H11蛋白基因对p38MAPK磷酸化的影响及其对烧伤后心肌损伤的保护作用

Effect of H11 protein genes on p38MAPK phosphorylation and its protective effects in myocardial injury after burn

目的探讨H11蛋白基因对p38丝裂原活化蛋白激酶（p38MAPK）磷酸化的影响及其对烧伤后心肌损伤的保护作用。方法选取36只雄性Wistar大鼠,建立大鼠重度烧伤模型,随机抽取24只纳入烧伤组,对烧伤组进行1～18 h的缺氧刺激,缺氧6～18 h的纳入D1组（12例）,缺氧1～6 h的纳入D2组（12例）,正常大鼠纳入D3组（12例）。烧伤处理后提取心肌细胞RNA,采用逆转录聚合酶链反应（RT-PCR）检测法分析烧伤后1 h、3 h、6 h、12 h、18 h各时间点H11蛋白基因mRNA表达情况,同时采用酶联免疫吸附实验（ELISA）检测缺氧刺激后5h D1、D2组血清p38MAPK磷酸化水平、核因子-κB（NF-κB）、肿瘤坏死因子-α（TNF-α）水平。结果 D1、D2组H11蛋白基因mRNA水平随时间延长较D3组显著增加（P〈0.05）,烧伤后6～12 h时H11蛋白基因mRNA水平达峰值;D1组刺激后p38MAPK磷酸化水平（0.21±0.12）μg/L、NF-κB（0.14±0.12）μg/L、TNF-α（0.28±0.04）μg/L水平显著低于D2组（P〈0.05）。结论 H11蛋白基因可抑制p38MAPK磷酸化,且能减轻烧伤后心肌损伤,具有保护心肌作用。

Objective： To investigate the effect of H11 protein genes on p38 mitogen activated protein kinase（ MAPK）phosphorylation and its protective effects in myocardial injury after burn. Methods： 36 male Wistar rats were selected to establish rat models of severe burn. 24 rats were randomly selected and included in burn group. 1 ～ 18 h hypoxic stimulation was performed in the burn group. Rats which were in hypoxic state for 6 ～ 18 h were included into group D1（ 12 cases）and which were in hypoxic state for 1 ～ 6 h were included into group D2（ 12 cases）. Normal rats were included into group D3（ 12 cases）. After burn management,myocardial cell RNA was extracted. The reverse transcription-polymerase chain reaction（ RT-PCR） detection method was used to analyze the expression of H11 protein gene mRNA at 1 h,3 h,6 h,12 h and 18 h after burn. Besides,the levels of p38 MAPK phosphorylation,nuclear factor kappa B（ NF-κB） and tumor necrosis factor alpha（ TNF-α） in group D1 and D2 were detected by enzyme-linked immunosorbent assay（ ELISA） at 5h after hypoxic stimulation. Results： The H11 protein gene mRNA levels in group D1 and D2 were significantly higher than those in group D3 with time expansion（ P〈0. 05）. 6 ～ 12 h after burn,H11 protein gene mRNA level reached the peak; After hypoxic stimulation,p38 MAPK phosphorylation level（ 0. 21 ± 0. 12） μg / L and levels of NF-κB（ 0. 14 ± 0. 12） μg / L and TNF-α（ 0. 28 ± 0. 04） μg / L in group D1 were significantly lower than those in group D2（ P〈0. 05）. Conclusion： H11 protein gene can inhibit p38 MAPK phosphorylation and alleviate myocardial injury after burn. It also can protect the myocardium.