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1.
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
2.
目前妊娠期糖尿病的发病机制尚不完全清楚,随着研究的进展,免疫炎症假说日益受到学者的重视.妊娠期糖尿病的发病与病人体内的促炎/抗炎细胞因子平衡失常有关,该平衡倾向于促炎细胞因子的分泌,而抗炎细胞因子在疾病的发生、发展中可能发挥保护作用,文章综述了促炎细胞因子(TNF-6、IL-6、IFN-γ、IL-2、IL-17、IL-1β)和抗炎细胞因子(IL-10、IL-37、IL-4)在妊娠期糖尿病发生、发展中的作用,以及它们在临床筛查和诊断中的应用,以期为妊娠期糖尿病的预测和防治提供参考. 相似文献
3.
目的:研究和探讨梓醇对脂多糖(LPS)诱导的牛子宫内膜上皮细胞和小鼠子宫内膜炎的保护机制。创新点:首次证明梓醇对LPS刺激的牛子宫内膜上皮细胞炎症和LPS诱导的小鼠子宫内膜炎具有保护作用,其保护机制与抑制Toll样受体4/核因子κB(TLR4/NF-κB)炎症信号通路有关。方法:通过LPS的诱导,分别建立牛子宫内膜上皮细胞体外炎症模型和小鼠子宫内膜体内炎症模型,设置不同梓醇作用浓度梯度,采用酶联免疫吸附测定法(ELISA)、实时荧光定量聚合酶链式反应(q RT-PCR)、蛋白免疫印迹(western blot)和免疫荧光技术检测TLR4/NF-κB信号通路及其下游炎症因子的表达。结论:梓醇可以显著抑制TLR4和p65 NF-κB信号通路的表达,降低炎性因子肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)和白细胞介素6(IL-6)的水平以及趋化因子CXCL8和CXCL5的表达,同时降低子宫组织髓过氧化物酶水平。通过在体内外炎症模型中加入梓醇,可以显著降低牛子宫内膜上皮细胞的炎症反应,有效保护小鼠体内子宫内膜的组织损伤。 相似文献
4.
采用白藜芦醇(Res)和富勒烯(C60)结合制备Res/C60纳米组装体并验证其安全性和抗炎性。通过扫描电子显微镜(SEM)、拉曼光谱和X射线衍射研究了纳米组装体形貌、粒径大小及波谱峰值特征。其次,通过紫外分光光度法研究了Res的药物缓释行为。最后,通过MTT法在人脐静脉内皮细胞(HUVECs)层面验证了Res/C60纳米组装体的安全性以及体外的抗炎性。研究发现,Res/C60纳米组装体呈短棒状,横截面平均直径为(379.74±33.47)nm。拉曼光谱和X射线衍射特征峰显示Res/C60纳米组装体被成功制备。Res/C60纳米组装体的聚氨酯(PU)涂层中Res药物在30 d内累积释放量可达60%左右。胆管支架涂层对HUVECs细胞毒性较低,且通过释放Res可显著抑制炎症因子的释放。 相似文献
5.
目的:以园蓝为研究材料,鉴定园蓝花青素提取物(GBBAEs)中的功能性成分的结构,建立脂多糖(LPS)诱导的体外炎症模型,并评价其抗炎作用和初步机制。创新点:首次探究了蓝莓花青素对建立的LPS诱导体外炎症模型的营养干预作用,并初步探究了发挥抗炎机制的作用通路。方法:将RAW 264.7细胞分为对照组(不作处理)和实验组(1μg/ml LPS刺激建模)。实验组进一步分为3个不同浓度组:400μg/ml GBBAEs组、800μg/ml GBBAEs组和1200μg/ml GBBAEs组。用酶联免疫吸附测定(ELISA)试剂盒检测一氧化氮(NO)、前列腺素E2(PGE2)、白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、干扰素γ(INF-γ)等炎症因子的释放量;用实时定量聚合酶链反应(RT-PCR)分析IL-1β、IL-6、TNF-α、环氧合酶-2(COX-2)及单核细胞趋化蛋白-1(MCP-1)的炎症相关基因m RNA的表达水平;用蛋白质印迹法(Western blot法)测定相关炎症蛋白COX-2和NF-κBp65表达水平。结论:试验结果表明,通过ELISA法测定GBBAEs可以显著性抑制NO、PGE2、IL-1β、IL-6、INF-γ等炎症因子的释放;RT-PCR分析阐明在LPS诱导的单核-巨噬细胞RAW 264.7中,GBBAEs可以显著性抑制IL-1β、IL-6、TNF-α、COX-2及MCP-1的炎症相关基因m RNA的表达水平。此外,Western blot法进一步显示GBBAEs对相关炎症蛋白COX-2和NF-κBp65表达具有明显抑制作用,进一步证实GBBAEs通过NF-κB机制通路来发挥抗炎作用。 相似文献
6.
