成年动物骨骼肌细胞原代培养及转染外源基因的初步研究

目的 观察损伤对成年动物骨骼肌卫星细胞增殖的影响,以获取基因治疗自体移植的载体。方法 以阳离子脂质体介导法将ｐＲＳＶ－ＬａｃＺ或ｐＲｃｈ－ＴＨ质粒转入培养肌细胞。结果 骨骼肌损伤后,分离出的肌卫星细胞及组织块游离成肌细胞数均显著增高;外源基因能在细胞内表达。结论 损伤可诱导成年骨骼的卫星细胞增殖并促进体外增减／肌细胞成活。     

成人骨骼肌细胞原代培养

本研究以获取基因治疗自体移植的载体为目的 ,观察了生长因子对成人骨骼肌卫星细胞增殖的影响。用手术中取得的成人骨骼肌进行体外组织块培养及酶消化培养 ,用成纤维细胞生长因子及表皮生长因子进行处理 ,作动态观察。结果证明 ,消化分离出的肌卫星细胞数量极少 ,培养不能成活 ;组织块培养肌卫星细胞的增殖与生长因子的作用有关 ,成纤维细胞生长因子及表皮生长因子处理组的增殖细胞数显著高于对照组。提示 ,生长因子可促进成年人骨骼肌卫星细胞增殖 ,但与其年龄及部位有一定关系。     

运动训练中骨骼肌细胞凋亡的研究进展

细胞凋亡是程序化细胞死亡过程 ,是受一系列基因控制的生理性细胞死亡方式。细胞凋亡是广泛存在于组织细胞的基本生物学现象 ,是细胞应答不同有害刺激或疾病而发生的一种特殊的细胞自杀行为。通过这种细胞自杀行为 ,机体消除损伤、衰老、突变的细胞 ,以维持生理平衡。这是完全不同于坏死的一种死亡途径 ,通常对机体是有利的。细胞凋亡存在于发育、成熟和衰老过程之中 ,同时也存在于多种现代疾病中 ,细胞凋亡异常与多种肌细胞疾病的发生机制有关。探讨肌细胞凋亡的机制对预防和处理运动导致的骨骼肌损伤 ,以及某些骨骼肌疾病的运动保健与功能…     

茶多酚对高糖所致人肾小球系膜细胞活性氧和TGF-β1表达的影响

目的 探讨高糖对人肾小球系膜细胞活性氧(ROS)和转化生长因子β1，(TGF-β1)表达的影响及茶多酚的干预作用。方法 培养人肾小球系膜细胞，分为正常对照组、高糖组、茶多酚组和茶多酚干预组，培养0、12、36h后，用分光光度比色法测定细胞上清液超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量，用半定量RT-PCR和免疫细胞化学法检测细胞内TGF-β1，mRNA和蛋白表达的变化。结果 高糖导致系膜细胞SOD活性下降和MDA含量升高，上调TGF-β1，mRNA和蛋白表达，随时间延长更为明显；茶多酚干预可拮抗高糖时系膜细胞的上述改变。结论 高糖能促进人肾小球系膜细胞ROS产生增加，上调TGF-β1 mRNA和蛋白表达，茶多酚能抑制高糖对系膜细胞的作用。     

运动对骨骼肌卫星细胞的影响及作用机制

骨骼肌卫星细胞在骨骼肌生长发育、损伤修复以及骨骼肌重塑等生理病理过程中具有重要的作用。适宜的运动训练可活化卫星细胞,促进卫星细胞增殖并向成肌细胞分化。本文就骨骼肌卫星细胞的起源、形态特征和特异性的标记以及运动训练调控骨骼肌卫星细胞活化、增殖、分化的作用机制进行综述。     

细胞因子对骨骼肌细胞和胰岛β细胞的作用

BACKGROUND: A variety of cytokines such as cytokines, growth factors and inflammatory proteins play an important role in the development of skeletal muscle. OBJECTIVE:To investigate the biological characteristics of a variety of cytokines and their effects on skeletal muscle cells and pancreatic β cells. METHODS: Relevant articles published from 2002 to 2015 were retrieved in CNKI and PubMed databases using the English keywords “cytokines, adiponectin, leptin, visfatin, skeletal muscle cells, pancreatic β cells”. Initially 253 literatures were obtained, and finally 53 eligible literatures were included based on the exclusion criteria. RESULTS AND CONCLUSION: As a fat-specific protein newly found, adiponectin can improve the insulin sensitivity by promoting glucose uptake, storage and utilization in skeletal muscle cells. The activation of muscle satellite cells and skeletal myoblast proliferation are both dependent on leptin, so leptin plays a vital role in the skeletal muscle cell growth and development. Visfatin, a pleiotropic cytokine, widely presents in the skeletal muscle, liver and bone marrow, and participates in the regulation of inflammation and immune function. Furthermore, visfatin contributes to glucose uptake and metabolism in the skeletal muscle, and makes considerable effects on the stress and signal transduction of skeletal muscle cells. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

