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Efficacy Evaluation of Ethyl Acetate Fraction of
增强出版Ipomoea muricatum in Prevention and Treatment of Alcoholic Gastric Ulcer- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 7, Pages: 105-114(2023)
- 作者机构：
河南中医药大学 中医药科学院，药学院，郑州 450046
- 作者简介：
- 基金信息：
DOI：10.13422/j.cnki.syfjx.20230141
CLC： R2-0;R33;R289;R318.14;R57Published：05 April 2023，
Published Online：12 January 2023，
Received：08 November 2022，
- 稿件说明：
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向西,郭辉,龚曼等.丁香茄乙酸乙酯部位防治酒精性胃溃疡的药效评价[J].中国实验方剂学杂志,2023,29(07):105-114.
XIANG Xi,GUO Hui,GONG Man,et al.Efficacy Evaluation of Ethyl Acetate Fraction of Ipomoea muricatum in Prevention and Treatment of Alcoholic Gastric Ulcer[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(07):105-114.
向西,郭辉,龚曼等.丁香茄乙酸乙酯部位防治酒精性胃溃疡的药效评价[J].中国实验方剂学杂志,2023,29(07):105-114. DOI： 10.13422/j.cnki.syfjx.20230141.
XIANG Xi,GUO Hui,GONG Man,et al.Efficacy Evaluation of Ethyl Acetate Fraction of Ipomoea muricatum in Prevention and Treatment of Alcoholic Gastric Ulcer[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(07):105-114. DOI： 10.13422/j.cnki.syfjx.20230141.
摘要
目的
2
探究丁香茄乙酸乙酯部位（IM-EA）防治酒精性胃溃疡（GU）的可行性，并采用网络药理学结合动物实验探究其发挥疗效的作用机制。
方法
2
将40只SD大鼠适应性喂养7 d后随机分为空白组、模型组、雷尼替丁组（2.7 mg·kg
-1
）、IM-EA低（DL，30 mg·kg
-1
）和高（DH，60 mg·kg
-1
）剂量组。预防性给药1周后，采用盐酸无水乙醇（150 mmol·L
-1
）复制酒精性GU大鼠模型，采用苏木素-伊红（HE）染色、高碘酸-希夫染色（PAS）初步评估IM-EA防治GU的疗效。根据ADMET属性筛选IM-EA先导化合物，采用SwissTarget平台获取IM-EA先导化合物的潜在作用靶标；通过DisGeNET、OMIM和GeneCards数据库收集GU疾病相关靶标，与IM-EA潜在作用靶标相映射获取IM-EA防治GU潜在作用靶标。采用STRING数据库构建靶标蛋白质-蛋白质相互作用（PPI）网络以筛选枢纽靶标，采用DAVID平台对交集靶标进行生物功能注释以探讨IM-EA防治GU的潜在作用机制，采用Autodock Vina软件进行计算机模拟初步验证。采用酶联免疫吸附测定法（ELISA）检测血清中肿瘤坏死因子-
α
（TNF-
α
）、白细胞介素-6（IL-6）含量和胃组织中前列腺素E
2
（PGE
2
）、基质金属蛋白酶-9（MMP-9）、超氧化物歧化酶（SOD）含量，采用蛋白免疫印迹法（Western blot）检测胃组织中丝裂原活化蛋白激酶（MAPK）信号通路的枢纽蛋白：Jun、细胞外调节蛋白激酶（ERK）和p38的相对表达量。
结果
2
动物实验结果表明，与空白组比较，模型组大鼠胃组织损伤明显，胃黏液分泌显著减少。与模型组比较，各给药组大鼠胃组织溃疡面积均显著减小（
P
<
0.01），胃组织病理学状态和胃黏液分泌得到改善，IM-EA防治GU确有疗效。经ADMET性质筛选获得16个IM-EA先导化合物，映射257个IM-EA防治GU潜在作用靶标，肿瘤坏死因子（TNF）、蛋白激酶B1（Akt1）、细胞肿瘤抗原p53（TP53）、表皮生长因子受体（EGFR）和ERK等靶标为PPI网络的枢纽节点。生物功能注释和分子对接结果提示MAPK信号通路可能在IM-EA防治GU的过程中起关键作用，与VEGF信号通路、磷脂酰肌醇3-激酶（PI3K）/Akt信号通路和NF-
κ
B信号通路在抗炎、抗氧化和损伤修复等方面协同生效。药理实验进一步验证发现，与空白组比较，模型组大鼠血清中IL-6含量极显著升高（
P
<
0.01），TNF-
α
具升高趋势；模型组大鼠胃组织中MMP-9含量极显著升高（
P
<
0.01），SOD含量明显降低（
P
<
0.05）。与模型组比较，IM-EA各给药组血清中TNF-
α
、IL-6含量、胃组织中MMP-9、PGE
2
含量均极显著降低（
P
<
0.01），胃组织中SOD含量极显著升高（
P
<
0.01）。与空白组比较，模型组大鼠胃组织中磷酸化（p）-p38、p-Jun、p-ERK显著上调（
P
<
0.01），p38、Jun显著上调（
P
<
0.01）；与模型组比较，IM-EA给药组胃组织中p-p38、p-Jun、p-ERK、p38则显著下调（
P
<
0.01），Jun、ERK相对表达量明显上调（
P
<
0.05）。
结论
2
丁香茄乙酸乙酯部位有较好的防治酒精性胃损伤的功效，可能通过MAPK通路介导抗炎、抗氧化和损伤修复等机制发挥作用。
Abstract
Objective
2
To investigate the feasibility of ethyl acetate fraction of
Ipomoea muricatum
（IM-EA） in the prevention and treatment of alcoholic gastric ulcer （GU） and explore its mechanism of action based on network pharmacology and experimental verification.
