Effect of modified Pentongling Formula on TRPV1/ERK1/2/p⁃CREB pathway in endometriosis model rats

LIU Xiaoqing; YU Siyun; XIA Yu; FU Jinrong

doi:10.16306/j.1008-861x.2024.03.008

您当前的位置：

首页 >

文章列表页 >

Effect of modified Pentongling Formula on TRPV1/ERK1/2/p⁃CREB pathway in endometriosis model rats

Experiment Research | 更新时间：2024-05-31

- Effect of modified Pentongling Formula on TRPV1/ERK1/2/p⁃CREB pathway in endometriosis model rats
- Academic Journal of Shanghai University of Traditional Chinese Medicine Vol. 38, Issue 3, Pages: 50-56(2024)
- 作者机构：
  
  1.上海中医药大学龙华临床医学院（上海　200032）
  2.上海中医药大学附属龙华医院妇科（上海　200032）
- 作者简介：
- 基金信息：
- DOI：10.16306/j.1008-861x.2024.03.008
  CLC：
- Published：25 May 2024，
  
  Received：18 May 2023，
  
  Revised：28 March 2024，
扫描看全文
刘晓庆,余思云,夏玉等.加味盆痛灵方对子宫内膜异位症模型大鼠TRPV1/ERK1/2/p⁃CREB通路的影响[J].上海中医药大学学报,2024,38(03):50-56.

LIU Xiaoqing,YU Siyun,XIA Yu,et al.Effect of modified Pentongling Formula on TRPV1/ERK1/2/p⁃CREB pathway in endometriosis model rats[J].Academic Journal of Shanghai University of Traditional Chinese Medicine,2024,38(03):50-56.
刘晓庆,余思云,夏玉等.加味盆痛灵方对子宫内膜异位症模型大鼠TRPV1/ERK1/2/p⁃CREB通路的影响[J].上海中医药大学学报,2024,38(03):50-56. DOI： 10.16306/j.1008-861x.2024.03.008.

LIU Xiaoqing,YU Siyun,XIA Yu,et al.Effect of modified Pentongling Formula on TRPV1/ERK1/2/p⁃CREB pathway in endometriosis model rats[J].Academic Journal of Shanghai University of Traditional Chinese Medicine,2024,38(03):50-56. DOI： 10.16306/j.1008-861x.2024.03.008.

摘要

目的

基于瞬时受体电位香草酸亚型1/细胞外信号调节激酶1/2/磷酸化环磷酸腺苷反应元件结合蛋白（TRPV1/ERK1/2/p-CREB）信号通路探讨加味盆痛灵方对子宫内膜异位症（EMs）疼痛模型大鼠的镇痛机制。

方法

采用自体内膜移植法建立SD大鼠EMs疼痛模型。造模成功的大鼠采用完全随机设计法分为模型组、TRPV1抑制剂组（CPZ组）、加味盆痛灵方组（JWPTL组）和TRPV1抑制剂+加味盆痛灵方组（CPZ+JWPTL组），每组8只；另设假手术组（

=8）。JWPTL组和CPZ+JWPTL组大鼠自造模成功后第1天起给予7.77 g/kg加味盆痛灵方灌肠，1次/d，其余大鼠灌肠给予等体积0.9% NaCl溶液，连续干预28 d。CPZ组和CPZ+JWPTL组自造模成功后第21天起鞘内注射辣椒平（CPZ），10 μL/d，1次/d；其余大鼠鞘内注射等体积助溶剂10%二甲基亚砜（DMSO），连续干预7 d。采用“Up＆Down”法测量大鼠的50%机械缩足反应阈值（PMWT），Western blot和RT-qPCR法检测脊髓组织中TRPV1、ERK1/2、p-CREB蛋白和mRNA的表达水平，ELISA法观察即刻早期基因c-fos蛋白的表达情况。

