High Concentration of Aspirin Induces Apoptosis in Rat Tendon Stem Cells via Inhibition of the Wnt/β-Catenin Pathway

Abstract

Background/Aims: Non-steroidal anti-inflammatory drugs (NSAIDs) are commonly used in clinical practice to relieve fever and pain. Aspirin, as a representative NSAID, has been widely used in the treatment of tendinopathy. Some reports have demonstrated that aspirin can induce apoptosis in cancer cells. However, evidence regarding aspirin treatment for tendinopathy, especially the effect of this treatment on tendon stem cells (TSCs), is lacking. Understanding the effect of aspirin on tendinopathy may provide a basis for the rational use of NSAIDs in clinical practice. The aim of our study was to determine whether aspirin induces apoptosis in rat TSCs via the Wnt/β-catenin pathway. Methods: First, we used flow cytometry and fluorescence to detect TSC apoptosis. Protein expression of the apoptosis-related caspase-3 pathway was investigated via western blot analysis. Next, we used western blotting to determine the effect of aspirin on the Wnt/β-catenin pathway. We used immunostaining to detect the levels of Bcl2, cleaved caspase-3, and P-β-catenin in the Achilles tendon. Finally, we used flow cytometry, fluorescence, and western blotting to investigate the aspirin-induced apoptosis of TSCs via the Wnt/β-catenin pathway. Results: Aspirin induced morphological apoptosis in rat TSCs via the mitochondrial/caspase-3 pathway and induced cellular apoptosis in the Achilles tendon. Apoptosis was partly reversed after adding the Wnt signaling activator Wnt3a and lithium chloride (LiCl, a GSK-3β inhibitor). Aspirin administration led to a dose-dependent increase in COX-2 expression. Apoptosis was promoted after adding the COX-2 inhibitor NS398. Conclusion: The Wnt/β-catenin pathway plays a vital role in aspirin-induced apoptosis by regulating mitochondrial/caspase-3 function. Elevating COX-2 levels may protect cells against apoptosis. More importantly, the results remind us to consider the apoptotic effect of aspirin on TSCs and tendon cells when aspirin is administered to treat tendinopathy. The relationship between the positive and negative effects of aspirin remains a subject for future study.

Tendon stem/progenitor cell ageing: Modulation and rejuvenation

“…Celastrol exerts beneficial effects on human TSPCs stemness and the vital role of HIF1α-Smad7 signaling in the process is elucidated[47]. Celecoxib inhibits the tenogenic differentiation of TSPCs but has no effects on cell proliferation[48], and a high concentration of aspirin induces apoptosis in TSPCs by delaying the activation of Wnt/β-catenin pathway[49]. All these factors might affect the quality of tendon healing by targeting TSPCs, regardless of whether the effects are positive or negative.…”

Aspirin inhibits adipogenesis of tendon stem cells and lipids accumulation in rat injury tendon through regulating PTEN/PI3K/AKT signalling

“…We performed the isolation and identification of rat TSCs as our previously studies . Briefly, rats in all groups were euthanized, and then, Achilles tendons were carefully collected.…”

“…We followed the methods as the previous studies. [12][13][14][15] Twentyfour male Sprague-Dawley rats were randomly divided into three groups: control group, injury model group and aspirin treatment group. Firstly, 30 μl collagenase I (10 mg/ml) were injected into both Achilles tendons of rats in injury group and aspirin treatment group to establish tendinopathy model.…”

“…We performed the isolation and identification of rat TSCs as our previously studies. 12,16 Briefly, rats in all groups were euthanized, and then, Achilles tendons were carefully collected. The samples were Isolated cells were cultured under conditions of 37°C with 5% CO 2 for 2 days, then washed in PBS to discard non-adherent cells.…”

Apoptosis of mesenchymal stem cells is regulated by Rspo1 via the Wnt/β‐catenin signaling pathway

“…Importantly, the Wnt/β-catenin pathway played a vital role in aspirin-induced apoptosis by regulating mitochondrial/caspase-3 function. 22 Wang et al 23 found that inhibition of the Wnt/β-catenin signaling pathway improved the therapeutic effect on transcatheter arterial chemoembolization by suppressing migration and invasion while promoting the apoptosis of transplanted hepatocellular carcinoma cells in rats. Moreover, Rspo1 activated the Wnt/β-catenin signaling pathway, which is involved in the development, proliferation, and differentiation of stem cells, as well as the repair of tissue damage.…”