Non-alcoholic steatohepatitis (NASH) is the progressive form of non-alcoholic fatty liver disease (NAFLD) which is the most common chronic liver disease worldwide. However, there are no FDA-approved drugs for treating NASH. The roles of hypoxia-inducible factor-1α (HIF1α) playing in NASH are not fully understood, and the mainstream view is that HIF1α knockout improved NASH. Here, in order to explore the clinical translation potential of HIF1α, we used HIF1α inhibitor KC7F2 to treat diet-induced NASH mice for 10 weeks. Surprisingly, it turned out that KC7F2 intensified liver lipid accumulation, inflammation, and fibrosis in NASH mice. Mechanistically, a most remarkable increase of expression of hepatokine Tsukushi (TSKU) was observed among twenty metabolism-related hepatokines in the liver of NASH mice after KC7F2 treatment. In vitro, we confirmed that inhibiting HIF1α or TSKU overexpression both significantly intensified lipid accumulation in hepatocytes, but didn’t increase expression of inflammation markers in LPS-induced macrophages or fibrosis markers in TGFβ-induced hepatic stellate cells. Moreover, Tsku knockdown reversed the HIF1α inhibitor-induced lipid accumulation in hepatocytes. Taken together, we first found that HIF1α inhibitor exacerbated NASH presumably via promoting hepatocyte lipid accumulation which was dependent on TSKU, indicating NASH may be a contraindication for HIF1α inhibitor rather than an indication.