open access

Vol 59, No 4 (2021)
Original paper
Submitted: 2021-09-09
Accepted: 2021-11-06
Published online: 2021-12-02
Get Citation

Tanshinone IIA attenuates high glucose-induced epithelial-to-mesenchymal transition in HK-2 cells through VDR/Wnt/β-catenin signaling pathway

Jingyi Zeng1, Xiaorong Bao1
DOI: 10.5603/FHC.a2021.0025
·
Pubmed: 34852178
·
Folia Histochem Cytobiol 2021;59(4):259-270.
Affiliations
  1. Department of Nephrology, Jinshan Hospital, Fudan University, Shanghai, China

open access

Vol 59, No 4 (2021)
ORIGINAL PAPERS
Submitted: 2021-09-09
Accepted: 2021-11-06
Published online: 2021-12-02

Abstract

Introduction. The progression of diabetic kidney disease (DKD) is closely related to renal tubular epithelial-
to-mesenchymal transition (EMT) and tubulointerstitial fibrosis. Tanshinone IIA (TSIIA), extracted from a
traditional Chinese medicine named Salvia miltiorrhiza, has been proved to have anti-fibrosis effects. The aim of this study was to investigate the effect of TSIIA on high glucose-induced EMT in human proximal tubular cells (HK-2 cells) and its possible mechanism.

Material and methods. The proliferation of cells exposed to different concentrations of glucose was measured by light microscopy and CCK-8 test. The cells were stimulated with 30 mM glucose and different concentrations of TSIIA (5 μM or 10 μM) for 48 h. Vitamin D receptor (VDR)-siRNA was used to transfect cells, and high glucose and TSIIA treatment were further used to treat cells. The expression of alpha smooth muscle actin (a-SMA) mRNA was detected by qPCR to ensure successful induction of EMT, and the expression of VDR mRNA was detected by qPCR to ensure successful transfection of VDR-siRNA. Protein expression of a-SMA, E-cadherin, VDR, b-catenin and glycogen synthase kinase 3b (GSK-3b) was detected by Western blot analysis.

Results. The results showed that high glucose concentration inhibited cell proliferation and promoted EMT in HK-2 cells. TSIIA could reverse high glucose-induced EMT by increasing the level of VDR protein and inhibiting the levels of b-catenin and GSK-3b proteins suggestive of a negative correlation between VDR and the Wnt/b-catenin pathway. After VDR-siRNA transfection and incubation of cells at high glucose concentration, the inhibitory effect of VDR on the expression of b-catenin and GSK-3b of Wnt pathway was suppressed and the b-catenin pathway was activated. When VDR level was restored by TSIIA, the inhibitory effect of VDR on the pathway was also restored and the activation of the pathway was suppressed.

Conclusions. TSIIA was able to attenuate high glucose-induced EMT in HK-2 cells by up-regulating VDR levels, which might be related to the inhibitory effect of VDR on the Wnt pathway.

Abstract

Introduction. The progression of diabetic kidney disease (DKD) is closely related to renal tubular epithelial-
to-mesenchymal transition (EMT) and tubulointerstitial fibrosis. Tanshinone IIA (TSIIA), extracted from a
traditional Chinese medicine named Salvia miltiorrhiza, has been proved to have anti-fibrosis effects. The aim of this study was to investigate the effect of TSIIA on high glucose-induced EMT in human proximal tubular cells (HK-2 cells) and its possible mechanism.

Material and methods. The proliferation of cells exposed to different concentrations of glucose was measured by light microscopy and CCK-8 test. The cells were stimulated with 30 mM glucose and different concentrations of TSIIA (5 μM or 10 μM) for 48 h. Vitamin D receptor (VDR)-siRNA was used to transfect cells, and high glucose and TSIIA treatment were further used to treat cells. The expression of alpha smooth muscle actin (a-SMA) mRNA was detected by qPCR to ensure successful induction of EMT, and the expression of VDR mRNA was detected by qPCR to ensure successful transfection of VDR-siRNA. Protein expression of a-SMA, E-cadherin, VDR, b-catenin and glycogen synthase kinase 3b (GSK-3b) was detected by Western blot analysis.

