Zheng Jun, Huang Yue-sheng, Huang Xiao-yuan, Fan Peng-ju, He Wei-feng, Zhang Xiao-rong
Department of Burns and Plastic Surgery, the First Affiliated Hospital of the University of South China, Hengyang 421001, China.
Zhonghua Shao Shang Za Zhi. 2013 Jun;29(3):267-71.
To study the effects of antisense p38α mitogen-activated protein kinase (hereinafter referred to as p38α) on myocardial cells exposed to hypoxia and burn serum.
Thirty adult SD rats were inflicted with 40% TBSA full-thickness burn on the back to obtain burn serum. The myocardial cells were isolated from 80 neonatal SD rats and cultured, then they were divided into 4 groups according to the random number table: normal control group (N, ordinary culture without any treatment), hypoxia+burn serum group (HB, exposed to hypoxia after being treated with 10% burn rat serum), hypoxia+burn serum+infection group (HBI, exposed to hypoxia and 10% burn rat serum after being infected with antisense p38α gene-carrying adenovirus), hypoxia+burn serum+empty vector infection group (exposed to hypoxia and 10% burn rat serum after being infected with adenovirus empty vector). At post hypoxia hour (PHH) 1, 3, 6, and 12, mRNA and protein expression levels of p38α in the latter 3 groups were determined by RT-PCR and Western blotting, cell viability was determined by methylthianolyldiphenyl-tetrazolium bromide assay, and lactate dehydrogenase (LDH) activity was assayed at the same time point. At PHH 1, 6, and 12, apoptosis rate of myocardial cells was assessed by annexin V staining method. The indexes of group N were determined with the methods mentioned-above. Three wells were set at each time point in each group. Data were processed with one-way analysis of variance and LSD- t test.
(1) At PHH 1, 3, and 6, the p38α mRNA level was higher in group HB than in group N and group HBI (with t values from 2.725 to 4.375, P values all below 0.05). (2) At PHH 1, 3, and 6, the p38α protein level was higher in group HB than those in group N and group HBI (with t values from 5.351 to 7.981, P values all below 0.01). (3) At PHH 3, 6, and 12, the cell viability in group HB (0.115 ± 0.007, 0.104 ± 0.006, 0.094 ± 0.005) was lower than that in group N (0.141 ± 0.014) and group HBI (0.136 ± 0.009, 0.124 ± 0.010, 0.112 ± 0.007, with t values from 2.357 to 6.812, P values all below 0.05). (4) The LDH activity was up-regulated in group HB as compared with that in group N and group HBI at each time point (with t values from 22.753 to 201.273, P values all below 0.01). (5) At PHH 1, 6, and 12, the apoptosis rate of myocardial cells in group HB [(5.4 ± 0.7)%, (8.7 ± 1.1)%, (13.6 ± 1.7)%] was higher than that of group N [(3.1 ± 0.3)%] and group HBI [(4.3 ± 0.5)%, (5.1 ± 0.7)%, (7.2 ± 0.9)%, with t values from 2.345 to 9.700, P < 0.05 or P < 0.01].
Antisense p38α can protect the myocardial cells from the injury of hypoxia and burn serum.
研究反义p38α丝裂原活化蛋白激酶（以下简称p38α）对缺氧及烧伤血清作用下心肌细胞的影响。
30只成年SD大鼠背部造成40%TBSA全层烧伤以获取烧伤血清。从80只新生SD大鼠分离培养心肌细胞，然后按随机数字表法分为4组：正常对照组（N组，常规培养未作任何处理）、缺氧+烧伤血清组（HB组，用10%烧伤大鼠血清处理后再缺氧）、缺氧+烧伤血清+感染组（HBI组，感染携带反义p38α基因的腺病毒后再缺氧及给予10%烧伤大鼠血清）、缺氧+烧伤血清+空载体感染组（感染腺病毒空载体后再缺氧及给予10%烧伤大鼠血清）。在缺氧后1、3、6及12小时（PHH），采用RT-PCR和Western印迹法检测后3组p38α的mRNA和蛋白表达水平，用噻唑蓝比色法检测细胞活力，并在同一时间点检测乳酸脱氢酶（LDH）活性。在PHH 1、6及12小时，采用膜联蛋白V染色法评估心肌细胞凋亡率。N组指标采用上述方法测定。每组每个时间点设3个孔。数据采用单因素方差分析和LSD-t检验处理。
（1）在PHH 1、3及6小时，HB组p38α mRNA水平高于N组和HBI组（t值为2.725至4.375，P值均<0.05）。（2）在PHH 1、3及6小时，HB组p38α蛋白水平高于N组和HBI组（t值为5.351至7.981，P值均<0.01）。（3）在PHH 3、6及12小时，HB组细胞活力（0.115±0.007、0.104±0.006、0.094±0.005）低于N组（0.141±0.014）和HBI组（0.136±0.009、0.124±0.010、0.112±0.007，t值为2.357至6.812，P值均<0.05）。（4）各时间点HB组LDH活性均高于N组和HBI组（t值为22.753至201.273，P值均<0.01）。（5）在PHH 1、6及12小时，HB组心肌细胞凋亡率[（5.4±0.7）%、（8.7±1.1）%、（13.6±1.7）%]高于N组[（3.1±0.3）%]和HBI组[（4.3±0.5）%、（5.1±0.7）%、（7.2±0.9）%，t值为2.345至9.700，P<0.05或P<0.01]。
反义p38α可保护心肌细胞免受缺氧及烧伤血清的损伤。
