康莱特注射液抗恶性肿瘤肝转移疗效及机制的实验研究

目的通过接种W256癌肉瘤建立Wistar大鼠肝癌转移模型,观察康莱特注射液与肝动脉结扎法对大鼠荷瘤脾重与肝癌转移灶的抑制效果,并测定其对瘤组织内基质金属蛋白酶(MMP-1)及肝细胞生长因子(HGF)表达的影响,探讨中西医结合防治肝癌转移的新途径。方法在Wistar大鼠脾下极注射W256癌肉瘤细胞悬液,建立肝癌转移模型。随机分4组:对照组、动脉结扎组、康莱特组、康莱特+动脉结扎组。计算各组瘤重抑制率与肝癌转移抑制率,采用原位杂交技术检测肿瘤细胞内MMP-1和HGF mRNA表达水平。结果康莱特和康莱特加动脉结扎组均显著抑制脾脏肿瘤生长并抑制肝脏瘤组织MMP-1 mRNA表达,康莱特组能明显抑制肝癌转移灶产生,各治疗组大鼠瘤细胞HGF mRNA表达水平均低于对照组。结论康莱特注射液可显著抑制原发灶生长与肝癌转移灶的形成,抑制瘤细胞MMP-1表达是其抗肝转移的重要机制;康莱特注射液结合动脉结扎可使瘤细胞MMP-1表达减少,提示可优化两者结合途径。     

康莱特注射液联合化疗治疗结直肠癌肝转移的疗效观察

目的探讨康莱特注射液联合化疗治疗结直肠癌肝转移的疗效与不良反应。方法入选晚期结直肠癌肝转移无法手术的患者78例,随机分为化疗组与联合组各39例。化疗组采用FOLFOX4化疗方案,联合组在此基础上加用康莱特注射液,治疗4个周期后观察临床疗效与不良反应情况。结果联合治疗组与单用化疗组总缓解率分别为35.9%和30.8%(P>0.05);CEA总缓解率分别为71.8%和64.1%(P>0.05);Karnofsky评分总有效率分别为69.2%和46.2%(P<0.05);疼痛总缓解率分别为66.7%和33.3%(P<0.01)。联合组消化道反应、肝功能异常、乏力的发生率显著降低(P<0.05)。结论康莱特联合化疗治疗晚期结直肠癌,可改善患者的临床症状,降低化疗不良反应。     

康莱特注射液治疗晚期肺癌疼痛的疗效观察

目的:观察康莱特注射液缓解晚期肺癌疼痛的疗效。方法:选取某院2006～2010年期间住院的晚期肺癌患者98例,分为治疗组50例(康莱特+吗啡缓释片)及对照组48例(单用吗啡缓释片),观察两组患者的止痛效果。结果:治疗组止痛显效率84%,观察组显效率62.5%,两组显效率比较差异有统计学意义(P<0.05)。结论:康莱特注射液在缓解晚期肺癌疼痛中具有较好的作用。     

康莱特注射液在Lewis肺癌中对肿瘤生长及EGFR蛋白表达的影响

目的：探讨康莱特注射液（KLT）对小鼠Lewis肺癌生长及对表皮生长因子受体（EGFR）蛋白表达的影响。方法：建立小鼠Lewis肺癌模型,将40只接种Lewis肺癌细胞的C57BL/6小鼠随机分成4组：模型对照组、KLT低剂量组（6.25mL/kg）、KLT中剂量组（12.5mL/kg）、KLT高剂量组（25mL/kg）,每组10只。小鼠连续腹腔注射KLT注射液14d,于接种后第16天处死小鼠,测量瘤重及体重,计算肿瘤抑制率,提取肿瘤组织总蛋白,通过Westernblot法检测肿瘤组织EGFR的蛋白表达情况。结果：KLT注射液能明显抑制Lewis肺癌小鼠肿瘤的生长,影响肿瘤组织的形态及组织学改变,并能降低肿瘤组织中EGFR的蛋白表达量。结论：KLT注射液可明显抑制小鼠Lewis肺癌的生长,其机制可能与抑制EGFR的蛋白表达有关。     

