Shift work is common worldwide and linked to deleterious cardiovascular effects that might be underlined by dyslipidemia. The aim of this systematic review and meta-analysis is to determine the impact of shiftwork on dyslipidemia. Searching in PubMed, Cochrane Library, Science Direct and Embase databases without language restriction on 15 February 2020, included studies that describe blood lipids levels or a risk measure in shift workers compared with fixed-day workers (controls). Differences by study-level characteristics were estimated using stratified meta-analysis by type of shift work, and meta-regression to examine relations between dyslipidemia and demographic, lifestyle and work characteristics. Estimates were pooled using random-effect meta-analysis. We included a total of 66 articles, representing 197,063 workers. Shift work globally increased the levels of triglycerides (overall SMD = 0.09; 95CI 0.05 to 0.13; p < 0.001), and globally decreased the levels of c-HDL (−0.08; 95CI −0.12 to −0.03; p = 0.001). Permanent night shift workers were an at-risk type of shift for dyslipidemia with significantly higher blood levels of total cholesterol (0.22; 95CI 0.01 to 0.42; p = 0.043) and triglycerides (0.18; 0.03 to 0.33; p = 0.017), and significantly lower blood levels of c-HDL (−0.16; 95CI −0.32 to 0.00; p = 0.05). Permanent night shift workers were more at-risk for total cholesterol than rotating 3 × 8 shift workers (Coefficient 0.22; 95CI 0.01 to 0.42; p = 0.038) and rotating 2 × 12 shift workers (0.24; 0.02 to 0.46; p = 0.037), and more at-risk for triglycerides than rotating day shift workers (0.21; 95CI 0.03 to 0.38; p = 0.023). Results were non-significant for c-LDL, nor depending on type of shifts. Shift work, and particularly permanent night shift, is associated with dyslipidaemia via elevated total cholesterol and triglycerides, and reduced HDL-cholesterol. Our current study provides a practical and valuable strengthening of the evidence-base required for preventive health initiatives and workplace reform. Display omitted •Shift work, involving 20% of the work force, is associated with deleterious cardiovascular effects that might be underlined by dyslipidemia.•Considering that 20% of the work force is subjected to shift work, the question of the best shifting rotation is of particular interest.•Analyzing a total of 66 articles and 197,063 workers, we showed that shift workers are associated with dyslipidemia.•We showed that shift work, and particularly permanent night shift work, increases the risk of dyslipidemia.•Promoting rotating shifts instead of permanent night shifts could be an effective preventive strategy to reduce cardiovascular risk of shift workers.