NADPH氧化酶对培养的小鼠血管平滑肌细胞Toll样受体4介导的炎症表型的影响

目的 探讨NADPH氧化酶对培养的小鼠血管平滑肌细胞(vascular smooth muscle cell,VSMC)Toll样受体4(Toll-like receptor 4,TLR4)介导的炎症表型中的作用.方法 分别采用NADPH氧化酶激动剂血小板衍生生长因子-BB(platelet-derived growth factor-BB,PDGF-BB)和抑制剂夹竹桃麻素(apocynin)处理培养的C57BL/6J和TLR4-/-小鼠胸主动脉VSMC.分别采用荧光探针二氯荧光素双醋酸盐染色法检测VSMC内活性氧(reactive oxygen species,ROS)含量,酶联免疫吸附法检测VSMC白细胞介素(interleukin,IL)-6、IL-1β和肿瘤坏死因子-α(tumor necrosis factor-o,TNF-α)表达,四甲基偶氮唑蓝染色法和Boyden小室检测VSMC的增殖和迁移.结果 PDGF-BB处理可显著增高C57BL/6J和TLR4-/-VSMC内ROS含量,而夹竹桃麻素可抑制ROS生成.PDGF-BB处理可使C57BL/6J VSMC IL-6[(52.69±3.49)ng/ml对(35.04±2.74) ng/ml,P=0.001]、IL-1β[(79.68±2.33) ng/ml对(62.38±0.54)ng/ml,P=0.000]和TNF-α[(218.35±5.42)ng/ml对(124.74±4.59) ng/ml,P=0.000]表达显著上调,增殖(1.69±0.53对1.04±0.40,P=0.000)和迁移(42.ll±4.05对1.69±0.53,P=0.000)能力均显著增强,而夹竹桃麻素预处理则可显著抑制VSMC IL-6[(42.11±4.05) ng/ml对(52.69±3.49) ng/ml,P=0.010]、IL-1β[(67.57± 1.36)ng/ml对(79.68±2.33) ng/ml,P=0.000]和TNF-α[(156.18±6.98) ng/ml对(218.35±5.42) ng/ml,P=0.000]表达以及增殖(1.23±0.42对1.69± 0.53,P=0.000)和迁移(42.11±4.05对52.69±3.49,P=0.000).TLR4-/-VSMC经PDGF-BB和夹竹桃麻素处理后,IL-6、IL-1β和TNF-α表达以及增殖和迁移能力均无显著变化.结论 NADPH氧化酶衍生的ROS参与了TLR4介导的VSMC炎症表型以及增殖和迁移,可能是其影响动脉粥样硬化发生和发展的重要机制.     

L’ingénierie cutanée pour le traitement des brûlures graves

Severe burned patients need definitive and efficient wound coverage. Outcome of massive burns has been improved by using cultured epithelial autografts (CEA). Despite fragility, percentages of success take, cost of treatment and long-term tendency to contracture, this surgical technique has been developed in few burn centres. First improvements were to combine CEA and dermis-like substitute. Cultured skin substitutes provide earlier skin closure and satisfying functional result. These methods have been used successfully in massive burns. Second improvement was to allow skin regeneration by using epidermal stem cells. Stem cells have capacity to differentiate into keratinocytes, to promote wound repair and to regenerate skin appendages. Human mesenchymal stem cells contribute to wound healing and were evaluated in cutaneous radiation syndrome. Skin regeneration and tissue engineering methods remain a complex challenge and offer the possibility of new treatment for injured and burned patients.     