动脉化疗栓塞联合艾迪注射液治疗肝转移瘤疗效分析

目的探讨动脉化疗栓塞与艾迪注射液联合治疗肝转移瘤的价值。方法选取26例不能外科切除的肝转移瘤患者,先经肝动脉化疗栓塞,然后行艾迪注射液静脉滴注,观察联合治疗后生存期、肿瘤局控及CEA值变化。结果联合治疗后所有病例均未出现严重并发症,平均生存期30.2个月。结论化疗栓塞联合艾迪注射液能有效灭活肿瘤组织,延长生存期,而治疗并发症较轻,值得进一步推广应用。     

康莱特注射液诱导胰腺癌Mia-Paca2细胞增殖及凋亡的实验研究

目的探讨康莱特注射液(KLT)对胰腺癌Mia-Paca2细胞增殖及凋亡的影响。方法通过CCK8法观察KLT对胰腺癌Mia-Paca2细胞的增殖影响,通过Hoechst 33258荧光染色法观察细胞凋亡改变。结果 KLT能明显抑制Mia-Paca2细胞的生长,对Mia-Paca2细胞的作用具有时间及剂量的依赖性,Hoechst染色法观察,随着KLT浓度增加,Mia-Paca2细胞呈现明显的凋亡特征。结论 KLT能明显地抑制Mia-Paca2细胞的增殖及促进细胞凋亡。     

胃癌肝转移的综合治疗分析

目的对患有胃癌的患者肝转移症状进行综合治疗与分析。方法共取30例患者进行疗效分析,平均分成三组,其中A组患者进行胃癌原发灶和肝转移灶同时切除的手术,B组患者对没有进行切除手术的患者进行局部治疗,在治疗的过程中使用对无水酒精瘤进行体内注射的方法以结合门静脉栓进行化疗的方法,C组患者在切除胃癌原发灶的基础上,对肝转移病灶患者不实施任何治疗措施。结果 A组的患者在1.5、3.5年时仍然生存的概率为56.8%、25.2%;B组患者1.5和3.5年患者仍然存活的概率为49.1%和243.%,两组患者经过对比可发现数据没有显著差异性(P>0.05),没有统计学意义。C组患者在1.5和3.5年时患者仍然存活的概率为15.0%和0%,其结果明显较B组较低,有明显差异性(P<0.05),具有统计学意义。结论胃癌肝转移的病死率较大,但是有效的综合治疗方式可以在一定程度延缓患者的病状,增加患者的生命时间,使得患者的生命质量得到了有效的提高,对与胃癌肝转移的患者,如果其转移病灶无法切除,应该积极的给予治疗,为患者的生命争取更多的时间。     

熊果酸抗实验性大鼠肝纤维化作用机制的研究

目的观察熊果酸对实验性大鼠肝纤维化的影响及可能的作用机理。方法将不同剂量的熊果酸10mg(kg/d)、20mg(kg/d)、40mg(kg/d)作用于二甲基亚硝胺(DMN)所致肝纤维化大鼠,在药物治疗4周后观察熊果酸对肝纤维化大鼠肝功能的影响;观察熊果酸对氧化指标SOD、MDA的影响;病理学方法观察熊果酸治疗后组织形态学的变化;RT-PCR方法检测熊果酸对MMP-1mRNA、TIMP-1mRNA表达变化。结果熊果酸可明显改善肝纤维化大鼠肝功能,并呈剂量依赖性;不同剂量熊果酸作用4周后能显著增加SOD表达,降低MDA表达;在病理学形态方面,熊果酸治疗组使肝组织结构不同程度改善;不同剂量熊果酸作用4周后RT-PCR方法检测治疗组MMP-1mRNA的表达较模型组相比明显升高,TIMP-1mRNA表达较模型组相比显著降低。结论熊果酸可明显改善肝纤维化大鼠肝功能,并呈剂量依赖性;熊果酸能显著增加SOD表达,降低MDA表达;熊果酸可上调MMP-1mRNA的表达,下调TIMP-1mRNA的表达;熊果酸治疗肝纤维化的作用机制可能与阻断氧化应激过程,抑制脂质过氧化,增加细胞外基质(ECM)的降解,减少ECM的沉积等机制有关。     

