Objective To explore the effect of alanyl-glutamine combined with hyperbaric oxygen therapy on peripheral blood T lymphocyte subsets， microRNA-124 （miR-124）， and miR-181c in young and middle-aged patients with ischemic stroke in the convalescent period.Methods A total of 128 young and middle-aged patients with ischemic stroke in the convalescent period in our hospital from July 2017 to July 2021 were selected as the study subjects， and they were divided into 4 groups according to the random number table： negative control group， control group A， control group B， and observation group， with 32 patients in each group. The negative control group received conventional treatment in the convalescent period of stroke. On this basis， the control group A was given alanyl-glutamine； the control group B was given hyperbaric oxygen therapy； the observation group was given alanyl-glutamine combined with hyperbaric oxygen therapy. All treatments lasted for 4 weeks. The neurological function ［National Institute of Health Stroke Scale （NIHSS） score］， ability of daily living ［Barthel index （BI）］， limb motor function ［Fugl-Meyer Motor Function Assessment score］， inflammatory factors ［（tumor necrosis factor alpha （TNF-α）， monocyte chemoattractant protein-1 （MCP-1）， and interleukin-12 （IL-12）］， peripheral blood T lymphocyte subsets （CD4⁺， CD8⁺， and CD4⁺/CD8⁺）， cerebral hemodynamics ［minimum blood flow velocity （V_min）， and minimum blood flow （Q_min） of the carotid artery］， miR-124， and miR-181c expression levels were compared among the 4 groups before treatment and after 2 and 4 weeks of treatment.Results After 2 and 4 weeks of treatment， the NIHSS score in the observation group was significantly lower than that in the control group A， control group B and negative control group （P<0.05）， and the BI and motor function scores of upper and lower limbs were significantly higher than those in the control group A， control group B， and negative control group （P<0.05）. After 2 and 4 weeks of treatment， serum TNF-α， MCP-1 and IL-12 levels in the observation group were significantly lower than those in the control group A， control group B and negative control group （P<0.05）； the CD4⁺ and CD4⁺/CD8⁺ levels were significantly higher than those in the control group A， control group B， and negative control group， and the CD8⁺ level was significantly lower than that in the control group A， control group B， and negative control group （P<0.05）. After 2 and 4 weeks of treatment， the CD4⁺ and CD4⁺/CD8⁺ levels in the control group A were significantly higher than those in the control group B and negative control group （P<0.05）， and the CD8⁺ level was significantly lower than that in the control group B and negative control group （P<0.05）. After 2 and 4 weeks of treatment， V_min and Q_min in the observation group were significantly higher than those in the control group A， control group B， and negative control group （P<0.05）， and V_min and Q_min in the control group B were significantly higher than those in the control group A and negative control group （P<0.05）. After 2 and 4 weeks of treatment， the expression levels of miR-124 and miR-181c in the observation group were significantly lower than those in the control group A， control group B， and negative control group （P<0.05）.Conclusions Alanyl-glutamine combined with hyperbaric oxygen therapy can further improve the cellular immune function of young and middle-aged patients with ischemic stroke in the convalescent period， down-regulate the expression levels of miR-124 and miR-181c， and significantly improve patients’ neurological function and ability of daily living.