结果

给药前，与假手术组比较，各造模组大鼠50% PMWT均明显降低（

0.05）；给药28 d后，CPZ组、JWPTL组和CPZ+JWPTL组50% PMWT较模型组显著升高（

0.05），CPZ+JWPTL组50% PMWT较CPZ组显著升高（

0.05）。模型组TRPV1、ERK1/2、CREB蛋白和mRNA的表达水平较假手术组显著升高（

0.01，

0.001），模型组c-fos蛋白表达水平较假手术组亦显著升高（

0.001）；与模型组比较，TRPV1抑制剂CPZ和加味盆痛灵方可不同程度下调TRPV1、ERK1/2、CREB蛋白和mRNA表达水平（

0.05，

0.01），且CPZ+JWPTL组效果最优。与模型组比较，JWPTL组和CPZ+JWPTL组c-fos蛋白表达水平明显降低（

0.001）。相关性分析显示，TRPV1、ERK1/2、CREB蛋白和mRNA表达水平及c-fos蛋白表达水平均与50% PMWT呈负相关。

结论

加味盆痛灵方可能通过干预TRPV1/ERK1/2/p-CREB通路及c-fos的表达缓解EMs疼痛。

Abstract

Objective： To explore the analgesic mechanism of modified Pengtongling Formula on endometriosis （EMs） pain rats based on the transient receptor potential vanillic acid subtype 1/extracellular signal regulated kinase 1/2/phosphorylated cyclic adenosine monophosphate responsive element binding protein （TRPV1/ERK1/2/p-CREB） signaling pathway.

Method

A rat EMs pain model was established by autologous endometrial transplantation. The successfully modeled rats were divided into model group， TRPV1 inhibitor group （CPZ group）， modified Pentongling Formula group （JWPTL group）， and TRPV1 inhibitor+modified Pentongling Formula group （CPZ+JWPTL group） by a completely randomized design method， with 8 rats in each group； another sham surgery group was set up （

=8）. The rats in JWPTL group and CPZ+JWPTL group were given 7.77 g/kg modified Pentongling Formula enema once a day from the first day after successful modeling， and the other rats were given equal volume 0.9% NaCl solution for continuous intervention for 28 d. CPZ group and CPZ+JWPTL group received intrathecal injection of capsazepine （CPZ）， 10 μL/d， once a day from the 21st day after successful modeling； The other rats were injected intrathecally with 10% dimethyl sulfoxide （DMSO）， which was treated continuously for 7 d. The "Up

Down" method was used to measure the 50% paw mechanical withdrawal threshold （PMWT） of rats. Western blot and RT-qPCR methods were used to detect the expression levels of TRPV1， ERK1/2， p-CREB proteins and mRNA in spinal cord tissues. ELISA method was used to observe the expression of the immediate early gene c-fos protein.

Results

Before administration， compared with the sham surgery group， the 50% PMWT of rats in each modeling group

was significantly reduced （

0.05）. After 28 d of administration， compared with the model group， the 50% PMWT of the CPZ group， JWPTL group， and CPZ+JWPTL group was significantly increase （

0.05）； Compared with the CPZ group， the 50% PMWT of the CPZ+JWPTL group was significantly increase （

0.05）. Compared with the sham surgery group， the protein and mRNA expression levels of TRPV1， ERK1/2 and CREB in the model group was significantly increased （

0.01，

0.001）， and the expression level of c-fos protein in the model group was also significantly increased （

0.001）； Compared with the model group， TRPV1 inhibitors CPZ and modified Pentongling Formula could down-regulate the protein and mRNA expression levels of TRPV1， ERK1/2 and CREB in different degrees （

0.05，

0.01）， and CPZ+JWPTL group has the best effect. Compared with the model group， the expression level of c-fos protein was significantly decreased. The JWPTL group and CPZ+JWPTL group （

0.001）. Correlation analysis showed that the expression levels of TRPV1， ERK1/2， CREB protein and mRNA as well as c-fos protein were negatively correlated with 50% PMWT.

Conclusion

Modified Pengtongling Formula may alleviate EMs pain by intervening in the TRPV1/ERK1/2/p-CREB pathway and the expression of c-fos.

关键词

加味盆痛灵方子宫内膜异位症疼痛TRPV1/ERK1/2/p-CREB通路c-fos

Keywords

modified Pentongling FormulaendometriosispainTRPV1/ERK1/2/p-CREB pathwayc-fos

references

TAYLOR H S，KOTLYAR A M，Flores VA. Endometriosis is a chronic systemic disease：clinical challenges and novel innovations［J］. Lancet， 2021， 397： 839-852.

MCNAMARA H C，FRAWLEY H C，DONOGHUE J F，et al. Peripheral，Central，and Cross Sensitization in Endometriosis-Associated Pain and Comorbid Pain Syndromes［J］. Front Reprod Health， 2021， 3： 729642.

付金荣，沈宇凤，张俊洁，等. 盆痛灵方灌肠治疗慢性盆腔疼痛异病同治的临床评价［J］. 上海中医药大学学报， 2018， 32（6）： 34-39.

