Mechanism of Modified Guizhi Fulingwan in Regulating Mitochondrial Apoptosis Pathway Through PTEN/PI3K/Akt Pathway to Prevent Colorectal Adenoma in Mice
10.13422/j.cnki.syfjx.20240721
- VernacularTitle:加味桂枝茯苓丸通过PTEN/PI3K/Akt通路调控线粒体凋亡途径预防小鼠结直肠腺瘤的作用机制
- Author:
Sujie JIA
1
;
Longhui LIU
1
;
Yifan ZHANG
1
;
Xiaomeng LANG
2
;
Jianping LIU
2
;
Xin KANG
2
;
Chaodi SUN
1
;
Jingyuan LIU
1
Author Information
1. Hebei University of Chinese Medicine， Shijiazhuang 050091，China
2. Hebei Provincial Hospital of Traditional Chinese Medicine， Shijiazhuang 050011，China
- Publication Type:Journal Article
- Keywords:
modified Guizhi Fulingwan;
colorectal adenoma;
phosphatase and tension homolog/phosphatidylinositol 3-kinase /protein kinase B （PTEN/PI3K/Akt）;
mitochondrial pathway;
apoptosis
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2024;30(19):10-20
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the effect and mechanism of modified Guizhi Fulingwan in preventing colorectal adenoma （CRA） in mice by regulating mitochondrial apoptosis pathway through the regulation of the phosphatase and tensin homolog （PTEN）/phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） pathway. MethodSixty SPF-grade male C57BL/6 mice were randomly divided into six groups： Normal group， model group， low， medium， and high dose groups of modified Guizhi Fulingwan （13， 26， 52 g·kg-1·d-1）， and positive control aspirin group （0.015 g·kg-1·d-1）. A mouse model of CRA was chemically induced using azoxymethane （AOM） and dextran sulfate sodium （DSS）. During the modeling process， mice received modified Guizhi Fulingwan or aspirin. Body weight of mice was measured weekly during the treatment. After 9 weeks， the number of adenomas formed was observed. Serum levels of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and interleukin-1β （IL-1β） were determined by enzyme-linked immunosorbent assay （ELISA）. Hematoxylin-eosin （HE） staining was used to observe the histopathologic changes in adenoma tissues. The expression of Cyclin D1 and proliferative nuclear antigen （Ki67） was detected by immunohistochemistry （IHC）. Terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） was used to assess the apoptosis in adenoma tissues. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to observe the mRNA and protein expression levels of PTEN， PI3K， Akt， phosphorylated PI3K （p-PI3K）， phosphorylated Akt （p-Akt）， B-cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax）， cytochrome C （Cyt C）， Caspase-9， and caspase-3. ResultCompared with the normal group， the model group showed no significant change in body weight from week 1 to week 2， but a significant decrease from week 3 to week 9 （P<0.05，P<0.01）. The colorectal length was significantly shortened， and the colorectal weight increased with visible varying sized tumor-like protrusions on the mucosal surface （P<0.01）. Serum levels of TNF-α， IL-6， and IL-1β were elevated （P<0.01）. Histopathology showed disordered epithelial gland structure， elongated nuclei with pathological mitosis， and numerous lymphocytic infiltrations in the lamina propria. The positive expression rates of Cyclin D1 and Ki67 were significantly increased （P<0.01）， while the apoptosis rate of adenoma cells was significantly decreased （P<0.01）. Expression levels of PI3K， Akt， Bcl-2 mRNA and proteins， as well as p-PI3K and p-Akt proteins， were significantly increased （P<0.01）， whereas PTEN， Bax， Cyt C， Caspase-9， and Caspase-3 mRNA and protein levels were significantly decreased （P<0.05， P<0.01）. Compared with the model group， all drug treatment groups showed an increase in body weight （P<0.01）， decreased intestinal weight， increased colorectal length， reduced number of adenomas significantly （P<0.05， P<0.01）， and significantly lowered serum levels of TNF-α， IL-6， and IL-1β （P<0.01）. Histopathology indicated improved glandular structure and reduced neutrophil infiltration in the mucosal lamina propria. The positive expression rates of Cyclin D1 and Ki67 significantly decreased （P<0.01）， while the apoptosis rate of adenoma cells significantly increased （P<0.05， P<0.01）. Expression levels of PI3K， Akt， Bcl-2 mRNA and proteins， and p-PI3K and p-Akt proteins were significantly reduced （P<0.05， P<0.01）， while PTEN， Bax， Cyt C， Caspase-9， and Caspase-3 mRNA and protein levels significantly increased （P<0.05， P<0.01）. The high-dose modified Guizhi Fulingwan group exhibited the most significant intervention effects. ConclusionModified Guizhi Fulingwan may prevent CRA in mice by regulating the PTEN/PI3K/Akt signaling pathway and inducing the mitochondrial apoptosis pathway.
