氢生理盐水对角叉菜胶诱导足肿胀的抗炎效应
发布时间：2018-11-10 09:59
【摘要】： 氧化应激在炎症的发生发展过程中发挥着重要作用,一方面它可以直接损伤机体,另一方面它可以通过分子间相互作用间接地对机体造成损害。在参与炎症过程的众多复杂因子中,活性氧(ROS)和活性氮(RNS)非常重要,它们包括羟自由基、超氧阴离子、过氧化氢、一氧化氮、亚硝酸阴离子。有大量的实验证明清除羟自由基和亚硝酸阴离子能够明显减轻炎症的严重程度。最近的研究表明,氢气能够选择性地清除羟自由基和亚硝酸阴离子。因此,氢气很有可能通过这一途径而具有抗炎的效应。但是,通过呼吸道吸入氢气的方法很不方便而且存在安全隐患,于是我们将氢气通过高压溶解于生理盐水中,并探索这种含有氢气的生理盐水是否具有抗炎效应。 本课题以角叉菜胶诱导小鼠足肿胀为实验模型,研究氢生理盐水对于该炎症模型是否具有抗炎效应,并以巨噬细胞为切入点研究氢生理盐水对炎性细胞的影响。 本实验通过足爪容积测量仪检测了小鼠足肿胀的程度;组织化学切片观察了炎症局部的中性粒细胞浸润程度;酶联免疫吸附试验检测了细胞培养上清中的TNF-α的含量;实时定量PCR检测了RAW264.7及小鼠原代腹腔巨噬细胞中TNF-α的mRNA水平。 研究工作取得的主要结果如下: 1.氢生理盐水能够显著抑制角叉菜胶诱导的足肿胀:以2.5ml/kg、5ml/kg、10ml/kg的氢生理盐水的量处理角叉菜胶诱导足肿胀的动物模型,发现处理组脚肿胀的程度明显低于对照组,而且5ml/kg这个剂量效果最好。 2.氢生理盐水能够显著抑制炎症局部中性粒细胞的浸润:将氢生理盐水处理组、非处理组及对照组的足爪做成石蜡切片,HE染色后观察中性粒细胞浸润程度,氢水处理组的中性粒细胞浸润明显比非处理组少。 3.氢生理盐水能够减少活化巨噬细胞的TNF-α的mRNA水平并能抑制巨噬细胞分泌TNF-α:氢生理盐水和100ng/ml的LPS共同处理巨噬细胞1h,1h后裂解细胞通过实时定量PCR检测TNF-α的mRNA水平,发现氢生理盐水能够抑制巨噬细胞TNF-α的mRNA的表达;氢生理盐水和100ng/ml的LPS共同处理RAW264.7细胞4h,4h后收集上清检测TNF-α,发现氢生理盐水能够显著抑制由LPS引起的TNF-α产生,而且这种效应具有剂量依赖性。 总之,氢生理盐水不论在角叉菜胶诱导的足肿胀动物模型上还是在LPS激活的巨噬细胞上都具有明显的抗炎效应,这一效应很可能是通过清除羟自由基、亚硝酸阴离子等自由基来实现的。我们推测,氢生理盐水凭借其有效性、安全性、方便性、廉价性,很可能成为我们以后治疗某些炎症疾病的药物。在今后的工作中,我们将深入研究氢生理盐水具有抗炎效应的机制,如炎症局部的自由基变化等。
[Abstract]:Oxidative stress plays an important role in the development of inflammation. On the one hand, it can directly damage the body, on the other hand, it can indirectly damage the body through intermolecular interaction. Among the many complex factors involved in inflammatory process, reactive oxygen species (ROS) and active nitrogen (RNS) are very important. They include hydroxyl radical, superoxide anion, hydrogen peroxide, nitric oxide, nitrite anion. A large number of experiments have shown that scavenging hydroxyl radicals and nitrite anions can significantly reduce the severity of inflammation. Recent studies have shown that hydrogen can selectively scavenge hydroxyl radicals and nitrite anions. Therefore, hydrogen may have an anti-inflammatory effect through this pathway. However, the method of inhaling hydrogen through respiratory tract is very inconvenient and has some potential safety problems. Therefore, we dissolve hydrogen into saline by high pressure, and explore whether the hydrogen containing saline has anti-inflammatory effect. In this study, the mouse foot swelling induced by carrageenin was used as the experimental model to study whether hydrophysiological saline has anti-inflammatory effect on the inflammatory model, and to study the effect of hydrogen physiological saline on inflammatory cells from macrophages. In this experiment, the degree of mouse foot swelling was detected by paw volume measuring instrument, the degree of neutrophil infiltration in inflammatory region was observed by histochemical section, the content of TNF- 伪 in supernatant of cell culture was detected by enzyme-linked immunosorbent assay (Elisa). The mRNA levels of TNF- 伪 in RAW264.7 and mouse primary peritoneal macrophages were measured by real time quantitative PCR. The main results of the research are as follows: 1. Hydrophysiological saline significantly inhibited paw swelling induced by carrageenin: the animal model of foot swelling was induced by carrageenin with 2.5 ml / kg of 5 ml / kg / kg of hydrophysiological saline and 10 ml / kg of hydrophysiological saline. It was found that the degree of foot swelling in the treatment group was significantly lower than that in the control group, and 5ml/kg was the best dose. 2. Hydrophysiological saline could significantly inhibit the infiltration of neutrophils in the inflammatory region. The paws of the hydrophysiological saline treated group, untreated group and control group were made into paraffin sections. The degree of neutrophil infiltration was observed after HE staining. The neutrophil infiltration in hydrogen water treatment group was significantly less than that in non-treated group. 3. HNS could reduce the mRNA level of TNF- 伪 in activated macrophages and inhibit the secretion of TNF- 伪 by macrophages: HNS and 100ng/ml LPS co-treated macrophages for 1 h. After 1 hour, the mRNA level of TNF- 伪 was detected by real-time quantitative PCR. It was found that hydrophysiological saline could inhibit the mRNA expression of TNF- 伪 in macrophages. The supernatant of RAW264.7 cells was collected for 4 h after co-treatment of HNS and LPS of 100ng/ml for 4 h. It was found that HNS could significantly inhibit the production of TNF- 伪 induced by LPS, and this effect was dose-dependent. In conclusion, both in carrageenin-induced foot swelling animal models and in LPS activated macrophages, hydrogen saline has a significant anti-inflammatory effect, probably by scavenging hydroxyl radicals. Nitrite anion and other free radicals to achieve. We speculate that hydrophysiological saline, because of its efficacy, safety, convenience and cheapness, is likely to become a drug for the treatment of certain inflammatory diseases in the future. In the future, we will further study the mechanism of anti-inflammatory effect of hydrophysiological saline, such as the changes of free radicals in inflammation.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R363
