丹参和红花提取物对脂多糖诱导的肝损伤的保护作用。

Protective effect of Salvia miltiorrhiza and Carthamus tinctorius extract against lipopolysaccharide-induced liver injury.

作者信息

Gao Li-Na, Yan Kuo, Cui Yuan-Lu, Fan Guan-Wei, Wang Yue-Fei

机构信息

Li-Na Gao, Kuo Yan, Yuan-Lu Cui, Guan-Wei Fan, Yue-Fei Wang, Research Center of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China.

出版信息

World J Gastroenterol. 2015 Aug 14;21(30):9079-92. doi: 10.3748/wjg.v21.i30.9079.

DOI:10.3748/wjg.v21.i30.9079

PMID:26290634

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4533039/

Abstract

AIM

To investigate the hepatoprotective effects and mechanisms of an extract of Salvia miltiorrhiza and Carthamus tinctorius in vivo.

METHODS

C57BL/6J mice were randomly assigned to five groups and intraperitoneally administered 0.9% saline, Salvia miltiorrhiza and Carthamus tinctorius extract [Danhong injection (DHI), 0.75 and 3 g/kg mixed extract] or reduced glutathione for injection (RGI, 300 mg/kg) for 30 min before exposure to lipopolysaccharide (LPS, 16 mg/kg). After intraperitoneal LPS stimulation for 90 min or 6 h, the mice were sacrificed by ether anaesthesia, and serum and liver samples were collected. Histological analysis (H&E) and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling (TUNEL) staining were performed. Alanine transferase (ALT), aspartate transaminase (AST), total bilirubin (TBil), glutathione-S-transferase (GST), malondialdehyde (MDA), tumour necrosis factor (TNF)-α, interleukin (IL)-6, and caspase-3 levels were measured. Bax, Bcl-2, P-IκBα, IκBα, P-NF-κB p65, and NF-κB p65 protein levels were determined by Western blot. TNF-α, IL-6, caspase-3, Bax and Bcl-2 mRNA expression was measured by real-time reverse transcription-polymerase chain reaction (RT-PCR).

RESULTS

Hematoxylin-eosin staining and TUNEL results suggested that DHI (3 g/kg) treatment alleviated inflammatory and apoptotic (P < 0.01) injury in the liver of mice. DHI treatment dose-dependently blunted the abnormal changes in biochemical parameters such as ALT (72.53 ± 2.83 for 3 g/kg, P < 0.01), AST (76.97 ± 5.00 for 3 g/kg, P < 0.01), TBil (1.17 ± 0.10 for 3 g/kg, P < 0.01), MDA (0.81 ± 0.36 for 3 g/kg, P < 0.01), and GST (358.86 ± 12.09 for 3 g/kg, P < 0.01). Moreover, DHI (3 g/kg) remarkably decreased LPS-induced protein expression of TNF-α (340.55 ± 10.18 for 3 g/kg, P < 0.01), IL-6 (261.34 ± 10.18 for 3 g/kg, P < 0.01), and enzyme activity of caspase-3 (0.93 ± 0.029 for 3 g/kg, P < 0.01). The LPS-induced mRNA expression of TNF-α, IL-6 and caspase-3 was also decreased by DHI. Western blot analysis revealed that DHI antagonised LPS-stimulated decrease of Bcl-2 and increase of Bax protein expression. Furthermore, DHI inhibited LPS-induced IκBα and NF-κB p65 phosphorylation.

CONCLUSION

DHI may be a multi-function protectant against acute hepatic injury in mice through its anti-inflammatory, anti-oxidative and anti-apoptotic activities.

摘要

目的

研究丹参和红花提取物在体内的肝保护作用及机制。

方法

将C57BL/6J小鼠随机分为五组，腹腔注射0.9%生理盐水、丹参和红花提取物[丹红注射液（DHI），0.75和3 g/kg混合提取物]或注射用还原型谷胱甘肽（RGI，300 mg/kg），在暴露于脂多糖（LPS，16 mg/kg）前30分钟给药。腹腔注射LPS刺激90分钟或6小时后，用乙醚麻醉处死小鼠，收集血清和肝脏样本。进行组织学分析（苏木精-伊红染色）和末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）染色。检测丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、总胆红素（TBil）、谷胱甘肽-S-转移酶（GST）、丙二醛（MDA）、肿瘤坏死因子（TNF）-α、白细胞介素（IL）-6和半胱天冬酶-3水平。通过蛋白质印迹法测定Bax、Bcl-2、磷酸化IκBα（P-IκBα）、IκBα、磷酸化核因子κB p65（P-NF-κB p65）和核因子κB p65蛋白水平。通过实时逆转录-聚合酶链反应（RT-PCR）检测TNF-α、IL-6、半胱天冬酶-3、Bax和Bcl-2 mRNA表达。

结果

苏木精-伊红染色和TUNEL结果表明，DHI（3 g/kg）处理减轻了小鼠肝脏的炎症和凋亡（P<0.01）损伤。DHI处理剂量依赖性地减弱了生化参数的异常变化，如ALT（3 g/kg组为72.53±2.83，P<0.01）、AST（3 g/kg组为76.97±5.00，P<0.01）、TBil（3 g/kg组为1.17±0.10，P<0.01）、MDA（3 g/kg组为0.81±0.36，P<0.01）和GST（3 g/kg组为358.86±12.09，P<0.01）。此外，DHI（3 g/kg）显著降低LPS诱导的TNF-α蛋白表达（3 g/kg组为340.55±10.18，P<0.01）、IL-6蛋白表达（3 g/kg组为261.34±10.18，P<0.01）和半胱天冬酶-3酶活性（3 g/kg组为0.93±0.029，P<0.01）。DHI还降低了LPS诱导的TNF-α、IL-6和半胱天冬酶-3 mRNA表达。蛋白质印迹分析显示，DHI拮抗LPS刺激引起的Bcl-2降低和Bax蛋白表达增加。此外，DHI抑制LPS诱导的IκBα和NF-κB p65磷酸化。