目的:评估安石榴苷对脂多糖诱导奶牛子宫内膜上皮细胞炎症损伤的保护作用,并初步探讨其作用机制。创新点:首次证明安石榴苷对脂多糖诱导奶牛子宫内膜上皮细胞炎症损伤具有保护作用,且此作用与核转录因子κB(NF-κB)和丝裂原活化蛋白激酶(MAPK)信号通路的抑制相关。方法:用不同浓度的脂多糖(1、10、30、50和100μg/ml)刺激奶牛子宫内膜上皮细胞3、6、9、12和18 h,筛选出建立炎症损伤的最佳作用浓度和时间。安石榴苷预处理细胞2 h后用脂多糖刺激12 h,用逆转录聚合酶链式反应(RT-PCR)检测炎症因子白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、白细胞介素8(IL-8)及肿瘤坏死因子α(TNF-α)的表达。用蛋白质免疫印迹试验(Western blotting)的方法检测核因子κB抑制蛋白α(IκBα)、磷酸化的p65、p38、c-Jun氨基末端激酶(JNK)和细胞外调节蛋白激酶(ERK)的表达水平。结论:MTT结果显示,30μg/ml脂多糖刺激奶牛子宫内膜上皮细胞12 h能够造成细胞活力下降和形态改变(图2和3)。RT-PCR结果显示,安石榴苷预处理后炎症因子显著降低(图4)。Western blotting结果显示,安石榴苷预处理能显著抑制IκBα降解以及p65、p38、JNK和ERK的磷酸化表达水平(图5和6)。综上所述,安石榴苷对脂多糖诱导奶牛子宫内膜上皮细胞炎症损伤具有保护作用,在治疗奶牛子宫内膜炎中具有重要价值。 相似文献
7.
目的:探讨解脲脲原体(Ureaplasma urealyticum)及其脂质相关膜蛋白(LAMPs)作用于人羊膜上皮细胞(HAECs)过程中白介素6(IL-6)、IL-8和肿瘤坏死因子α(TNF-α)的变化情况,阐明Toll样受体2(TLR2)的调控机制,明确解脲脲原体潜在的致病性。创新点:从解脲脲原体诱导炎症反应的分子机制入手,提出TLR2信号通路在其中的关键作用。方法:经TX-114处理萃取解脲脲原体获得LAMPs,将LAMPs和解脲脲原体分别感染HAECs,用酶联免疫吸附试验(ELISA)测定炎症细胞因子(IL-6、IL-8和TNF-α);采用实时聚合酶链反应(real-time PCR)测定TLR2 mRNA水平,用蛋白质印迹(Western blot)检测TLR2的表达量;经TLR2阻断剂(anti-h TLR2-IgA)处理后,测定相应炎症细胞因子。结论:解脲脲原体LAMPs能诱导HAECs的TLR2表达上调和炎症因子增加,从而发生炎症反应;TLR2受阻断后,炎症因子表达减少,炎症水平下降。TLR2在解脲脲原体LAMPs感染HAECs过程起关键作用。 相似文献
8.
王晗毓 杨沅浩 张潇 王彦 樊慧 石锦峰 谭雪莲 徐宝石 强静超 潘恩壮 储铭仪 董自波 董井泉 《Journal of Zhejiang University. Science. B》2023,(2):185-194
本研究基于脂多糖（LPS）诱导的小鼠脓毒症模型,探讨莲心碱（LIE）对脓毒症中脾损伤的潜在保护作用。脓毒症常伴有炎症、氧化应激和细胞凋亡,并将导致身体器官功能障碍,对脾脏损伤尤甚。将30只小鼠随机分为5组（n=6）：对照组、LPS(10 mg/kg)、LIE(10 mg/kg)+LPS、LIE(20 mg/kg)+LPS和LIE(40 mg/kg)+LPS。脾脏损伤通过苏木精–伊红染色（H&E）确定;采用定量聚合酶链反应（q PCR）检测脾脏组织中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IL-10、IL-1β和一氧化氮合酶（i NOS）的转录水平。同时对包括丙二醛（MDA）、过氧化氢酶（CAT）、超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）在内的氧化应激指标进行测定;采用TUNEL法检测细胞凋亡水平。结果显示,LIE可减轻脾脏组织病理学损伤并抑制细胞凋亡,显著降低促炎因子TNF-α、IL-6、IL-1β和i NOS的转录水平,并增加抗炎因子IL-10的表达。此外,LIE预处理后可降低MDA脂质过氧化指标,增强CAT、SOD和GSHPx的抗氧化活性... 相似文献
9.
