Synergistic Effect of Baicalin and Wogonoside on NF-<italic style="font-style: italic">κ</italic>B Signaling Pathway Based on Medium-efficiency Equation

Zheng-wen ZHU; Qing JIANG; Yu LUO; Yu-sheng LIANG; Jia-si WU; Si-yu SU; Xian-li MENG; Yong ZENG; Ping WANG

doi:10.13422/j.cnki.syfjx.20202138

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Synergistic Effect of Baicalin and Wogonoside on NF-κB Signaling Pathway Based on Medium-efficiency Equation

Pharmacology | 更新时间：2020-12-10

- Synergistic Effect of Baicalin and Wogonoside on NF-κB Signaling Pathway Based on Medium-efficiency Equation
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 21, Pages: 84-91(2020)
- 作者机构：
  
  成都中医药大学，成都 611137
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20202138
  CLC： R2-0;R289;R285.5
- Received：22 May 2020，
  
  Published Online：27 August 2020，
  
  Published：05 November 2020
- 稿件说明：
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朱正文,蒋晴,罗煜等.基于中效方程的黄芩苷与汉黄芩苷调控NF-κB信号通路的协同作用[J].中国实验方剂学杂志,2020,26(21):84-91.

ZHU Zheng-wen,JIANG Qing,LUO Yu,et al.Synergistic Effect of Baicalin and Wogonoside on NF-κB Signaling Pathway Based on Medium-efficiency Equation[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(21):84-91.
朱正文,蒋晴,罗煜等.基于中效方程的黄芩苷与汉黄芩苷调控NF-κB信号通路的协同作用[J].中国实验方剂学杂志,2020,26(21):84-91. DOI： 10.13422/j.cnki.syfjx.20202138.

ZHU Zheng-wen,JIANG Qing,LUO Yu,et al.Synergistic Effect of Baicalin and Wogonoside on NF-κB Signaling Pathway Based on Medium-efficiency Equation[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(21):84-91. DOI： 10.13422/j.cnki.syfjx.20202138.

摘要

目的

利用中效原理定量分析黄芩苷与汉黄芩苷合用的抗炎协同药效学机理。

方法

通过脂多糖（LPS）100 μg·L

-1

刺激RAW264.7细胞构建体外炎症细胞模型；实验设置正常组，模型组，穿心莲内酯组（10 μmol·L

-1

），黄芩苷组（2.06，4.13，8.25，16.5，33，66，132 μmol·L

-1

），汉黄芩苷组（2.94，5.88，11.75，23.5，47，94，188 μmol·L

-1

）和黄芩苷-汉黄芩苷联合组［（2.06+2.94）（4.13+5.88）（8.25+11.75）（16.5+23.5）（33+47）（66+94）（132+188） μmol·L

-1

］；采用酶联免疫吸附测定（ELISA）分别检测药物干预50 min和4 h后细胞培养上清中的肿瘤坏死因子-

（TNF-

）和白细胞介素-6（IL-6）含量；采用硝酸盐还原酶（Griess）法检测药物干预24 h后细胞培养上清中一氧化氮（NO）的含量；蛋白免疫印迹法（Western blot）分别检测药物干预2，12 h后细胞中磷酸化核转录因子-

B p65（p-NF-

B p65）和诱导型一氧化氮合酶（iNOS）的活化水平；绘制作用率/非作用率（

）与剂量的关系表。

结果

与正常组比较，模型组RAW264.7细胞中p-NF-

B p65蛋白和iNOS蛋白表达均明显增加（

0.05，

0.01），细胞因子TNF-

，IL-6和NO的表达均显著增加（

0.01）；与模型组比较，各给药组高剂量可抑制p-NF-

B p65蛋白的表达（

0.05），汉黄芩苷组和联合组可浓度依赖性下调iNOS蛋白的表达（

0.01），黄芩苷组无明显差异。各给药组高剂量均可显著抑制NO的生成（

0.05），对IL-6生成无明显抑制作用。汉黄芩苷组和联合组可显著抑制TNF-

的生成（

0.05），黄芩苷组无明显差异；在实验剂量下，

与剂量关系表显示，联合组p-NF-

B p65活化和IL-6生成的

小于黄芩苷组和汉黄芩苷组，联合组iNOS，TNF-

和NO生成的

在高剂量时大于黄芩苷组和汉黄芩苷组。

结论

黄芩苷和汉黄芩苷对NF-

B通路中不同靶标作用强度差异较大，汉黄芩苷是二者合用的主体药效物质，二者合用对不同靶点表现为不同程度的协同或拮抗作用。

Abstract

Objective

Quantitative analysis of anti-inflammatory synergistic pharmacodynamics mechanism of baicalin and wogonoside by medium efficiency principle.

Method

inflammatory cell model was constructed by stimulating RAW264.7 cells by lipopolysaccharide （LPS） 100 μg·L

-1

in vitro

. The experiment was performed in the normal group， the model group， the andrographolide group （10 μmol·L

-1

）， the baicalin group （2.06，4.13，8.25，16.5，33，66，132 μmol·L

-1

） and the wogonoside group （2.94，5.88，11.75，23.5，47，94，188 μmol·L

-1

） and the baicalin-wogonoside combination group ［（2.06+2.94）（4.13+5.88）（8.25+11.75）（16.5+23.5）（33+47）（66+94）（132+188） μmol·L

-1

］. The levels of tumor necrosis factor-

（TNF-

） and interleukin-6 （IL-6） in the cell culture supernatants after drug intervention for 50 min and 4 h were detected by enzyme-linked immunosorbent assay （ELISA） method. The level of nitric oxide （NO） in the cell culture supernatant after drug intervention for 24 h were detected by Griess method. Western blot was used to detect the activation levels of phosphorylation of nuclear factor-

B p65（p-NF-

B p65） and inducible nitric oxide synthase（iNOS） in cells after drug intervention for 2 h and 12 h. The

-dose profile of each indicator was drawn to observe the increase or decrease of effect.

Result

Compared with normal group， the expression of p-NF-

B p65， iNOS and cytokines including TNF-

， IL-6 and NO （

0.05，

0.01） in the model group were significantly up-regulated. Compared with the model group， each group at high doses could inhibit the phosphorylation of NF-

B p65 protein（

0.05），the baicalin group and the combined group could down-regulate the expression of iNOS protein in a concentration-dependent manner（

0.01） and the baicalin group had no obvious inhibitory effect. each administration group at high dose could significantly inhibit the production of NO（

0.05），but each group had no inhibitory effect on IL-6 production. The baicalin group and the combined group could significantly Inhibit the production of TNF-

（

0.05） and there was no significant difference between the baicalin group and the model group. At the experimental dose， the

-dose table showed that the

value of p-NF-

B p65 and IL-6 in the combined group was not greater than the baicalin group and the wogonoside group. The

value of iNOS， TNF-

and NO in the combined group is higher than the baicalin group and the wogonoside group.

Conclusion

The baicalin and wogonoside have different effects on different targets in the NF-

B pathway. The wogonoside is the main pharmacological substance in this combination and the combination shows different degrees of synergy or antagonism effects on different targets.

关键词

Keywords

references

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Synergistic Effect of Baicalin and Wogonoside on NF-κB Signaling Pathway Based on Medium-efficiency Equation

DOI：10.13422/j.cnki.syfjx.20202138

摘要

Abstract

关键词

Keywords

references