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1.
通过建立人脐静脉内皮细胞(HUVEC)体外培养,采用电镜、免疫组化等技术观察肿瘤坏死因子-α(TNF-α)对HUVEC的存活率、形态和凋亡相关蛋白的影响,从而探讨TNF-α致血管内皮细胞(VEC)损伤的分子机制以及临床意义.  相似文献   

2.
颈动脉粥样硬化形成及不稳定性斑块脱落是造成颈动脉及脑内动脉狭窄或闭塞最主要的因素。白细胞分化抗原40及其配体(CD40-CD40L)的高度表达,会激发免疫及炎症反应,导致动脉粥样斑块内局部炎症细胞浸润,促发斑块破裂,进而发生缺血性脑卒中。因此及时了解颈动脉内膜和斑块的稳定情况,探索预测斑块破损的指标,进行及时干预,对于预防缺血性脑卒中的发生具有重要意义。  相似文献   

3.
目的:应用双向电泳及质谱技术分析对人脐静脉内皮细胞(HUVEC)蛋白质组的影响,探讨引起心血管毒性的潜在机制。创新点:多项流行病学调查表明,与人类心血管疾病有密切关系,而血管内皮细胞是血管的第一道防线。本项目应用蛋白质组技术研究对血管内皮细胞损伤的作用,并从DNA损伤的角度探讨了心血管毒性机制,具有较好的创新性。研究结果可为的危害分析提供基础数据,同时为的防治提供新的依据及思路。方法:培养HUVEC细胞,分为正常组(正常培养的HUVEC细胞)、处理组(50、100μg/mlPM_(2.5)处理HUVEC细胞24h)。双向电泳技术建立各组细胞蛋白质组图谱,质谱技术鉴定差异表达的蛋白质,流式细胞术分析细胞凋亡情况,酶联免疫吸附法(ELISA)检测DNA损伤产物8-OHd G的水平,荧光标记技术分析DNA双链断裂的形成,并用免疫印迹法(Western blotting)检测DNA损伤相关蛋白的表达。结论:经处理HUVEC后,31个蛋白表达发生了显著性的变化(图2,表1),其中8个蛋白质参与了DNA的损伤与RNA的编缉,7个蛋白质与细胞凋亡有关(表2)。进一步实验表明:能够促细胞凋亡(图5),提高DNA损伤产物8-OHdG的含量(图6),促进DNA双链断裂位点的形成(图7),调节损伤修复相关蛋白(Mer11A、Rad50和Rad51)的表达(图8),抑制超氧化物歧化酶(SOD)的活性,增加HUVEC细胞内活性氧(ROS)和丙二醛(MDA)的水平(图9)。综上所述,能够通过调节一系列蛋白的表达,加重HUVEC细胞氧化应激水平,增加DNA损伤,促进细胞凋亡,造成内皮细胞损伤,从而导致心脑血管事件的发生。  相似文献   

4.
目的:应用全转录组芯片研究缺氧/复氧诱导下人脐静脉内皮细胞(HUVEC)的转录组轮廓。创新点:血管内皮细胞(VEC)缺氧/复氧损伤被视定为许多生理和病理过程中导致器官功能障碍的重要驱动因素。然而,其详细病理生理机制和基因表达谱信息尚未阐明。本研究首次应用全转录组芯片技术研究VEC缺氧/复氧诱导下的转录组轮廓。方法:采用缺氧孵育3h后复氧1h的HUVEC为缺氧/复氧组,同时常氧孵育的HUVEC为常氧对照组。应用含58 339条探针的全转录组芯片检测每组三个样本。对差异表达基因进行生信分析和功能验证。结论:本研究发现372个有意义的差异表达基因探针。相关基因涵盖多种途径和功能,例如氧自由基的产生、钙超载、炎症、糖脂代谢、内皮细胞增殖、分化、细胞骨架及通透性调节、细胞裂解、凋亡和血管生成。另外,实验进一步表明,差异表达基因pleckstrin同源样域家族A成员1(PHLDA1)的m RNA和蛋白质表达结果与微阵列结果一致。STRING分析发现,PHLDA1可能与差异表达基因SLC38A3、SLC5A5、Lnc-SLC36A4-1和Lnc-PLEKHJ1-1具有物理性和/或功能性相互作用,这有望揭示VEC在缺氧/复氧环境下长链非编码RNA(lnc RNA)的相关机制。  相似文献   

