Exploring the Effect of Medication-Containing Serum of <italic style="font-style: italic">Pingwei Capsules</italic> （平胃胶囊） on MNNG-Induced Inflammatory Cancer Transformation of Human Gastric Mucosal Epithelial Cells Based on Raf/MEK/ERK Pathway

WANG Xia; WANG Lijuan; NIU Xiaoying; FAN Zekun; NIU Yuanyuan; MAO Lanfang; WANG Longde

doi:10.13288/j.11-2166/r.2024.10.013

您当前的位置：

首页 >

文章列表页 >

Exploring the Effect of Medication-Containing Serum of Pingwei Capsules （平胃胶囊） on MNNG-Induced Inflammatory Cancer Transformation of Human Gastric Mucosal Epithelial Cells Based on Raf/MEK/ERK Pathway

更新时间：2024-05-13

- Exploring the Effect of Medication-Containing Serum of Pingwei Capsules （平胃胶囊） on MNNG-Induced Inflammatory Cancer Transformation of Human Gastric Mucosal Epithelial Cells Based on Raf/MEK/ERK Pathway
- Journal of Traditional Chinese Medicine Vol. 65, Issue 10, Pages: 1056-1062(2024)
- 作者机构：
  
  1.甘肃中医药大学，甘肃省兰州市定西东路35号，730000
  2.敦煌医学与转化教育部重点实验室
  3.甘肃中医药大学附属医院
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2024.10.013
  CLC：
- Received：05 December 2023，
  
  Revised：2024-01-27，
  
  Published：17 May 2024
- 稿件说明：
移动端阅览
汪霞,王丽娟,牛小英等.基于Raf/MEK/ERK通路探讨平胃胶囊含药血清对MNNG诱导的人胃黏膜上皮细胞炎癌转化的影响[J].中医杂志,2024,65(10):1056-1062.

WANG Xia,WANG Lijuan,NIU Xiaoying,et al.Exploring the Effect of Medication-Containing Serum of Pingwei Capsules （平胃胶囊） on MNNG-Induced Inflammatory Cancer Transformation of Human Gastric Mucosal Epithelial Cells Based on Raf/MEK/ERK Pathway[J].Journal of Traditional Chinese Medicine,2024,65(10):1056-1062.
汪霞,王丽娟,牛小英等.基于Raf/MEK/ERK通路探讨平胃胶囊含药血清对MNNG诱导的人胃黏膜上皮细胞炎癌转化的影响[J].中医杂志,2024,65(10):1056-1062. DOI： 10.13288/j.11-2166/r.2024.10.013.

WANG Xia,WANG Lijuan,NIU Xiaoying,et al.Exploring the Effect of Medication-Containing Serum of Pingwei Capsules （平胃胶囊） on MNNG-Induced Inflammatory Cancer Transformation of Human Gastric Mucosal Epithelial Cells Based on Raf/MEK/ERK Pathway[J].Journal of Traditional Chinese Medicine,2024,65(10):1056-1062. DOI： 10.13288/j.11-2166/r.2024.10.013.

