Anti-apoptotic effect of adipose tissue-derived stromal vascular fraction in denervated rat muscle

Abstract

Background Recovery of muscle function after peripheral nerve injury is often poor, and this can be attributed to muscle fiber atrophy and cell death. In the current study, we have investigated the effects of stromal vascular fraction (SVF) on muscle cell apoptosis and its potential to preserve muscle tissue following denervation. Methods Rat gastrocnemius muscle was denervated by sciatic nerve transection. At 2 and 4 weeks after injury, muscles were examined histologically and apoptosis was measured using TUNEL assay and PCR array for a range of apoptotic genes. Additionally, an in vitro TNF-α apoptosis model was established using SVF cells co-cultured indirectly with primary rat myoblasts. Annexin V and TUNEL were used together with Western blotting to investigate the signaling pathways. Results Denervated muscles showed significantly higher TUNEL reactivity at 2 and 4 weeks following nerve injury, and an increased expression of caspase family genes, mitochondria-related apoptotic genes, and tumor necrosis factor family genes. In cultured rat primary myoblasts, Annexin V labeling was significantly increased at 12 h after TNF-α treatment, and this was followed by a significant increase in TUNEL reactivity at 48 h. Western blotting showed that caspase-7 was activated/cleaved as well as the downstream substrate, poly (ADP-ribose) polymerase (PARP). Co-culture of myoblasts with SVF significantly reduced all these measures of apoptosis. Bax and Bcl-2 levels were not changed suggesting that the TNF-α-induced apoptosis occurred via mitochondria-independent pathways. The protective effect of SVF was also shown in vivo; injections of SVF cells into denervated muscle significantly improved the mean fiber area and diameter, as well as reduced the levels of TUNEL reactivity. Conclusions This study provides new insights into how adipose tissue-derived cells might provide therapeutic benefits by preserving muscle tissue.

Laser-activated autologous adipose tissue-derived stromal vascular fraction restores spinal cord architecture and function in multiple sclerosis cat model

“…Previous studies have shown that the SVF secretome contains many growth factors that can stimulate cell proliferation and prevent the expression of apoptotic factors, such as Bax; this finding was reported by [79,80]. Our study recorded a significantly higher expression of Bax in the EB-treated group than in the control and SVF cotreated groups, which agrees with [23] in MS and [80] in denervated muscle.…”

BNIP3 mediates the different adaptive responses of fibroblast-like synovial cells to hypoxia in patients with osteoarthritis and rheumatoid arthritis

“…Prepared enough TdT incubation buffer for all experiments according to Recombinant TdT enzyme, FITC-12-dUTP Labeling Mix and Equilibration Buffer at the ratio of 1 μL:5 μL:50 μL. Added 56 μL of dT incubation buffer to the samples, incubated at 37 °C for 1 h in the dark, washed and then added 500 μL of 10 μg/mL DAPI solution to room temperature for 8 min to stain the cells (El-Habta et al 2021 ).…”

Effect of umbilical cord mesenchymal stem cells on hypoxia-inducible factor-1 alpha (HIF-1α) production in arteriovenous fistula (AVF) animal model: A preliminary study

“…The mechanism through which UC-MSCs exert their anti-IH effects is multi-faceted. As these stem cells have the ability to differentiate into various cell types, such as smooth muscle cells, endothelial cells, and fibroblasts [ 17 , 18 ], they can promote tissue repair and inhibit the development of IH when introduced into the site of AVF creation [ 19 ]. One possible mechanism is that the transplanted stem cells release factors or cytokines that promote angiogenesis and vasculogenesis, thereby enhancing the formation of new blood vessels in the AVF.…”