Objective To explore the relationship between the levels of serum nucleotide-binding oligomerization domain-like receptor 4（ NLRC4） and Krüpple-like factor 5（ KLF5） in patients with acute ischemic stroke（ AIS） and the progression of the disease and short-term prognosis. Methods A total of 126 patients with AIS and 126 healthy volunteers who underwent physical examinations at Zibo Central Hospital from January 2020 to January 2023 were selected as the study group and the control group， respectively. The groups were classified based on the National Institutes of Health Stroke Scale（ NIHSS） assessment criteria. Among the 126 AIS patients， 46 were in the mild group（ NIHSS score 0-5）， 59 in the moderate group（ NIHSS score 5-20）， and 21 in the severe group（ NIHSS score 20-46）. The prognosis of the patients was evaluated based on the modified Rankin Scale（ mRS） criteria， 81 patients were classified into the good prognosis group （mRS score＜3）， and 45 into the poor prognosis group（ mRS score≥3）. The levels of NLRC4 and KLF5 were detected by enzyme-linked immunosorbent assay（ ELISA）. Spearman and Pearson correlation analyses were used to investigate the correlations between the serum levels of NLRC4 and KLF5 in AIS patients and the infarction area， high-density lipoprotein cholesterol（ HDL-C）， time from onset to hospital admission， and disease severity. Multivariate Logistic regression was used to explore the influencing factors of poor prognosis in AIS patients. The receiver operating characteristic（ ROC） curve was used to evaluate the predictive value of NLRC4 and KLF5 for the prognosis of patients. Results The serum levels of NLRC4（ 9.64±1.53） ng/ml and KLF5（ 5.36±1.20） ng/ml in the study group were significantly higher than those in the control group NLRC4（ 6.31±1.25） ng/ml， KLF5 （1.85±0.31） ng/ml， with statistically significant differences（ t=18.919， 31.789， all P＜0.05）. The serum levels of NLRC4 and KLF5 in severe AIS patients were higher than those in moderate and mild AIS patients， with statistically significant differences（ F=113.423， 126.402， all P ＜ 0.05）. In the poor prognosis group， 62.2% （28/45） of patients had an infarction area ＞ 4 cm2， 64.4%（ 29/45） had an NIHSS score＞20 at admission， and the time from onset to hospital admission was（ 4.51±0.78） h， NLRC4 level was（ 10.58±1.25） ng/ml， and KLF5 level was（ 6.63±1.61） ng/ml， which were all higher than those in the good prognosis group 38.3%（ 31/81）， 40.7%（ 33/81），（ 3.46±0.45） h，（ 9.12±1.36） ng/ml，（ 4.66±1.28） ng/ml， with statistically significant differences （χ2/t=6.665，6.503，9.594， 5.940， 7.536， all P ＜ 0.05）. The HDL-C level in the poor prognosis group （1.59±0.15） mmol/L was lower than that in the good prognosis group（ 1.84±0.17） mmol/L， with a statistically significant difference（ t=8.240， P＜0.05）. The serum levels of NLRC4 and KLF5 in AIS patients were positively correlated with the infarction area， time from onset to hospital admission， and disease severity（ r=0.625， 0.586， 0.618， 0.637， 0.644， 0.622， all P＜0.05）， and negatively correlated with the HDL-C level（ r=-0.654， -0.619，all P＜ 0.05）. Infarction area ＞ 4 cm2 （OR=2.456， 95%CI： 1.332-4.527，P＜ 0.001）， long time from onset to hospital admission（ OR=2.457， 95%CI： 1.403-4.304，P=0.002）， high NLRC4 level（ OR=4.319， 95%CI： 1.892-9.857，P＜0.001）， and high KLF5 level（ OR=3.998， 95%CI： 1.959-8.160， P＜0.001） were identified as risk factors affecting the prognosis of AIS patients， while high HDL-C level（ OR=0.427， 95%CI： 0.251-0.726，P=0.002） was a protective factor. The area under the curve（ AUC） for predicting the prognosis of AIS patients by serum NLRC4 and KLF5 was 0.816 and 0.838， respectively. The AUC for predicting the prognosis of AIS patients by the combination of NLRC4 and KLF5 was 0.931， which was superior to that of NLRC4 and KLF5 alone（ ZNLRC4=2.675， ZKLF5=2.211，all P ＜ 0.05）. Conclusions NLRC4 and KLF5 levels in the serum of AIS patients increase with the severity of the disease and are closely related to the prognosis of patients.