5.
目的:应用全转录组芯片研究缺氧/复氧诱导下人脐静脉内皮细胞(HUVEC)的转录组轮廓。创新点:血管内皮细胞(VEC)缺氧/复氧损伤被视定为许多生理和病理过程中导致器官功能障碍的重要驱动因素。然而,其详细病理生理机制和基因表达谱信息尚未阐明。本研究首次应用全转录组芯片技术研究VEC缺氧/复氧诱导下的转录组轮廓。方法:采用缺氧孵育3h后复氧1h的HUVEC为缺氧/复氧组,同时常氧孵育的HUVEC为常氧对照组。应用含58 339条探针的全转录组芯片检测每组三个样本。对差异表达基因进行生信分析和功能验证。结论:本研究发现372个有意义的差异表达基因探针。相关基因涵盖多种途径和功能,例如氧自由基的产生、钙超载、炎症、糖脂代谢、内皮细胞增殖、分化、细胞骨架及通透性调节、细胞裂解、凋亡和血管生成。另外,实验进一步表明,差异表达基因pleckstrin同源样域家族A成员1(PHLDA1)的m RNA和蛋白质表达结果与微阵列结果一致。STRING分析发现,PHLDA1可能与差异表达基因SLC38A3、SLC5A5、Lnc-SLC36A4-1和Lnc-PLEKHJ1-1具有物理性和/或功能性相互作用,这有望揭示VEC在缺氧/复氧环境下长链非编码RNA(lnc RNA)的相关机制。  相似文献   

6.
合成了一种新的环状金属配体4-甲氧甲酰基-6-(4-甲基苯基)-2,2‘-联吡啶(HL)及它的单核与双核Pt(Ⅱ)配合物[Pt(L)PPh3](ClO4)(1)与[Pt2L2(μ-dppm)](ClO4)2(2)(dppm=二(二苯基磷)-甲烷),并研究了它们的结构及光物理性质.配合物2的晶体结构分析表明,中心金属离子Pt(Ⅱ)呈扭曲平面正方形构型,桥配体dppm连接两个金属中心,0.3375 nm的Pt---Pt距离表明双核配合物中存在金属-金属相互作用.两配合物在～450 nm处的肩峰归属于金属到配体的电荷转移(MLCT)吸收,在固体及溶液中均观测到强烈的光致磷光发射.配合物1在固态时620 nm的低能发射归属为3(π-π)跃迁,并暗示配合物1晶体结构中存在分子间配体-配体相互作用,然而在溶液中仅观察到3MLCT发射光谱,但配合物2在固态及溶液中都观察到明显的金属和金属相互作用到配体的电荷转移(3MMLCT)发射.  相似文献   

7.
通过水热法成功合成一个新的含N杂环二羧酸配体和草酸根离子的稀土配合物,其结构式为{[Gd2(Hpimda)2(μ4-C2O4).2H2O].2H2O}n(H3pimda为2-(吡啶-4-基)-1H-4,5-咪唑二羧酸),并对该配合物进行了表征.结构分析表明该配合物呈现草酸根离子作为柱子的三维框架结构,H3pimda配体把Gd(Ⅲ)离子连接成二维层状结构,进一步堆积形成一维通道,其中容纳着自由水和配位水分子.磁性研究表明,一维链内相邻的两个顺磁中心Gd(Ⅲ)离子之间存在着弱的反铁磁相互作用.  相似文献   