目的:研究高压氧治疗降低瘢痕疙瘩切除联合放疗后复发率的效果及作用机制。创新点:将高压氧治疗应用于瘢痕疙瘩的辅助治疗。方法:(1)将240例瘢痕疙瘩患者随机分成两组(高压氧治疗组和非高压氧治疗组),高压氧治疗组接受瘢痕疙瘩切除术、放疗及高压氧治疗;非高压氧治疗接受瘢痕疙瘩切除及放疗。(2)收集两组复发病人二次手术切下的瘢痕疙瘩标本及正常皮肤标本,通过免疫组化染色观察三组组织标本因子白细胞介质-6(IL-6)、缺氧诱导因子-1α(HIF-1α)、肿瘤坏死因子-α(TNF-α)、核因子-κB(NF-κB)和血管内皮生长因子(VEGF)的表达量。结论:高压氧治疗通过提高瘢痕疙瘩组织氧含量及减少炎症反应来降低瘢痕疙瘩切除联合放疗后的复发率。 相似文献
10.
目的:动态观察四季变化对实验性脑梗死大鼠免疫炎性因子的的影响。方法:分别在春分、夏至、秋分、冬至日应用免疫印迹杂交法检测实验性脑梗死大鼠脑组织中NF-κB表达,应用放射免疫法检测血清中IL-1β、IL-2、及IL-6的水平,应用酶联免疫吸附法检测脑组织中IFN-γ的水平。结果:与春分组以及秋分组比较,夏至组以及冬至组IL-1β、IL-2、IL-6、IFN-γ及NF-κB表达均有不同程度升高(P<0.05或P<0.01)。结论:免疫炎性因子存在春秋低冬夏高的季节节律,季节对脑梗死的影响是通过免疫炎性因子网络产生的。 相似文献
11.
Zhang SZ Qian H Wang Z Fan JL Zhou Q Chen GM Li R Fu S Sun J 《Journal of Zhejiang University. Science. B》2010,11(11):889-894
Long-term preservation and easy transportation of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) will facilitate
their application in medical treatment and bioengineering. A pilot study on the freeze-drying of hBM-MSCs was carried out.
hBM-MSCs were loaded with trehalose. The glass transition temperature of the freeze-drying suspension was measured to provide
information for the cooling and primary drying experiment. After freeze-drying, various rehydration processes were tested.
The highest recovery rate of hBM-MSCs was (69.33±13.08)%. Possible methods to improve freeze-drying outcomes are discussed.
In conclusion, the present study has laid a foundation for the freeze-drying hBM-MSCs. 相似文献
12.
Tan Z Su ZY Wu RR Gu B Liu YK Zhao XL Zhang M 《Journal of Zhejiang University. Science. B》2011,12(1):18-27
Objective
Human embryonic stem cells (hESCs) have recently been reported as an unlimited source of mesenchymal stem cells (MSCs). The present study not only provides an identical and clinically compliant MSC source derived from hESCs (hESC-MSCs), but also describes the immunomodulative effects of hESC-MSCs in vitro and in vivo for a carbon tetrachloride (CCl4)-induced liver inflammation model. 相似文献13.
Objective: To investigate the infection of human embryo fibroblast cell line HF cells by CMV as well as the effects of CMV on β-actin mRNA and microfilaments. Methods: HF cells shape was observed after the infection of CMV. RT-PCR assay was used to detect the mRNA expression of CMV immediate early (IE) gene, β-actin and GAPDH genes of HF cells infected by CMV. CMV particles and cell microfilaments were detected with electron microscope. Results: Shape of HF cell changed after the infection by CMV. HF cells infected by CMV could express IE mRNA and the expression of β-actin mRNA decreased in a time- and titer-dependent manner compared with the uninfected HF cells whose expression of GAPDH mRNA did not change much. CMV particles were found with electron microscope in the cells. Microfilaments were ruptured and shortened after the infection of CMV. Conclusion: CMV can not only infect human embryo fibroblast cells line HF cells and replicate in the cells, but can also affect the expression of β-actin mRNA and the microfilaments. 相似文献
14.
INTRODUCTION Cell encapsulation in biocompatible and semipermeable polymeric membranes is an effective method for immunoprotection, regardless of the type of recipient (allograft, xenograft, etc.). The semipermeable nature of the membrane prevents high molecular weight molecules, antibodies and other immunologic moieties from coming into contact with the encapsulated cells and destroying them as foreign invaders, but permits the entry of nutrients and oxy- gen and the exit of therapeutic p… 相似文献
15.
