Background and Purpose Inflammatory processes induced by leukocytes are crucially involved in the pathophysiology of acute ischemic stroke. This study aimed to elucidate the inflammatory mechanism of long non-coding RNA (lncRNA) H19-mediated regulation of C1QTNF6 by sponging miR-29b in leukocytes during ischemic stroke. Experimental Approach H19 and miR-29b expression in leukocytes of patients with ischemic stroke and middle cerebral artery occlusion rats was measured through real-time PCR. Moreover, we performed in vivo and in vitro experiments to determine the impact of H19 and miR-29b on C1QTNF6 expression in leukocytes after ischemic injury. Key Results There was H19 and C1QTNF6 upregulation, as well as miR-29b downregulation, in neutrophils of patients with stroke. Moreover, miR-29b could bind C1QTNF6 mRNA to repress its expression while H19 could sponge miR-29b to maintain C1QTNF6 expression. Moreover, C1QTNF6 overexpression promoted the release of IL-1β and TNF-α in leukocytes and further exacerbates blood-brain barrier disruption. Conclusion and Implications Our findings confirm the inflammation-modulatory effect of the H19/miR-29b/C1QTNF6 axis on cerebral ischemic injury.