Results. The results showed that high glucose concentration inhibited cell proliferation and promoted EMT in HK-2 cells. TSIIA could reverse high glucose-induced EMT by increasing the level of VDR protein and inhibiting the levels of b-catenin and GSK-3b proteins suggestive of a negative correlation between VDR and the Wnt/b-catenin pathway. After VDR-siRNA transfection and incubation of cells at high glucose concentration, the inhibitory effect of VDR on the expression of b-catenin and GSK-3b of Wnt pathway was suppressed and the b-catenin pathway was activated. When VDR level was restored by TSIIA, the inhibitory effect of VDR on the pathway was also restored and the activation of the pathway was suppressed.

Conclusions. TSIIA was able to attenuate high glucose-induced EMT in HK-2 cells by up-regulating VDR levels, which might be related to the inhibitory effect of VDR on the Wnt pathway.

Get Citation

Keywords

tanshinone IIA; HK-2 cells; epithelial-to-mesenchymal transition; VDR; Wnt/β-catenin pathway

About this article
Title

Tanshinone IIA attenuates high glucose-induced epithelial-to-mesenchymal transition in HK-2 cells through VDR/Wnt/β-catenin signaling pathway

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 59, No 4 (2021)

Article type

Original paper

Pages

259-270

Published online

2021-12-02

Page views

2314

Article views/downloads

378

DOI

10.5603/FHC.a2021.0025

Pubmed

34852178

Bibliographic record

Folia Histochem Cytobiol 2021;59(4):259-270.

Keywords

tanshinone IIA
HK-2 cells
epithelial-to-mesenchymal transition
VDR
Wnt/β-catenin pathway

