Synergism of calycosin and bone marrow-derived mesenchymal stem cells to combat podocyte apoptosis to alleviate adriamycin-induced focal segmental glomerulosclerosis

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 (MSCs^CA) 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 MSCs^CA 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 MSCs^CA-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 MSCs^CA-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 MSC^CA treatment more significantly than by MSCs or CA alone. When Smad3 was overexpressed in MPC5 cells, MSCs^CA could not fulfill their potential to inhibit podocyte apoptosis.

CONCLUSION

MSCs^CA enhance the protection of MSCs against ADR-induced podocyte apoptosis. The underlying mechanism may be related to MSCs^CA-targeted inhibition of p-Smad3 in podocytes.

Key Words: Calycosin, Mesenchymal stem cells, Focal segmental glomerulosclerosis, Apoptosis, Smad3

Core Tip: Calycosin (CA)-pretreated mesenchymal stem cells (MSCs^CA) 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 MSCs^CA on podocyte apoptosis, suggesting that MSCs^CA 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.

• Citation: Hu QD, Tan RZ, Zou YX, Li JC, Fan JM, Kantawong F, Wang L. Synergism of calycosin and bone marrow-derived mesenchymal stem cells to combat podocyte apoptosis to alleviate adriamycin-induced focal segmental glomerulosclerosis. World J Stem Cells 2023; 15(6): 617-631
• URL: https://www.wjgnet.com/1948-0210/full/v15/i6/617.htm
• DOI: https://dx.doi.org/10.4252/wjsc.v15.i6.617