赵 欢, 傅 豪, 解营利, 杨 涛, 张 明, 张海燕, 徐 晖. NLRP3炎症小体在大鼠严重烫伤早期心肌内的表达及意义[J]. 心脏杂志, 2016, 28(5): 508-511.
    引用本文: 赵 欢, 傅 豪, 解营利, 杨 涛, 张 明, 张海燕, 徐 晖. NLRP3炎症小体在大鼠严重烫伤早期心肌内的表达及意义[J]. 心脏杂志, 2016, 28(5): 508-511.
    Severe burn induces NLRP3 inflammasome activation in rat myocardium[J]. Chinese Heart Journal, 2016, 28(5): 508-511.
    Citation: Severe burn induces NLRP3 inflammasome activation in rat myocardium[J]. Chinese Heart Journal, 2016, 28(5): 508-511.

    NLRP3炎症小体在大鼠严重烫伤早期心肌内的表达及意义

    Severe burn induces NLRP3 inflammasome activation in rat myocardium

    • 摘要: 目的 探讨严重烫伤大鼠早期心肌组织内NLRP3炎症小体信号通路的表达及意义。方法 采用大鼠95℃热水浴18 s,制成40%总体表面积Ⅲ度烫伤模型,将24只健康雄性SD大鼠按随机数表法分为对照组、烫伤组、干预组。烫伤组大鼠立即腹腔注射10 ml生理盐水补液;干预组大鼠注射10 ml生理盐水+(BAY11-7082) 10 μl(0.2 mg/ml,10 mg/kg);对照组动物背部置于25℃温水浴18 s模拟烫伤过程。伤后8 h分别取各组大鼠心肌组织及血清,采用蛋白免疫印迹法和实时荧光定量RT-PCR法检测心肌组织中NLRP3、半胱氨酸天冬氨酸蛋白酶(caspase)-1蛋白表达水平和NLRP3、caspase-1、IL-1β、TNF-α mRNA表达水平,同时观察HE染色组织大体形态,酶联免疫吸附试验法检测血清肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)含量。结果 ①烫伤组大鼠心肌组织内NLRP3、caspase-1蛋白表达量显著高于对照组(P<0.01);与烫伤组比较,干预组大鼠心肌组织内NLRP3、caspase-1明显降低(P<0.01);②烫伤组大鼠心肌组织中NLRP3、caspase-1、IL-1β和TNF-α的mRNA表达量分别为3.15±0.46,2.47±0.24,3.26±0.22和2.17±0.32,显著高于对照组的1.00±0.09,1.00±0.06,1.00±0.08和1.00±0.07(均P<0.01);干预组大鼠心肌组织中NLRP3、caspase-1、IL-1β和TNF-α的mRNA表达量分别为1.68±0.37,1.55±0.17,1.21±0.15和1.38±0.23,显著低于烫伤组(均P<0.01);③对照组大鼠心肌组织呈不规则圆柱形、排列整齐、横纹清晰,结构完整。烫伤组大鼠部分心肌纤维排列紊乱,细胞和间质可见水肿,见点片状溶解灶及炎性细胞浸润。BAY11-7082干预组部分心肌纤维排列紊乱,心肌细胞和间质水肿、心肌纤维溶解及炎细胞浸润程度均较烫伤组减轻;④烫伤组大鼠血清CK-MB、LDH含量分别为(2753±825)U/L,(2618±238)U/L,显著高于对照组的(247±76)U/L,(721±59)U/L(均P<0.01);干预组大鼠血清CK-MB、LDH含量分别为(1267±248)U/L,(1982±183)U/L,显著低于单纯烫伤组(均P<0.01)。结论 NLRP3炎症小体在严重烫伤大鼠早期心肌组织内活性增高,使用BAY11-7082干预后,可抑制NLRP3炎症小体的活化,减轻心肌组织损伤,降低心肌炎症反应。

       

      Abstract: AIM To study the expression and effect of NLRP3 inflammasome in severely burned rat myocardium. METHODSTwenty-four rats were randomly divided into three groups: sham group, burn group and BAY11-7082 treatment group. Animals were subjected to full-thickness burn injury of a 40% total body surface area via exposure to water maintained at 95°C for 18 sec. Rats in burn group received an intraperitoneal injection of 10 ml normal saline for resuscitation and rats in treatment group received 10 ml normal saline and 10 μl BAY11-7082 (0.2 mg/ml, 10 mg/kg). Rats in the sham group were immersed in room temperature water for 18 sec. Rats were sacrificed 8 h after burn and myocardial tissues and serums were sampled. Protein expressions of NLRP3 and caspase-1 were examined by Western blot and mRNA levels of NLRP3, caspase-1, IL-1β and TNF-α were tested by RT-PCR. Histomorphological images were also observed by H&E staining and levels of CK-MB and LDH were detected by ELISA. RESULTSProtein expressions of myocardial NLRP3 and caspase-1 in the burn group were significantly higher than in the sham group (P<0.01). When treated with BAY11-7082, expressions of NLRP3 and caspase-1 were significantly decreased (P<0.01). mRNA values of myocardial NLRP3, caspase-1, IL-1β and TNF-α in the burn group were 3.15±0.46, 2.47±0.24, 3.26±0.22 and 2.17±0.32, respectively, much higher than 1.00±0.09, 1.00±0.06, 1.00±0.08 and 1.00±0.07 in the sham group (P<0.01). When treated with BAY11-7082, mRNA values changed to 1.68±0.37, 1.55±0.17, 1.21±0.15 and 1.38±0.23, respectively, significantly lower compared with those in the burn group (P<0.01). In the sham group, myocardial fibers were in a well-arranged irregular stripe shape and the transverse striation was distinct. In the burn group, part of the myocardial fibers were arranged in a disordered manner with edema of the myocardial interstitium. Part of the myocardial fibers dissolved and inflammatory cells were infiltrated. In the treatment group, myocardial fibers were disordered and myocardial interstitial edema and inflammatory cell infiltration were alleviated. The values of CK-MB and LDH in serum of in burned rats were (2753±825) U/L and (2618±238) U/L, respectively, much higher than (247±76) U/L and (721±59) U/L in the sham rats (P<0.01). When treated with BAY11-7082, the values changed to (1267±248) U/L and 1982±183 U/L, respectively, significantly lower compared to those in the burn group (P<0.01). CONCLUSIONNLRP3 inflammasome is activated in severely burned rat myocardium. When treated with BAY11-7082, the activity of NLRP3 is inhibited, with myocardial injury alleviated and myocardial inflammatory response relieved.

       

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