康莱特肝动脉灌注对移植性肝肿瘤大鼠肝功能的影响

目的　研究康莱特注射液肝动脉灌注对移植性肝肿瘤大鼠肝功能的影响。方法　复制大鼠移植性肝肿瘤模型,经肝动脉灌注给药方法,以化疗药、超液化碘油等作对照,测定血清ALT,AST,ALP,GGT,TBIL,DBIL,TP,Alb等水平,评价康莱特作为介入药物治疗大鼠肝肿瘤时对肝功能的影响。结果　大鼠经肝动脉灌注治疗后,单纯康莱特灌注对大鼠ALT及AST水平无明显影响,所有含碘化油组大鼠血清ALT及AST水平均较对照组或康莱特组升高(P<0. 05)。与丝裂霉素 /碘化油组或碘化油组比较,康莱特 /碘化油组大鼠血清ALT水平均明显较低,有显著性差异(P<0. 05)。其他各项血清生化指标经统计学分析,各组之间均无明显差异(P>0. 05)。结论　单纯康莱特灌注对大鼠肝功能无明显影响,康莱特 /碘化油肝动脉灌注对肝功能影响明显低于MMC/碘化油组。     

康莱特肝动脉灌注对移植性肝肿瘤大鼠肝功能的影响

目的　研究康莱特注射液肝动脉灌注对移植性肝肿瘤大鼠肝功能的影响。方法　复制大鼠移植性肝肿瘤模型,经肝动脉灌注给药方法,以化疗药、超液化碘油等作对照,测定血清ALT,AST,ALP,GGT,TBIL,DBIL,TP,Alb等水平,评价康莱特作为介入药物治疗大鼠肝肿瘤时对肝功能的影响。结果　大鼠经肝动脉灌注治疗后,单纯康莱特灌注对大鼠ALT及AST水平无明显影响,所有含碘化油组大鼠血清ALT及AST水平均较对照组或康莱特组升高(P<0.05)。与丝裂霉素/碘化油组或碘化油组比较,康莱特/碘化油组大鼠血清ALT水平均明显较低,有显著性差异(P<0.05)。其他各项血清生化指标经统计学分析,各组之间均无明显差异(P>0.05)。结论　单纯康莱特灌注对大鼠肝功能无明显影响,康莱特/碘化油肝动脉灌注对肝功能影响明显低于MMC/碘化油组。     

腹膜转移胃癌的临床病理特征和相关机制研究

目的:探讨发生腹膜转移胃癌的临床病理学特征和胃癌发生腹膜转移的机制.方法:收集164例临床资料和随访资料完整的胃癌患者,将其分成腹膜转移组41例和无腹膜转移组123例,对2组一般资料进行比较,并行缺氧诱导因子-1 a(HIF-1a)、黏着斑激酶(FAK)和基质金属蛋白酶-9( MMP-9)的免疫组化染色.结果:2组Lauren分型、浸润深度及TNM分期差异有统计学意义(P＜0.01),其中弥漫型胃癌(DGC)和肠型胃癌(IGC)患者的腹膜转移率(29.37％vs10.53％)差异亦有统计学意义(P＜0.01).Kaplan-Meier生存分析显示腹膜转移组的生存率低于无腹膜转移组(P＜0.01),Cox回归模型显示TNM分期和腹膜转移是影响胃癌患者生存率的危险因素.腹膜转移组HIF-la、FAK和MMP-9的表达均高于无腹膜转移组(P＜0.05或P＜0.01).结论:缺氧可能在胃癌的腹膜转移中发挥了重要的作用.     

白介素-10对肝纤维化大鼠星状细胞EGF及HGF表达的影响

目的探讨IL10干预对实验性肝纤维化大鼠肝星状细胞EGF、HGF表达的影响及其抗纤维化作用的可能机制。方法60只♂SD大鼠随机分为正常对照组(N组,8只)、肝纤维化模型组(C组,28只)和IL10干预组(I组,24只),分别于CCl4造模第7周及11周采用链霉蛋白酶E、Ⅳ型胶原酶经门静脉灌注分离肝星状细胞。半定量RTPCR法检测各组新分离肝星状细胞中EGF、HGFmRNA表达水平;SP免疫细胞化学法检测各组原代培养3天后肝星状细胞EGF的表达。并于上述两个时间段分别将各组肝组织行HE染色判断炎症和肝纤维化程度。结果成功建立大鼠肝纤维化模型和分离大鼠肝星状细胞。与正常组相比,纤维化模型组EGF、HGF的mRNA水平显著升高(P<001),经IL10干预后则明显降低(P<001)。对EGF而言,IL10组表达量高于正常组(P<005);对HGF而言,IL10组皆低于正常组(P<005)。肝纤维化模型组EGFmRNA水平在第11周较第7周有进一步的升高(P<005)。SP免疫细胞化法检测各组EGF的蛋白表达情况与上述mRNA结果相平行。结论EGF、HGF随肝纤维化进程逐渐升高,IL10可抑制纤维化大鼠肝星状细胞EGF、HGF的表达,具有抗纤维化作用。     

