The aim of this study was to identify the association polymorphism (rs11536889) in the 3′‐untranslated region (3′‐UTR) of Toll‐like receptors 4 (TLR4) and the risk for ventilator‐associated pneumonia (VAP). miRNA database online and luciferase assays were used to validate TLR4 as the target gene of miR‐1236. Enzyme‐linked immunosorbent assay analysis and western blot were used to analyze the level of TLR4 in different genotype groups. In the present study, miR‐1236 was predicted to bind to the rs11536889 G allele rather than the rs11536889 C allele, which was further confirmed by the luciferase activity suppressed by a fragment of 3′‐UTR containing the rs11536889 G allele induced by lipopolysaccharide (LPS) and interleukin‐6 (IL‐6). Bronchial epithelial cells isolated from participants genotyped as GG, GC, and CC, with no remarkable difference in TLR4 messenger RNA (mRNA) levels were observed among these genotype groups. After stimulating by LPS, a TLR4 ligand, the CC‐genotyped cells expressed higher levels of IL‐8, IL‐6, and tumor necrosis factor alpha (TNF‐α) on their surfaces than cells with the other genotypes. Finally, the western blot analysis results showed that the expression level of IL‐8, IL‐6, and TNF‐α protein was much higher in the CC group than the GC and GG groups subsequent to stimulation by LPS, and the IL‐8, IL‐6, and TNF‐α protein levels in the GC were grouped much lower compared with the GG group. These findings indicated the regulatory association of miR‐1236 with TLR4 and the abnormal expression of TLR4 caused by the presence of rs11536889 in the 3′‐UTR of mRNA, which interfere with its interaction with the miR‐1236, contributing to the risk of VAP.