联合应用NGF 和胰岛素对糖尿病大鼠烫伤创面表皮干细胞的影响

目的:通过局部联合应用神经生长因子和胰岛素对糖尿病大鼠深II度烫伤创面表皮干细胞标记物β1整合素和角蛋白19(K19)表达的影响,探讨神经生长因子和胰岛素联合应用于糖尿病烫伤创面治疗后对创面愈合的影响。方法:雄性wistar大鼠腹腔注射链脲佐菌素(STZ)建立糖尿病模型60只,1月后在大鼠背部造成深II度烫伤。将大鼠随机分为糖尿病对照组(B)、胰岛素治疗组(C)、神经生长因子治疗组(D)、神经生长因子联合胰岛素治疗组(E),每组15只。另取15只正常雄性wistar大鼠作为正常对照组(A)。观察伤后3、7、11、15、21 d各组创面愈合情况,检测创面β1整合素和角蛋白19(K19)的表达并计算创面愈合率。结果:E组创面愈合率自第7天起较A、B、C、D组增加,为[(25.33±2.32)%,(P<0.05)];A、C、D组创面愈合率较B组增加分别为[(22.51±1.78)%,(16.68±1.95)%,(18.29±1.70)%,(P<0.05)]。E组整合素β1和角蛋白19(K19)表达自伤后第7至21天各时相点显著增加,(P<0.05)。结论:糖尿病大鼠深II度烫伤创面局部联合应用神经生长因子和胰岛素可促进表皮干细胞的增殖与分化从而加速创面的愈合。     

Effect of oleic and linoleic acids on the inflammatory phase of wound healing in rats

Inflammation is a crucial step for the wound healing process. The effect of linoleic and oleic acids on the inflammatory response of the skin during the healing process and on the release of pro-inflammatory cytokines by rat neutrophils in vitro was investigated. A wound in the dorsal surface of adult rats was performed and fatty acids were then topically administered. Both oleic and linoleic acids increased the wound healing tissue mass. The total protein and DNA contents of the wounds were increased by the treatment with linoleic acid. The treatments with oleic and linoleic acids did not affect vascular permeability. However, the number of neutrophils in the wounded area and air pouches was increased and the thickness of the necrotic cell layer edge around the wound was decreased. A dose-dependent increase in vascular endothelial growth factor-alpha (VEGF-alpha) and interleukin-1beta (IL-1beta) by neutrophils incubated in the presence of oleic and linoleic acid was observed. Oleic acid was able to stimulate also the production of cytokine-induced neutrophil chemoattractant in inflammation 2 alpha/beta (CINC-2alpha/beta). This pro-inflammatory effect of oleic and linoleic acids may speed up the wound healing process.     

Migration of mesenchymal cell fated to blastema is necessary for fish fin regeneration

Urodeles and fish have higher regeneration ability in a variety of tissues and organs than do other vertebrate species including mammals. Though many studies have aimed at identifying the cellular and molecular basis for regeneration, relatively little is known about the detailed cellular behaviors and involved molecular basis. In the present study, a small molecule inhibitor was used to analyzed the role of phosphoinositide 3-kinase (PI3K) signaling during regeneration. We showed that the inhibitor disrupted the formation of blastema including the expression of characteristic genes. The failure of blastema formation was due to the impaired migration of mesenchymal cells to the distal prospective blastema region, although it had a little affect on cell cycle activation in mesenchymal cells. Moreover, we found that the epidermal remodeling including cell proliferation, distal cell migration and Akt phosphorylation was also affected by the inhibitor, implying a possible involvement of epidermis for proper formation of blastema. From these data, we propose a model in which distinct signals that direct the cell cycle activation, mesenchymal cell migration and epidermal remodeling coordinate together to accomplish the correct blastema formation and regeneration.     

枯草芽胞杆菌活菌制剂促进烧伤创面愈合的实验研究

目的　探讨枯草芽胞杆菌活菌制剂对烧伤创面愈合的影响。方法　通过大鼠深Ⅱ°烧伤模型,使用枯草芽胞杆菌活菌制剂,观察烧伤创面愈合过程中,成纤维细胞增殖周期的变化,以及测定羟脯氨酸（OHP）的含量,同时记录烧伤创面愈合时间,从而评价该制剂对烧伤创面愈合的影响。结果　应用枯草芽胞杆菌活菌制剂可促进成纤维细胞分裂、增殖,胶原含量提高,创面愈合时间明显缩短。结论　枯草芽胞杆菌活菌制剂具有促进烧伤创面愈合的作用。     

HGF/SF Induces Mesothelial Cell Migration and Proliferation by Autocrine and Paracrine Pathways

