柴胡加龙骨牡蛎汤对心肌梗死合并焦虑模型小鼠核转录因子-κB介导的炎症反应的影响

Effect of Chaihu Jia Longgu Muli Decoction on Nuclear Transcription Factor κB Mediated Inflammatory Response in Mice with Myocardial Infarction and Anxiety

目的:观察柴胡加龙骨牡蛎汤对心肌梗死合并焦虑模型小鼠心功能及核转录因子-κB(nuclear transcription factor-κB,NF-κB)介导的炎症反应的影响,探究其对梗死心肌的保护作用及机制。方法:采用冠状动脉前降支结扎结合不确定空瓶刺激法建立C57BL/6J小鼠心肌梗死合并焦虑模型。将造模成功的小鼠随机分为模型组、中药组(柴胡加龙骨牡蛎汤颗粒剂),假手术组只穿线不结扎,每组8只,每天灌胃给药1次,连续给药7 d。通过超声心动图评价小鼠心功能,旷野实验评价小鼠活动能力,高架十字迷宫评价小鼠焦虑状态,HE染色观察心脏病理改变,马松(Masson)染色观察心肌纤维化程度,酶联免疫吸附法(ELISA)检测外周血白细胞介素(interleukin,IL)-1β、肿瘤坏死因子(tumor necrosis factor,TNF)-α、IL-6的含量,采用RT-PCR定量检测心肌组织中IL-1β、TNF-α、NF-κB p65 mRNA表达水平,蛋白印迹法(WB)检测心肌组织NF-κB p65的蛋白表达水平。结果:与模型组比较,中药组能够提高小鼠左心室射血分数,改善心功能(P<0.01);与假手术组比较,模型组小鼠在旷野实验中的运动能力减弱且倾向于停留在高架十字迷宫闭合臂(P<0.01);与模型组比较,中药组小鼠在旷野实验中的运动能力提高并倾向于停留在高架十字迷宫开放臂(P<0.01)。病理染色结果显示:与模型组比较,中药组小鼠的心肌坏死和纤维化程度减轻;与模型组比较,中药组小鼠血清IL-1β、TNF-α、IL-6含量均降低(P<0.01),心肌组织中IL-1β、TNF-αmRNA表达量也呈降低趋势(P<0.01);中药组小鼠心肌组织中NF-κB p65的mRNA和蛋白表达降低(P<0.01)。结论:柴胡加龙骨牡蛎汤能够改善心肌梗死合并焦虑模型小鼠的心肌损伤,增强左心室功能,减轻炎症反应,其机制可能与NF-κB信号介导的炎症反应相关。

Objective: To observe the effect of Chaihu Jia Longgu Muli Decoction on the cardiac function and the inflammatory response mediated by nuclear transcription factor-κB( NF-κB) in the model mice of myocardial infarction with anxiety and to explore the protective effect and mechanism of Chaihu Jia Longgu Muli Decoction on the infarcted myocardium. Methods: The model of myocardial infarction with anxiety in C57 BL/6 J mice was established by ligation of anterior descending branch of coronary artery combined with uncertain empty bottle stimulation. The mice were randomly divided into two groups: model group and traditional Chinese medicine group( Chaihu Jia Longgu Muli Decoction granules). The sham operation group only wore thread without ligation,8 mice in each group were administrated by gavage once a day for 7 days. The cardiac function of mice was evaluated by echocardiography,the activity of mice was evaluated by field experiment,the anxiety state of mice was evaluated by elevated cross maze,the pathological changes of heart were observed by HE staining,the degree of myocardial fibrosis was observed by Masson staining,the levels of interleukin( IL)-1β and tumor necrosis factor( TNF)-α,IL-6 in peripheral blood were detected by enzymelinked immunosorbent assay( ELISA). The expression of IL-1β,TNF-α,NF-κB p65 mRNA in myocardium was detected by RT PCR,and the expression of NF-κB p65 protein in myocardium was detected by Western blotting( WB). Results: Compared with the model group,the Chinese medicine group can improve the left ventricular ejection fraction and improve the heart function( P <0. 01);compared with the sham operation group,the model group’s motor ability in the field experiment is weakened and tends to stay in the closed arm of the elevated cross maze( P < 0. 01);compared with the model group,the motor ability of mice in the traditional Chinese medicine group was improved and tended to stay in the open arm of the elevated cross maze( P < 0. 01). The results of pathological staining showed that: compared with the model group,the degree of myocardial necrosis and fibrosis of the mice in the traditional Chinese medicine group decreased;compared with the model group,the contents of IL-1β,TNF-α and IL-6 in the serum of the mice in the traditional Chinese medicine group decreased( P < 0. 01),and the expression of IL-1β,TNF-αmRNA in the myocardial tissue of the mice in the traditional Chinese medicine group also decreased( P < 0. 01);the expression of NF-κB p65 and protein in the myocardium of mice in the traditional Chinese medicine group decreased( P < 0. 01). Conclusion:Chaihu Jia Longgu Muli Decoction can improve myocardial injury,enhance left ventricular function and reduce inflammatory response in mice with myocardial infarction and anxiety. Its mechanism may be related to NF-κB signal mediated inflammatory response.