人参皂苷Rg1调控TLR4/NF-κB信号通路抗重症肺炎大鼠心肌组织损伤的机制探讨  

Mechanism of ginsenoside Rg1 regulating TLR4/NF-κB signaling pathway against myocardial tissue injury in rats with severe pneumonia

作　　者：孙治霞[1] 索红亮[2] 李华[1] 李宗尚 施光远 SUN Zhi-xia;SUO Hong-liang;LI Hua;LI Zong-shang;SHI Guang-yuan(Department of Critical Medicine,Henan Provincial Hospital of Traditional Chinese Medicine,Zhengzhou 450002,China;Department of Cardiology,Henan Provincial Hospital of Traditional Chinese Medicine,Zhengzhou 450002,China)

机构地区：[1]河南省中医院重症医学科,450002 [2]河南省中医院心病科,450002

出　　处：《天津医药》2021年第3期271-276,共6页Tianjin Medical Journal

基　　金：国家中医临床研究基地科研专项课题(2019JDZX046);河南省科技发展计划项目(172102310538)。

摘　　要：目的探讨人参皂苷Rg1对重症肺炎大鼠心肌组织损伤的影响及其机制。方法将40只大鼠以气管注入肺炎克雷伯菌液法构建重症肺炎模型后,将其均分为模型组和低、中、高剂量药物组;另取10只未处理大鼠作为对照组。低、中、高剂量药物组大鼠以2.5、5和10 mg/kg人参皂苷Rg1灌胃,模型组和对照组大鼠以等量生理盐水灌胃,每日1次,持续14 d。苏木精-伊红染色法观察大鼠心肌组织病理形态并行积分统计;采用心脏彩超检测大鼠心功能指标,包括左室舒张压(LVDP)、左室舒张末压(LVEDP)、左室最大舒张速率(-dP/dtmax)和左室最大收缩速率(+dP/dtmax)值;酶联免疫吸附测定(ELISA)法检测血清中B型钠尿肽(BNP)、肌钙蛋白I(cTnI)和肌酸激酶同工酶MB(CKMB)含量;实时荧光定量PCR(qPCR)法检测心肌组织中白细胞介素(IL)-6、IL-1β和肿瘤坏死因子-α(TNF-α)mRNA表达水平;蛋白免疫印迹法(Western blot)检测心肌组织中凋亡相关蛋白活化的含半胱氨酸的天冬氨酸蛋白酶3(cleaved caspase-3)和非活性态(procaspase-3)、Bcl-2、Bcl-2相关X蛋白(Bax)、Toll样受体4(TLR4)、髓样分化因子88(MyD88)、核因子-κB(NF-κB)p65和磷酸化(p)-NF-κB p65蛋白表达水平。结果对照组大鼠心肌组织结构正常,模型组大鼠心肌组织中心肌细胞排列紊乱且肿胀变形,肌纤维部分断裂且伴随有炎性细胞浸润;相对模型组,低、中、高剂量药物组大鼠心肌组织中上述细胞形态变化明显改善。与对照组比较,模型组大鼠心肌组织病理积分,LVEDP水平,BNP、cTnI、CK-MB含量和IL-6、IL-1β、TNF-αmRNA水平,cleaved caspase-3/procaspase-3和Bax/Bcl-2比值以及TLR4、MyD88、p-NF-κB p65蛋白表达水平均明显升高,而LVDP、-dP/dtmax和+dP/dtmax水平明显降低(P<0.05);以人参皂苷Rg1灌胃干预后可呈剂量依赖性逆转建模引起的上述变化(P<0.05)。结论人参皂苷Rg1可通过减轻炎症反应和心肌细胞凋亡保护重症肺Objective To investigate the effect and mechanism of ginsenoside Rg1 on myocardial tissue injury in rats with severe pneumonia.Methods A total of 40 rats were injected with Klebsiella pneumoniae solution into trachea to construct severe pneumonia model.The model rats were divided into model group and low,medium and high dose drug groups.Another 10 untreated rats were taken as control group.The rats in the low,medium and high dose drug groups were given ginsenoside Rg1 with 2.5,5 and 10 mg/kg by gavage,while the rats in the model group and the control group were given the same amount of normal saline,once a day for 14 days.Hematoxylin eosin stain method was used to observe the pathological morphology of myocardial tissue in rats and integral statistics was performed.The left ventricular diastolic pressure(LVDP),left ventricular end-diastolic pressure(LVEDP),maximal descending rate of left ventricular pressure(-dP/dtmax)and maximal developing rate of left ventricular pressure(+dP/dtmax)of rats were measured by color Doppler ultrasound.The serum contents of B-type natriuretic peptide(BNP),cardiac troponin I(cTnI)and MB isoenzyme of creatine kinase(CK-MB)were detected by enzyme-linked immunosorbent assay(ELISA).The mRNA expression levels of interleukin(IL)-6,IL-1βand tumor necrosis factor-α(TNF-α)were detected by real-time fluorescence quantitative PCR(qPCR).The protein expression levels of apoptosis-related protein cleaved cysteiny l aspartate specific proteinase 3(cleaved caspase-3)and inactive state(procaspase-3),Bcl-2 and Bcl-2 associated X protein(Bax),Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),nuclear factor-κB(NF-κB)p65 and phosphorylated(p)-NF-κB p65 in myocardial tissue were detected by Western blot assay.Results The myocardial tissue structure was normal in the control group,and the central muscle cells of the model group were disorderly arranged and swelled,and the muscle fibers were partially broken and accompanied with inflammatory cell infiltration.Compared with the model

关 键 词：肺炎 心肌 人参皂甙类 TOLL样受体4 NF-ΚB 细胞凋亡 

分 类 号：R563.1[医药卫生—呼吸系统] R542.2[医药卫生—内科学]