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生脉注射液抗脓毒症作用及对小鼠肠黏膜屏障的保护机制。

Mechanism of Shengmai Injection on Anti-Sepsis and Protective Activities of Intestinal Mucosal Barrier in Mice.

机构信息

Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, 100193, China.

出版信息

Chin J Integr Med. 2022 Sep;28(9):817-822. doi: 10.1007/s11655-021-3292-y. Epub 2021 Jul 9.

Abstract

OBJECTIVE

To study the mechanism of Shengmai Injection (SMI, ) on anti-sepsis and protective activities of intestinal mucosal barrier.

METHODS

The contents of 11 active components of SMI including ginsenoside Rb1, Rb2, Rb3, Rd, Re, Rf, Rg1, Rg2, ophioposide D, schisandrol A and schisantherin A were determined using ultra-performance liquid chromatography. Fifty mice were randomly divided into the blank, the model, the low-, medium- and high-dose SMI groups (0.375, 0.75, 1.5 mL/kg, respectively) by random number table, 10 mice in each group. In SMI group, SMI was administrated to mice daily via tail vein injection for 3 consecutive days, while the mice in the blank and model groups were given 0.1 mL of normal saline. One hour after the last SMI administration, except the blank group, the mice in other groups were intraperitoneally injected with lipopolysaccharide (LPS) saline solution (2 mL/kg) at a dosage of 5 mL/kg for development of endotoxemia mice model. The mice in the blank group were given the same volume of normal saline. Inflammatory factors including interferon-γ (INF-γ), tumor necrosis factor-α (TNF-α), interleukin (IL)-2 and IL-10 were measured by flow cytometry. Myosin light-chain kinase (MLCK), nuclear factor κB (NF-κB) levels, and change of Occludin proteins in jejunum samples were analyzed by Western blot.

RESULTS

The decreasing trends of INF-γ, TNF-α and IL-2 were found in serum of SMI treatment groups. In SMI-treated mice, the content of Occludin increased and MLCK protein decreased compared with the model group (P<0.05 or P<0.01). The content of cellular and nuclear NF-κB did not change significantly (P>0.05).

CONCLUSION

SMI may exert its anti-sepsis activity mainly through NF-κB-pro-inflammatory factor-MLCK-TJ cascade.

摘要

目的

研究生脉注射液(SMI)对内毒素血症及肠黏膜屏障保护作用的机制。

方法

采用超高效液相色谱法测定 SMI 中 11 种活性成分,包括人参皂苷 Rb1、Rb2、Rb3、Rd、Re、Rf、Rg1、Rg2、ophioposide D、五味子甲素和五味子乙素的含量。将 50 只小鼠随机分为空白组、模型组、低剂量 SMI 组(0.375 mL/kg)、中剂量 SMI 组(0.75 mL/kg)和高剂量 SMI 组(1.5 mL/kg),每组 10 只。SMI 组小鼠连续 3 天经尾静脉注射给药,每天 1 次,空白组小鼠给予 0.1 mL 生理盐水。末次给药 1 h 后,除空白组外,其余各组小鼠均按 5 mL/kg 剂量腹腔注射脂多糖(LPS)生理盐水溶液(2 mL/kg),制备内毒素血症小鼠模型。空白组小鼠给予相同体积的生理盐水。采用流式细胞术检测干扰素-γ(INF-γ)、肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-2 和 IL-10 等炎症因子,Western blot 法分析肌球蛋白轻链激酶(MLCK)、核因子κB(NF-κB)水平及空肠组织中Occludin 蛋白的变化。

结果

与模型组比较,SMI 治疗组血清中 INF-γ、TNF-α 和 IL-2 水平呈下降趋势。与模型组比较,SMI 治疗组小鼠肠组织中 Occludin 含量增加,MLCK 蛋白表达减少(P<0.05 或 P<0.01),细胞及核内 NF-κB 含量无明显变化(P>0.05)。

结论

SMI 发挥抗内毒素血症作用可能主要通过 NF-κB-促炎因子-MLCK-TJ 级联。

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