[关键词]
[摘要]
【目的】 观察电针内关穴减轻脂多糖(LPS)诱导的脓毒症模型小鼠心功能障碍的作用机制。【方法】 将 48 只雄性 C57BL/6小鼠随机分为4组,即正常对照组、生理盐水+电针组、脓毒症模型组、脓毒症+电针组。脓毒症模型组及脓毒症+ 电针组小鼠给予腹腔注射LPS(5 mg/kg)诱导脓毒症心肌损伤模型。LPS注射后立即对小鼠实施电针治疗。于LPS处理后6 h, 采用彩色超声心动图监测小鼠左心室短轴收缩末期内径(LVIDs)和短轴缩短率(FS)。酶联免疫吸附分析(ELISA)检测血清炎 症因子肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)水平,蛋白免疫印迹(Western Blot)法检测心脏组织中Rho激酶(ROCK) 1、ROCK2蛋白表达水平及肌球蛋白磷酸酶底物 1(MYPT1)、肌球蛋白轻链 2(MLC2)磷酸化水平。【结果】 与正常对照组比 较,脓毒症模型组小鼠 LVIDs增加(P < 0.05),FS下降(P < 0.05),血清中 TNF-α、IL-6含量均升高(P < 0.001),心脏组织 MYPT1磷酸化水平显著升高(P < 0.05);与脓毒症模型组比较,脓毒症+电针组LVIDs降低(P < 0.05),FS升高(P < 0.05),血清 中TNF-α、IL-6含量下降(P < 0.05或P < 0.01),MYPT1磷酸化水平显著降低(P < 0.05)。各组ROCK1、ROCK2蛋白表达水 平以及MLC2的磷酸化水平无显著性差异(P > 0.05)。【结论】 电针内关穴可能通过抑制心脏组织RhoA/ROCK信号通路活化, 减轻脓毒症模型小鼠全身和局部炎性反应水平,改善心功能,产生心肌保护效应。
[Key word]
[Abstract]
Objective To observe the mechanism of action of electroacupuncture at Neiguan(PC6)points for alleviating lipopolysaccharide(LPS)-induced myocadial dysfunction in sepsis model mice. Methods Forty-eight male C57BL / 6 mice were randomly divided into four groups, namely normal control group, saline + electroacupuncture group, sepsis model group, and sepsis + electroacupuncture group. The rats in the sepsis model group and sepsis+electroacupuncture group were given intraperitoneal injection of LPS(5 mg/kg)to induce the sepsis model. The mice were treated with electroacupuncture immediately after injection. At hour 6 after the LPS intervention,the left ventricular short-axis end-systolic diameter(LVIDs)and fractional shortening(FS)of the left ventricular were monitored by color doppler echocardiography. The levels of inflammatory factors such as tumor necrosis factor alpha (TNF - α) and interleukin 6 (IL-6) in serum were detected by enzyme-linked immunosorbent assay(ELISA),and the protein expression levels of Rho-kinase(ROCK)1 and ROCK2,and phosphorylation levels of myosin phosphatase substrate 1(MYPT1)and myosin light chain 2(MLC2)in cardiac tissues were detected by Western Blot. Results Compared with the normal control group,the LVIDs in mice in the sepsis model group was increased(P < 0.05),FS was decreased(P < 0.05),the levels of TNF-α and IL-6 in serum were all elevated(P < 0.001), and the level of phosphorylation of MYPT1, a substrate of ROCK, in cardiac tissues was significantly elevated(P < 0.05);compared with the sepsis model group,the LVIDs in the sepsis+electroacupuncture group was decreased(P < 0.05),FS was increased(P < 0.05),the levels of TNF-α and IL-6 in serum were decreased(P < 0.05 or P < 0.01), and the phosphorylation level of MYPT1 was significantly reduced (P < 0.05), and the protein expression levels of ROCK1 and ROCK2 as well as the phosphorylation level of MLC2 had no significant differences(P > 0.05). Conclusion Electroacupuncture at Neiguan points may reduce systemic and local inflammatory reaction levels in mice with sepsis model to improve cardiac function through inhibiting the activation of Rho / ROCK signaling pathway in cardiac tissues, thus producing myocardial protection efficacy .
[中图分类号]
R631;R245.3
[基金项目]
国家自然科学基金委员会青年项目(编号:82205245);广东省自然科学基金面上项目(编号:2023A1515011143);广州市基础 与应用基础研究项目(编号:2023A04J0421)