摘要

目的

从快速加速纤维肉瘤/丝裂原活化的细胞外信号调节激酶/细胞外信号调节激酶（Raf/MEK/ERK）通路探讨平胃胶囊治疗慢性萎缩性胃炎的可能作用机制。

方法

15只SD大鼠随机分为空白组5只、平胃胶囊组10只，空白组大鼠给予1 ml/100 g的生理盐水灌胃，平胃胶囊组大鼠给予平胃胶囊混悬液0.63 g/（kg·d）灌胃，连续3天后采集血清。采用N-甲基-N-硝基-亚硝基胍（MNNG）诱导人胃黏膜上皮细胞GES-1建立癌前病变细胞模型。将造模成功细胞分为对照组（10%胎牛血清）、空白血清组（10%胎牛血清+10%空白血清）、含药血清组（平胃胶囊含药血清），采用CCK-8法筛选平胃胶囊含药血清干预体积分数及时间。将人胃黏膜上皮细胞GES-1分为正常组、模型组、空白血清组、含药血清组、U0126组、联合组，每组3个复孔。除空白组外其余各组细胞造模成功后，正常组和模型组加入等体积的胎牛血清，空白血清组加入等体积的空白血清，含药血清组加入筛选确认的体积分数的平胃胶囊含药血清，U0126组给予10 μmol/L的丝裂原活化的细胞外信号调节激酶1（MEK1）抑制剂U0126，联合组给予等体积的平胃胶囊含药血清+10 μmol/L的U0126。培养筛选确认的时间后，ELISA法检测细胞上清中白细胞介素6（IL-6）水平，免疫荧光检测细胞中IL-6、MEK1的表达，RT-qPCR法检测细胞IL-6、Raf、MEK1、ERK mRNA表达，Western Blot法检测细胞IL-6、Raf、MEK1、ERK1/2蛋白表达。

结果

选取5.35%的体积分数、干预48 h的平胃胶囊含药血清进行后续实验。与正常组比较，模型组、空白血清组细胞上清中IL-6含量，细胞中IL-6、Raf、MEK1、ERK mRNA及IL-6、Raf、MEK1、ERK1/2蛋白表达均升高（

＜0.01）。与模型组比较，含药血清组、U0126组、联合组上述各指标均下降（

＜0.05或

＜0.01）。与含药血清组比较，联合组细胞中IL-6、MEK1表达，IL-6、Raf、MEK1、ERK mRNA表达，IL-6、Raf、MEK1、ERK1/2蛋白表达均降低（

＜0.05或

＜0.01）。与U0126组比较，联合组细胞中IL-6表达降低，IL-6、MEK1、ERK1/2蛋白表达降低（

＜0.05或

P＜

0.01）。

结论

平胃胶囊含药血清可能通过抑制Raf/MEK/ERK通路改善GES-1细胞的炎癌转化，从而发挥治疗慢性萎缩性胃炎的作用。

Abstract

Objective

To explore the possible mechanism of

Pingwei Capsules

（平胃胶囊） for chronic atrophic gastritis from rapidly accelerated fibrosarcoma / mitogen-activated protein kinase /extracellular-signal-regulated kinase

（Raf/MEK/ERK） pathway that influences the activation of fibrosarcoma protein/mitogen.

Methods

Fifteen SD rats were randomly divided into 5 rats in the blank group and 10 rats in

Pingwei Capsules

group. The rats in the blank group were given 1 ml/100 g of saline by gavage， and the rats in

Pingwei Capsules

group were given 0.63 g/（kg·d） of

Pingwei Capsule

suspension by gavage， and serum was collected for 3 consecutive days. N-methyl-N'-nitro-N-nitrosoguanidine （MNNG） was used to induce human gastric mucosal epithelial cells GES-1 to establish a precancerous lesion cell model. The successful cells were divided into control group （10% fetal bovine serum）， blank serum group （10% fetal bovine serum plus 10% blank serum）， and medication-containing serum group （serum with medication of

Pingwei Capsule

）， and the volume fraction and time of intervention of

Pingwei Capsule

-containing serum were screened by CCK-8 assay. Human gastric mucosal epithelial cells GES-1 were divided into normal group， model group， blank serum group， medication-containing serum group， U0126 group， and combined group， with 6 replicate wells in each group. After successful modelling of the cells in all groups except the blank group， an equal volume of fetal bovine serum was added to the normal and model groups， an equal volume of blank serum was added to the blank serum group， a screening volume fraction of

Pingwei Capsule

-containing serum was added to

Pingwei Capsule

group， a 10 μmol/L mitogen-activated extracellular signal regulated kinase 1 （MEK1） inhibitor U0126 was administered in the U0126 group， an equal dose of

Pingwei Capsule

-containing serum plus 10 μmol/L of U0126 was administered to the combined group. After the selected incubation time， the level of interleukin 6 （IL-6） was detected in the cells by ELISA， the expression of IL-6 and MEK1 was detected by immunofluorescence， a

nd the expression of IL-6， Raf， MEK1， and ERK mRNA was detected by RT-qPCR， and the expression of IL-6， Raf， MEK1， and ERK mRNA in the cells was detected by Western blot.