本文编号：2322116
[Abstract]:Oxidative stress plays an important role in the development of inflammation. On the one hand, it can directly damage the body, on the other hand, it can indirectly damage the body through intermolecular interaction. Among the many complex factors involved in inflammatory process, reactive oxygen species (ROS) and active nitrogen (RNS) are very important. They include hydroxyl radical, superoxide anion, hydrogen peroxide, nitric oxide, nitrite anion. A large number of experiments have shown that scavenging hydroxyl radicals and nitrite anions can significantly reduce the severity of inflammation. Recent studies have shown that hydrogen can selectively scavenge hydroxyl radicals and nitrite anions. Therefore, hydrogen may have an anti-inflammatory effect through this pathway. However, the method of inhaling hydrogen through respiratory tract is very inconvenient and has some potential safety problems. Therefore, we dissolve hydrogen into saline by high pressure, and explore whether the hydrogen containing saline has anti-inflammatory effect. In this study, the mouse foot swelling induced by carrageenin was used as the experimental model to study whether hydrophysiological saline has anti-inflammatory effect on the inflammatory model, and to study the effect of hydrogen physiological saline on inflammatory cells from macrophages. In this experiment, the degree of mouse foot swelling was detected by paw volume measuring instrument, the degree of neutrophil infiltration in inflammatory region was observed by histochemical section, the content of TNF- 伪 in supernatant of cell culture was detected by enzyme-linked immunosorbent assay (Elisa). The mRNA levels of TNF- 伪 in RAW264.7 and mouse primary peritoneal macrophages were measured by real time quantitative PCR. The main results of the research are as follows: 1. Hydrophysiological saline significantly inhibited paw swelling induced by carrageenin: the animal model of foot swelling was induced by carrageenin with 2.5 ml / kg of 5 ml / kg / kg of hydrophysiological saline and 10 ml / kg of hydrophysiological saline. It was found that the degree of foot swelling in the treatment group was significantly lower than that in the control group, and 5ml/kg was the best dose. 2. Hydrophysiological saline could significantly inhibit the infiltration of neutrophils in the inflammatory region. The paws of the hydrophysiological saline treated group, untreated group and control group were made into paraffin sections. The degree of neutrophil infiltration was observed after HE staining. The neutrophil infiltration in hydrogen water treatment group was significantly less than that in non-treated group. 3. HNS could reduce the mRNA level of TNF- 伪 in activated macrophages and inhibit the secretion of TNF- 伪 by macrophages: HNS and 100ng/ml LPS co-treated macrophages for 1 h. After 1 hour, the mRNA level of TNF- 伪 was detected by real-time quantitative PCR. It was found that hydrophysiological saline could inhibit the mRNA expression of TNF- 伪 in macrophages. The supernatant of RAW264.7 cells was collected for 4 h after co-treatment of HNS and LPS of 100ng/ml for 4 h. It was found that HNS could significantly inhibit the production of TNF- 伪 induced by LPS, and this effect was dose-dependent. In conclusion, both in carrageenin-induced foot swelling animal models and in LPS activated macrophages, hydrogen saline has a significant anti-inflammatory effect, probably by scavenging hydroxyl radicals. Nitrite anion and other free radicals to achieve. We speculate that hydrophysiological saline, because of its efficacy, safety, convenience and cheapness, is likely to become a drug for the treatment of certain inflammatory diseases in the future. In the future, we will further study the mechanism of anti-inflammatory effect of hydrophysiological saline, such as the changes of free radicals in inflammation.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R363
【引证文献】
相关博士学位论文 前1条
1 以敏;桃仁改善不同病因所致血液循环障碍的药效及相关分子机制研究[D];广西医科大学;2012年
相关硕士学位论文 前1条
1 杨丽珍;不同制备工艺葛根芩连汤抗感染作用的比较研究[D];黑龙江中医药大学;2011年
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