Method
2
Forty SD rats were randomly divided into a control group， a model group， a ranitidine group （2.7 mg·kg
-1
）， and low- and high-dose IM-EA groups （30，60 mg·kg
-1
） after adaptive feeding for 7 days. The GU model was replicated by hydrochloric acid in absolute ethanol （150 mmol·L
-1
） in rats after prophylactic administration for one week. Hematoxylin-eosin （HE） staining and periodic acid-Schiff （PAS） staining were used to preliminarily evaluate the efficacy of IM-EA in the prevention and treatment of GU. Lead compounds of IM-EA were screened out by ADMET， and the SwissTarget platform was used to identify the potential targets for these compounds. GU-related targets were collected through DisGeNET， OMIM， and GeneCards databases， which were mapped to potential IM-EA targets to obtain the potential targets of IM-EA against GU. The STRING database was used to construct the protein-protein interaction （PPI） network to screen the hub targets， and the DAVID platform was used to annotate the biological functions of common targets to explore the underlying mechanism of IM-EA against GU. Autodock Vina software was used for the preliminary verification of the computer simulation. The serum levels of tumor necrosis factor （TNF）-
α
and interleukin （IL）-6 and the content of prostaglandin E
2
（PGE
2
）， matrix metalloproteinase-9 （MMP-9）， and superoxide dismutase （SOD） in the gastric tissues were determined by enzyme-linked immunosorbent assay （ELISA）. The relative expression levels of core proteins in the mitogen-activated protein kinase （MAPK） signaling pathway， such as Jun oncoprotein， extracellular signal-regulated kinase （ERK）， and p38， in the gastric tissues were detected by Western blot.
Result
2
As revealed by the results of animal experiments， compared with the control group， the model group showed significantly damaged gastric tissues and reduced secretion of gastric mucus. Compared with the model group， the groups with drug intervention showed reduced ulcer areas in the gastric tissues （
P
<
0.01） and improved gastric histopathological status and gastric mucus secretion， suggesting that IM-EA was effective in the prevention and treatment of GU. Sixteen lead compounds of IM-EA were screened out by ADMET， and 257 potential targets of IM-EA against GU were obtained. The hub nodes in the PPI network included targets of TNF-
α
， protein kinase B1 （Akt1）， tumor protein 53 （TP53）， epidermal growth factor receptor （EGFR）， and ERK. Biological functional annotation and molecular docking results suggested that the MAPK signaling pathway potentially played a key role in the prevention and treatment of GU by IM-EA， which was synergistic with the vascular endothelial growth factor （VEGF） signaling pathway， phosphoinositide 3-kinase （PI3K）/Akt signaling pathway， and nuclear factor （NF）-
κ
B signaling pathway in anti-inflammation， anti-oxidation， and damage repair. The pharmacological experiment results showed that compared with the control group， the model group showed increased serum IL-6 content （
P
<
0.01）， an increasing trend of TNF-
α
content， increased MMP-9 content in the gastric tissues （
P
<
0.01）， and decreased SOD content （
P
<
0.05）. Compared with the model group， the IM-EA groups showed decreased TNF-
α
and IL-6 levels in the serum and PGE
2
and MMP-9 levels in the gastric tissues （
P
<
0.01）， and increased SOD content in the gastric tissues （
P
<
0.01）. Compared with the control group， the model group showed up-regulated expression of p-p38， p-Jun， and p-ERK in the gastric tissues （
P
<
0.01） and up-regulated p38 and Jun （
P
<
0.01）. Compared with the model group， the IM-EA groups showed down-regulated p-p38， p-Jun， p-ERK， and p38 in the gastric tissues （
P
<
0.01） and up-regulated relative expression of Jun and ERK （
P
<
0.05）.
Conclusion
2
IM-EA has a remarkable effect in the prevention and treatment of alcoholic gastric injury， which may be achieved through the mechanisms of anti-inflammation， anti-oxidation， and wound repair mediated by the MAPK signaling pathway.
关键词
丁香茄胃溃疡网络药理学作用机制丝裂原活化蛋白激酶（MAPK）信号通路
Keywords
Ipomoea muricatumgastric ulcernetwork pharmacologymechanism of actionmitogen activated protein kinase （MAPK） signaling pathway
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