硫化氢对烧伤大鼠体外培养皮肤巨噬细胞MAPK信号通路中p-ERK、p-p38、p-JNK的影响

发布时间：2018-04-18 22:00

本文选题：硫化氢 + 烧伤　；参考：《青海大学》2017年硕士论文

【摘要】：目的通过建立烧伤模型(大鼠),提取烧伤后皮肤基底的巨噬细胞,加入外源性H2S,用免疫印迹(Western blotting,WB)法测定体外培养巨噬细胞中的p-ERK、p-p38、p-JNK的蛋白含量的变化,探究硫化氢对体外培养皮肤巨噬细胞p-ERK、p-p38、p-JNK的影响,探究H2S参与创面愈合的一些机制。方法本实验的研究对象是烧伤大鼠创面皮肤基底提取的巨噬细胞。在30只大鼠中选取5只,正常饲养;其余25只做烫伤处理,从5只正常大鼠中随机选取1只大鼠作为正常组,25只烧伤大鼠中随机选取12只分为烧伤组、格列本脲(GLBN)组、Na HS+GLBN和Na HS组,每组3只大鼠。其中烧伤组和正常组加入等量PBS,格列本脲组加入格列本脲(调整终浓度至200umol/L),分别作用1、6、12小时。Na HS组加入Na HS(调整终浓度至200umol/L),分别作用1、6、12小时。Na HS+GLBN组加入Na HS+GLBN(二者调整终浓度均为200umol/L),作用1小时、6小时、12小时。在达到作用时间后通过免疫印迹法分别测定巨噬细胞细胞中p-PERK、p-p38、p-JNK的蛋白含量。结果(1)p-ERK蛋白相对表达量比较(P0.05):(1)烧伤组与空白对照组p-ERK量做相应检验,分析结果有意义。(2)在同一时间,Na HS组与各组别p-ERK相对量进行相应分析,认为分析结果有意义。(3)Na HS+GLBN组与GLBN组p-ERK蛋白量进行相应分析,结果有一定的差别,有统计意义。(4)GLBN组与烧伤组p-ERK蛋白相对量进行相应分析,分析所得到的结果有意义。(2)p-JNK蛋白相对表达量比较(P0.05):各组间比较:(1)烧伤组和空白组p-JNK蛋白表达量进行对比,结果经相应方法检验后,得出两组结果有差异,其结果有意义。(2)在同一时相点,Na HS组与其他组p-JNK表达量之间进行相应统计检验,结果有意义。(3)Na HS+GLBN组与GLBN组p-JNK蛋白表达量结果比较,有统计学意义。(4)GLBN组与烧伤组p-JNK蛋白量之间进行相应检验,得出的结果有差异。(3)p-p38蛋白表达量之间比较(p0.05):(1)烧伤组与空白对照组p-p38相对表达量经过相应的检验后,认为结果有一定的意义。(2)同一时间比较,Na HS组和其他各组别p-p38蛋白相对量经比较后认为差别之间比较有意义。(3)Na HS+GLBN组与GLBN组p-p38蛋白表达量结果比较有统计意义。(4)GLBN组与烧伤组行相应对比,p-p38蛋白相对量进行相应的检验分析,认为结果有意义。结论外源性硫化氢可以影响烧伤后大鼠体外培养皮肤巨噬细胞MAPK传导通路,表现为促进p-ERK的表达,抑制p-JNK和p-p38的表达。
[Abstract]:Objective to establish a burn model (rat model) by extracting macrophages from skin base after burn, adding exogenous H _ 2s, and using Western blotting method to determine the protein content of p-ERK _ (38) P _ (38) P _ (-JNK) in cultured macrophages in vitro.To explore the effect of hydrogen sulfide on the skin macrophage p-ERKN p-p38 p-JNK in vitro and to explore the mechanism of H2S involved in wound healing.Methods macrophages were extracted from the skin of burn rats.Five of the 30 rats were fed normally, the remaining 25 rats were scalded, and one of the 5 normal rats was randomly selected as the normal group. 12 of the 25 burn rats were randomly divided into burn group.There were 3 rats in each group in NaHS GLBN and NaHS groups.The burn group and the normal group added the same amount of PBSs, the glibenclamide group added glibenclamide (adjust the final concentration to 200umol / L), and the NaHS group added NaHS (adjusted final concentration to 200umolp / L), respectively. The NaHS GLBN group added NaHS GLBNs.Both adjusted final concentrations were 200umol / L ~ (-1), 1 h ~ 6 h / L ~ (-1) and 12 h ~ (-1).The protein contents of p-PERKN p-p38 + p-JNK in macrophages were determined by Western blotting after the action time was reached.Results the relative expression of p-ERK protein was compared between the burn group and the blank control group (P0.05: 1) the p-ERK content of the burn group and the blank control group was tested. The results were significant. 2) at the same time, the relative amount of p-ERK between the Na-HS group and the different groups was analyzed.The results showed that the p-ERK protein content in the HS GLBN group and the GLBN group was significantly higher than that in the GLBN group, and there was some difference between the two groups. There was statistical significance in the analysis of the relative amount of p-ERK protein between the GLBN group and the burn group.The results showed that the relative expression of p-JNK protein was compared between the burn group and the blank group. The results showed that there were differences between the two groups.The results were significant. (2) the corresponding statistical test was carried out between the p-JNK expression of NaHS group and other groups at the same time point. The results showed that the expression of p-JNK protein in Na-HS GLBN group and GLBN group was significantly higher than that in GLBN group, and the expression of p-JNK protein in Na-HS group was compared with that in GLBN group at the same time.There was a significant difference in the p-JNK protein expression between the burn group and the burn group. The results showed that the relative expression of p-p38 in the burn group and the blank control group was compared with that in the control group, and the relative expression of p-p38 in the burn group was compared with that in the blank control group.The relative amount of p-p38 protein was compared with that of burn group.Think the result makes sense.Conclusion exogenous hydrogen sulfide can affect the MAPK conduction pathway of skin macrophages cultured in vitro in burn rats, which can promote the expression of p-ERK and inhibit the expression of p-JNK and p-p38.
【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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