Mesenchymal stem cells therapy ameliorates esophageal dysmotility in gastroesophageal reflux disease by modulating smooth muscle contractility, alleviating senescence by targeting the DNMT3a-HIF1α axis

Figure 5 Mesenchymal stem cells ameliorate tumor necrosis factor-α-accelerated cellular senescence, protein dysregulation, and calcium signaling in human esophageal smooth muscle cells. This figure investigates the effects of mesenchymal stem cells on human esophageal smooth muscle cells under tumor necrosis factor-α exposure. A: Β-galactosidase staining images indicating cellular senescence; B: Western blot images and quantification for myosin light chain kinase, phosphorylated-myosin light chain kinase, alpha-smooth muscle actin, myosin phosphatase target subunit 1, phosphorylated-myosin phosphatase target subunit 1, anoctamin 1 and calcium voltage-gated channel subunit alpha1 C protein levels; C: Fluorescence images of Fluo-3, AM staining and quantification of intracellular Ca²⁺ intensity. Error bars represent SEM. ^aP < 0.05 for gastroesophageal reflux disease model group vs control group; ^cP < 0.001 for mesenchymal stem cell group vs gastroesophageal reflux disease model group; ^dP < 0.05 for mesenchymal stem cell group vs gastroesophageal reflux disease model group; ^fP < 0.001 for mesenchymal stem cell group vs gastroesophageal reflux disease model group; ^gP < 0.001 for si-ANO1/(+)Cacna1C group vs mesenchymal stem cell group. TNF: Tumor necrosis factor; NS: Not significant; Ctr: Control; GM: Gastroesophageal reflux disease model; MSC: Mesenchymal stem cell; α-SMA: Alpha-smooth muscle actin; MYLK: Myosin light chain kinase; p-MYLK: Phosphorylated-myosin light chain kinase; MYPT1: Myosin phosphatase target subunit 1; p-MYPT1: Phosphorylated-myosin phosphatase target subunit 1; ANO1: Anoctamin 1; CACNA1C: Calcium voltage-gated channel subunit alpha1 C; si-NC: Negative control small interfering RNA.