8.
目的:放创复合伤是一种以血管损伤和促炎细胞因子缺乏为特征的难愈性创伤。瘦素(leptin)的直接应用在血管生成和炎症中起着重要作用。本研究构建了一种可持续稳定的leptin表达系统——leptin修饰的人胎盘来源间充质干细胞(HPMSCs/leptin),并探究其对经X射线辐照后的人脐静脉内皮细胞(HUVECs)的成血管潜能及周围炎症的影响和潜在机制。创新点:可持续稳定的leptin表达系统(HPMSCs/leptin)促进受X射线辐照后HUVECs的成血管潜能及外周炎症反应,有助于解决放创复合伤伤口愈合过程中血管损伤和促炎因子缺乏的问题。方法:利用慢病毒载体将leptin基因转染HPMSCs获得HPMSCs/leptin。采用X射线单次照射HUVECs,剂量为20 Gy。细胞迁移侵袭实验技术(Transwell)检测照射后HUVECs的迁移情况。在Transwell体系的基础上,建立HPMSCs与受辐照HUVECs共培养体系。CCK-8比色法测定细胞增殖。酶联免疫吸附法(ELISA)检测促炎细胞因子(粒细胞-巨噬细胞集落刺激因子(GM-CSF)、白细胞介素-1α(IL-1α)、IL-6和IL-8)的分泌。实时荧光定量聚合酶链式反应(RT-qPCR)检测促血管生成因子(VEGF和bFGF)mRNA的表达。蛋白免疫印迹法(westernblot)检测核因子κB(NF-κB)和JAK/STAT信号通路的相关分子表达。结论:可持续稳定的leptin表达系统(HPMSCs/leptin)具有更好的细胞增殖、迁移和成血管潜能。HPMSCs/leptin单独培养和HPMSCs/leptin与受辐照HUVECs共培养体系中,促炎细胞因子的分泌增加与NF-κB和JAK/STAT信号通路的相互作用有关。HPMSCs/leptin可能促进X射线照射后HUVECs的成血管潜能和外周炎症反应。  相似文献   

9.
目的:放创复合伤是一种以血管损伤和促炎细胞因子缺乏为特征的难愈性创伤。瘦素(leptin)的直接应用在血管生成和炎症中起着重要作用。本研究构建了一种可持续稳定的leptin表达系统——leptin修饰的人胎盘来源间充质干细胞(HPMSCs/leptin),并探究其对经X射线辐照后的人脐静脉内皮细胞(HUVECs)的成血管潜能及周围炎症的影响和潜在机制。创新点:可持续稳定的leptin表达系统(HPMSCs/leptin)促进受X射线辐照后HUVECs的成血管潜能及外周炎症反应,有助于解决放创复合伤伤口愈合过程中血管损伤和促炎因子缺乏的问题。方法:利用慢病毒载体将leptin基因转染HPMSCs获得HPMSCs/leptin。采用X射线单次照射HUVECs,剂量为20 Gy。细胞迁移侵袭实验技术(Transwell)检测照射后HUVECs的迁移情况。在Transwell体系的基础上,建立HPMSCs与受辐照HUVECs共培养体系。CCK-8比色法测定细胞增殖。酶联免疫吸附法(ELISA)检测促炎细胞因子(粒细胞-巨噬细胞集落刺激因子(GM-CSF)、白细胞介素-1α(IL-1α)、IL-6和IL-8)的分泌。实时荧光定量聚合酶链式反应(RT-q PCR)检测促血管生成因子(VEGF和b FGF)m RNA的表达。蛋白免疫印迹法(westernblot)检测核因子κB(NF-κB)和JAK/STAT信号通路的相关分子表达。结论:可持续稳定的leptin表达系统(HPMSCs/leptin)具有更好的细胞增殖、迁移和成血管潜能。HPMSCs/leptin单独培养和HPMSCs/leptin与受辐照HUVECs共培养体系中,促炎细胞因子的分泌增加与NF-κB和JAK/STAT信号通路的相互作用有关。HPMSCs/leptin可能促进X射线照射后HUVECs的成血管潜能和外周炎症反应。  相似文献   

