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1.
目的 观察膜型基质金属蛋白酶-1(MT1-MMP)对人肝癌细胞株浸润能力的影响,并初步探讨其作用机制.方法 将稳定表达MT1-MMP基因转染至HepG2细胞中,应用Western印迹法检测MT1-MMP蛋白表达;明胶酶谱分析法检测MMP-2酶原活性;体外侵袭实验检测细胞株浸润能力.结果 重组质粒转染株MT1-MMP蛋白表达水平明显高于空质粒组和未转染组(P<0.01).明胶酶谱分析实验发现,MT1-MMP组同时检测到72000酶原形式和64000活性形式的MMP-2,另两组只检测到72000酶原形式的MMP-2.体外侵袭实验结果显示,MT1-MMP组细胞穿过基质胶(Matrigel)的细胞数目明显多于其他两组(P<0.01).结论 MT1-MMP能显著增强肝癌细胞株的浸润能力,其机制主要是通过激活MMP-2-酶原,降解肿瘤周围的基质成分实现的. 相似文献
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TIMP-2、MMP-2和MT1-MMP在活化肝星状细胞中的表达及对细胞外基质合成分泌的影响 总被引:1,自引:0,他引:1
目的 研究金属蛋白酶组织抑制剂-2(TIMP-2)、基质金属蛋白酶-2(MMP-2)和膜型基质金属蛋白酶-1(MT1-MMP)在活化肝星状细胞(HSCs)中的表达,观察其对细胞外基质(ECM)合成分泌的影响.方法 原代分离培养大鼠HSCs活化后,分别给予40~160 pmol化学合成经修饰抗TIMP-2 siRNA进行干预,检测培养细胞上清液透明质酸(HA)、Ⅲ型前胶原(PCⅢ)和羟脯氨酸(Hyp)的含量,采用荧光实时定量PCR法检测TIMP-2、MMP-2、MT1-MMP、MMP-13、COL Ⅰ和COL Ⅲ mRNA的表达,western印迹检测TIMP-2、MT1-MMP和MMP-13蛋白表达及明胶酶谱法检测MMP-2蛋白表达.结果 应用化学合成经修饰抗TIMP-2 siRNA后,TIMP-2、MMP-2、MT1-MMP、COL Ⅰ和COL Ⅲ的表达明显降低,而MMP-13的表达则明显增加,培养细胞上清液中HA、PCⅢ和Hyp的含量也明显减少.结论 TIMP-2通过MT1-MMP介导MMP-2的活化,抑制TIMP-2的表达,MT1-MMP和MMP-2的表达随之降低,而HSCs合成分泌ECM也相应减少. 相似文献
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狼疮肾炎肾间质浸润与肾损害进展及肾功能相关性的研究 总被引:1,自引:1,他引:1
目的:观察狼疮肾炎肾间质浸润的白细胞类型与肾功能损害进展的关系。方法:检测40例狼疮肾炎患者24h尿蛋白定量、血肌酐,随即进行肾活检及免疫组织化学检测,21例患者进行二次肾活检。结果:狼疮肾炎患者首次肾活检时显示T淋巴细胞与单核细胞所占比例差异无显著性,而第2次肾活检时以单核细胞为主。首次肾活检时间质浸润的淋巴细胞CD4^ /CD8^ 较正常对照组增加,第2次肾活检时CD^ /CD8^ 低于正常对照组。第2次肾活检时显著间质浸润的发生率由首次活检时的47.5%升至100%。浸润白细胞总数与血肌酐水平呈正相关,其中T细胞、中性粒细胞亦与血肌酐呈正相关,单核细胞与血肌酐无显著相关性。结论:重复肾活检可发现更多病例肾间质损害,狼疮肾炎肾间质的主要浸润细胞类型呈疾病进展阶段依赖性,CD4^ T细胞在间质损伤机制中起始发作用,CD8^ T细胞的作用可能是继发的,长期的蛋白尿可能对CD8^ 细胞的浸润有激发作用。 相似文献
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卵巢浆液性腺癌组织中MMP-2、MT1-MMP的表达变化及意义 总被引:1,自引:0,他引:1
目的观察卵巢浆液性腺癌中的基质金属蛋白酶-2(MMP-2)、膜型基质金属蛋白酶-1(MT1-MMP)的表达变化。方法采用免疫组化PV6000法检测50例卵巢浆液性腺癌、30例卵巢交界性浆液性腺瘤和30例卵巢良性浆液性腺瘤组织中的MMP-2、MT1-MMP。结果MMP-2和MT1-MMP在卵巢浆液性腺癌和交界性浆液性腺瘤中的表达阳性率高于良性浆液性腺瘤(P〈0.05)。卵巢良性浆液性腺瘤组织中的MMP-2、MT1-MMP与病理分级、淋巴结转移、临床分期和预后有关(P均〈0.0.5)。卵巢浆液性腺癌中MMP-2、MT1-MMP的表达呈正相关(L=0.61,P〈0.01)。结论卵巢浆液性腺癌中MMP-2、MT1-MMP表达上调。二者在卵巢浆液性腺癌的发生、浸润和转移中过程中协同发挥重要作用,可作为判断肿瘤恶性程度和预后的重要指标。 相似文献
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目的 研究胚肺成纤维细胞对肺癌H460细胞膜型基质金属蛋白酶-1(MTl-MMP)、基质金属蛋白酶-2(MMP-2)表达的影响.方法 采用Western blot方法检测各组MT1-MMP的表达.取其上清,采用酶联免疫吸附法检测各组细胞培养液中活性MMP-2的浓度.结果 H460细胞、胚肺成纤维细胞单独培养时均有MT1-MMP表达,但混合培养后MT1-MMP表达增强(P<0.05).H460细胞、胚肺成纤维细胞单独培养时MMP-2均有分泌,混合培养MMP-2分泌增强(P<0.05).结论 胚肺成纤维细胞和肺癌H460细胞相互作用能通过上调MT1-MMP、MMP-2的表达从而促进肺癌的侵袭和转移,这可能为肺癌侵袭转移的一个重要机制. 相似文献
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CD40-CD40L及基质蛋白酶在大鼠动脉粥样硬化中的表达 总被引:1,自引:0,他引:1
目的:为探讨粘附分子CD40、CD40配体(CD40 Ligand,CD40L)及基质蛋白酶在动脉粥样硬化中的表达;方法:利用高脂饮食制作大鼠动脉粥样硬化模型(AS组,n=6),与正常饮食大鼠模型(N组,n=6)作对照,流式细胞技术法检测外周血中CD40及CD40L,Zymography法检测外周血中基质蛋白酶-2(MMP-2)及基质蛋白酶-9(MMP-9)的表达情况,免疫组织化学法观察CD40、CD40L及MMP-2、MMP-9在主动脉弓表达情况;并对CD40、CD40L与MMP-2、MMP-9做相关性分析;结果:动脉粥样硬化模型大鼠外周血CD40、CD40L、MMP-2、MMP-9表达均高于正常饮食组大鼠,免疫组织化学法显示CD40、CD40L在主动脉弓内膜上表达,MMP-2、MMP-9在主动脉弓内皮细胞及内皮细胞间质中表达,而且AS组表达强于N组;CD40、CD40L与MMP-2、MMP-9有相关性;结论:提示动脉粥样硬化的形成可能与CD40、CD40L及基质蛋白酶的异常表达有关。 相似文献