Objective: To prepare microencapsulated cells releasing human tissue inhibitor of metalloproteinase-2 (TIMP-2), and investigate
their biological characteristics in vitro. Methods: Chinese hamster ovary (CHO) cells were stably transfected with a human
TIMP-2 expression vector, encapsulated in barium alginate microcapsules and cultured in vitro. Morphological appearance of
the microcapsules was observed under a light microscope. Cell viability was assessed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide) assay. Enzyme linked immunosorbent assay (ELISA) and reverse zymography were used to confirm the release of biologically
active TIMP-2 from the microcapsules. Cryopreservation study of the microencapsulated cells was carried out using dimethyl
sulfoxide (DMSO) as preservative agent. Results: The microcapsules appeared like a sphere with diameter of 300–600 μm. The
surface of the capsule wall was clearly smooth. The microencapsulated cells survived well and kept proliferating over the
6 weeks observed. No significant difference in TIMP-2 secretion was found between encapsulated and unencapsulated cells. Reverse
zymography confirmed the bioactivity of MMP (matrix metalloproteinase) inhibition of TIMP-2. The cryopreservation process
did not damage the microcapsule morphology nor the viability of the cells inside. Conclusion: Microencapsulated engineered
CHO cells survive at least 6 weeks after preparation in vitro, and secrete bioactive TIMP-2 freely from the microcapsules. 相似文献
16.
Abdul Hamid HAFIZAH Zakaria ZAITON Amom ZULKHAIRI Adenan MOHD ILHAM Megat Mohd Nordin NOR ANITA Abdullah Mahdy ZALEHA 《Journal of Zhejiang University. Science. B》2010,11(5):357-365
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
17.
18.
Objective: To investigate the infection of human embryo fibroblast cell line HF cells by CMV as well as the effects of CMV
on β-actin mRNA and microfilaments. Methods: HF cells shape was observed after the infection of CMV. RT-PCR assay was used
to detect the mRNA expression of CMV immediate early (IE) gene, β-actin and GAPDH genes of HF cells infected by CMV. CMV particles
and cell microfilaments were detected with electron microscope. Results: Shape of HF cell changed after the infection by CMV.
HF cells infected by CMV could express IE mRNA and the expression of β-actin mRNA decreased in a time-and titer-dependent
manner compared with the uninfected HF cells whose expression of GAPDH mRNA did not change much. CMV particles were found
with electron microscope in the cells. Microfilaments were ruptured and shortened after the infection of CMV. Conclusion:
CMV can not only infect human embryo fibroblast cells line HF cells and replicate in the cells, but can also affect the expression
of β-actin mRNA and the microfilaments.
Project (No. 001103058) supported by the Key Program of Science and Technology Bureau of Zhejiang Province, China 相似文献
19.
Zhi Yang Miao Liu Yin-gang Zhang Xiong Guo Peng Xu 《Journal of Zhejiang University. Science. B》2009,10(3):188-192
Objective: We investigated the effects of intermittent negative pressure on osteogenesis in human bone marrow-derived stroma cells (BMSCs) in vitro. Methods: BMSCs were isolated from adult marrow donated by a hip osteoarthritis patient with prosthetic replacement and cultured in vitro. The third passage cells were divided into negative pressure treatment group and control group. The treatment group was induced by negative pressure intermittently (pressure: 50 kPa, 30 rain/times, and twice daily). The control was cultured in conventional condition. The osteogenesis of BMSCs was examined by phase-contrast mi-croscopy, the determination of alkaline phosphatase (ALP) activities, and the immunohistochemistry of collagen type I. The mRNA expressions of osteoprotegerin (OPG) and osteoprotegerin ligand (OPGL) in BMSCs were analyzed by real-time poly-merase chain reaction (PCR). Results: BMSCs showed a typical appearance of osteoblast after 2 weeks of induction by intermit-tent negative pressure, the activity of ALP increased significantly, and the expression of collagen type 1 was positive. In the treatment group, the mRNA expression of OPG increased significantly (P<0.05) and the mRNA expression of OPGL decreased significantly (P<0.05) after 2 weeks, compared with the control. Conclusion: Intermittent negative pressure could promote os-teogenesis in human BMSCs in vitro. 相似文献
20.
Yan-hong Hu Xiu-rong Huang Ming-xin Qi Bu-yuan Hou 《Journal of Zhejiang University. Science. B》2012,13(5):402-407