Effect of Yishen Huayu Prescription on Autophagy of Transdifferentiated TCMK-1 Cells Based on SIRT1/FoxO1 Pathway
10.13422/j.cnki.syfjx.20242037
- VernacularTitle:基于SIRT1/FoxO1通路探讨益肾化瘀方对转分化TCMK-1细胞自噬的影响
- Author:
Qingru LI
1
;
Linqi ZHANG
2
;
Binyi LI
3
;
Zihao GE
2
Author Information
1. The Eighth Clinical Medical College of Guangzhou University of Chinese Medicine，Foshan 528000，China
2. The First Affiliated Hospital of Henan University of Chinese Medicine，Zhengzhou 450000，China
3. Shenzhen Bao'an Authentic Traditional Chinese Medicine Therapy Hospital，Shenzhen 518100，China
- Publication Type:Journal Article
- Keywords:
Yishen Huayu prescription;
TCMK-1 cell;
silent information regulator 1 （SIRT1）/acetylated forkhead box protein O1 （FoxO1） pathway;
transdifferentiation;
autophagy
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2025;31(12):91-99
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the protective effect and underlying mechanisms of Yishen Huayu prescription （YSHYP） on transdifferentiation of mouse renal tubular epithelial cells （TCMK-1） induced by transforming growth factor-β1 （TGF-β1）. MethodsA transdifferentiation model was established by treating TCMK-1 cells with 10 μg·L-1 TGF-β1. Experimental groups were established using 2 μmol·L-1 silent information regulator 1 （SIRT1） inhibitor EX527. These included the blank group， model group， YSHYP group （treated with 10% YSHYP-medicated serum）， valsartan group （treated with 10% valsartan-medicated serum）， EX527+TGF-β1 group， EX527+YSHYP group， and EX527 group. Immunofluorescence was used to detect the protein localization of α-smooth muscle actin （α-SMA）， E-cadherin， and microtubule-associated protein light chain 3 （LC3）. Western blot and Real-time polymerase chain reaction （Real-time PCR） were used to assess the expression of proteins and mRNA related to transdifferentiation， autophagy， and associated signaling pathways. ResultsThe results from Real-time PCR and Western blot indicate that compared with those in the blank group， expression levels of α-SMA， ubiquitin-binding protein p62 （p62）， and acetylated forkhead box protein O1（Ac-FoxO1） were significantly increased in the model group， EX527+TGF-β1 group， and EX527 group （P<0.01）. Compared with that in the model group， the expression of α-SMA and p62 were significantly downregulated in the YSHYP and valsartan groups （P<0.05， P<0.01）. Ac-FoxO1 protein levels were significantly reduced in the YSHYP group （P<0.05）， while the valsartan group showed no significant changes in Ac-FoxO1 levels. Compared with the YSHYP group， the valsartan group showed significant differences in p62 mRNA， α-SMA， and p62 protein expression （P<0.05）. Compared with those in the blank group， LC3， Beclin1， SIRT1， and forkhead box protein O1 （FoxO1） expression levels were significantly decreased in the model group， EX527+TGF-β1 group， and EX527 group （P<0.01）. In the model and EX527+TGF-β1 groups， E-cadherin expression levels were significantly reduced （P<0.01）， while the EX527 group showed no statistically significant change. Compared with the model group， E-cadherin， LC3， Beclin1， SIRT1， and FoxO1 expression levels were significantly increased in both the YSHYP and valsartan groups （P<0.01， P<0.05）. Compared with the YSHYP group， the valsartan group exhibited significant differences in LC3， SIRT1， and FoxO1 mRNA expression （P<0.05， P<0.01）. Immunofluorescence results were consistent with those of Western blot and Real-time PCR. ConclusionYSHYP may protect TCMK-1 cells by activating the SIRT1/FoxO1 pathway， thereby promoting autophagy and restoring the autophagy flux to reduce the extent of transdifferentiation of TCMK-1 cells.
