Abstract：
To explore the effect of carbachol on myocardial injury in septic rats, and to further study its influence on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway.
A total of 48 healthy male Sprague-Dawley rats were randomly divided into sham group (n=16), model group (n=16), and carbachol group (n=16). The rat model of sepsis was established via cecal ligation and puncture. Carbachol was intraperitoneally injected (10 μg/kg) immediately after operation in carbachol group, and no cecal ligation was performed in sham group. At 48 h after operation, the survival rate of rats in each group was recorded, the activity of plasma creatine kinase-MB (CK-MB) was detected, and the cardiac function in each group was determined. Moreover, the heart was isolated, and the myocardial tissues were taken to detect the apoptosis level using the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) apoptosis kit. The content of inflammatory factors in myocardial tissues was determined using enzyme-linked immunosorbent assay (ELISA) kits, and the expression levels of apoptosis-related proteins and the PI3K/AKT signaling pathway-related proteins were detected via Western blotting.
Carbachol could significantly raise the survival rate of septic rats (p<0.01), remarkably decrease the activity of CK-MB (p<0.01), markedly reduce the left ventricular internal diameter at end-systole (LVIDs), and markedly increase the left ventricular ejection fraction (LVEF, %) and left ventricular fractional shortening (LVFS, %). Besides, carbachol could evidently lower the apoptosis level of myocardial cells of septic rats (p<0.01), reduce the content of inflammatory factors including tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and IL-6 (p<0.01), notably decrease the expression of Caspase-3 in myocardial tissues (p<0.01), remarkably increase the expression of Bcl-2/Bax (p<0.01), and distinctly inhibit the expressions of phosphorylated (p-)PI3K, p-AKT, Nod-like receptor protein 3 (NLRP3), and Caspase-1 (p<0.01).
Carbachol can reduce the release of inflammatory factors in myocardial cells, the expression of apoptotic proteins and the apoptosis of myocardial cells, and improve the cardiac function and survival rate of septic rats by inhibiting the PI3K/AKT signaling pathway.
A total of 48 healthy male Sprague-Dawley rats were randomly divided into sham group (n=16), model group (n=16), and carbachol group (n=16). The rat model of sepsis was established via cecal ligation and puncture. Carbachol was intraperitoneally injected (10 μg/kg) immediately after operation in carbachol group, and no cecal ligation was performed in sham group. At 48 h after operation, the survival rate of rats in each group was recorded, the activity of plasma creatine kinase-MB (CK-MB) was detected, and the cardiac function in each group was determined. Moreover, the heart was isolated, and the myocardial tissues were taken to detect the apoptosis level using the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) apoptosis kit. The content of inflammatory factors in myocardial tissues was determined using enzyme-linked immunosorbent assay (ELISA) kits, and the expression levels of apoptosis-related proteins and the PI3K/AKT signaling pathway-related proteins were detected via Western blotting.
Carbachol could significantly raise the survival rate of septic rats (p<0.01), remarkably decrease the activity of CK-MB (p<0.01), markedly reduce the left ventricular internal diameter at end-systole (LVIDs), and markedly increase the left ventricular ejection fraction (LVEF, %) and left ventricular fractional shortening (LVFS, %). Besides, carbachol could evidently lower the apoptosis level of myocardial cells of septic rats (p<0.01), reduce the content of inflammatory factors including tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and IL-6 (p<0.01), notably decrease the expression of Caspase-3 in myocardial tissues (p<0.01), remarkably increase the expression of Bcl-2/Bax (p<0.01), and distinctly inhibit the expressions of phosphorylated (p-)PI3K, p-AKT, Nod-like receptor protein 3 (NLRP3), and Caspase-1 (p<0.01).
Carbachol can reduce the release of inflammatory factors in myocardial cells, the expression of apoptotic proteins and the apoptosis of myocardial cells, and improve the cardiac function and survival rate of septic rats by inhibiting the PI3K/AKT signaling pathway.