FU J R，SHEN Y F，ZHANG J J，et al. Clinical evaluation on chronic pelvic pain of different diseases treated with Pentongling Formula coloclysis［J］. Academic Journal of Shanghai University of Traditional Chinese Medicine， 2018， 32（6）： 34-39.

温欢欢，付金荣，徐英，等. 盆痛灵方对子宫内膜异位症模型大鼠TRPV1表达的影响［J］. 中国中西医结合杂志， 2020， 40（9）： 1070-1074.

WEN H H，FU J R，XU Y，et al. Effect of Pentongling Formula on the Expression of TPPV1 in Endometriosis Model Rats［J］. Chinese Journal of Integrated Traditional and Western Medicine， 2020， 40（9）： 1070-1074.

章仕雯，付金荣，徐英，等. 盆痛灵方对子宫内膜异位症模型大鼠镇痛作用机制的研究［J］. 中国中西医结合杂志， 2021， 41（12）： 1461-1467.

ZHANG S W，FU J R，XU Y，et al. Analgesic Mechanism of Pentonglin Recipe on Model Rats with Endometriosis［J］. Chinese Journal of Integrated Traditional and Western Medicine， 2021， 41（12）： 1461-1467.

葛曼，付金荣，唐虹，等. 加味盆痛灵对子宫内膜异位症模型大鼠血清及腹腔液IL-1β、NGF的影响［J］. 上海中医药杂志，2013， 47（3）： 79-81.

GE M，FU J R，TANG H，et al. Effect of ＂Jiawei Pentongling＂ on IL-1β and NGF in serum and peritoneal fluid in EMs rats［J］. Shanghai Journal of Traditional Chinese Medicine， 2013， 47（3）： 79-81.

ZHEN W，ZHEN H，WANG Y，et al. Mechanism of ERK/CREB pathway in pain and analgesia［J］. Front Mol Neurosci， 2023， 16： 1156674.

BERKLEY KI，DMITRIEVA N，CURTIS K S，et al. Innervation of ectopic endometrium in a rat model of endometriosis［J］. Proc Natl Acad Sci U S A， 2004， 101（30）： 11094-11098.

汤家铭，陈民利. 医学实验动物学［M］. 北京：中国中医药出版社，2012： 7.

TANG J M，CHEN M L. Medical Experimental Zoology［M］. Beijing：China Press of Traditional Chinese Medicine， 2012： 7.

CHAPLAN S R，BACH F W，POGREL J W，et al. Quantitative assessment of tactile allodynia in the rat paw［J］. J Neurosci Methods， 1994， 53（1）： 55-63.

夏玉，李宣儒，余思云，等.基于PI3K/Akt信号通路探讨加味盆痛灵方对子宫内膜异位症模型大鼠的镇痛作用机制［J］. 上海中医药大学学报， 2023， 37（1）： 54-61.

XIA Y，LI X R，YU S Y，et al. Exploring mechanism of modified Pentongling Formula on analgesic in endometriosis model rats based on PI3K/Akt signaling pathway［J］. Academic Journal of Shanghai University of Traditional Chinese Medicine， 2023， 37（1）： 54-61.

唐虹，葛曼，付金荣. 付金荣论治子宫内膜异位症疼痛经验撷要［J］. 山西中医， 2013， 29（7）： 6-7.

TANG H，GE M，FU J R. Summary of Fu Jinrong's Experience in Treating Pain in Endometriosis［J］. Shanxi Journal of Traditional Chinese Medicine， 2013， 29（7）： 6-7.

AGHAZADEH TABRIZI M，BARALDI P G，BARALDI S，et al. Medicinal Chemistry，Pharmacology，and Clinical Implications of TRPV1 Receptor Antagonists［J］. Med Res Rev， 2017， 37（4）： 936-983.

余思云，刘晓庆，付金荣，等. 加味盆痛灵对子宫内膜异位症疼痛大鼠背根神经节神经元动作电位的影响［J］. 中国中西医结合杂志， 2023： 1-6.

YU S Y，LIU X Q，FU J R，et al. Effect of Jiawei Pentongling on Action Potential of Dorsal Root Ganglion Neurons in Rats with Endometriosis Pain［J］. Chinese Journal of Integrated Traditional and Western Medicine， 2023： 1-6.