10.
目的:研究环氧合酶-2(COX-2)对卵巢癌细胞迁移和耐药性的影响及其机制。创新点:本研究发现COX-2对卵巢癌发生发展有一定的促进作用,可以通过上皮间质转化(EMT)途径促进卵巢癌细胞的迁移和顺铂(CDDP)耐药。其抑制剂塞来昔布(CXB)能起到协同抗癌的效果。方法:CCK-8检测CXB和CDDP对SKOV3和ES2细胞的毒性作用。划痕实验评估COX-2对卵巢癌细胞迁移的作用。蛋白质免疫印迹(western blot)和聚合酶链式反应(PCR)检测EMT相关基因和蛋白的表达水平。结论:COX-2可以通过EMT促进卵巢癌细胞迁移和CDDP耐药;CXB可以起到抑制作用,与CDDP协同抗癌。COX-2可以作为卵巢癌治疗的一个潜在靶点。  相似文献   

11.
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12.
１ＩｎｔｒｏｄｕｃｔｉｏｎＨｙｐｅｒ－ＩｇＭｓｙｎｄｒｏｍｅ（ＨＩＭ）ｉｓａｒａｒｅｉｍｍｕｎｏｄｅｆｉｃｉｅｎｃｙｃｈａｒａｃｔｅｒｉｚｅｄｂｙａｎｉｎｃｒｅａｓｅｄｓｕｓｃｅｐｔｉｂｉｌｉｔｙｔｏｒｅｃｕｒｒｅｎｔｉｎｆｅｃｔｉｏｎｓａｎｄｍａｒ...  相似文献   

13.
Endothelial cell death due to increased reactive oxygen species(ROS) may contribute to the initial endothelial injury,which promotes atherosclerotic lesion formation.Piper sarmentosum(PS),a natural product,has been shown to have an antioxidant property,which is hypothesized to inhibit production of ROS and prevent cell injury.Thus,the present study was designed to determine the effects of PS on the hydrogen peroxide(H2O2)-induced oxidative cell damage in cultured human umbilical vein endothelial cells(HUVECs).In this experiment,HUVECs were obtained by collagenase perfusion of the large vein in the umbilical cord and cultured in medium M200 supplemented with low serum growth supplementation(LSGS).HUVECs were treated with various concentrations of H2O2(0-1000 μmol/L) and it was observed that 180 μmol/L H2O2 reduced cell viability by 50% as denoted by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay.Using the above concentration as the positive control,the H2O2-induced HUVECs were concomitantly treated with various concentrations(100,150,250 and 300 μg/ml) of three different extracts(aqueous,methanol and hexane) of PS.Malondialdehyde(MDA),superoxide dismutase(SOD),catalase(CAT) and glutathione peroxidase(GPX) levels showed a significant increase(P0.05) in HUVECs compared to the negative control.However,PS extracts showed a protective effect on HUVECs from H2O2-induced cell apoptosis with a significant reduction in MDA,SOD,CAT and GPX levels(P0.05).Furthermore,PS had exhibited ferric reducing antioxidant power with its high phenolic content.Hence,it was concluded that PS plays a beneficial role in reducing oxidative stress in H2O2-induced HUVECs.  相似文献   

14.
The article "Cationic liposome-mediated trans-fection of CD40 ligand gene inhibits hepatic tumor growth of hepatocellular carcinoma in mice" [doi:10. 1631/jzus.B0820178] by Jiang et a1.(2009) in a recent issue of the Journal of Zhejiang University SCIENCE B was highly thought provoking. The authors have clearly demonstrated the efficacy of CD40 ligand gene therapy in inhibiting the growth of hepatocellu-lar carcinomas. The findings of Jiang et al.(2009) are highly important as they further support and cor-roborate the rapidly expanding role of CD40 ligand gene therapy in the management of systemic malig-nancies besides hepatocellular carcinomas.  相似文献   