Results

The 5.35% volume fraction， 48 h intervention of

Pingwei Capsule

-containing serum was selected for subsequent experiments. Compared with the normal group， the IL-6 content in cell supernatants and the expression of IL-6， Raf， MEK1， ERK mRNA and ERK1/2 proteins in cells increased in the model group and blank serum group （

＜0.01）. Compared with the model group， all of the above indexes were improved in medication-containing serum group， U0126 group， and combined group （

＜0.05 or

＜0.01）. Compared with medication-containing serum group， the expression of IL-6， MEK1 expression， the expression of IL-6， Raf， MEK1 and ERK mRNA， and the expression of IL-6， Raf， MEK1 and ERK1/2 proteins reduced in the cells of combined group （

＜0.05 or

＜0.01）. Compared with the U0126 group， IL-6 expression reduced and IL-6， MEK1 and ERK1/2 protein expression reduced in cells of combined group （

＜0.05 or

＜0.01）.

Conclusion

The

Pingwei Capsule

-containing serum may play a role in the treatment of chronic atrophic gastritis by improving the inflammation-cancer transformation of GES-1 cells through inhibiting the Raf/MEK/ERK pathway.

关键词

Keywords

references

李军祥，陈誩，吕宾，等 . 慢性萎缩性胃炎中西医结合诊疗共识意见（2017年）［J］. 中国中西医结合消化杂志， 2018 ， 26 （ 2 ）： 121 - 131 .

CORREA P ， SHIAO Y . Phenotypic and genotypic events in gastric carcinogenesis ［J］. Cancer Research ， 1994 ， 54 （ 7 Suppl ）： 1941s - 1943s .

刘伟，倪家慧，张丹，等 . 中医药调控胃“炎癌转化”癌前微环境的思考与策略［J］. 中华中医药杂志， 2023 ， 38 （ 4 ）： 1431 - 1435 .

WU PK ， BECKER A ， PARK JI . Growth inhibitory signaling of the Raf/MEK/ERK pathway ［J］. Int J Mol Sci ， 2020 ， 21 （ 15 ）： 5436 .

杨丹芬，谢圆媛，康睿 . 血管紧张素Ⅱ通过Raf/MEK/ERK通路对PDGF诱导的气道上皮细胞转分化的影响［J］. 河北医药， 2022 ， 44 （ 8 ）： 1141 - 1146 .

杨少军，汪龙德，张晶，等 . 平胃胶囊治疗慢性萎缩性胃炎癌前病变45例临床研究［J］. 中医杂志， 2014 ， 55 （ 20 ）： 1745 - 1749 .

王琦，杨少军 . 平胃胶囊治疗慢性萎缩性胃炎癌前病变的临床研究［J］. 时珍国医国药， 2013 ， 24 （ 11 ）： 2685 - 2686 .

王四兵，汪龙德，刘晓燕，等 . 平胃胶囊治疗慢性萎缩性胃炎伴肠化生45例［J］. 中国中医药信息杂志， 2013 ， 20 （ 9 ）： 75 - 76 .

何子才，庞国学，汪龙德 . 平胃胶囊治疗慢性萎缩性胃炎疗效分析［J］. 实用中医药杂志， 2014 ， 30 （ 8 ）： 689 - 689，690 .

赵伟，孙国志 . 不同种实验动物间用药量换算［J］. 畜牧兽医科技信息， 2010 （ 5 ）： 52 - 53 .

刘雪，孙向红，戚欣 . MNNG诱导GES-1细胞恶性转化细胞模型的建立［J］. 齐鲁医学杂志， 2013 ， 28 （ 2 ）： 115 - 116，120 .

孙振卿，闫慧明，贾彬，等 . 从形态及功能研究MNNG诱导GES-1永生化细胞的恶性转化［J］. 中国现代医学杂志， 2012 ， 22 （ 22 ）： 14 - 18 .