胚鼠大脑皮层神经元培养方法的改进

目的建立一种简单、高效的胚鼠大脑皮层神经元原代培养的方法。方法17d胎鼠大脑皮质,直接机械切碎后经200目滤网研磨过滤,得到细胞悬液进行培养。结果神经元生长良好。相邻细胞的突起交错形成网络,鉴定神经元纯度为（94．7±5．6）％。结论本方法简单可行。可以获得高纯度的胚鼠大脑皮层神经元。     

人腔前卵泡体外培养研究

目的探讨FSH对人腔前卵泡(preantralfollicle)体外生长的影响.方法培养液中加入不同浓度的FSH,观察卵泡存活天数和卵泡发育增大.结果卵泡体外存活天数对照组为2.80±1.69天,Ⅰ、Ⅱ、Ⅲ组(FSH浓度为0.5IU/ml、1IU/ml、2IU/ml)分别为5.36±0.63、5.47±2.50、8.13±4.19天,与对照组相比,分别为P＜0.05、P＜0.01、P＜0.001.卵泡增大比率对照组为20.00%,Ⅰ、Ⅱ、Ⅲ组分别为35.71%、60.00%、81.25%,与对照组相比分别为P＞0.05、P＜0.05、P＜0.01.结论FSH能使卵泡体外存活天数延长,卵泡发育增大,并大致呈正相关关系.     

Growth Properties and Receptor Expression in Vascular Smooth Muscle Cells from Hypertensive Rats

The objective of this study was to evaluate the growth properties and receptor expression in aorta-ring derived smooth muscle cells (SMCs) cultured from control (WKY) and spontaneously hypertensive rats (SHR). SHR-SMCs exhibited a 3–4 day lag period before migrating. In addition, SHR-SMCs had a significantly higher growth rate, shorter population doubling time and higher saturation density level characteristics that were retained at higher passage levels. O-adrenergic and angiotensin (AII) receptors were measured using iodocyanopindolol (ICYP) and [3H]-All, respectively. All receptor expression was similar in both WKY and SHR-SMC cultures. WKY-SMCs exhibited little ICYP binding (Bmax 8.27± 2.0 fmol/mg) while SHR-SMC binding capacity was 8 fold higher (Bmax 65± 9.2 fmol/mg). In addition, the responsiveness of the P-receptor, as assessed by adenylyl cyclase stimulation, was similar for WKY and SHR-SMCs. These data suggest that factors regulating SMC receptor expression in vitro are selective since All and adrenergic receptor densities exhibit different responses to hypertension.     

镁对血管内皮细胞缺氧再复氧损伤的保护作用

目的探讨镁离子对人脐静脉血管内皮细胞缺氧再复氧损伤的保护作用及其可能机制。方法取人脐静脉分离内皮细胞培养传代 ,至第三代分为对照组、缺氧复氧组和硫酸镁组。用缺氧再复氧诱导血管内皮细胞损伤。镁离子组浓度分别为 (2、4、8、11、16mmol/L)。对照组常规培养 ,硫酸镁组加入不同浓度的镁离子预处理后缺氧再复氧。分别测定细胞培养液中一氧化氮 (NO) ,丙二醛 (MDA)及内皮素(ET-1)的含量。结果缺氧复氧组NO含量较对照组明显降低 (x缺氧 ±svsx对照 ±s,P<0.01) ,MDA与ET -1含量显著上升 (x缺氧 ±svsx对照 ±s,P<0.01) ,硫酸镁组NO含量较缺氧复氧组明显升高(x硫酸镁±svsx缺氧 ±s,P<0.05) ,MDA和ET -1含量显著降低 (x硫酸镁±svsx缺氧±s,P<0.05) ,且一定范围内呈剂量依赖关系。结论缺氧再复氧能造成VECs损伤 ,镁离子能保护缺氧再复氧造成的VECs损伤 ,其机制可能与抗脂质过氧化、稳定细胞膜及促进自由基清除有关。     