Authors

Jingyi Zeng
Xiaorong Bao

References (42)
  1. Beshay ON, Ewees MG, Abdel-Bakky MS, et al. Resveratrol reduces gentamicin-induced EMT in the kidney via inhibition of reactive oxygen species and involving TGF-β/Smad pathway. Life Sci. 2020; 258: 118178.
  2. Zhou T, Luo M, Cai W, et al. Runt-Related Transcription Factor 1 (RUNX1) Promotes TGF-β-Induced Renal Tubular Epithelial-to-Mesenchymal Transition (EMT) and Renal Fibrosis through the PI3K Subunit p110δ. EBioMedicine. 2018; 31: 217–225.
  3. Hu HH, Cao G, Wu XQ, et al. Wnt signaling pathway in aging-related tissue fibrosis and therapies. Ageing Res Rev. 2020; 60: 101063.
  4. Xie H, Miao N, Xu D, et al. FoxM1 promotes Wnt/β-catenin pathway activation and renal fibrosis via transcriptionally regulating multi-Wnts expressions. J Cell Mol Med. 2021; 25(4): 1958–1971.
  5. Chen Yu, Chen X, Ji YR, et al. PLK1 regulates hepatic stellate cell activation and liver fibrosis through Wnt/β-catenin signalling pathway. J Cell Mol Med. 2020; 24(13): 7405–7416.
  6. Kadoya H, Satoh M, Nishi Y, et al. Klotho is a novel therapeutic target in peritoneal fibrosis via Wnt signaling inhibition. Nephrol Dial Transplant. 2020; 35(5): 773–781.
  7. Lee DW, Lee WJ, Cho J, et al. Inhibition of Wnt signaling pathway suppresses radiation-induced dermal fibrosis. Sci Rep. 2020; 10(1): 13594.
  8. Yu M, Wu H, Wang J, et al. Vitamin D receptor inhibits EMT via regulation of the epithelial mitochondrial function in intestinal fibrosis. J Biol Chem. 2021; 296: 100531.
  9. Cong, L., Mechanism of vitamin D receptor modulating Wnt/β-catenin signaling pathway in the proliferation and invasion of gastric cancer. 2016, Shandong University.
  10. Commission, C.P., Pharmacopoeia of the People’s Republic of China. One Edition ed. 2020, Beijing: China: Pharmaceutical Science and Technology Press.
  11. Li Y, Gu B, Liu J, et al. Research progress of Tanshinone IIA. Shizhen Guo Yi Guo Yao. 2010; 21(7): 1770–1772.
  12. Li Z, Zou J, Cao D, et al. Pharmacological basis of tanshinone and new insights into tanshinone as a multitarget natural product for multifaceted diseases. Biomed Pharmacother. 2020; 130: 110599.
  13. Feng J, Chen HW, Pi LJ, et al. Protective effect of tanshinone IIA against cardiac hypertrophy in spontaneously hypertensive rats through inhibiting the Cys-C/Wnt signaling pathway. Oncotarget. 2017; 8(6): 10161–10170.
  14. Li ZY, Huang GD, Chen L, et al. Tanshinone IIA induces apoptosis via inhibition of Wnt/β‑catenin/MGMT signaling in AtT‑20 cells. Mol Med Rep. 2017; 16(5): 5908–5914.
  15. Chen SJ, Lv LL, Liu BC, et al. Crosstalk between tubular epithelial cells and glomerular endothelial cells in diabetic kidney disease. Cell Prolif. 2020; 53(3): e12763.
  16. Li X, Zhang F, Qu L, et al. Identification of YAP1 as a novel downstream effector of the FGF2/STAT3 pathway in the pathogenesis of renal tubulointerstitial fibrosis. J Cell Physiol. 2021; 236(11): 7655–7671.
  17. Chan SC, Zhang Y, Shao A, et al. Mechanism of Fibrosis in -Related Autosomal Dominant Tubulointerstitial Kidney Disease. J Am Soc Nephrol. 2018; 29(10): 2493–2509.
  18. Zhang Bo, Ru F, Chen X, et al. Autophagy attenuates renal fibrosis in obstructive nephropathy through inhibiting epithelialtomesenchymal transition. Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2021; 46(6): 601–608.
  19. Jonckheere S, Adams J, De Groote D, et al. Epithelial-Mesenchymal Transition (EMT) as a Therapeutic Target. Cells Tissues Organs. 2021 [Epub ahead of print]: 1–26.
  20. Bruner HC, Derksen PWB. Loss of E-Cadherin-Dependent Cell-Cell Adhesion and the Development and Progression of Cancer. Cold Spring Harb Perspect Biol. 2018; 10(3).
  21. Ding H, Chen J, Qin J, et al. TGF-β-induced α-SMA expression is mediated by C/EBPβ acetylation in human alveolar epithelial cells. Mol Med. 2021; 27(1): 22.
  22. Wang YP, Wang QY, Li CH, et al. COX-2 inhibition by celecoxib in epithelial ovarian cancer attenuates E-cadherin suppression through reduced Snail nuclear translocation. Chem Biol Interact. 2018; 292: 24–29.
  23. Liu B, Li X, Li C, et al. miR-25 mediates metastasis and epithelial-mesenchymal-transition in human esophageal squamous cell carcinoma via regulation of E-cadherin signaling. Bioengineered. 2019; 10(1): 679–688.