厄贝沙坦对高糖刺激的大鼠肾小球系膜细胞结缔组织生长因子及膜型基质金属蛋白酶-1表达的影响

目的研究血管紧张素Ⅱ受体1拮抗剂厄贝沙坦对高糖刺激的大鼠肾小球系膜细胞结缔组织生长因子(CTGF)和膜型基质金属蛋白酶-1(MT1-MMP)表达的影响。方法体外培养的大鼠肾小球系膜细胞,分别给予高糖和厄贝沙坦干预,采用RT-PCR及Western blot法分别检测CTGF和MT1-MMP的mRNA及蛋白的表达,用酶联免疫吸附法(ELISA)检测培养上清中Ⅳ型胶原的含量。结果与对照组相比,高糖组各时间点系膜细胞CTGF表达明显上调,Ⅳ型胶原的分泌增加,且二者随时间持续增高;而MT1-MMP的表达则随时间呈明显下降趋势。厄贝沙坦能够抑制高糖引起的上述变化。结论高糖可诱导肾小球系膜细胞CTGF表达增加,抑制MT1-MMP的表达。厄贝沙坦抑制肾小球系膜细胞基质分泌的作用可能部分通过抑制CTGF,增加MT1-MMP的表达而实现。     

山慈菇提取液对乳腺癌大鼠肿瘤组织血管内皮生长因子和基质金属蛋白酶-9表达的影响

目的研究山慈菇提取液对乳腺癌大鼠肿瘤组织中血管内皮生长因子(VEGF)和基质金属蛋白酶-9(MMP-9)表达水平的影响。方法选取健康雌性未孕SD大鼠为研究对象,用含有二甲基苯蒽的芝麻油建立乳腺癌模型。将造模成功的SD大鼠随机分为模型组、对照组、实验A组和实验B组,每组各16只;另取16只健康大鼠为空白组。模型组和空白组均予以灭菌0.9%Na Cl 1m L,qd,灌胃;对照组予以0.6 mg·m L^-1枸橼酸他莫昔芬1 m L,qd,灌胃;实验A、B组分别予以1.5,3.0 mg·m L^-1山慈菇提取液1 m L,qd,灌胃,5组大鼠均干预6周。比较5组大鼠的抑瘤率、肿瘤组织中VEGF和MMP-9表达水平。结果治疗后,对照组、实验A组和实验B组的抑瘤率分别为70.74%,21.51%和59.88%,两两比较,差异均有统计学意义(均P<0.05)。治疗后,空白组、模型组、对照组、实验A组和实验B组肿瘤组织中VEGF分别为(5454.68±754.37),(35485.58±4638.36),(9975.75±1243.44),(24564.35±3028.43)和(14346.54±1976.56)DPI,肿瘤组织中MMP-9分别为(3353.26±406.34),(29584.37±3856.56),(8143.32±1043.82),(19473.46±2315.47)和(10865.47±1324.43)DPI,两两比较,差异均有统计学意义(均P<0.05)。结论山慈菇提取液能有效抑制乳腺癌大鼠肿瘤生长,且呈剂量依赖性,其可能与抑制肿瘤组织中VEGF及MMP-9的表达有关。     

