角质细胞生长因子-2对烟雾吸入所致肺损伤修复作用的实验研究

发布时间：2019-01-26 10:47

【摘要】：目的观察角质细胞生长因子-2对烟雾吸入所致肺损伤的修复作用,并研究其相关机制。方法新西兰大白兔120只制成烟雾吸入性损伤模型,随机平均分为五组:0mg/kg组:致伤兔不作处理;PBS组:致伤后4小时,雾化吸入磷酸缓冲盐溶液(PBS)5ml,每日1次,连续7日;1mg/kg组:致伤后4小时,雾化吸入KGF-2(1mg/Kg,溶于5ml PBS),每日1次,连续7日;2mg/kg组:致伤后4小时,雾化吸入KGF-2(2mg/K g,溶于5ml PBS),每日1次,连续7日;5mg/kg组:致伤后4小时,雾化吸入KGF-2(5mg/K g,溶于5ml PBS),每日1次,连续7日;每组动物分别于治疗1d、3d、5d、7d后,在各时相点行动脉血气分析,放血处死,收集标本,Western blot法测定肺组织中肺表面活性蛋白A(SP-A)、血管内皮生长因子(VEGF)及Bcl-2蛋白水平,Real time PCR检测肺组织中肺表面活性蛋白A、血管内皮生长因子及Bcl-2蛋白mRNA水平,TUNEL检测细胞凋亡情况,并观察肺组织病理改变。结果(1)与0mg/kg组比较,PBS组烟雾吸入性损伤兔的PaO2、PaCO2均无显著差异(P0.05);致伤后,生存时间对PaO2、PaCO2均有影响(P0.01),随着生存时间延长,PaO2、PaCO2总体变化具有上升趋势;雾化吸入KGF-2对烟雾吸入性损伤兔PaCO2无明显影响(P0.05),对PaO2有影响(P0.01),能够提高PaO2,以5mg/kg组与各组比较差异均显著(P0.05),且KGF-2与时间有交互作用(P0.05),5mg/kg组,在第7d提高氧分压最明显。(2)与0mg/kg组比较,PBS组动物肺组织中SP-A、VEGF、Bcl-2含量及相关mRNA表达均无显著差异(P0.05);致伤后,生存时间对SP-A、VEGF、Bcl-2含量及相关mRNA表达均有影响(P0.01),随着生存时间延长,SP-A、VEGF、Bcl-2含量及相关mRNA表达总体变化均具有增加趋势;雾化吸入KGF-2对致伤动物肺组织中SP-A、VEGF、Bcl-2含量及相关mRNA表达均有影响(P0.01),能够提高肺组织中SP-A、VEGF、Bcl-2含量及相关mRNA表达,以5mg/kg组与各组比较差异均显著(P0.05),且KGF-2与时间有交互作用(P0.01),5mg/kg组,在第7d提高肺组织中SP-A、VEGF、Bcl-2含量及相关mRNA表达最明显。(3)与0mg/kg组比较,PBS组动物肺组织细胞凋亡指数无显著差异(P0.05);致伤后,生存时间对肺组织细胞凋亡指数有影响(P0.01),随着生存时间延长,肺组织细胞凋亡指数总体变化具有降低趋势;雾化吸入KGF-2对动物肺组织细胞凋亡指数有影响(P0.01),能够降低肺组织细胞凋亡指数,以5mg/kg组与各组比较差异均显著(P0.05),且KGF-2与时间有交互作用(P0.01),5mg/kg组,在第7d降低肺组织细胞凋亡指数最明显。(4)肺组织病理检查显示,与伤后第1天相比,各组动物第7天肺部损伤表现均较前减轻,各组相比,以5mg/Kg组动物损伤减轻最为明显。结论KGF-2对烟雾吸入所致肺损伤具有修复作用。其修复作用可能与KGF-2增加烟雾吸入性损伤的肺表面活性蛋白A合成,促进肺新生血管形成,以及抑制肺损伤细胞凋亡这三方面机制有关。
[Abstract]:Objective to investigate the repair effect of keratinocyte growth factor 2 (KGF 2) on lung injury induced by smoke inhalation and its related mechanism. Methods 120 New Zealand white rabbits were randomly divided into five groups: 0mg/kg group: the injured rabbits were not treated; In PBS group, 4 hours after injury, (PBS) 5 ml of phosphate buffer solution was inhaled once a day for 7 days, while in 1mg/kg group, KGF-2 (1 mg / kg) dissolved in 5ml PBS), once daily for 7 days, 4 hours after injury. In 2mg/kg group, KGF-2 (2mg/K g, dissolved in 5ml PBS), once a day for 7 days) was inhaled 4 hours after injury. In 5mg/kg group, KGF-2 (5mg/K g, dissolved in 5ml PBS), once a day for 7 days) was inhaled 4 hours after injury. The animals in each group were treated for 1 day, 3 days, 5 days and 7 days later. Arterial blood gas analysis was performed at each time point, and blood was put to death. The lung surfactant protein A (SP-A) in lung tissue was determined by, Western blot method. The levels of vascular endothelial growth factor (VEGF) and Bcl-2 protein were detected by, Real time PCR, and the levels of pulmonary surfactant protein A, vascular endothelial growth factor (VEGF) and Bcl-2 protein mRNA in lung tissue were detected by, Real time PCR. Apoptosis was detected by TUNEL, and the pathological changes of lung tissue were observed. Results (1) there was no significant difference