ip) of TCDD. Only Nrf2-null mice lost body weight after TCDD treatment; however, blood levels of ALT were not markedly changed in either genotype, indicating a lack of extensive necrosis. Compared to livers of TCDD-treated WT mice, livers of TCDD-treated Nrf2-null mice had: 1) degenerated hepatocytes, lobular inflammation, marked fat accumulation, and higher mRNA expression of inflammatory and fibrotic genes; 2) depletion of glutathione, elevation in lipid peroxidation and marker of DNA damage; 3) attenuated induction of phase-II enzymes Nqo1, Gsta1/2, and Ugt2b35 mRNAs, but higher induction of cytoprotective Ho-1, Prdx1, Trxr1, Gclc, and Epxh1 mRNAs; 4) higher mRNA expression of Fgf21 and triglyceride-synthesis genes, but down-regulation of bile-acid-synthesis genes and cholesterol-efflux transporters; and 5) trend of induction/activation of c-jun and NF-kB. Additionally, TCDD-treated Nrf2-null mice had impaired adipogenesis in white adipose tissue. In conclusion, Nrf2 protects livers of mice against oxidative stress, DNA damage, and steatohepatitis induced by TCDD-mediated sustained activation of the AhR. The aggravated hepatosteatosis in TCDD-treated Nrf2-null mice is due to increased lipogenesis in liver and impaired lipogenesis in white adipose tissue.     

The adaptor Lnk (SH2B3): an emerging regulator in vascular cells and a link between immune and inflammatory signaling

A better knowledge of the process by which inflammatory extracellular signals are relayed from the plasma membrane to specific intracellular sites is a key step to understand how inflammation develops and how it is regulated. This review focuses on Lnk (SH2B3) a member, with SH2B1 and SH2B2, of the SH2B family of adaptor proteins that influences a variety of signaling pathways mediated by Janus kinase and receptor tyrosine kinases. SH2B adaptor proteins contain conserved dimerization, pleckstrin homology, and SH2 domains. Initially described as a regulator of hematopoiesis and lymphocyte differentiation, Lnk now emerges as a key regulator in hematopoeitic and non hematopoeitic cells such as endothelial cells (EC) moderating growth factor and cytokine receptor-mediated signaling. In EC, Lnk is a negative regulator of TNF signaling that reduce proinflammatory phenotype and prevent EC from apoptosis. Lnk is a modulator in integrin signaling and actin cytoskeleton organization in both platelets and EC with an impact on cell adhesion, migration and thrombosis. In this review, we discuss some recent insights proposing Lnk as a key regulator of bone marrow-endothelial progenitor cell kinetics, including the ability to cell growth, endothelial commitment, mobilization, and recruitment for vascular regeneration. Finally, novel findings also provided evidences that mutations in Lnk gene are strongly linked to myeloproliferative disorders but also autoimmune and inflammatory syndromes where both immune and vascular cells display a role. Overall, these studies emphasize the importance of the Lnk adaptor molecule not only as prognostic marker but also as potential therapeutic target.     

Miltirone induces cell death in hepatocellular carcinoma cell through GSDME-dependent pyroptosis

Pyroptosis is a form of programmed cell death, and recently described as a new molecular mechanism of chemotherapy drugs in the treatment of tumors. Miltirone, a derivative of phenanthrene-quinone isolated from the root of Salvia miltiorrhiza Bunge, has been shown to possess anti-cancer activities. Here, we found that miltirone inhibited the cell viability of either HepG2 or Hepa1-6 cells, and induced the proteolytic cleavage of gasdermin E (GSDME) in each hepatocellular carcinoma (HCC) cell line, with concomitant cleavage of caspase 3. Knocking out GSDME switched miltirone-induced cell death from pyroptosis to apoptosis. Additionally, the induction effects of miltirone on GSDME-dependent pyroptosis were attenuated by siRNA-mediated caspase three silencing and the specific caspase three inhibitor Z-DEVD-FMK, respectively. Miltirone effectively elicited intracellular accumulation of reactive oxygen species (ROS), and suppressed phosphorylation of mitogen-activated and extracellular signal-regulated kinase (MEK) and extracellular regulated protein kinases 1/2 (ERK1/2) for pyroptosis induction. Moreover, miltirone significantly inhibited tumor growth and induced pyroptosis in the Hepa1-6 mouse HCC syngeneic model. These results provide a new insight that miltirone is a potential therapeutic agent for the treatment of HCC via GSDME-dependent pyroptosis.