血清VEGF、VEGF-C及MMP-9在肺癌患者淋巴结转移的作用与意义

目的联合检测术前肺癌患者血清中VEGF、VEGF-C及MMP-9的水平,探讨其作为预测肺癌淋巴转移、预后和手术疗效监测指标的可行性。方法应用酶联免疫吸附法（ELISA）联合检测65例NSCLC患者术前的血清VEGF、VEGF-C及MMP-9的表达。结果有淋巴转移组的血清VEGF、VEGF-C、MMP-9水平明显高于无淋巴转移组,有显著差异性（P〈0.01）,相对于检测单个蛋白因子或两项蛋白因子,联合检测3项蛋白因子对淋巴结转移的诊断更有价值。结论联合检测血清VEGF、VEGF-C及MMP-9水平有助于提高对NSCLC淋巴结转移判断的准确率,手术前观察肺癌患者血清VEGF、VEGF-C及MMP-9水平将有助于判断疗效,监测预后和指导肺癌术后的多学科综合治疗。     

Transinactivation of the epidermal growth factor receptor tyrosine kinase and focal adhesion kinase phosphorylation by dietary flavonoids: effect on invasive potential of human carcinoma cells

Focal adhesion kinase (FAK), a member of a growing family of structurally distinct protein tyrosine kinases (PTK), has been linked to specific phosphorylation events, and the elevation of FAK activity in human carcinoma cells correlated with increased invasive potential. Transactivation of the epidermal growth factor receptor (EGFR) tyrosine kinase activity is proposed to stimulate cell migration and the subsequent activation of downstream signaling pathways. Quercetin (Qu) and luteolin (Lu), are potent PTK inhibitors as well as putative chemopreventive agents. The present work, we demonstrate that Qu and Lu at concentration of 20 microM transinactivated EGFR tyrosine kinase activity with marked reduction in phosphotyrosyl level of 170, 125, 65, 60 and 42 kDa cellular proteins, and induced apoptosis in MiaPaCa-2 cells. The 125 kDa protein was further identified as a FAK by immunoprecipitation and immunoblotting analyses. Tumor cells treated with Lu or Qu dampened the phosphorylation of FAK. In addition, our data clearly demonstrated that tumor cells responded to Qu and Lu by parallel reductions in the levels of phosphorylated FAK and the secreted matrix metalloproteinase (MMP) that may lead to the suppression of invasive potential and cell migration in vitro. While the molecular mechanism of FAK regulation of MMP secretion in tumor cells remains unclear, our results suggested that blockade of the EGFR-signaling pathway may contributed to the net effect. As suggested in the current study, targeting EGFR and FAK with the objective of modulating their regulatory pathways could offer prospects for the treatment of EGFR-responsive cancers in the future.     

Regulation of smooth muscle cell growth, function and death in vitro by activated mast cells--a potential mechanism for the weakening and rupture of atherosclerotic plaques

The fibrous cap of a lipid-containing atherosclerotic plaque consists of collagen produced by arterial smooth muscle cells (SMCs) of synthetic phenotype. A thick cap protects the lipid-rich core, whereas a thin cap predisposes it to rupture, with ensuing acute clinical complications, such as myocardial infarction. Among the pathological mechanisms leading to plaque weakening and rupture, one possibility is loss of the matrix-synthesizing SMCs. Indeed, caps of ruptured coronary plaques contain a reduced number of SMCs. In contrast, in such lesions, the number of activated inflammatory cells, such as mast cells, is increased, suggesting that they may regulate the SMC number. We have shown that heparin proteoglycans secreted by activated mast cells can efficiently inhibit proliferation of SMCs in vitro and reduce their ability to produce collagen. Chymase, a neutral serine protease secreted by activated mast cells, can also inhibit SMC-mediated collagen synthesis by a transforming growth factor-beta-dependent and -independent mechanism, and moreover, cause degradation of the collagen matrix by activating latent interstitial collagenase (MMP-1). Furthermore, chymase can induce SMC apoptosis by degrading the extracellular matrix component fibronectin necessary for SMC adhesion, with subsequent disruption of focal adhesions and loss of outside-in survival signaling. Thus, activated mast cells may participate in the weakening and rupture of atherosclerotic plaques by secreting mediators, such as heparin proteoglycans and chymase, which affect the growth, function and death of arterial SMCs.     