骨骼肌卫星细胞对心肌梗死的治疗

细胞移植是目前对于急性心肌梗死治疗的热门话题,骨骼肌卫星细胞移植是最优选的心肌移植干细胞之一。本文阐述骨骼肌卫星细胞的特点,移植治疗心肌梗死的可能机制,体内移植时间,移植方法和标记方法,总结其目前在临床上的应用,指出目前关于其移植存在的问题,并对其应用做出展望。     

麦芽酚可抑制活性氧引起的SH-SY5Y细胞凋亡

近年来研究表明，氧化应激在阿尔茨海默病等神经退行性疾病发生发展中具有重要作用。本文研究了麦芽酚对过氧化氢(H2O2)引起的人神经瘤细胞株(SH-SY5Y)凋亡时细胞形态、磷脂酰丝氨酸(PS)外翻和DNA损伤的保护作用。结果发现麦芽酚浓度为2mmol／L，作用2h后，可以保护细胞免受H2O2引起的氧化损伤。实验结果表明麦芽酚可作为一种抗氧化剂，有效保护H2O2引起的SH-SY5Y的凋亡。     

大鼠骨骼肌卫星细胞的原代培养和鉴定

目的采用组织块培养技术探索大鼠骨骼肌卫星细胞的原代培养方法。方法以成年SPF级Sprague-Dawley大鼠为研究对象,采用组织块培养法获取大鼠骨骼肌卫星细胞,并与C2C12成肌细胞进行比较,对两种细胞进行形态学研究及采用免疫荧光和免疫组织化学法测定两种细胞α-actin蛋白和Desmin蛋白的表达及分布,从而对骨骼肌卫星细胞进行鉴定。结果通过组织块培养法获取的细胞增殖旺盛,分化良好。免疫细胞荧光和免疫组织化学实验结果显示,α-actin蛋白和Desmin蛋白在两种细胞胞浆中均有分布。结论用组织块培养法获取的骨骼肌卫星细胞具有良好的增殖与分化能力,用此种方法可培养出高纯度的骨骼肌卫星细胞。     

骨骼肌卫星细胞的培养及其在骨骼肌组织工程研究中的应用

近年来,随着组织工程技术的不断发展,生命科学、材料科学及制造科学的相互渗透,使得体外构建人体组织和器官的功能性替代物成为可能.骨骼肌卫星细胞作为组织工程的种子细胞在体外扩增至一定数量后与生物可降解三维支架材料结合,植入患者体内来修复缺损及恢复生理功能.目前对肌卫星细胞的体外培养已进行了较多研究,而对于肌卫星细胞的体外生长、增殖、鉴定,以及与组织工程三维支架的相互作用和生物功能的研究尚处于发展阶段,文中将对这些方面的研究进展作一综述.     

Reactive oxygen species and antioxidants in apoptosis of esophageal cancer cells induced by As2O3

To explore the relationship between the reactive oxygen species (ROS) and apoptosis in esophageal carcinoma cells (SHEE85) induced by arsenic trioxide (As2O3), we focused on changes of apoptosis, ROS, and antioxidants. Apoptosis of SHEE85 was confirmed by means of DNA fragmentation stained by Hoechst 33342, Sub-G1 cells scored by flow cytometry and ultrastructure of cells by electron microscopy. To evaluate the level of ROS, the chemiluminescent method was used for measuring the production of superoxide anion (O(-)*2). Lipid peroxide (malondialdehyde, MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were measured respectively by the photometry method. In the cells treated with As2O3 at a concentration of 5.0 micromol/l for 2-24 h, the content of cellular O(-)*2 and MDA was increased, but SOD and GSH-Px were significantly lower in the process of apoptosis in SHEE85. As2O3 at concentration of 0.5 micromol/l did not cause cell apoptosis but promoted cell proliferation. These results suggest that As2O3 at a high dosage (5 micromol/l) causes cell apoptosis and at a low dosage (0.5 micromol/l) causes cell proliferation. The essential mechanisms of cell apoptosis induced by As2O3 may be related to the increase of ROS and decrease of anti-oxidation. ROS and antioxidants participate in the apoptotic pathway of esophageal carcinoma cells.     