15.
Objective: To establish normally conditionally-immortalized human umbilical vein endothelial cells (HUVECs) by ectopic expression of the human telomerase catalytic enzyme (hTERT) and simian virus 40 large T (SV40 LT) antigen. Methods: Primary HUVECs were transfected with recombinant retrovirus containing hTERT or SV40 LT respectively. Subsequently drug resistant cell clones were screened and expanded for further studies. Endothelial cell biomarkers were confirmed by examination. Results: The morphological phenotype of the transfected cells was similar to the non-transfected cells. Von Willebrand factor, hTERT and SV40 LT could be detected in transfected HUVECs. Moreover, higher telomerase activity in transfected cells was maintained for over 50 population doublings compared with only low level of endogenous telomerase transiently at early population doublings in primary HUVECs. When exposed to TNF-α(tumor necrosis factor-α), the expression of E-selectin in transfected cells was significantly up-regulated, but no alteration of endothelial lipase was found. Conclusion: Ectopic coexpression of hTERT and SV40 LT can effectively immortalize HUVECs without tumorigenicity in vitro. Immortalized HUVECs may be an ideal target of further molecular function studies.  相似文献   

16.
Homoharringtonine (HHT) has currently been used successfully in the treatment of acute and chronic myeloid leukemias and has been shown to induce apoptosis of different types of leukemic cells in vitro. Emerging evidence suggests that angiogenesis may play an important role in hematological malignancies, such as leukemia. How ever, whether HHT can relieve leukemia by anti-angiogenesis is still unknown. We investigated the anti-angiogenesis potential of HHT with the human umbilical vein endothelial cell line (ECV304) and leukemic cell line (K562) in vitro. Cellular proliferation was determined by MTT assay and apoptosis was analyzed by flow cytometry. The mRNA expression of vascular endothelial growth factor (VEGF) was assessed by RT-PCR and VEGF protein production was detected by Western blot. Inhibition of cell proliferation and induction of apoptosis by HHT were discovered in ECV304 cells, and appeared in a dose- and time-dependent manner. Also, treatment with HHT caused down-regulation of VEGF mRNA expression in K562 cells in similar dose- and time-dependent manner and inhibition of VEGF protein production in K562 cells in response to the enhancing concentration of HHT. The results demonstrated that HHT could also induce apoptosis in endothelium and down-regulate VEGF expression in K562 cells. In conclusion, we believe HHT has anti-angiogenesis potential and speculate that HHT might exert its anti-leukemia effects via reduction of angiogenesis.  相似文献   

17.
INTRODUCTION Homoharringtonine (HHT) is a cephalotoxin alkaloid with anti-leukemic activity and had been used successfully in the treatment of acute and chronic myeloid leukemias (O払rien et al., 1995; 1999; Feldman et al., 1992). The principal mecha-nism of action by HHT is the inhibition of protein synthesis in a dose- and time-dependent manner by binding to ribosome and inhibiting polypeptide chain elongation (Tujebajeva et al., 1989; Zhou et al., 1995). HHT had been shown to indu…  相似文献   

