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1.陕西中医药大学 基础医学院,陕西 咸阳 712046
2.陕西中医药大学 附属医院,陕西 咸阳 712000
[第一作者] 闫曙光,博士,副教授,从事中医药治疗溃疡性结肠炎的基础与临床研究,Tel:029-38185122,E-mail:ysg2002.student@sina.com
*魏海梁,副主任医师,从事中西医结合治疗消化道疾病的基础与临床研究,Tel:029-33321832,E-mail:86888694@qq.com
收稿日期:2019-06-16,
网络出版日期:2019-10-09,
纸质出版日期:2020-02-20
移动端阅览
闫曙光, 惠毅, 李倩, 等. 黄连-干姜提取物对溃疡性结肠炎小鼠结肠上皮TLR4/NF-
Shu-guang YAN, Yi HUI, Qian LI, et al. Effect of Coptidis Rhizoma-Zingiberis Rhizoma Extract on Colonic Epithelium TLR4/NF-κB Signaling Pathway in Mice with Ulcerative Colitis[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(4): 70-75.
闫曙光, 惠毅, 李倩, 等. 黄连-干姜提取物对溃疡性结肠炎小鼠结肠上皮TLR4/NF-
Shu-guang YAN, Yi HUI, Qian LI, et al. Effect of Coptidis Rhizoma-Zingiberis Rhizoma Extract on Colonic Epithelium TLR4/NF-κB Signaling Pathway in Mice with Ulcerative Colitis[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(4): 70-75. DOI: 10.13422/j.cnki.syfjx.20200202.
目的:
2
观察中药黄连、干姜的主要成分黄连素和6-姜烯酚对溃疡性结肠炎小鼠结肠上皮细胞炎症信号通路Toll样受体4(TLR4)/核转录因子-
κ
B(NF-
κ
B)的影响。
方法:
2
50只昆明种小鼠随机分为正常组,模型组,黄连素组(100 mg·kg
-1
),6-姜烯酚组(100 mg·kg
-1
),6-姜烯酚合黄连素组(200 mg·kg
-1
),每组10只。采用2%葡聚糖硫酸钠口服2周建立溃疡性结肠炎小鼠模型,各组给予相应剂量的药物灌胃,正常组和模型组给予等量生理盐水,给药20 d后取血清及结肠组织样本,酶联免疫吸附测定法(ELISA)检测血清白细胞介素-1
β
(IL-1
β
),肿瘤坏死因子-
α
(TNF-
α
)含量,实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)分别检测结肠上皮组织TLR4,NF-
κ
B p65 mRNA和蛋白的表达。
结果:
2
与正常组比较,模型组小鼠结肠上皮组织TLR4,NF-
κ
B p65 mRNA和蛋白表达量显著升高(
P
<
0.01),血清IL-1
β
,TNF-
α
含量显著升高(
P
<
0.01);与模型组比较,6-姜烯酚组,黄连素组,6-姜烯酚合黄连素组小鼠结肠上皮组织TLR4,NF-
κ
B p65 mRNA和蛋白表达量显著降低(
P
<
0.01),血清IL-1
β
,TNF-
α
含量显著降低(
P
<
0.01);三组间比较,6-姜烯酚合黄连素组作用力最强(
P
<
0.01)。
结论:
2
6-姜烯酚、黄连素均能够抑制结肠组织炎症,减轻炎症损伤,治疗溃疡性结肠炎,黄连素和6-姜烯酚联合应用有显著的协同增效作用,其机制与抑制TLR4/NF-
κ
B通路的过度活化进而调节肠道非可控性炎症有关。
Objective:
2
To observe the effect of berberine and 6-shogaol
main components of Coptiae Rhizoma and Zingiberis Rhizoma
on the inflammatory signaling pathway of Toll-like receptors 4 (TLR4)/nuclear factor kappa B (NF-
κ
B) in colonic epithelial cells of mice with ulcerative colitis.
Method:
2
Fifty Kunming mice were randomly divided into normal group
model group
berberine group (100 mg·kg
-1
)
6-shogaol group (100 mg·kg
-1
)
and 6-shogaol combined with berberine group (200 mg·kg
-1
)
with 10 mice in each group. A mouse model of ulcerative colitis was established through oral administration with 2% dextroan sulfate for two weeks. Each group was given corresponding drugs by gavage
while normal group and model group were given equal amount of normal saline. Serum and colon tissue samples were taken 20 days after administration. Enzyme-linked immunosorbent method was used to detect serum interleukin-1
β
(IL-1
β
)
tumor necrosis factor-
α
(TNF-
α
) expressions
and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) and Western blot method were used to detect TLR4
NF-
κ
B p65 mRNA and protein expressions in colon epithelial tissue.
Result:
2
Compared with the normal group
relative expressions of TLR4 and NF-
κ
B p65 mRNA and protein were increased in the model group (
P
<
0.01)
and the contents of serum IL-1
β
and TNF-
α
were increased (
P
<
0.01). Compared with the model group
relative expressions of TLR4 and NF-
κ
B p65 mRNA and protein were significantly decreased in 6-shogaol group
berberine group and 6-shogaol combined with berberine group (
P
<
0.01)
and the contents of serum IL-1
β
and TNF-
α
were significantly decreased (
P
<
0.01). Among the three groups
6-shogaol combined with berberine group had the strongest effect (
P
<
0.01).
Conclusion:
2
Both 6-shogaol and berberine can inhibit colonic inflammation
reduce inflammatory damage and treat ulcerative colitis. The combined application of 6-shogaol and berberine has a significant synergism effect. The mechanism is related to the excessive activation of TLR4/NF-
κ
B pathway and the regulation of non-controllable intestinal inflammation.
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