in PaO2,PaCO2 between PBS group and 0mg/kg group (P0.05). After injury, the survival time had an effect on PaO2,PaCO2 (P0.01). With the prolongation of survival time, the overall change of PaO2,PaCO2 showed an upward trend. Aerosol inhalation of KGF-2 had no significant effect on PaCO2 in rabbits with smoke inhalation injury (P0.05), but had an effect on PaO2 (P0.01), and could improve PaO2, significantly in 5mg/kg group compared with each group (P0.05). There was interaction between KGF-2 and time (P0.05). In 5mg/kg group, the increase of oxygen partial pressure was the most obvious on the 7th day. (2) compared with 0mg/kg group, SP-A,VEGF, in lung tissue of PBS group was more obvious than that of PBS group. There was no significant difference in Bcl-2 content and related mRNA expression (P0.05). After injury, survival time had an effect on SP-A,VEGF,Bcl-2 content and related mRNA expression (P0.01). With the prolongation of survival time, the content of SP-A,VEGF,Bcl-2 and the expression of related mRNA increased. Aerosol inhalation of KGF-2 had an effect on the expression of SP-A,VEGF,Bcl-2 and related mRNA in lung tissue of injured animals (P0.01), and could increase the content of SP-A,VEGF,Bcl-2 and the expression of related mRNA in lung tissue. The difference between 5mg/kg group and each group was significant (P0.05), and there was interaction between KGF-2 and time (P0.01). In 5mg/kg group, the SP-A,VEGF, in lung tissue was increased on the 7th day. Bcl-2 content and related mRNA expression was the most obvious. (3) compared with 0mg/kg group, the apoptosis index of lung tissue in PBS group had no significant difference (P0.05). After injury, survival time had an effect on the apoptosis index of lung tissue (P0.01), and with the prolongation of survival time, the overall change of apoptosis index of lung tissue showed a decreasing trend. Aerosol inhalation of KGF-2 had an effect on the apoptosis index of lung tissue (P0.01), and could decrease the apoptosis index of lung tissue. The difference between 5mg/kg group and each group was significant (P0.05). KGF-2 had interaction with time (P0.01). In 5mg/kg group, the apoptosis index of lung tissue was significantly decreased on the 7th day. (4) the pathological examination of lung tissue showed that compared with the first day after injury, the apoptosis index of lung tissue was significantly decreased in 5mg/kg group. On the 7th day, lung injury was alleviated in all groups, especially in 5mg/Kg group. Conclusion KGF-2 can repair lung injury induced by smoke inhalation. The repair effect of KGF-2 may be related to the increase of pulmonary surfactant A synthesis in smoke inhalation injury, the promotion of pulmonary angiogenesis, and the inhibition of apoptosis of lung injury cells.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R563

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