重组人角质细胞生长因子-2对烟雾吸入性损伤兔肺保护作用及其机制的研究

发布时间：2018-06-13 13:20

本文选题：重组人角质细胞生长因子-2 + 烟雾吸入性损伤　；参考：《南昌大学》2017年博士论文

【摘要】：目的:研究重组人角质细胞生长因子-2(rhKGF-2)对烟雾吸入性损伤兔肺组织保护作用及其可能机制。方法:新西兰大白兔120只制成烟雾吸入性损伤模型,随机分成五组,每组24只:0mg/kg组(对照组):致伤兔不作处理;PBS组:致伤后4h,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4h,雾化吸入rhKGF-2(1mg/Kg,溶于5ml PBS),每日1次,连续7日;2mg/kg组:致伤后4h,雾化吸入rhKGF-2(2mg/Kg,溶于5ml PBS),每日1次,连续7日;5mg/kg组:致伤后4h,雾化吸入rhKGF-2(5mg/Kg,溶于5ml PBS),每日1次,连续7日;每组分别于治疗1d、3d、5d、7d天随机抽取一只动物,抽动脉血行血气分析,放血处死动物,取肺泡灌洗液测定IL-6、IL-8、IL-10、MCP-1水平,取气管及肺组织行病理检查,并测定肺组织内NF-κB、表面活性蛋白A(SP-A)、血管内皮生长因子(VEGF)、磷酯酰肌醇3-激酶(P13K)、蛋白激酶B(Akt)及Bcl-2蛋白水平的蛋白及基因表达水平。TUNEL检测肺组织细胞凋亡情况。结果:(1)各组动物伤后1天,PaO2均处于较低水平,随着时间的延长,PaO2有升高的趋势,但各时点间无明显差异(P0.05);各剂量治疗组均可以明显提高PaO2(P〈0.01),但对PH、PaCO2、HCO3等无明显影响(P0.05)。(2)病理检查结果表明,伤后1天,气管粘膜肿胀、上皮细胞脱落,粘膜下见大量炎性细胞浸润;肺部水肿、出血,肺泡内可见大量炎性渗出,肺间质及肺泡内可见大量炎性细胞存在。(3)雾化吸入rhKGF-2后,肺泡灌洗液中IL-6、IL-8水平下降,以5mg组较为显著,IL-10升高不明显,各治疗组对MCP-1影响不大(P0.05)。(4)雾化吸入rhKGF-2后,NF-κB的蛋白水平及基因表达均出现下降,以5mg组为佳。(5)雾化吸入rhKGF-2后,肺组织中SP-A、VEGF蛋白逐渐升高,以5mg组动物明显,但Akt、PI3K的蛋白在治疗期间未见明显改变(P0.05)。(6)雾化吸入rhKGF-2后,肺组织中SP-A、VEGF、PI3K、Akt RNA表达明显增强,以5mg组动物明显;SP-A、VEGF、PI3K表达增强与剂量相关,但Akt RNA表达明显增强与剂量的高低无明显关系。(7)雾化吸入rhKGF-2后,肺组织中Bcl-2表达明显;细胞凋亡减少,2mg组、5mg组动物肺组织中Bcl-2的含量升高,但各时点1mg组动物与对照组比较无明显差异(P0.05);肺组织中Bcl-2 RNA表达增强,但组间差异不明显。结论:雾化吸入rhKGF-2可以改善烟雾吸入性损伤兔的氧合,减轻肺部炎症反应,并促进气道的修复,减少细胞凋亡,高剂量组表现更佳。其作用机制可能与下调NF-κB、激活PI3K/Akt通路及上调Bcl-2的表达有关。
[Abstract]:Aim: to study the protective effect of recombinant human keratinocyte growth factor-2 (rhKGF-2) on lung tissue in rabbits with smoke inhalation injury and its possible mechanism. Methods: 120 New Zealand white rabbits were randomly divided into five groups: 24 rabbits in each group (n = 24): 0 mg / kg / kg group (control group: PBS group: 4 h after injury, PBS5 ml of phosphate buffer solution was inhaled once a day. For 7 days, 1 mg / kg group: 4 hours after injury, aerosolized inhalation of rhKGF-2mg / kg, dissolved in 5ml, once a day, 7 days after 2 mg / kg: 4 h after injury, 2 mg / kg of rhKGF-2mg / kg, dissolved in 5ml once a day, 5 mg / kg for 7 consecutive days: 4 h after injury, 5 mg / kg of rhKGF-2mg / kg dissolved in 5ml PBSN, 1 time per day. For 7 consecutive days, one animal was randomly selected from each group on the 1st day, 3rd day, 5d and 7d after treatment. Blood gas analysis was performed on arterial blood, blood was put to death, IL-6, IL-8, IL-10 and MCP-1 levels were measured in alveolar lavage fluid, and the trachea and lung tissues were taken for pathological examination. The protein and gene expression levels of NF- 魏 B, surface-active protein Agn-SP-An, vascular endothelial growth factor (VEGF), phosphatidylinositol 3-kinase (P13KN), protein kinase B (Akt) and Bcl-2 protein in lung tissue were determined. Tunel was used to detect the apoptosis of lung tissue. Results (1) PaO2 was at a low level 1 day after injury, and the PaO2 increased with time. However, there was no significant difference between different time points (P < 0.05), and the Pao _ 2P < 0.01 ~ (P < 0.01) was significantly increased in each dose group, but there was no significant effect on PHPA _ (CO _ 2) HCO _ (3). The pathological examination showed that the tumefaction of tracheal mucosa and the exfoliation of epithelial cells were observed on the 1st day after injury. A large number of inflammatory cells infiltrated in the submucous membrane, pulmonary edema, hemorrhage, massive inflammatory exudation in the alveoli, the presence of a large number of inflammatory cells in the interstitium and alveoli of lung, and the level of IL-6 IL-8 in the alveolar lavage fluid decreased after inhalation of rhKGF-2 by atomization. The level of protein and gene expression of NF- 魏 B decreased after inhaled rhKGF-2, and the SP-AVEGF protein in lung tissue increased gradually after inhalation of rhKGF-2. After inhaled rhKGF-2, the expression of SP-Ag-VEGFPI3K mRNA was significantly increased in 5mg group, but the expression of SP-AVEGFPI3K protein was significantly increased in 5mg group. The increase of SP-AVEGFPI3K expression in 5mg group was correlated with the dose of rhKGF-2, and the expression of SP-AVEGFPI3K protein was significantly increased in 5mg group after inhalation of rhKGF-2, and the expression of SP-AVEGFPI3K was significantly increased in 5mg group. However, there was no significant relationship between the increase of Akt mRNA expression and the dose of rhKGF-2) after inhalation of rhKGF-2, the expression of Bcl-2 in lung tissue was obvious, and the expression of Bcl-2 in lung tissue was increased in the group of reduced apoptosis and 5 mg of apoptosis. However, there was no significant difference between the 1mg group and the control group at all time points, but the expression of Bcl-2 mRNA was increased in the lung tissue, but there was no significant difference between the two groups. Conclusion: aerosol inhalation of rhKGF-2 can improve oxygenation, alleviate pulmonary inflammation, promote airway repair and reduce apoptosis in rabbits with smoke inhalation injury. The mechanism may be related to down-regulation of NF- 魏 B, activation of PI3K / Akt pathway and up-regulation of Bcl-2 expression.
【学位授予单位】：南昌大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R563

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