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目的 观察细胞外基质金属蛋白酶诱导剂(EMMPRIN/CD147)对类风湿关节炎(RA)患者滑膜成纤维细胞(FLS)合成基质金属蛋白酶(MMPs)的作用。方法 手术切除的RA患者的滑膜组织,分离成FLS,传代培养。明胶酶谱法测FLS的MMPs含量将FLS与高表达EMMPRIN/CD147的单核细胞株(THP)-1共培养,观察THP-1细胞对FLS MMPs表达量的影响,同时观察EMMPRIN/CD147拮抗肽激活蛋白(AP)-9对FLS MMPs表达的影响。结果 THP-1细胞表达高水平的CD147,而MMP-2、MMP-9的表达水平很低;分离的FLS表达低水平的CD147和一定水平的MMP-2、MMP-9。二者混合培养后,FLS表达MMP-2、MMP-9水平显著升高,随着THP-1细胞数的增加,MMP-2、MMP-9的合成量也增加THP-1对FLS合成MMP-2、MMP-9的刺激作用可被CD147拮抗肽AP-9所抑制。结论 CD147对RA患者FLS合成MMP-2、MMP-9有刺激作用,而这种刺激作用呵被CD147拮抗肽AP-9所抑制。 相似文献
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目的探讨肾癌及癌旁肾组织中基质金属蛋白酶-2,9(MMP-2、MMP-9)及血管细胞黏附分子1(VCAM-1)的表达差异。方法采用免疫组化法检测MMP-2、MMP-9及VCAM-1在肾癌及癌旁肾组织中的表达,应用明胶酶图法检测MMP-2及MMP-9在肾癌及癌旁肾组织中明胶酶的活性。结果肾癌组织中MMP-2、MMP-9及VCAM-1的表达阳性率较癌旁肾组织显著升高(P<0.01),肾癌组织中MMP-2及MMP-9的明胶酶活性较癌旁肾组织也呈增高趋势(P<0.01)。结论 MMP-2、MMP-9及VCAM-1在肾癌组织中表达升高,可能促进肾癌的发病及进展。 相似文献
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《中国糖尿病杂志》2016,(10)
目的观察厄贝沙坦是否对抗糖尿病大鼠肺损伤,探讨大鼠肺组织胶原和基质金属蛋白酶(MMPs)通路中相关因子的变化。方法雄性SD大鼠随机分为对照组(NC)、T2DM组、厄贝沙坦+糖尿病组(Ir+T2DM)。8周后测定各组FPG、TG和TC,HE和Masson染色观察肺组织形态、纤维化改变。ELISA测定肺组织MMP-2、MMP-9、TIMP-1和TIMP-2含量,Western blot检测MT1-MMP蛋白的表达。结果与NC组比较,T2DM组FPG、TG和TC水平增高,肺泡结构大部分消失,胶原增生,肺组织MMP-2、MMP-9水平下降,TIMP-1、TIMP-2水平升高,MT1-MMP蛋白表达较NC组增加(P0.05或P0.01)。与T2DM组比较,Ir+T2DM组肺组织形态和纤维增生改善,MMP-2、MMP-9水平增加,TIMP-1、TIMP-2水平降低,MT1-MMP蛋白表达降低(P0.05)。结论 T2DM可诱导肺脏损伤,厄贝沙坦通过调节MMPs信号通路中的相关因子,减轻肺损伤。 相似文献
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The immunoneuroendocrine role of melatonin 总被引:19,自引:0,他引:19
Georges J. M. Maestroni 《Journal of pineal research》1993,14(1):1-10
Abstract: A tight, physiological link between the pineal gland and the immune system is emerging from a series of experimental studies. This link might reflect the evolutionary connection between self-recognition and reproduction. Pinealectomy or other experimental methods which inhibit melatonin synthesis and secretion induce a state of immunodepression which is counteracted by melatonin. In general, melatonin seems to have an immunoenhancing effect that is particularly apparent in immunodepressive states. The negative effect of acute stress or immunosuppressive pharmacological treatments on various immune parameters are counteracted by melatonin. It seems important to note that one of the main targets of melatonin is the thymus, i.e., the central organ of the immune system. The clinical use of melatonin as an immunotherapeutic agent seems promising in primary and secondary immunodeficiencies as well as in cancer immunotherapy. The immunoenhancing action of melatonin seems to be mediated by T-helper cell-derived opioid peptides as well as by lymphokines and, perhaps, by pituitary hormones. Melatonin-induced-immuno-opioids (MHO) and lymphokines imply the presence of specific binding sites or melatonin receptors on cells of the immune system. On the other hand, lymphokines such as -γ-interferon and interleukin-2 as well as thymic hormones can modulate the synthesis of melatonin in the pineal gland. The pineal gland might thus be viewed as the crux of a sophisticated immunoneuroendocrine network which functions as an unconscious, diffuse sensory organ. 相似文献