MADDERN J，GRUNDY L，CASTRO J，et al. Pain in Endometriosis［J］. Front Cell Neurosci， 2020， 14： 590823.

VALTSCHANOFF J G， RUSTIONI A， GUO A， et al. Vanilloid receptor VR1 is both presynaptic and postsynaptic in the superficial laminae of the rat dorsal horn［J］. J Comp Neurol，2001， 436（2）： 225-235.

WOOLF C J. Central sensitization： implications for the diagnosis and treatment of pain［J］. Pain， 2011， 152（3 Suppl）： S2-S15.

GAO Y J， JI R R. c-Fos and pERK， which is a better marker for neuronal activation and central sensitization after noxious stimulation and tissue injury？［J］. Open Pain J， 2009， 2： 11-17.

SHEN C P， TSIMBERG Y， SALVADORE C， et al. Activation of Erk and JNK MAPK pathways by acute swim stress in rat brain regions［J］. BMC Neurosci， 2004， 5： 36.

梁文娜，阮璐薇，张鹏横，等. 柴胡疏肝散调节围绝经期肝郁证大鼠ERK1/2-CREB-BDNF信号通路的机制［J］. 中华中医药杂志， 2022， 37（6）： 3147-3152.

LIANG W N，RUAN L W，ZHANG P H，et al. Mechanism of Chaihu Shugan Powder on ERK1/2-CREB-BDNF signaling pathway in perimenopausal liver-stagnation pattern rats

J］. China Journal of Traditional Chinese Medicine and Pharmacy，2022， 37（6）： 3147-3152.

杨翠珠，李诗琪，张润恒，等. 石菖蒲挥发油对炎性痛大鼠脊髓中ERK1/2/CREB/c-fos信号通路的影响［J］. 神经解剖学杂志， 2022， 38（1）： 15-20.

YANG C Z，LI S Q，ZHANG Y H，et al. Effects of volatile oil from Acori graminei Rhizoma on ERK1/2/CREB/c-fos signaling pathway in the spinal cord of rats with inflammatory pain［J］. Chinese Journal of Neuroanatomy， 2022， 38（1）： 15-20.

KAWASAKI Y，KOHNO T，ZHUANG Z Y，et al. Ionotropic and metabotropic receptors， protein kinase A，protein kinase C，and Src contribute to C-fiber-induced ERK activation and cAMP response element-binding protein phosphorylation in dorsal horn neurons，leading to central sensitization［J］. J Neurosci， 2004， 24（38）： 8310-8321.

汪逸纯. 基于COX-2/PGE2信号通路探讨温肾消癥汤治疗子宫内膜异位症模型小鼠疼痛的机制［D］. 南京：南京中医药大学， 2022.

WANG Y C. Mechanism of Wenshen Xiaozheng Decoction relieving pain in endometriosis mouse model based on COX-2/PGE2 signaling pathway［D］. Nanjing： Nanjing University of Chinese Medicine， 2022.

徐秀琦，李光，张广钦. 天然产物中镇痛活性成分对TRPV1通道的调控作用［J］. 药学研究， 2022， 41（4）： 259-263.

XU X Q，LI G，ZhANG G Q. Regulation of analgesic active components in natural products on TRPV1 channel［J］. Journal of Pharmaceutical Research， 2022， 41（4）： 259-263.

更多指标>

Views

下载量

CSCD

CNKI被引量

Alert me when the article has been cited

提交

Tools

Publicity Resources

Exploring mechanism of modified Pentongling Formula on analgesic in endometriosis model rats based on PI3K/Akt signaling pathway

Academic thought and clinical experience of Chen's gynecology in Shanghai

Discussion on the effect mechanisms of acupuncture combined with Xishao Decoction in the treatment of pain from the theory of yin-yang coordination

Effects of Tongjingning Granule on expressions of ICAM-1,MMP-9 and VEGF in eutopic and ectopic endometrium of rats with endometriosis

Effects of Hongteng Formula on mesenchymal-epithelial transformation in rats with endometriosis

Related Author

LI Xuanru

FU Jinrong

XU Weina

CHEN Yijia

YOU Min

WU Yimei

CHEN Shuangjia

SHI Zhaofeng

Related Institution

Department of Gynecology， Longhua Hospital， Shanghai University of Traditional Chinese Medicine

Zhoujiadu Community Health Service Center， Pudong New Area

Pujin Community Health Service Center， Minhang District

Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine

Dongzhimen Hospital, Beijing University of Chinese Medicine

⁰