Mesothelial repair differs from that of other epithelial-like surfaces as healing does not occur solely by centripetal in-growth of cells as a sheet from the wound margins. Mesothelial cells lose their cell-cell junctions, divide, and adopt a fibroblast-like morphology while scattering across and covering the wound surface. These features are consistent with a cellular response to hepatocyte growth factor/scatter factor (HGF/SF). In this study, we examined the ability of mesothelial cells to secrete HGF/SF and investigated its possible role as an autocrine regulator of mesothelial cell motility and proliferation. We found that human primary mesothelial cells expressed HGF/SF mRNA and secreted active HGF/SF into conditioned medium as determined by ELISA and in a scattering bioassay. These cells also expressed the HGF/SF receptor, Met, as shown by RT-PCR and by Western blot analysis and immunofluorescence. Incubation of mesothelial cells with neutralizing antibodies to HGF/SF decreased cell migration to 25% of controls, whereas addition of HGF/SF disrupted cell-cell junctions and induced scattering and enhanced mesothelial cell migration. Furthermore, HGF/SF showed a small but significant mitogenic effect on all mesothelial cell lines examined. In conclusion, HGF/SF is produced by mesothelial cells and induces both motility and proliferation of these cells. These data are consistent with HGF/SF playing an autocrine role in mesothelial healing.     

Expression of the ipomoelin gene from sweet potato is regulated by dephosphorylated proteins, calcium ion and ethylene

A wound‐inducible cDNA, ipomoelin (IPO) was isolated from the subtraction library of sweet potato (Ipomoea batatas cv. Tainung 57) and used as a molecular probe to investigate the transduction pathway of wounding signal within plant cells. Following mechanical wounding of the leaves of sweet potato, IPO mRNA accumulation peaked at 6 h and then continuously declined. However, IPO gene expression in the apical unwounded leaves began at 6 h after wounding and continued for a further 10 h. Besides mechanical wounding, methyl jasmonate (MeJA) was identified as a signal transducer leading to the accumulation of IPO mRNA. Treatment with salicylic acid reduced the production of IPO mRNA, further supporting the involvement of the octadecanoid pathway in the signal transduction of wounding in sweet potato. In addition, ethylene was involved in the signal pathway and induced the expression of the IPO gene. Furthermore, the application of okadaic acid, a protein phosphatase inhibitor, blocked the accumulation of IPO mRNA induced by MeJA or ethylene, indicating that activation of the IPO gene by both MeJA and ethylene was via dephosphorylated proteins. The presence of a calcium ion chelator or channel blockers also inhibited the expression of the IPO gene after wounding. However, investigation by confocal scanning microscopy further pointed out that mechanical wounding rather than the application of MeJA induced the accumulation of the calcium ion. These results may indicate that the calcium ion is also involved in the activation of IPO mRNA. In addition, wounding signals the accumulation of calcium ion first and then stimulates the biosynthesis of MeJA in sweet potato. Hence, the reaction sequence of signal transducers, including the calcium ion, MeJA and protein kinase/phosphatase, in the wounding signalling pathway of sweet potato is suggested in this report.     

B.B.R对严重烧伤大鼠肠道屏障功能的保护作用

目的 :探索 B.B.R(复合生态制剂 )对严重烧伤早期肠道屏障功能的保护作用 ,为防治肠源性感染寻找新途径。方法 :选用健康 Wistar大鼠 13 0只 ,体重 180～ 2 2 0 g,雌雄各半 ,随机分为 B.B.R治疗组 ,BFL(单一双歧杆菌制剂 )治疗组 ,烧伤对照组 (BC)和正常对照组 (NC) ,建立 3 0 % °烫伤肠源性感染的动物模型 ,按时分批活杀取材 ,检测细菌易位率、盲肠膜菌群、血浆内毒素和肠粘膜 s Ig A的含量。结果 :B.B.R和 BFL 组 ,伤后 3 d肠道细菌易位率分别为 10 %、2 0 .7% ,与 BC组 (3 3 % )比较明显降低了细易位率。血浆内毒素 BC组明显高于 NC组 (P>0 .0 5 ) ,而 B.B.R组与 NC组比较 ,差异无显著性 (P<0 .0 1)。BC组盲肠中双歧杆菌数量与 BC组比较明显减少 ,而 B.B.R、BFL 组与 NC组比较差异无显著性 (P>0 .0 5 ) ;NC组肠道中酵母菌和大肠埃希菌与 NC比较明显增加 ,B.B.R组差异无显著性 ,肠粘液 s Ig A水平与上述指标有类似变化。结论 :3 0 % °烫伤大鼠肠道内容物双歧杆菌明显下降 ,大肠埃希菌、酵母菌迅速过度生长 ,导致肠道微生态失衡 ,应用 B.B.R治疗后 ,促进肠粘膜机械屏障功能的恢复 ,减少细菌和内毒素的易位 ,调整肠道微生态平衡 ,提高了肠道局部和全身免疫功能