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1.河南中医药大学,郑州 450046
2.中国中医科学院 中药研究所,北京 100700
Received:10 April 2022,
Published Online:25 August 2022,
Published:20 January 2023
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崔兵迪,郭辉,龚曼等.基于Akt/NF-κB信号通路探讨牛蒡根水提物对盐酸/乙醇诱导的急性胃溃疡大鼠的保护作用[J].中国实验方剂学杂志,2023,29(02):1-9.
CUI Bingdi,GUO Hui,GONG Man,et al.Protective Effect of Aqueous Extract of Arctium lappa Roots on HCl/EtOH-induced Acute Gastric Ulcer in Rats Based on Akt/NF-κB Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(02):1-9.
崔兵迪,郭辉,龚曼等.基于Akt/NF-κB信号通路探讨牛蒡根水提物对盐酸/乙醇诱导的急性胃溃疡大鼠的保护作用[J].中国实验方剂学杂志,2023,29(02):1-9. DOI: 10.13422/j.cnki.syfjx.20220850.
CUI Bingdi,GUO Hui,GONG Man,et al.Protective Effect of Aqueous Extract of Arctium lappa Roots on HCl/EtOH-induced Acute Gastric Ulcer in Rats Based on Akt/NF-κB Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(02):1-9. DOI: 10.13422/j.cnki.syfjx.20220850.
目的
2
基于蛋白激酶B(Akt)/核转录因子-
κ
B(NF-
κ
B)信号通路研究牛蒡根水提物(ALR-AE)对盐酸/乙醇(HCl/EtOH)诱导的大鼠急性胃溃疡的作用机制。
方法
2
雄性大鼠随机分为5组,分别为正常组、模型组、雷尼替丁组(35 mg·kg
-1
)和ALR-AE低、高剂量组(50、100 mg·kg
-1
),各给药组连续灌胃给药3 d,每天2次;末次给药30 min后,模型组及各给药组给予HCl/EtOH(以60% EtOH为溶剂,使HCl浓度为150 mmol·L
-1
)诱导大鼠急性胃溃疡。取各组胃组织样品,采用电子成像技术扫描溃疡面,ImageJ 1.8.0软件计算溃疡抑制率;分别采用苏木素-伊红(HE)和高碘酸-希夫(PAS)染色观察病理变化及糖蛋白的分布;比色法测定组织中氧化应激因子丙二醛(MDA)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)的水平;酶联免疫吸附试验(ELISA)检测组织样品中肿瘤坏死因子-
α
(TNF-
α
)、白细胞介素 (IL)-6、IL-1
β
的水平;通过蛋白质免疫印迹法(Western blot)检测各组组织中磷酸化(p)-Akt/Akt、p-NF-
κ
B p65/NF-
κ
B p65、p-NF-
κ
B抑制蛋白
α
(I
κ
B
α
)/I
κ
B
α
、p-I
κ
B激酶
α
(IKK
α
)/IKK
α
和蛋白水平。
结果
2
与正常组比较,模型组胃组织损伤严重,胃溃疡面积显著增大(
P
<
0.01);MDA、TNF-
α
、IL-6和IL-1
β
水平显著升高(
P
<
0.01),GSH-Px和SOD水平显著降低(
P
<
0.01),Akt、NF-
κ
B p65、IKK
α
和I
κ
B
α
的磷酸化水平显著升高(
P
<
0.01);与模型组比较,ALR-AE可显著降低HCl/EtOH诱导的胃组织损伤程度、提高溃疡抑制率(
P
<
0.01),剂量依赖性地降低MDA、TNF-
α
、IL-6和IL-1
β
的水平(
P
<
0.05,
P
<
0.01),显著升高GSH-Px和SOD的水平(
P
<
0.01),ALR-AE低剂量组可明显抑制Akt的磷酸化水平(
P
<
0.05),ALR-AE高剂量组可明显抑制Akt、NF-
κ
B p65、IKK
α
、I
κ
B
α
的磷酸化水平(
P
<
0.05,
P
<
0.01)。
结论
2
ALR-AE对HCl/EtOH诱导的大鼠胃溃疡有显著的保护作用,其作用机制可能与抑制Akt/NF-
κ
B信号通路介导的炎症介质表达及降低氧化应激水平有关。
Objective
2
To investigate protective effect of
Arctium lappa
root aqueous extract (ALR-AE) on hydrochloric acid/ethanol (HCl/EtOH)-induced acute gastric ulcer in rats based on protein kinase B/nuclear transcription factor-
κ
B (Akt/NF-
κ
B) signaling pathway.
Method
2
Rats were randomly divided into 5 groups, namely normal group, model group, ranitidine group (35 mg·kg
-1
), ALR-AE low dose group (50 mg·kg
-1
, ALR-AE-L group) and ALR-AE high dose group (100 mg·kg
-1
, ALR-AE-H group). Different doses of ALR-AE were orally administered twice daily for three consecutive days before the animals were subjected to HCl/EtOH (60% ethanol in 150 mmol·L
-1
HCl) to induce acute gastric ulcer. For the gastric tissue samples, the ulcer surface was recorded by electronic imaging technique, and then the ulcer inhibition rate was calculated using ImageJ 1.8.0, hematoxylin-eosin (HE) and periodic acid-Schiff (PAS) staining were used to observe the pathological changes and mucoprotein distribution, respectively. The levels of oxidative stress factors of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in rat gastric tissues were determined by colorimetric method, the levels of pro-inflammatory mediators of tumor necrosis factor-
α
(TNF-
α
), interleukin-6 (IL-6) and IL-1
β
were determined by enzyme linked immunosorbent assay (ELISA), protein levels of phosphorylation and non-phosphorylation of Akt, NF-
κ
B p65, NF-
κ
B inhibitor protein
α
(I
κ
B
α
) and I
κ
B kinase
α
(IKK
α
) were evaluated by Western blot.
Result
2
Compared with the normal group, the gastric tissue of the model group was severely damaged, and the area of gastric ulcer were significantly enlarged (
P
<
0.01), the levels of MDA, TNF-
α
, IL-6 and IL-1
β
in gastric tissue were significantly increased (
P
<
0.01), levels of GSH-Px and SOD were significantly decreased (
P
<
0.01), and phosphorylation levels of Akt, NF-
κ
B p65, IKK
α
and I
κ
B
α
in gastric tissue were significantly increased (
P
<
0.01). Compared with the model group, ALR-AE significantly attenuated HCl/EtOH-induced gastric tissue damage, significantly increased ulcer inhibition rate (
P
<
0.01), and dose-dependently reduced the levels of MDA, TNF-
α
, IL-6 and IL-1
β
(
P
<
0.05,
P
<
0.01), and elevated GSH-Px and SOD levels (
P
<
0.01), ALR-AE-L group could significantly inhibit the phosphorylation levels of Akt (
P
<
0.05), and ALR-AE-H group could significantly inhibit phosphorylation levels of Akt, NF-
κ
B p65, IKK
α
and I
κ
B
α
(
P
<
0.05,
P
<
0.01).
Conclusion
2
ALR-AE has a significant protective effect on HCl/EtOH-induced acute gastric ulcers in rats, and its mechanism may be related to the inhibition of inflammatory mediator expression and reduction of oxidative stress levels mediated by Akt/NF-
κ
B signaling pathway.
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