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Changes in connexin43, the gap junction protein of astrocytes, during development of the rat pineal gland 总被引:2,自引:0,他引:2
Abstract: The abundance of gap junctions between rat pineal astrocytes formed by connexin43 (Cx43) was studied during development. Levels and distribution of Cx43 were measured by immunoblotting and indirect immunofluorescence, respectively. The amount of Cx43 in cells located within the gland was low until about the 7th postnatal day and increased to adult values between the 14th and 21st days postpartum. Although astrocytes, recognized by their vimentin immunoreactivity, were scarce before birth, they were abundant by the 7th postnatal day suggesting that the low levels of Cx43 found at this age corresponded to a low expression of this protein. Localization of the immunoreactivity to Cx43 and vimentin showed a close correlation, indicating that mature or immature pineal astrocytes form gap junctions made of Cx43. Since Cx43 levels attained their adult values at about the time the innervation and the functional state of the gland reached maturity (2–3 weeks after birth), it is proposed that astrocyte gap junctions are involved in the function of the adult rat pineal gland. 相似文献
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Martinez-Cecilia D Arjona-Sanchez A Gomez-Alvarez M Torres-Tordera E Luque-Molina A Valenti-Azcarate V Briceno-Delgado J Padillo FJ Lopez-Cillero P Rufian-Pena S 《World journal of gastroenterology : WJG》2008,14(12):1949-1951
Duodenal diverticula are a relatively common condition. They are asymptomatic, unless they become complicated, with perforation being the rarest but most severe complication. Surgical treatment is the most frequently performed approach. We report the case of a patient with a perforated duodenal diverticulum, which was diagnosed early and treated conservatively with antibiotics and percutaneous drainage of secondary retroperitoneal abscesses. We suggest this method could be an acceptable option for the management of similar cases, provided that the patient is in good general condition and without septic signs. 相似文献
