目的 研究伏立诺他(SAHA)对严重烫伤大鼠心脏的保护作用及机制。方法 雄性SD大鼠48只,随机分为假烫组、烫伤组和SAHA组,每组16只,烫伤组和SAHA组采用沸水水浴法制作50％TBSA Ⅲ度烫伤动物模型,伤后即刻腹腔内分别注射0.25 ml生理盐水或SAHA(7.5 mg/kg,溶于0.25 ml生理盐水),假烫组以37 ℃水浴代替沸水。每组根据烫伤后3、6 h两个观察时间点,又分为两个亚组,每亚组8只大鼠,分别于烫伤后3 h和6 h两个时间点取大鼠腹主动脉血,测定血浆肌酸激酶同工酶(CK-MB)水平,然后处死动物取心肌组织,干/湿重法测定心肌组织含水率,TUNEL方法检测心肌凋亡细胞率,caspase-3活性试剂盒测定心肌组织内caspase-3活性。结果 烫伤后3、6 h,烫伤组血浆内CK-MB[(2552.9±145.3)U/L,(5418.7±292.9)U/L]水平较假烫组[(585.6±46.1)U/L]明显升高,心肌组织含水率[(77.65±1.75)%,(80.23±1.82)%]较假烫组[(70.12±1.99)%]明显升高,心肌细胞凋亡率[(16.1±0.8)%,(21.5±1.2)%]较假烫组[(0.5±0.1)%]明显升高、caspase-3活性[(0.716±0.052)μmol/mg,(0.912±0.063)μmol/mg]较假烫组[(0.435±0.041)μmol/mg]明显升高,差异均具有统计学意义(均P＜0.05);而给予SAHA处理后,CK-MB水平[(1642.4±157.2)U/L,(3475.3±187.5)U/L]、心肌组织含水率[(72.31±1.95)%,(75.21±2.00)%]、心肌细胞凋亡率[(11.6±0.6)%,(15.1±0.9)%]、caspase-3活性[(0.557±0.052)μmol/mg,(0.535±0.034)μmol/mg]较烫伤组明显下降,差异均有统计学意义(均P＜0.05)。结论 在严重烫伤大鼠治疗中,SAHA能显著降低血浆内CK-MB水平及心肌组织含水率,保护心脏功能,可能与减少心肌细胞凋亡、降低心肌组织内caspase-3活性有关。
Abstract
Objective To study the protective effect of suberoylanilide hydroxamic acid(SAHA) on the heart of rats with severe scald injury.Methods Forty-eight male SD rats were randomly divided into the sham scald group, the scald group and the SAHA group (n=16). Rats in the scald group and SAHA group were subjected to 50％TBSA third-degree full-thickness scald injury with boiled water before they received an intraperitoneal injection of 0.25 ml of 0.9% normal saline(NS)or SAHA (7.5 mg/kg, dissolved in 0.25 ml of NS) . 37 ℃ water instead of boiled water was used for rats in the sham scald group. According to time points for observation, each scald group was divided into two subgroups: the 3-hour group and 6-hour group after scald(n=8). At 3h or 6h after burns, blood in each group was withdrawn from the abdominal aorta to detect the level of CK-MB before the rats were sacrificed. Myocardial tissues were harvested, the water content of which was detected by dry/wet weight method. The rate of myocardial apoptosis in each group was calculated using the TUNEL method while the activity of caspase-3 was measured by detection kits.Results At 3 h and 6 h after scald, the level of CK-MB in the scald group[(2552.9±145.3)U/L,(5418.7±292.9)U/L] was more significantly increased than in the sham scald group[(585.6±46.1)U/L], so were the water content of myocardial tissues [(77.65±1.75)%,(80.23±1.82)%]vs.[(70.12±1.99)%], rate of myocardial apoptosis [(16.1±0.8)%,(21.5±1.2)%] vs. [(0.5±0.1)%], and the activity of caspase-3 [(0.716±0.052)μmol/mg;(0.912±0.063)μmol/mg] vs. [(0.435±0.041)μmol/mg], and the difference was statistically significant (all P<0.05). After SAHA treatment, the level of CK-MB [(1642.4±157.2) U/L, (3475.3±187.5) U/L], water content of myocardial tissues [(72.31±1.95)%, (75.21±2.00)%],rate of myocardial apoptosis[(11.6±0.6)%,(15.1±0.9)%],and activity of caspase-3[(0.557±0.052)μmol/mg,(0.535±0.034)μmol/mg]were significantly lower than in the scald group.Conclusions In rats with fatal scald, SAHA can reduce the level of CK-MB in plasma and the water content of myocardial tissues while protecting cardiac function,which may be related to the decrease of the apoptosis of myocardial cells and the activity of caspase-3 in myocardium.
关键词
烫伤 /
伏立诺他 /
组蛋白去乙酰化酶抑制剂 /
凋亡
Key words
scald /
suberoylanilide hydroxamic acid /
histone deacetylase inhibitors /
apoptosis
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基金
国家自然科学基金(81471872);承德市科学技术研究与发展计划项目(201904A106)
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