硫化氢对烧伤大鼠体外培养皮肤巨噬细胞分泌TNF-α和IL-1β的影响

发布时间：2018-06-07 15:17

本文选题：硫化氢 + 巨噬细胞　；参考：《青海大学》2017年硕士论文

【摘要】：目的探讨硫化氢对烧伤大鼠创面皮肤巨噬细胞体外培养分泌炎症介质TNF-α和IL-1β的影响方法探究对象选取经过烧伤处理的大鼠背部创面皮肤巨噬细胞。取健康SD大鼠30只(雌雄不限),其中5只正常喂养,余下25只实验大鼠行烧伤造模后喂养。随机选择正常喂养大鼠中的1只设为正常对照组,烧伤造模处理大鼠中的12只设为烧伤实验组。烧伤实验组再随机分成4组、每组3只,即烧伤对照组,Na HS(外源性H2S供体)组1h、6h、12h,格列本脲组(KATP通道阻滞剂)1h、6h、12h,Na HS+格列本脲组1h、6h、12h。烧伤实验大鼠于背部全部制作成深II°5%TBSA烧伤损伤(创面病理切片证实)的模型组,正常对照组大鼠不予任何处理,分离正常对照组大鼠背部皮肤及烧伤大鼠创面处皮肤基底巨噬细胞进行体外培养。在培养过程中,正常对照组与烧伤对照组巨噬细胞加入PBS液,Na HS组中加入浓度为200μmol/L的Na HS,格列本脲(Gli)组中加入浓度为200μmol/L的Gli,Na HS+Gli组中加入浓度为200μmol/L的Na HS和浓度为200μmol/L的Gli,于加入试剂之后的1h、6h、12h共3个时间点采取ELISA法检测巨噬细胞体外培养上清液中TNF-α与IL-1β的含量。结果于各个相同时间点检测,烧伤对照组巨噬细胞各时间点TNF-α与IL-1β的生成量均明显高于正常对照组各对应时间点生成量,差异明显存在统计学意义(P0.05);Na HS组巨噬细胞各时间点TNF-α与IL-1β的生成量均低于烧伤对照组各对应时间点生成量,差异明显存在统计学意义(P0.05);格列本脲(Gli)组巨噬细胞各时间点TNF-α与IL-1β的生成量均高于烧伤对照组各对应时间点生成量,差异明显存在统计学意义(P0.05);Na HS组巨噬细胞各时间点TNF-α与IL-1β的生成量均低于Gli组各对应时间点生成量,差异明显存在统计学意义(P0.05);Na HS组巨噬细胞各时间点TNF-α与IL-1β的生成量均低于Na HS+Gli组各对应时间点生成量,差异明显存在统计学意义(P0.05)。Gli组巨噬细胞各时间点TNF-α与IL-1β的生成量均高于Na HS+Gli组各对应时间点生成量,差异存在统计学意义(P0.05)。结论微量外源性H2S可减少烧伤创面巨噬细胞过量分泌炎症介质TNF-α和IL-1β,可降低烧伤后创面炎症反应程度。H2S凭借KATP通道途径在巨噬细胞内发挥其生理调节功能,格列本脲可凭借抑制KATP通道阻碍H2S产生生物学效应。
[Abstract]:Objective to investigate the effects of hydrogen sulfide on the secretion of inflammatory mediators TNF- 伪 and IL-1 尾 by skin macrophages of burn rats in vitro. Thirty healthy SD rats (male and female) were selected. 5 of them were fed normally. The remaining 25 rats were fed with burn model. One of the normal feeding rats was randomly selected as the normal control group, and 12 of the burn model rats were set up as the burn experimental group. Burn experimental group was divided into 4 groups at random, 3 rats in each group, namely burn control group (external H 2S donor) group (1 h, 6 h, 12 h), glibenclamide group (1 h, 6 h, 12 h) and glibenclamide group (1 h, 6 h, 12 h). All the experimental rats were made into the model group of deep II 掳5%TBSA burn injury (confirmed by pathological section of the wound) in the back, while the normal control group did not receive any treatment. Basal macrophages were isolated from the dorsal skin of normal control rats and burn rats. In the process of cultivation, The macrophages of normal control group and burn control group were added to PBS solution NaHS group with 200 渭 mol/L NaHS, glibenclamide Glix group added 200 渭 mol/L Gligna Na HS Gli group and 200 渭 mol/L NaHS group and 200 渭 mol/L group respectively. The concentration of TNF- 伪 and IL-1 尾 in the supernatant of macrophages cultured in vitro was determined by ELISA method at 3 time points (1 h, 6 h and 12 h after reagents). Results the levels of TNF- 伪 and IL-1 尾 in macrophages in burn control group were significantly higher than those in normal control group at the same time points. There was significant difference in the amount of TNF- 伪 and IL-1 尾 in macrophages at different time points in P0.05An Na HS group, which was lower than that in burn control group at different time points. The difference was statistically significant (P 0.05), and the amount of TNF- 伪 and IL-1 尾 in the glibenclamide group was higher than that in the burn control group at each time point, and the content of TNF- 伪 and IL-1 尾 in the Glib group was higher than that in the burn control group at each time point. There was significant difference in the amount of TNF- 伪 and IL-1 尾 in macrophages at each time point in P0.05Na-Na HS group, which was lower than that in Gli group at different time points. There was significant difference in the amount of TNF- 伪 and IL-1 尾 in macrophages at each time point in P0.05Na-Na HS group, which was lower than that in Na HS Gli group at different time points. There was significant difference in the amount of TNF- 伪 and IL-1 尾 in macrophages at each time point in P0.05Gli group, which was significantly higher than that in Na HS Gli group at each time point. The difference was statistically significant (P 0.05). Conclusion Trace exogenous H2S can reduce the excessive secretion of inflammatory mediators TNF- 伪 and IL-1 尾 by burn wound macrophages, and reduce the degree of inflammatory response of burn wounds. H2S can play its physiological regulation function in macrophages by means of KATP channel pathway. Glibenclamide can inhibit the biological effect of H 2S by inhibiting KATP channel.
【学位授予单位】：青海大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R644

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