摘要：
探讨卡巴胆碱对脓毒症大鼠心肌损伤的影响,并进一步研究其对磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)信号通路的影响。
将48只健康雄性SD大鼠随机分为假手术组(n=16),模型组（n=16），和卡巴胆碱组(n=16)。采用盲肠结扎穿刺术建立脓毒症大鼠模型。卡巴胆碱组术后即刻腹腔注射卡巴胆碱(10μg/kg),假手术组不进行盲肠结扎。术后48h,记录各组大鼠的存活率,检测血浆肌酸激酶同工酶（CK-MB）的活性，并测定各组的心功能。此外，心脏是孤立的,并使用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）凋亡试剂盒检测心肌组织的凋亡水平。采用酶联免疫吸附试验(ELISA)试剂盒测定心肌组织中炎症因子的含量,免疫印迹法检测细胞凋亡相关蛋白和PI3K/AKT信号通路相关蛋白的表达水平。
卡巴胆碱能显著提高脓毒症大鼠的存活率(p<0.01),显著降低CK-MB的活性（p<0.01），收缩末期左心室内径（LVIDs）明显减小，并显著增加左心室射血分数(LVEF，%)和左心室缩短分数(LVFS，%).此外,卡巴胆碱能明显降低脓毒症大鼠心肌细胞凋亡水平(p<0.01),降低炎症因子包括肿瘤坏死因子-α（TNF-α）的含量，白细胞介素-1β（IL-1β）和IL-6（p<0.01），显著降低心肌组织中Caspase-3的表达(p<0.01),Bcl-2/Bax的表达显着增加（p<0.01），并明显抑制磷酸化（p-）PI3K的表达，p-AKT,Nod样受体蛋白3（NLRP3），和Caspase-1(p<0.01)。
卡巴胆碱可以减少心肌细胞炎症因子的释放，凋亡蛋白的表达和心肌细胞的凋亡，通过抑制PI3K/AKT信号通路改善脓毒症大鼠心功能和存活率。
将48只健康雄性SD大鼠随机分为假手术组(n=16),模型组（n=16），和卡巴胆碱组(n=16)。采用盲肠结扎穿刺术建立脓毒症大鼠模型。卡巴胆碱组术后即刻腹腔注射卡巴胆碱(10μg/kg),假手术组不进行盲肠结扎。术后48h,记录各组大鼠的存活率,检测血浆肌酸激酶同工酶（CK-MB）的活性，并测定各组的心功能。此外，心脏是孤立的,并使用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）凋亡试剂盒检测心肌组织的凋亡水平。采用酶联免疫吸附试验(ELISA)试剂盒测定心肌组织中炎症因子的含量,免疫印迹法检测细胞凋亡相关蛋白和PI3K/AKT信号通路相关蛋白的表达水平。
卡巴胆碱能显著提高脓毒症大鼠的存活率(p<0.01),显著降低CK-MB的活性（p<0.01），收缩末期左心室内径（LVIDs）明显减小，并显著增加左心室射血分数(LVEF，%)和左心室缩短分数(LVFS，%).此外,卡巴胆碱能明显降低脓毒症大鼠心肌细胞凋亡水平(p<0.01),降低炎症因子包括肿瘤坏死因子-α（TNF-α）的含量，白细胞介素-1β（IL-1β）和IL-6（p<0.01），显著降低心肌组织中Caspase-3的表达(p<0.01),Bcl-2/Bax的表达显着增加（p<0.01），并明显抑制磷酸化（p-）PI3K的表达，p-AKT,Nod样受体蛋白3（NLRP3），和Caspase-1(p<0.01)。
卡巴胆碱可以减少心肌细胞炎症因子的释放，凋亡蛋白的表达和心肌细胞的凋亡，通过抑制PI3K/AKT信号通路改善脓毒症大鼠心功能和存活率。