18.
Objective  This study is to determine the effect of the natural product parthenolide, a sesquiterpene lactone isolated from extracts of the herb Tanacetum parthenium, on the proliferation of vascular smooth muscle cells (VSMCs). Methods  Rat aortic VSMCs were isolated and cultured in vitro, and treated with different concentrations of parthenolide (10, 20 and 30 μmol/L). [3H]thymidine incorporation was used as an index of cell proliferation. Cell cycle progression and distribution were determined by flow cytometric analysis. Furthermore, the expression of several regulatory proteins relevant to VSMC proliferation including IκBα, cyclooxygenase-2 (Cox-2), p21, and p27 was examined to investigate the potential molecular mechanism. Results  Treatment with parthenolide significantly decreased the [3H]thymidine incorporation into DNA by 30%∼56% relative to control values in a dose-dependent manner (P<0.05). Addition of parthenolide also increased cell population at G0/G1 phase by 19.2%∼65.7% (P<0.05) and decreased cell population at S phase by 50.7%∼84.8% (P<0.05), which is consistent with its stimulatory effects on p21 and p27. In addition, parthenolide also increased IκBα expression and reduced Cox-2 expression in a time-dependent manner. Conclusion  Our results show that parthenolide significantly inhibits the VSMC proliferation by inducing G0/G1 cell cycle arrest. IκBα and Cox-2 are likely involved in such inhibitory effect of parthenolide on VSMC proliferation. These findings warrant further investigation on potential therapeutic implications of parthenolide on VSMC proliferation in vivo. Project (No. 491020-W50315) supported by the Foundation of the Health Bureau of Zhejiang, China  相似文献   

19.
Objective: To observe the effect of Yangxueqingnao particles on rat vascular smooth muscle cell (VSMC) proliferation induced by lysophosphatidic acid (LPA). Methods: The amount of3H-TdR (3H-thymidine) admixed in cultured rat VSMC was measured and mitogen-activated protein kinase (MAPK) activity and lipid peroxidation end product malondialdehyde (MDA)content of the VSMC were assayed. Results: 1×10-9, 1×10-8, 1×10-7 mol/L LPA in a concentration dependent manner, induced the amount of 3H-TdR admixed, MAP kinase activity, and MDA content of the cultured rat VSMC to increase. However, 5%, 10%,and 15% Yangxueqingnao serum preincubation resulted in a decrease of 23.0%, 42.0%, and 52.0% (P＜0.01) respectively in the amount of 3H-TdR admixed, a decline in VSMC MAP kinase activity of 13.9% (P＜0.05), 29.6% (P＜0.01), and 48.9% (P＜0.01)respectively, and also, a decrease in MDA content of VSMC of 19.4%, 24.7%, and 43.2% (P＜0.01) respectively, in the 1×10-7mol/L LPA-treated VSMC. Conclusions: LPA activates the proliferation and lipid peroxidation of VSMC in a concentration dependent manner. The LPA-induced VSMC proliferation is related to the activity of MAP kinases, enzymes involved in an intracellular signalling pathway. The results of the present study showed that Yangxueqingnao particles can effectively inhibit LPA-induced VSMC proliferation, MAP kinase activation, and reduce lipid peroxidative lesion.  相似文献   

20.
Ds-echinoside A (DSEA), a non-sulfated triterpene glycoside, was isolated from the sea cucumber Pearsonothuria graeffei. In vitro and in vivo investigations were conducted on the effects of DSEA on tumor cell adhesion, migration, invasion, and angiogenesis. In this study, we found that DSEA inhibited the proliferation of human hepatocellular liver carcinoma cells Hep G2, with a half-maximal inhibitory concentration (IC50) of 2.65 μmol/L, and suppressed Hep G2 cell adhesion, migration, and invasion in a dose-dependent manner. DSEA also reduced tube formation of human endothelial cells ECV-304 on matrigel in vitro and attenuated neovascularization in the chick embryo chorioallantoic membrane (CAM) assay in vivo. Immunocytochemical analysis revealed that DSEA significantly decreased the expression of matrix metalloproteinase-9 (MMP-9), which plays an important role in the degradation of basement membrane in tumor metastasis and angiogenesis. DSEA also increased the protein expression level of tissue inhibitor of metalloproteinase-1 (TIMP-1), an important regulator of MMP-9 activation. From the results of Western blotting, the expressions of nuclear factor-kappa B (NF-κB) and vascular endothelial growth factor (VEGF) were found to be remarkably reduced by DSEA. These findings suggest that DSEA exhibits a significant antimetastatic activity through the specific inhibition of NF-κB-dependent MMP-9 and VEGF expressions.  相似文献   

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