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©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
Synergism of calycosin and bone marrow-derived mesenchymal stem cells to combat podocyte apoptosis to alleviate adriamycin-induced focal segmental glomerulosclerosis
Qiong-Dan Hu, Rui-Zhi Tan, Yuan-Xia Zou, Jian-Chun Li, Jun-Ming Fan, Fahsai Kantawong, Li Wang
Qiong-Dan Hu, Rui-Zhi Tan, Fahsai Kantawong, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand
Qiong-Dan Hu, Rui-Zhi Tan, Yuan-Xia Zou, Jian-Chun Li, Li Wang, Research Center of Integrated Traditional Chinese and Western Medicine, The Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China
Qiong-Dan Hu, Department of Nephrology, The Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China
Yuan-Xia Zou, Jian-Chun Li, Molecular Imaging and Therapy Research Unit, Department of Radiologic Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand
Jun-Ming Fan, Department of Nephrology, The Affiliated Hospital of Chengdu Medical College, Chengdu 610500, Sichuan Province, China
Author contributions: Wang L and Fahsai K contributed equally to this article as co-corresponding authors; Wang L and Fahsai K designed the study and supervised the laboratory experiments; Hu QD conducted the experiments and drafted the manuscript; Tan RZ assisted with the experiments; Zou YX and Li JC collected the samples; Fan JM and Wang L contributed new reagents and analytic tools; and all authors read and approved the final manuscript.
Supported by the National Natural Science Foundation of China (General Program), No. 82205002; Science and Technology Project of Sichuan Province, No. 2022YFS0621, No. 21ZDYF0348, and No. 2022NSFSC1459; Luzhou-Southwest Medical University Science and Technology Strategic Cooperation Project, No. 2021LZXNYD-P04; and Southwest Medical University of Affiliated Traditional Medicine Hospital Project, No. 2022-CXTD-03.
Institutional animal care and use committee statement: All animal experiments were reviewed and approved by the Animal Ethics Committee of Southwest Medical University (Approval No. 20221031-012).
Conflict-of-interest statement: The authors have no conflicts of interest to declare.
Data sharing statement: All data are available from the corresponding author upon reasonable request Wangli120@swmu.edu.cn.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
https://creativecommons.org/Licenses/by-nc/4.0/ Corresponding author: Li Wang, PhD, Professor, Research Center of Integrated Traditional Chinese and Western Medicine, The Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University, No. 182 Chunhui Road, Longmatan District, Luzhou 646000, Sichuan Province, China. wangli120@swmu.edu.cn
Received: March 29, 2023
Peer-review started: March 29, 2023
First decision: April 25, 2023
Revised: April 28, 2023
Accepted: May 25, 2023
Article in press: May 25, 2023
Published online: June 26, 2023
BACKGROUND
Bone marrow-derived mesenchymal stem cells (MSCs) show podocyte-protective effects in chronic kidney disease. Calycosin (CA), a phytoestrogen, is isolated from Astragalus membranaceus with a kidney-tonifying effect. CA preconditioning enhances the protective effect of MSCs against renal fibrosis in mice with unilateral ureteral occlusion. However, the protective effect and underlying mechanism of CA-pretreated MSCs (MSCsCA) on podocytes in adriamycin (ADR)-induced focal segmental glomerulosclerosis (FSGS) mice remain unclear.
AIM
To investigate whether CA enhances the role of MSCs in protecting against podocyte injury induced by ADR and the possible mechanism involved.
METHODS
ADR was used to induce FSGS in mice, and MSCs, CA, or MSCsCA were administered to mice. Their protective effect and possible mechanism of action on podocytes were observed by Western blot, immunohistochemistry, immunofluorescence, and real-time polymerase chain reaction. In vitro, ADR was used to stimulate mouse podocytes (MPC5) to induce injury, and the supernatants from MSC-, CA-, or MSCsCA-treated cells were collected to observe their protective effects on podocytes. Subsequently, the apoptosis of podocytes was detected in vivo and in vitro by Western blot, TUNEL assay, and immunofluorescence. Overexpression of Smad3, which is involved in apoptosis, was then induced to evaluate whether the MSCsCA-mediated podocyte protective effect is associated with Smad3 inhibition in MPC5 cells.
RESULTS
CA-pretreated MSCs enhanced the protective effect of MSCs against podocyte injury and the ability to inhibit podocyte apoptosis in ADR-induced FSGS mice and MPC5 cells. Expression of p-Smad3 was upregulated in mice with ADR-induced FSGS and MPC5 cells, which was reversed by MSCCA treatment more significantly than by MSCs or CA alone. When Smad3 was overexpressed in MPC5 cells, MSCsCA could not fulfill their potential to inhibit podocyte apoptosis.
CONCLUSION
MSCsCA enhance the protection of MSCs against ADR-induced podocyte apoptosis. The underlying mechanism may be related to MSCsCA-targeted inhibition of p-Smad3 in podocytes.
Core Tip: Calycosin (CA)-pretreated mesenchymal stem cells (MSCsCA) enhanced the protective effect of MSCs against adriamycin (ADR)-induced podocyte injury in vitro and in vivo by inhibiting apoptosis, accompanied by more reversal of the upregulated expression of p-Smad3 after ADR induction. Smad3 overexpression eliminated the inhibitory effect of MSCsCA on podocyte apoptosis, suggesting that MSCsCA inhibit podocyte apoptosis by targeting p-Smad3. These results broaden our understanding of the potential of MSCs pretreated with herbal extract and provide new theories for possible therapeutic mechanisms for ADR-induced focal segmental glomerulosclerosis.