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Abstract: Herein we documented the response of pineal melatonin production to electrolytes known to be effective on pineal function in view of a possible circadian stage dependence. We studied the release of melatonin by perifused rat pineal glands at 2 different circadian stages corresponding to the middle of the light and dark periods, i.e., respectively, 7 and 19 HALO (Hours After Light Onset, L:D = 12:12). The initial efflux rates were, as expected, much higher in the perifusates of glands removed from rats sacrificed during the dark phase than of those removed during the light phase. After 3 hr of perifusion, melatonin release reached similar levels which were found constant up to the 8th hr of perifusion, whatever the circadian stage. Perifusion of the glands with physiological concentrations for the rat of calcium (5.2 mmol/1) and magnesium (1.34 mmol/1) resulted in a stimulatory effect on the pineal glands removed from rats sacrificed in the middle of the dark period (19 HALO), whereas no effects were observed on the pineal glands removed from rats sacrificed during the light (7 HALO). Lithium (0.28 and 0.55 mmol/1) was ineffective on melatonin release in pineal glands removed 7 and 19 HALO. Our results show differences in the initial efflux rates of melatonin and in the response of perifused pineal glands to calcium and magnesium according to the circadian stage. 相似文献
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H.P.J.M. Noteborn P.P. van Balen A.A. van der Gugten I.C. Hart I. Ebels C.A. Salemink 《Journal of pineal research》1993,14(1):11-22
Abstract: The use of antisera raised against bovine growth hormone (GH) and ovine prolactin (PRL) enabled the detection of related immunoreactive (ir) sequences of proteins in ovine pineal tissue. The isolation of PRL-like ir-material was accomplished using a 0.25 M ammonium sulphate (pH 5.5) extraction followed by ethanol precipitation, whereas the resulting 2.0 M ammonium sulphate (pH 7.0) precipitate contained a GH-like immunoreactivity. Gel chromatography of the GH-like immunoreactivity (Sephadex G-100) indicated the presence of several GH-like fragments ranging in the Mr range of 7,000 to 55,000. Analyses of the PRL-like ir-material found in pineal tissue on HPLC using a TSK 545-DEAE column led to the resolution into a single peak of immunoreactivity. A single peak of activity was also observed following chromatofocusing and hydrophobic interaction chromatography of the ir-peak from the TSK 545-DEAE column. The PRL-like ir-material inhibited the binding of [125I]ovine PRL-S14 to anti-ovine PRL antibodies without showing an affinity for binding to anti-rat PRL or anti-bovine GH antibodies. Scatchard analysis of the binding of pineal PRL-like ir-material and pituitary ovine PRL-S14 to liver membranes from day-20 pregnant rats revealed similar affinity constants (Ka of 4.7 ± 0.2 × 109 M-1). In addition, the replication of Nb 2 Node rat lymphoma cells was stimulated by pineal PRL-like ir-material, an effect known to be specific for lactogenic hormones. The pineal PRL-like immunoreactivity appeared on sodium dodecyl sulfate polyacrylamide gels as a single major band of Mr 24,000. The functional status of PRL-and GH-like ir-material in the ovine pineal remains to be determined, but evidence is presented that the overall protein synthesis rate of the rat pineal responded to circulating concentrations of PRL. 相似文献
