Objective To investigate the alterations in the serum levels of neurogulin-1 （NRG-1）， chitinase 1 （CHIT1）， gap connexin 43 （Cx43）， and methyl-CpG-binding protein 2 （MECP2） in patients with acute cerebral infarction （ACI）， as well as their associations with the severity and prognosis of the disease. Methods A total of 355 patients with ACI admitted to Wuhan First Hospital from September 2023 to November 2024 were selected as the case group. These patients were divided into the severe group （49 cases）， the moderate group （126 cases）， and the mild group （180 cases） according to their condition. All patients underwent ultra-early intravenous thrombolysis upon admission. After treatment， they were observed for 3 months and then divided into the poor-prognosis group （81 cases） and the good-prognosis group （274 cases） based on the prognosis. Additionally， 355 healthy individuals who underwent physical examinations at Wuhan First Hospital during the same period were selected as the control group at a ratio of 1∶1. The basic data and serum levels of NRG-1， CHIT1， Cx43， and MECP2 were compared among groups. Multivariate Logistic regression analysis was employed to analyze the risk factors of poor prognosis in patients with ACI. The receiver operating characteristic curve （ROC） was plotted to obtain the area under the curve （AUC）， and the predictive value of serum NRG-1， CHIT1， Cx43， and MECP2 for poor prognosis in patients with ACI was analyzed. Results When compared with the control group， the serum level of NRG-1 in the case group was significantly lower， whereas the serum levels of CHIT1， Cx43， and MECP2 were significantly higher （P < 0.05）. The serum level of NRG - 1 gradually declined among the mild， moderate， and severe groups， while the serum levels of CHIT1， Cx43， and MECP2 gradually rose among these three groups （P < 0.05）. The proportions of patients with a score of the National Institutes of Health Stroke Scale （NIHSS） ≥ 15 points at admission， large-area infarction， a time interval of 3 ~ 4.5 h from onset to thrombolysis， and the serum levels of CHIT1， Cx43， and MECP2 in the poor-prognosis group were higher than those in the good-prognosis group （P < 0.05）， and the serum level of NRG-1 was lower than that in the good-prognosis group （P < 0.05）. Multivariate Logistic regression analysis indicated that the serum levels of CHIT1， Cx43， and MECP2 were risk factors for poor prognosis in patients with ACI （P < 0.05）， while the serum level of NRG-1 was a protective factor （P < 0.05）. ROC analysis demonstrated that the AUC value of the combined detection of serum NRG-1， CHIT1， Cx43， and MECP2 in predicting poor prognosis of patients with ACI was 0.931， which was higher than that of the single detection of each index （0.796， 0.801， 0.791， 0.805， P < 0.05）. Conclusions The serum level of NRG-1 in patients with acute cerebral infarction （ACI） was low， whereas the serum levels of CHIT1， Cx43， and MECP2 were high. These four factors were associated with the patients' disease severity and poor prognosis. Simultaneously， the combined detection of serum NRG-1， CHIT1， Cx43， and MECP2 exhibited a higher predictive value for the poor prognosis of ACI patients.