The present study was conducted to investigate the effects of chicory water-soluble extract (CWE) on performance, egg quality, gut microflora, serum and yolk cholesterol in laying hens. A total of 300 Brown Nick laying hens were divided randomly into 5 groups with 6 replicates of 10 hens each, and fed diets containing 0 (control), 0.5% (CWE0.5), 1.0% (CWE1.0) and 1.5% (CWE1.5) chicory water-soluble extract, respectively, as well as 1.0% inulin (I1.0) during 4 weeks’ trial. The results showed that there were no significant differences in laying rate, average egg weight, feed intake, feed/egg, cracked egg rate, yolk weight, eggshell thickness and Haugh unit compared with the control (P>0.05). Supplementing diets with CWE or inulin significantly reduced cholesterol concentration (mg/g yolk) and total cholesterol content (mg/yolk) in egg-yolk (P<0.05). Compared with control, atherogenic index in serum was reduced by CWE1.5 (P<0.05). Dietary CWE or inulin supplementation significantly reduced the number of E. coli and pH in caecum of laying hens (P<0.05). Caecum Bifidobacteria counts were increased in CWE1.5 group and I1.0 group (P<0.05). Egg-yolk cholesterol content and the number of E. coli were decreased linearly (r=-0.957 and r=-0.977, respectively, P<0.05) in accordance with increasing of dietary CWE level. It could be concluded that the incorporation of CWE or inulin in laying hens’ diets could reduce egg-yolk cholesterol and caecum harmful bacteria (E. coli) counts with no any negative effects on laying hens’ performance and egg quality in this trial (P<0.05). Therefore, the dietary supplementation of CWE or inulin might lead to the development of low-cholesterol eggs as demanded by health-conscious consumers.