Curcumin ameliorates intrahepatic angiogenesis and capillarization of the sinusoids in carbon tetrachloride-induced rat liver fibrosis

Neoangiogenesis and the development of an abnormal angio-architecture in the liver are strongly linked with progressive fibrogenesis. This study aimed to evaluate the ability of curcumin to protect liver fibrosis-associated angiogenesis and capillarization of the sinusoids in experimental rats. Liver fibrosis was induced by intraperitoneal injection of carbon tetrachloride (CCl₄) with or without curcumin for 6 weeks. The results suggest that curcumin treatment markedly attenuated CCl₄-induced liver fibrosis, as assessed by histology and hydroxyproline content, and inhibited hepatic stellate cell activation. Curcumin ameliorated hepatic angiogenesis, as assessed by measuring microvessel density using Von Willebrand factor staining and by examining the expression of the endothelial cell markers CD31 and vascular endothelial growth factor receptor (VEGFR)-2 in the livers. Pathologic remodeling of liver sinusoidal capillarization, as assessed by electron-microscopic analysis of Disse's space and by evaluation of the levels of basement membrane protein expression, was also attenuated by curcumin administration. The intrahepatic gene or protein expression of hypoxia-inducible factor-1α, VEGFR-1, placental growth factor, and cyclooxygenase-2 decreased with treatment with curcumin in fibrotic rats. In conclusion, curcumin ameliorates hepatic angiogenesis and sinusoidal capillarization in CCl₄-induced rat liver fibrosis through suppressing multiple proangiogenic factors.     

New aspects of the role of hydroxyeicosatetraenoic acids in cell growth and cancer development

Lipoxygenase (LOX) pathway leads to the formation of leukotrienes and also catalyses the conversion of arachidonic acid (AA) to hydroperoxyeicosatetraenoic acids that are then reduced to hydroxyeicosatetraenoic acids (HETE) by glutathione peroxidase. There are four mammalian LOXs that produce 5-, 8-, 12- and 15-HETE, respectively. Cytochrome P-450 isozymes are also capable of metabolising AA to HETEs either by bis-allylic oxidation (lipoxygenase-like reaction) to generate 5-, 8-, 9-, 11-, 12- and 15-HETE; or by ?/?-1 hydroxylation to yield 16-, 17-, 18-, 19- and 20-HETEs.It is now widely recognised that HETEs have important physiological and pathological functions that modulate ion transport, renal and pulmonary functions, vascular tone and reactivity, and inflammatory and growth responses. They can be released during the action of growth factors and cytokines, reaching physiological concentrations higher than that of prostanoids and modulating the functions of these factors. Their effects can occur through receptor or non-receptor mechanisms. Recent reviews have summarised the effects of HETEs in vascular homeostasis or lung and renal physiology. The present review focuses on the emerging effects of HETEs on cell signalling and physiological cell growth. It also discusses current observations regarding the role of HETEs in apoptosis, angiogenesis, the proliferation of cancer cells and metastasis, which constitute a potential area for successful therapeutic intervention.     

Prevention of in vitro hepatic stellate cells activation by the adenosine derivative compound IFC305

We have previously shown that adenosine and the aspartate salt of adenosine (IFC305) reverse pre-established CCl₄-induced cirrhosis in rats. However, their molecular mechanism of action is not clearly understood. Hepatic stellate cells (HSC) play a pivotal role in liver fibrogenesis leading to cirrhosis, mainly through their activation, changing from a quiescent adipogenic state to a proliferative myofibrogenic condition. Therefore, we decided to investigate the effect of IFC305 on primary cultured rat HSC. Our results reveal that this compound suppressed the activation of HSC, as demonstrated by the maintenance of a quiescent cell morphology, including lipid droplets content, inhibition of α-smooth muscle actin (α-SMA) and collagen α1(I) expression, and up-regulation of MMP-13, Smad7, and PPARγ expression, three key antifibrogenic genes. Furthermore, IFC305 was able to repress the platelet-derived growth factor (PDGF)-induced proliferation of HSC. This inhibition was independent of adenosine receptors stimulation; instead, IFC305 was incorporated into cells by adenosine transporters and converted to AMP by adenosine kinase. On the other hand, addition of pyrimidine ribonucleoside as uridine reversed the suppressive effect of IFC305 on the proliferation and activation of HSC, suggesting that intracellular pyrimidine starvation would be involved in the molecular mechanism of action of IFC305. In conclusion, IFC305 inhibits HSC activation and maintains their quiescence in vitro; these results could explain in part the antifibrotic liver beneficial effect previously described for this compound on the animal model.