汽化吸入PFC对肺损伤猪肾上皮细胞凋亡的影响(1)
【摘要】 目的 通過对肺灌洗造成小猪肺损伤模型行全氟化碳(PFC)汽化吸入， 了解其对肺损伤小猪肾小管上皮细胞凋亡的影响。方法 取健康小猪20只， 随机分为VMV组(PFC汽化吸入， 10只)、CMV组(常规机械通气， 8只)和假手术组(2只)。取各组肾皮髓交界处肾脏组织， 采用电镜观察凋亡细胞超微结构， DNA断裂的原位末端标记法(TUNEL)法检测细胞凋亡指数， 免疫组织化学法测定凋亡基因B细胞淋巴瘤/白血病-2(bcl-2)、bcl-2相关X蛋白(BAX)蛋白的表达， 半定量逆转录-聚合酶链反应(RT-PCR)法测定BAX、bcl-2核糖核酸(mRNA)表达。结果 TUNEL结果：肾的阳性细胞主要分布在近端肾小管， 肾小球未见凋亡细胞。VMV组细胞凋亡率为(20.2±2.9)%， 明显低于CMV组的(80.2±4.8)%， 差异有统计学意义(t=32.868， P=0.000<0.05)。肾BAX、bcl-2蛋白表达结果：VMV组的肾BAX蛋白表达(12.6±6.8)明显低于CMV组的(62.2±3.6)， bcl-2蛋白表达(36.8±2.1)明显高于CMV组的(6.2±2.3)， 差异均有统计学意义(P<0.05)。肾BAX、bcl-2 mRNA表达结果：VMV组的肾BAX mRNA表达(7.10±0.32)明显低于CMV组的(9.29±1.06)， bcl-2 mRNA表达(10.29±2.51)， 明显高于CMV组的(5.18±1.08)， 差异均有统计学意义(t=6.230、5.350， P<0.05)。假手术组细胞凋亡数极少， BAX、bcl-2表达均很低。结论 汽化吸入PFC可减少急性肺损伤小猪肾小管上皮细胞凋亡， 其作用机制可能与抑制BAX基因的蛋白表达及升高bcl-2基因表达有关。
【关键词】 全氟化碳;急性肺损伤;凋亡;机械通气;肾脏
DOI：10.14163/j.cnki.11-5547/r.2019.17.108
Effect of vaporization inhalation of PFC on apoptosis of renal tubular epithelial cells in pigs with lung injury QIN Xin， QIN Lei， LIU You-ning. Department of Respiratory Medicine， Second Hospital of Shanxi Medical University， Taiyuan 030001， China
【Abstract】 Objective To understand the effect of vaporization inhalation of perfluorocarbon (PFC) for model of pig lung injury caused by lung lavage on apoptosis of renal tubular epithelial cells in lung injury. Methods A total of 20 healthy pigs were randomly divided into VMV group (vaporization inhalation of PFC，10 pigs)， CMV group (conventional mechanical ventilation， 8 pigs) and sham operation group (2 pigs). Kidney tissue at the junction of renal cortex and medulla was taken from each group. Ultrastructure of apoptotic cells was observed by electron microscopy， apoptotic index was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling(TUNEL)， expression of apoptotic genes B cell lymphoma/leukemia-2 (bcl-2) and bcl-2 related X protein (BAX) was determined by immunohistochemistry， and expression of BAX and Bcl-2 ribonucleic acid (mRNA) was determined by Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Results TUNEL results： The positive cells of the kidney were mainly distributed in the proximal tubules， and no apoptotic cells were found in the glomeruli. VMV group had obviously lower apoptosis rate as (20.2±2.9)% than (80.2±4.8)% in CMV group， and the difference was statistically significant (t=32.868， P=0.000<0.05). Renal BAX， bcl-2 protein expression results： VMV group had obviously lower BAX protein expression as (12.6±6.8) than (62.2±3.6) in CMV group， and obviously higher bcl-2 protein expression as (36.8±2.1) than (6.2±2.3) in CMV group. Their difference was statistically significant (P<0.05). Renal BAX and bcl-2 mRNA expression results：VMV group had obviously lower BAX mRNA expression as (7.10±0.32) than (9.29±1.06) in CMV group， and obviously higher bcl-2 mRNA expression as (10.29±2.51) than (5.18±1.08) in CMV group. Their difference was statistically significant (t=6.230， 5.350， P<0.05). The apoptotic cells in sham operation group were very few and the expressions of BAX and Bcl-2 were very low. Conclusion Vaporized inhalation of PFC can reduce the apoptosis of renal tubular epithelial cells in piglets with acute lung injury. Its mechanism may be related to inhibiting the expression of BAX gene and increasing the expression of Bcl-2 gene., http://www.100md.com(秦鑫 秦蕾 刘又宁)
【关键词】 全氟化碳;急性肺损伤;凋亡;机械通气;肾脏
DOI：10.14163/j.cnki.11-5547/r.2019.17.108
Effect of vaporization inhalation of PFC on apoptosis of renal tubular epithelial cells in pigs with lung injury QIN Xin， QIN Lei， LIU You-ning. Department of Respiratory Medicine， Second Hospital of Shanxi Medical University， Taiyuan 030001， China
【Abstract】 Objective To understand the effect of vaporization inhalation of perfluorocarbon (PFC) for model of pig lung injury caused by lung lavage on apoptosis of renal tubular epithelial cells in lung injury. Methods A total of 20 healthy pigs were randomly divided into VMV group (vaporization inhalation of PFC，10 pigs)， CMV group (conventional mechanical ventilation， 8 pigs) and sham operation group (2 pigs). Kidney tissue at the junction of renal cortex and medulla was taken from each group. Ultrastructure of apoptotic cells was observed by electron microscopy， apoptotic index was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling(TUNEL)， expression of apoptotic genes B cell lymphoma/leukemia-2 (bcl-2) and bcl-2 related X protein (BAX) was determined by immunohistochemistry， and expression of BAX and Bcl-2 ribonucleic acid (mRNA) was determined by Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Results TUNEL results： The positive cells of the kidney were mainly distributed in the proximal tubules， and no apoptotic cells were found in the glomeruli. VMV group had obviously lower apoptosis rate as (20.2±2.9)% than (80.2±4.8)% in CMV group， and the difference was statistically significant (t=32.868， P=0.000<0.05). Renal BAX， bcl-2 protein expression results： VMV group had obviously lower BAX protein expression as (12.6±6.8) than (62.2±3.6) in CMV group， and obviously higher bcl-2 protein expression as (36.8±2.1) than (6.2±2.3) in CMV group. Their difference was statistically significant (P<0.05). Renal BAX and bcl-2 mRNA expression results：VMV group had obviously lower BAX mRNA expression as (7.10±0.32) than (9.29±1.06) in CMV group， and obviously higher bcl-2 mRNA expression as (10.29±2.51) than (5.18±1.08) in CMV group. Their difference was statistically significant (t=6.230， 5.350， P<0.05). The apoptotic cells in sham operation group were very few and the expressions of BAX and Bcl-2 were very low. Conclusion Vaporized inhalation of PFC can reduce the apoptosis of renal tubular epithelial cells in piglets with acute lung injury. Its mechanism may be related to inhibiting the expression of BAX gene and increasing the expression of Bcl-2 gene., http://www.100md.com(秦鑫 秦蕾 刘又宁)