Jin J L, Zhang H, Liu Q, Jiang H H, Liu X X, Tang X Z
Department of Critical Care Medicine, Shenzhen Hospital (Futian) of Guangzhou University of Chinese Medicine, Shenzhen 518034, China.
Shenzhen Emergency Center, Shenzhen 518035, China.
Zhonghua Yi Xue Za Zhi. 2020 Sep 22;100(35):2779-2784. doi: 10.3760/cma.j.cn112137-20200227-00477.
To investigated whether berberine could ameliorate septic cardiomyopathy in a rat model of sepsis and it's mechanisms. SD rats were divided into 3 groups: sepsis group (LPS group), rats were intraperitoneal injected of LPS (10 mg/kg); Berberine intervention group (Ber group), Ber (50 mg/kg, one time per day) was gavage fed 3 days before intraperitoneally injection of lipopolysaccharides (LPS); control group (Con group), rats were gavage fed with double distilled water (2 ml/100 g, one time per day) 3 days before intraperitoneal injection of normal saline (1 ml/100 g). LPS group and the Ber group was further divided into 3 subgroups (6), and the follow-up experiments were conducted at 6 h, 24 h and 48 h after LPS injection (of which 48 h subgroup rats were gavage fed with Ber/saline at 24 h). Left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), and the maximum rate of change of left ventricular pressure (±dp/dtmax) were monitored, the level of cardiac troponin T (cTnT), tumor necrosis factor (TNF)-α and interleukin (IL)-1β was detected by ELISA method, HE staining of myocardial tissues was done to observe myocardial injury; Western blotting method was used to detect the expression of toll-like receptor 4(TLR4) protein in rat myocardial tissue, the level of myocardial cell nucleus protein p65 was detected to reflects the degree of NF-κB activation. The correlation of factors was analyzed with Pearson correlation analysis. Pre-treatment with berberine stabilized cardiac hemodynamics and improved the systolic function and diastolic function in the heart of LPS-induced rats, as evidenced by the partial recovery of the reduced±dp/dtmax and LVSP, as well as the decreased LVEDP. Compared with the LPS group, the Ber group showed improved myocardial injury, as demonstrated by decreased cTnT at each time point. HE staining results showed that berberine decreased inflammatory cell infiltration and LPS-induced cell swelling. These effects were observed early at 6 hours, severe at 24 hours, and become more serious at 48 hours after LPS injection. Further, TLR4 and NF-κB p65 subunits, which were the two key factors of the TLR4/NF-κB signaling, were upregulated in the LPS group and attenuated in the Ber group. Consistently, the expression levels of the downstream cytokines TNF-α and IL-1β were lower in the Ber group than those in the LPS group (all 0.05). Myocardial injury markers were positively correlated with the markers of TLR4/NF-κB signals and the downstream host inflammatory factors (all 0.05). Berberine can improve myocardial injury and cardiac function in sepsis rats, the mechanism is considered to be related to that it can inhibit the activation of TLR4/NF-κB signaling pathway induced by LPS and further reducing the production of TNF-α and IL-1β.
为研究小檗碱能否改善脓毒症大鼠模型中的脓毒症性心肌病及其机制。将SD大鼠分为3组：脓毒症组（LPS组），大鼠腹腔注射LPS（10 mg/kg）；小檗碱干预组（Ber组），在腹腔注射脂多糖（LPS）前3天，每天一次灌胃给予Ber（50 mg/kg）；对照组（Con组），在腹腔注射生理盐水（1 ml/100 g）前3天，每天一次灌胃给予双蒸水（2 ml/100 g）。LPS组和Ber组再进一步分为3个亚组（每组6只），并在注射LPS后6 h、24 h和48 h进行后续实验（其中48 h亚组大鼠在24 h时灌胃给予Ber/生理盐水）。监测左心室收缩压（LVSP）、左心室舒张末期压力（LVEDP）以及左心室压力最大变化率（±dp/dtmax），采用ELISA法检测心肌肌钙蛋白T（cTnT）、肿瘤坏死因子（TNF）-α和白细胞介素（IL）-1β水平，对心肌组织进行HE染色观察心肌损伤；采用Western blotting法检测大鼠心肌组织中Toll样受体4（TLR4）蛋白的表达，检测心肌细胞核蛋白p65水平以反映NF-κB激活程度。采用Pearson相关分析对各因素的相关性进行分析。小檗碱预处理可稳定心脏血流动力学，改善LPS诱导大鼠心脏的收缩功能和舒张功能，表现为降低的±dp/dtmax和LVSP部分恢复，以及LVEDP降低。与LPS组相比，Ber组心肌损伤改善，各时间点cTnT均降低。HE染色结果显示，小檗碱可减少炎症细胞浸润和LPS诱导的细胞肿胀。这些作用在注射LPS后6小时早期出现，24小时时严重，48小时时更严重。此外，TLR4/NF-κB信号通路的两个关键因子TLR4和NF-κB p65亚基在LPS组中上调，而在Ber组中减弱。一致地，Ber组下游细胞因子TNF-α和IL-1β的表达水平低于LPS组（均P<0.05）。心肌损伤标志物与TLR4/NF-κB信号标志物及下游宿主炎症因子均呈正相关（均P<0.05）。小檗碱可改善脓毒症大鼠的心肌损伤和心脏功能，其机制可能与抑制LPS诱导的TLR4/NF-κB信号通路激活，进而减少TNF-α和IL-1β的产生有关。