人工蛹虫草子实体对小鼠脾淋巴细胞增殖的影响

目的：观察人工蛹虫草子实体及其不同提取物对正常小鼠脾淋巴细胞增殖的影响，分析其影响免疫功能的有效提取部位。方法：以不同浓度提取物与脾细胞共孵育72h，采用MTT法测定淋巴细胞的增殖，观察提取物对淋巴细胞增殖的影响。另以10mg·kg^-1，30mg·kg^-1剂量人工蛹虫草子实体的水提取物及微粉化原药水溶液小鼠ig给药11d，制备脾淋巴细胞悬液，培养72h后测定细胞增殖。结果：在体外。水提取物有显著增强脾淋巴细胞增殖效应，且与ConA和LPS有协同促增殖效应，水提醇沉提取物可促进正常淋巴细胞增殖，而对ConA、LPS活化后的增殖无明显影响；甲醇提取物对淋巴细胞增殖有抑制作用。体内给予水提物和子实体微粉水溶液两个剂量也均显示促进淋巴细胞的自然增殖作用（P〈0．05），但对ConA或LPS活化后的淋巴细胞无协同作用。结论：人工蛹虫草子实体含有免疫促进和免疫抑制的成分。其水溶性和中等浓度的醇溶性成分有免疫促进作用。甲醇提取物抑制淋巴细胞增殖。     

体外转染SV40 T抗原对鼠内皮细胞周期素D1、P53和P21表达的影响

目的:探讨SV40 T抗原促进鼠内皮细胞增殖动力改变的机制,为基因治疗转化细胞提供理论依据。方法:用含有SV40 T抗原的表达质粒(pRNS-1)转染大鼠主动脉内皮细胞,经G418加压筛选,获得阳性克隆后扩增培养。在光镜下观察转染前后细胞的形态和生长特性;流式细胞仪检测细胞增殖周期;PCR检测外源基因的整合;免疫组化蛋白水平检测SV40 T抗原的表达;Western blot检测周期素(Cyclin)D1、P53和P21的表达。结果:转染后的鼠内皮细胞体外生存期延长,大量细胞由G1期进入S期,对血清和生长因子的依赖性降低。与转染前的细胞相比,转染后的鼠内皮细胞表达更多的P53和Cyclin D1,P21表达量降低。结论:SV40 T抗原可能通过调节Cyclin D1、P53和P21的表达来影响鼠内皮细胞的增殖。     

针灸对荷瘤小鼠瘤细胞和红细胞膜脂流动性的影响

目的：探讨针灸抗肿瘤的机理。方法：针刺、艾灸荷瘤小鼠的“气海”“足三里”穴,检测处理后组瘤细胞和红细胞膜脂流动性。结果：处理后各治疗组腹水瘤细胞膜脂流动性明显低于模型组（P＜0．01）,各治疗组红细胞膜脂流动性明显高于模型组（P＜0．01）,结论：针灸治疗使肿瘤细胞生物学活性降低,使机体生物学活性升高,从而在一定程度上揭示了针灸肿瘤的生物学机制。     

Effects of metabolic fragments of [Arg]-vasopressin on nerve growth in cultured hippocampal neurons

The effects of metabolic fragments of [Arg⁸]-vasopressin (AVP), [pGlu⁴, Cyt⁶]AVP (AVP_4–9), and desglycinamide-[pGlu⁴, Cyt⁶]AVP (AVP_4–8) on the growth of hippocampal neurons in culture were investigated in comparison with those of AVP. AVP_4–9 caused a significant increase in filopodial length following 96 h of exposure at concentrations higher than 300 nM. AVP_4–9 was more potent than AVP. AVP_4–8 also induced an increase in filopodial length, but this effect was less than that of AVP. The selective V₁ agonist [Phe², Ile³, Orn⁸]-vasopressin caused a significant increase in filopodial length, whereas the selective V₂ agonist [deamino-Cys¹, -Arg⁸]-vasopressin showed no such effect. OPC-21268, a vasopressin V₁ antagonist, blocked AVP and AVP fragment-induced increases in filopodial length. However, the V₂ antagonist OPC-31260 showed no such effect. A23187, a representative Ca ionophore, also increased filopodial length, and the A23187-induced increase in filopodial length was potentiated by AVP and AVP fragments. These results indicated that AVP_4–9 and AVP_4–8 increased filopodial length in cultured hippocampal neurons by activating V₁ receptors. Both phenomena induced by AVP_4–9 and AVP_4–8 were associated with intracellular calcium mobilization.