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Gary S. Goldberg B.A. Bruce A. Orkin M.D. Lee E. Smith M.D. 《Diseases of the colon and rectum》1994,37(5):439-443
PURPOSE: Individuals who are seropositive for the human immunodeficiency virus are at high risk for opportunistic infection and anorectal disorders. Little prospective information is available regarding anorectal pathogens in these patients. METHODS: One hundred sixty-three HIV-seropositive patients presented to the colorectal clinic between 1989 and 1992. Forty-seven (29 percent) patients were thought to have an infectious process and were prospectively studied using a standardized multiculture protocol. RESULTS: Mean age was 33 (range, 19–59) years. All were male; high-risk behavior accounted for 87 percent of HIV transmissions. Presenting complaints included anorectal pain (79 percent), pus per anum (28 percent), and blood per anum (26 percent). Examination revealed perianal tenderness (60 percent), condyloma (38 percent), perianal ulcers (38 percent), and anal fissures (34 percent). Sixty-six sets of cultures were performed; 28 patients had one set, 15 had two sets, and 4 had three sets. Thirty-two of these 47 patients (68 percent) had positive cultures including herpes (50 percent), cytomegalovirus (25 percent),Neisseria gonorrhoeae
(16 percent), chlamydia (16 percent), acidfast bacilli (2 percent), and others (9 percent). Six of 32 patients with positive cultures had more than one organism cultured. Sixteen (50 percent) patients with positive cultures were treated medically, 8 (25 percent) were treated surgically and 8 (25 percent) were treated with both modalities. Sixty-one procedures were performed on 17 patients for condylomata. Eighteen patients had 20 procedures for abscesses, 50 percent of whom had positive cultures for other than common bowel flora; all improved. Fourteen patients underwent 33 procedures for perianal fistulas.Mycobacterium fortuitum
was cultured from one patient who required 13 procedures for abscesses and fistulas. Forty-five (96 percent) patients were followed for an average of 12.5 months ±2.9 SEM (range, 1–94 months). Symptoms were improved or resolved in 22 of 32 (69 percent) patients with positive cultures and in 11 of 13 (84 percent) with negative cultures. CONCLUSIONS: Specific pathogens may often be identified in human immunodeficiency virus-seropositive patients with anorectal disorders if aggressively sought. Although patients without specific pathogens identified may be expected to improve with planned empiric treatment, positive identification allows more directed therapy. 相似文献