Reactive oxygen species and adhesion formation: clinical implications in adhesion prevention

Postoperative adhesion formation is a major clinical problem.It has been demonstrated that the pneumoperitoneum used duringlaparoscopy is a cofactor in adhesion formation. Reactive oxygenspecies (ROS) are produced in a hyperoxic environment and duringthe ischaemia/reperfusion process. ROS activity is deleteriousfor cells, which protect themselves by an antioxidant systemknown as ROS scavengers. ROS activity can increase by up-regulationof ROS themselves or by down-regulation of ROS scavengers. Recentdata also point to a role for ROS in adhesion formation sincethe administration of ROS scavengers decreases adhesion formationin several animal models. ROS activity increases during bothlaparotomy and laparoscopy. During laparoscopy, the pneumoperitoneumdetermines ischaemia at the time of insuflation and reperfusionat the time of deflation. During laparotomy, the environmenthas a 150 mmHg partial pressure of oxygen (pO₂), which is muchhigher than the intracellular pO₂ (5–40 mmHg). This canexplain the increase in ROS activity. The aim of this debateis to open a discussion about the importance of ROS activity,besides the known players and mechanisms involved, in adhesionformation and in adhesion prevention.     

Polykaryocyte formation induced by VSV in mouse L cells.

Infection of mouse L cells with VSV leads to the formation of polykaryocytes about 4 to 12 h p.i. When anti-VSV immune serum was added during the course of infection, progression of cell fusion was soon suppressed. Cycloheximide completely suppressed the cell fusion when the drug was added within 1 h p.i., while the cell fusion was not suppressed at all when the drug was added at and after 3 h. Early polykaryocyte formation, 'fusion from without', was observed only at a low level in cells infected at very high multiplicities. The development of cell fusion induced by VSV was found to be different in several cell types, although all these cells produced a rather high yield of virus: L and C-243-3 mouse cell lines showed a high level of polykaryocytosis (80 to 100%), BHK and RK-13 cells responded at low level, and PS and Vero cells showed no cell fusion in response to VSV infection. In PS cells, however, cell fusion occurred when VSV-infected L cells were co-cultivated. From these observations, the mechanism of cell fusion induced by VSV was discussed.     

Reactive oxygen species inhibited by titanium oxide coatings

Titanium is a successful biomaterial that possesses good biocompatibility. It is covered by a surface layer of titanium dioxide, and this oxide may play a critical role in inhibiting reactive oxygen species, such as peroxynitrite, produced during the inflammatory response. In the present study, titanium dioxide was coated onto silicone substrates by radio-frequency sputtering. Silicone coating with titanium dioxide enhanced the breakdown of peroxynitrite by 79%. At physiologic pH, the peroxynitrite donor 3-morpholinosydnonimine-N-ethylcarbamide (SIN-1) was used to nitrate 4-hydroxyphenylacetic acid (4-HPA) to form 4-hydroxy-3-nitrophenyl acetic acid (NHPA). Titanium dioxide-coated silicone inhibited the nitration of 4-HPA by 61% compared to aluminum oxide-coated silicone and 55% compared to uncoated silicone. J774A.1 mouse macrophages were plated on oxide-coated silicone and polystyrene and stimulated to produce superoxide and interleukin-6. Superoxide production was measured by the chemiluminescent reaction with 2-methyl-6-[p-methoxyphenyl]-3,7-dihydroimidazo[1,2-a]pyrazin-3-one (MCLA). Titanium dioxide-coated silicone exhibited a 55% decrease in superoxide compared to uncoated silicone and a 165% decrease in superoxide compared to uncoated polystyrene. Titanium dioxide-coated silicone inhibited IL-6 production by 77% compared to uncoated silicone. These results show that the anti-inflammatory properties of titanium dioxide can be transferred to the surfaces of silicone substrates.     

Reactive oxygen species signaling in plants

The evolution of aerobic metabolism such as respiration and photosynthesis resulted in the generation of reactive oxygen species (ROS). A common property of all ROS types is that they can cause oxidative damage to proteins, DNA, and lipids. This toxicity of ROS explains the evolution of complex arrays of nonenzymatic and enzymatic detoxification mechanisms in plants. However, increasing evidence indicates that plants also make use of ROS as signaling molecules for regulating development and various physiological responses. In this review, novel insights into the mechanisms of how plants sense and respond to ROS are discussed in the context of the biological effects and functions of ROS in plants.     