  24. Zhan S, Liu Z, Zhang M, et al. Overexpression of B7-H3 in α-SMA-Positive Fibroblasts Is Associated With Cancer Progression and Survival in Gastric Adenocarcinomas. Front Oncol. 2019; 9: 1466.
  25. Na TY, Schecterson L, Mendonsa AM, et al. The functional activity of E-cadherin controls tumor cell metastasis at multiple steps. Proc Natl Acad Sci U S A. 2020; 117(11): 5931–5937.
  26. Li XR, Jin JJ, Yu Y, et al. PET-CT radiomics by integrating primary tumor and peritumoral areas predicts E-cadherin expression and correlates with pelvic lymph node metastasis in early-stage cervical cancer. Eur Radiol. 2021; 31(8): 5967–5979.
  27. Masuda T, Nakashima T, Namba M, et al. Inhibition of PAI-1 limits chemotherapy resistance in lung cancer through suppressing myofibroblast characteristics of cancer-associated fibroblasts. J Cell Mol Med. 2019; 23(4): 2984–2994.
  28. Wang YN, Zhao SL, Su YY, et al. Astragaloside IV attenuates high glucose-induced EMT by inhibiting the TGF-β/Smad pathway in renal proximal tubular epithelial cells. Biosci Rep. 2020; 40(6).
  29. Wang WW, Liu YL, Wang MZ, et al. Inhibition of Renal Tubular Epithelial Mesenchymal Transition and Endoplasmic Reticulum Stress-Induced Apoptosis with Shenkang Injection Attenuates Diabetic Tubulopathy. Front Pharmacol. 2021; 12: 662706.
  30. Cao L, Huang B, Fu X, et al. Effects of tanshinone IIA on the regulation of renal proximal tubular fibrosis. Mol Med Rep. 2017; 15(6): 4247–4252.
  31. Fu D, Senouthai S, Wang J, et al. FKN Facilitates HK-2 Cell EMT and Tubulointerstitial Lesions via the Wnt/β-Catenin Pathway in a Murine Model of Lupus Nephritis. Front Immunol. 2019; 10: 784.
  32. Yiu WH, Li Ye, Lok SWY, et al. Protective role of kallistatin in renal fibrosis via modulation of Wnt/β-catenin signaling. Clin Sci (Lond). 2021; 135(3): 429–446.
  33. Xiong Y, Zhou L. The Signaling of Cellular Senescence in Diabetic Nephropathy. Oxid Med Cell Longev. 2019; 2019: 7495629.
  34. Shi M, Tian P, Liu Z, et al. MicroRNA-27a targets Sfrp1 to induce renal fibrosis in diabetic nephropathy by activating Wnt/β-Catenin signalling. Biosci Rep. 2020; 40(6).
  35. Guo Q, Zhong W, Duan A, et al. Protective or deleterious role of Wnt/beta-catenin signaling in diabetic nephropathy: An unresolved issue. Pharmacol Res. 2019; 144: 151–157.
  36. Ai K, Zhu X, Kang Ye, et al. miR-130a-3p inhibition protects against renal fibrosis in vitro via the TGF-β1/Smad pathway by targeting SnoN. Exp Mol Pathol. 2020; 112: 104358.
  37. Perretta-Tejedor N, Muñoz-Félix JM, Düwel A, et al. Cardiotrophin-1 opposes renal fibrosis in mice: Potential prevention of chronic kidney disease. Acta Physiol (Oxf). 2019; 226(2): e13247.
  38. Higgins DF, Ewart LM, Masterson E, et al. BMP7-induced-Pten inhibits Akt and prevents renal fibrosis. Biochim Biophys Acta Mol Basis Dis. 2017; 1863(12): 3095–3104.
  39. Xu S, He L, Ding K, et al. Tanshinone IIA Ameliorates Streptozotocin-Induced Diabetic Nephropathy, Partly by Attenuating PERK Pathway-Induced Fibrosis. Drug Des Devel Ther. 2020; 14: 5773–5782.
  40. Wang DT, Huang RH, Cheng X, et al. Tanshinone IIA attenuates renal fibrosis and inflammation via altering expression of TGF-β/Smad and NF-κB signaling pathway in 5/6 nephrectomized rats. Int Immunopharmacol. 2015; 26(1): 4–12.
  41. Zhang F.X. The role of VDR on epithelial-mesenchymal transition induced by high glucose of Mice Podocyte. 2016, Zhengzhou University.
  42. Guo J, Lu C, Zhang F, et al. VDR Activation Reduces Proteinuria and High-Glucose-Induced Injury of Kidneys and Podocytes by Regulating Wnt Signaling Pathway. Cell Physiol Biochem. 2017; 43(1): 39–51.

Regulations

Important: This website uses cookies. More >>

The cookies allow us to identify your computer and find out details about your last visit. They remembering whether you've visited the site before, so that you remain logged in - or to help us work out how many new website visitors we get each month. Most internet browsers accept cookies automatically, but you can change the settings of your browser to erase cookies or prevent automatic acceptance if you prefer.

By "Via Medica sp. z o.o." sp.k., ul. Świętokrzyska 73, 80–180 Gdańsk

tel.:+48 58 320 94 94, faks:+48 58 320 94 60, e-mail:  viamedica@viamedica.pl