摘要
目的探讨血必净注射液通过介导丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)信号通路减轻大鼠脓毒症诱导的心肌损伤(sepsis induced myocardial injury,SIMI)的作用机制。方法采用盲肠结扎穿孔术(cecal-ligation and puncture,CLP)构建脓毒症大鼠模型,并将SPF级成年雄性SD大鼠按照随机数字表法分为:假手术组、模型组和血必净组,每组6只。假手术组仅开腹分离盲肠,不结扎,不穿刺;模型组及血必净组开腹分离盲肠,结扎并穿刺;术后给予假手术组和模型组0.9%氯化钠溶液治疗,血必净组给予血必净注射液治疗,均给药2d。观察大鼠在模型制备成功后的一般状况;苏木精-伊红染色检测心肌组织结构改变情况;采用超声心动图检测心功能改变情况;生物化学检测大鼠心肌酶改变情况;采用酶联免疫吸附(enzyme-linked immunosorbent assay,ELISA)检测血清中白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)水平;采用蛋白质印迹法测定心肌组织中p-ERK 1/2、ERK 1/2、p-P38、P38、p-JNK和JNK蛋白水平。结果超声心动图、心肌酶、ELISA检测结果显示,与模型组相比,血必净组左心室短轴缩短率和左心室射血分数、SOD水平升高[(50.82±8.27)%比(30.53±9.22)%,(81.52±7.56)%比(55.92±10.91)%,(549.41±25.97)ng/ml比(391.77±68.97)ng/ml,P均<0.05];血必净组心肌肌钙蛋白I、乳酸脱氢酶、肌酸激酶、IL-6、TNF-α、MDA、p-ERK 1/2、p-P38、p-JNK水平均低于模型组[(0.32±0.05)ng/ml比(0.44±0.08)ng/ml,(673.58±62.96)U/L比(905.58±134.74)U/L,(784.49±345.64)U/L比(1200.25±220.44)U/L,(122.72±16.52)pg/ml比(162.08±20.25)pg/ml,(153.39±18.95)pg/ml比(203.1±28.84)pg/ml,(286.39±42.45)nmol/L比(443.53±69.38)nmol/L,(1.57±0.09)%比(1.94±0.06)%,(0.39±0.04)%比(1.18±0.09)%,(0.4±0.05)%比(1.19±0.04)%,P均<0.05]。结论血必净注射液可明显抑制SIMI炎症反应和氧化应激,促进细胞增殖,改善心功能,可能与抑制MAPK信号通路有关。
Objective To explore the mechanism of Xuebijing injection in reducing sepsis induced myocardial injury(SIMI)in rats by mediating mitogen-activated protein kinase(MAPK)signaling pathway.Method Sepsis rat model was constructed by cecal-ligation and puncture(CLP),and adult male SD rats with specific pathogen free(SPF)grade were divided into sham group,CLP group and Xuebijing group according to random number table method,with 6 rats in each group.In the sham operation group,the cecum was separated by laparotomy without ligation or puncture.The ceca of model group and Xuebijing group were separated by laparotomy,ligation and puncture.After operation,sham operation group and model group were treated with 0.9%sodium chloride solution,and Xuebijing group was treated with Xuebijing injection for 2 days.The general condition of the rats after the successful preparation of the model was observed.The changes of myocardial tissue structure were detected by hematoxylin-eosin staining.The changes of cardiac function were detected by echocardiography.The changes of myocardial enzymes were detected by biochemistry.Enzyme-linked immunosorbent assay(ELISA)was used to detect serum interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),superoxide dismutase(SOD),and malondialdehyde(MDA)levels.The protein levels of p-ERK 1/2,ERK 1/2,p-P38,P38,p-JNK and JNK in myocardial tissue were determined by the western blot.Result Compared with the model group,the left ventricular short axis shortening rate,left ventricular ejection fraction and SOD level in Xuebijing group were increased[(50.82±8.27)%v.s.(30.53±9.22)%,(81.52±7.56)%v.s.(55.92±10.91)%,(549.41±25.97)ng/ml v.s.(391.77±68.97)ng/ml,all P<0.05].The levels of cardiac troponin I,lactate dehydrogenase,creatine kinase,IL-6,TNF-α,MDA,p-ERK 1/2,p-P38 and p-JNK in Xuebijing group were lower than those in model group[(0.32±0.05)ng/ml v.s.(0.44±0.08)ng/ml,(673.58±62.96)U/L v.s.(905.58±134.74)U/L,(784.49±345.64)U/L v.s.(1200.25±220.44)U/L,(122.72±16.52)pg/ml v.s.(162.08±20.25)pg/ml,(153.39±18.95)pg/ml v.s.(203.1±28.84)pg/ml,(286.39±42.45)nmol/L v.s.(443.53±69.38)nmol/L,(1.57±0.09)%v.s.(1.94±0.06)%,(0.39±0.04)%v.s.(1.18±0.09)%,(0.4±0.05)%v.s.(1.19±0.04)%,all P<0.05].Conclusion Xuebijing injection can significantly inhibit SIMI inflammation and oxidative stress,promote cell proliferation and improve cardiac function,which may be related to the inhibition of MAPK signaling pathway.
作者
田娜
赵继军
张俊飞
王圣
马磊
杨立山
Tian Na;Zhao Jijun;Zhang Junfei;Wang Sheng;Ma Lei;Yang Lishan(Clinical Medical College of Ningxia Medical University,Ninxia Yinchuan 750004,China;Department of Emergency,General Hospital of Ningxia Medical University,Ninxia Yinchuan 750004,China)
出处
《创伤与急诊电子杂志》
2024年第3期153-162,共10页
Journal of Trauma and Emergency(Electronic Version)
基金
国家自然科学基金(82205116)
宁夏自然科学基金项目(2023AAC03618)。
关键词
脓毒症
心肌损伤
血必净注射液
氧化应激
炎症
丝裂原活化蛋白激酶
Sepsis
Myocardial injury
Xuebijing injection
Oxidative stress
Inflammation
Mitogen-activated protein kinase
作者简介
通信作者:杨立山,Email:nyfyyls@126.com。