DNA damage induced by tumour necrosis factor-alpha in L929 cells is mediated by mitochondrial oxygen radical formation.

Treatment of L929 cells with tumour necrosis factor-alpha (TNF-alpha) plus actinomycin D induced DNA damage (indicated by the appearance of a sub-G1 peak due to extracellular leakage of low molecular weight DNA following DNA fragmentation) before significant cell lysis occurred. The DNA damage occurred in parallel with a decrease of the intracellular total glutathione content and an increase of intracellular reactive oxygen intermediates (ROI), as indicated by increased dihydrorhodamine 123 oxidation. Because the inhibition of mitochondrial respiration suppressed the increase of dihydrorhodamine 123 oxidation and DNA damage as well as the decrease in the total glutathione content, it was suggested that increased mitochondrial formation of ROI was responsible for DNA damage after TNF treatment. Deferoxamine (a ferric iron chelator) and dithiothreitol (a sulfhydryl reagent) both prevented DNA damage and cell killing, indicate that hydroxyl radicals generated from O2- and H2O2 produced by the mitochondria in a process catalysed by iron contributed to DNA damage and that this pathway may be involved in TNF-alpha-induced cytotoxicity. An inhibitor of poly(ADP)-ribose polymerase (3-aminobenzamide), worsened DNA damage, but was protective against cell lysis, suggesting that DNA repair subsequent to injury was more important than DNA damage per se in development of TNF-alpha cytotoxicity.     

Reactive oxygen species and aldehyde dehydrogenase activity in Hodgkin lymphoma cells

Tumor cells with tumorigenic potential might be limited to a small population of cells, called cancer-initiating cells (CICs). CICs efficiently form colonies in vitro, yield both CIC and non-CIC populations, maintain reactive oxygen species (ROS) at low levels, show high aldehyde dehydrogenase (ALDH) activity, and are mostly in a quiescent state of the cell cycle. CICs of Hodgkin lymphoma (HL) are small in size, with low levels of ROS. The relationship between ROS level and ALDH activity in CICs was examined in HL cell lines. ROS-low and ALDH-high populations formed colonies in semi-solid cultures more efficiently than ROS-high and ALDH-low populations. ALDH-high populations yielded both ALDH-low and -high populations, whereas ALDH-low populations rarely yielded an ALDH-high population. The number of cells in a quiescent state was significantly greater in ROS-low than in ROS-high cells, whereas that of ALDH-high and ALDH-low cells was comparable to each other. These findings show that ALDH-high and ROS-low cells share CIC-like potential, but they differ in their cell cycle status, suggesting that CICs are comprised of cells with heterogeneous characteristics.     

人视网膜色素上皮细胞诱导型一氧化氮合酶和活性氧在高糖状态下的表达

背景：高血糖导致的自由基损伤是糖尿病视网膜病变发病机制的中心环节。 目的：观察高糖对体外培养的人视网膜色素上皮细胞的氧化损伤作用以及高糖对人视网膜色素上皮细胞诱导型一氧化氮合酶和活性氧表达的影响。 方法：将培养人视网膜色素上皮细胞,分为对照组、高糖组和甘露醇组,分别用含5.5 mmol/L葡萄糖,33 mmol/L葡萄糖及5.5 mmol/L葡萄糖和27.5 mmol/L甘露醇的DMEM培养液培养。采用相差倒置显微镜观察细胞生长形态,采用免疫荧光染色研究诱导型一氧化氮合酶和3-硝基酪氨酸蛋白表达的变化,用氯甲基二氯二氢荧光素二乙酯荧光染色检测视网膜色素上皮细胞中活性氧的产生量。 结果与结论：与对照组相比,应用含33 mmol/L葡萄糖的DMEM培养基处理视网膜色素上皮细胞48 h可见细胞胞体变薄,形态表现多样,不规则细胞增多;高糖培养的视网膜色素上皮细胞诱导型一氧化氮合酶和3-硝基酪氨酸蛋白表达增加,活性氧产生明显增多。说明高浓度葡萄糖培养可造成人视网膜色素上皮细胞氧化损伤,使细胞形态发生变化,并导致细胞中3-硝基酪氨酸产生增多。