闫慧明，孙振卿，贾彬，等 . 研究MNNG诱导GES-1发生恶性转化后PTEN蛋白的变化意义［J］. 中国医疗前沿， 2011 ， 6 （ 23 ）： 6 ， 50 .

徐星榕，白菊萍，边芸，等 . CK、MUC5ac、Ki67表达与胃癌癌前病变的关系及对胃癌早期诊断的价值［J］. 国际检验医学杂志， 2023 ， 44 （ 12 ）： 1522 - 1525 .

李欣瑜，刘俊宏，户艳玲，等 . 国医大师周信有教授辨治消化性溃疡的经验总结［J］. 时珍国医国药， 2023 ， 34 （ 4 ）： 966 - 967 .

郭金惠，武彩娥 . 胃癌前病变时血浆白细胞介素2和白细胞介素6的变化［J］. 标记免疫分析与临床， 1996 （ 1 ）： 58 .

DEGIRMENCI U ， WANG M ， HU J . Targeting aberrant RAS/RAF/MEK/ERK signaling for cancer therapy ［J］. Cells ， 2020 ， 9 （ 1 ）： 198 .

ULLAH R ， YIN Q ， SNELL AH ， et al . RAF-MEK-ERK pathway in cancer evolution and treatment ［J］. Semin Cancer Biol ， 2022 ， 85 （ 10 ）： 123 - 154 .

YOSIFOV DY ， REUFSTECK C ， KONSTANTINOV SM ， et al . Interleukin-6， osteopontin and Raf/MEK/ERK signaling modulate the sensitivity of human myeloma cells to alkylphosphocholines ［J］. Leuk Res ， 2012 ， 36 （ 6 ）： 764 - 772 .

LI S ， TIAN J ， ZHANG H ， et al . Down-regulating IL-6/GP130 targets improved the anti-tumor effects of 5-fluorouracil in colon cancer ［J］. Apoptosis ， 2018 ， 23 （ 5 ）： 356 - 374 .

郭敏，王晓鸽，杨国红，等 . 健脾活瘀方抑制IL-6/JAK1/STAT3信号通路抗胃癌前病变的机制研究［J］. 中医药信息， 2020 ， 37 （ 3 ）： 44 - 49 .

查看原文

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Exploration on the Pathogenesis of Chronic Atrophic Gastritis from the Perspectives of Stasis，Toxins，and Constraints

A Randomized Controlled Trial of Jianpi Jiedu Formulation （健脾解毒方） in the Treatment of Kimura-Takemoto Closed-Type Atrophic Gastritis with Spleen Deficiency and Damp-Heat Syndrome

SHAN Zhaowei's Experience in Treating Chronic Atrophic Gastritis with the Three-Unblocking Method

Prevention and Treatment of Inflammation-Cancer Transformation in Chronic Atrophic Gastritis Based on Collateral Disease Theory

Clinical Study on the Treatment of 70 Cases Chronic Atrophic Gastritis with Intestinal Metaplasia Using Xianglian Huazhuo Granules （香连化浊颗粒）：A Randomized，Double-Blind，Placebo-Controlled Trial

Related Author

Tai ZHANG

Beihua ZHANG

Xiangxue MA

Fengyun WANG

Ping WANG

Xudong TANG

GU Bingyan

QIN Danping

Related Institution

China Academy of Chinese Medical Sciences

Xiyuan Hospital，China Academy of Chinese Medical Sciences

Graduate School，China Academy of Chinese Medical Sciences

Huzhou Central Hospital

Zhejiang Provincial Hospital of Traditional Chinese Medicine

Address：No.16, Nanxiao street, Dongzhimen, Dongcheng District, Beijing Postal code：100700
Tel：010-64035632 Email：jtcmcn@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备09064830号-19 京公网安备11010102007558号
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰

Exploring the Effect of Medication-Containing Serum of Pingwei Capsules （平胃胶囊） on MNNG-Induced Inflammatory Cancer Transformation of Human Gastric Mucosal Epithelial Cells Based on Raf/MEK/ERK Pathway

DOI：10.13288/j.11-2166/r.2024.10.013

摘要

Abstract

关键词

Keywords

references