Besides, no significant differences were observed in the UREA and TG concentrations among the three treatments ( 0.05). Open in a separate window Figure 2 Effects of dietary supplemented withMCE on serum biochemical parameters of broiler chickens. cholesterol, total cholesterol, glucose, immunoglobulin A, immunoglobulin M, and Cyclopamine complements (C3, C4) concentrations in the MCE group were significantly higher than in the CON group ( 0.05). Dietary MCE or aureomycin supplementation significantly reduced the hepatic contents of 8-hydroxy-2′-deoxyguanosine, malondialdehyde, interleukin (IL)-1, IL-6, NLRs family pyrin domain containing 3 (NLRP3), and caspase-1 in the liver ( 0.05). Moreover, MCE or aureomycin supplementation significantly inhibited mRNA expressions of Toll-like receptor 4, myeloid differentiation factor 88, nuclear factor-B, and NLRP3, as well as the expression ratio of Bax to Bcl-2 mRNA ( 0.05). Therefore, our study suggested that dietary supplementation with 0.6 mg/kg MCE containing protopine and allocryptopine improved growth performance and benefited liver health in broiler chickens possibly through inhibiting caspase-1-induced pyroptosis by inactivating TLR4/MyD88/NF-B/NLRP3 signaling pathway, and provided support for the application of MCE containing protopine and allocryptopine as an alternative to antibiotics in the feed industry. belongs to the family and is mainly distributed in central and southeastern China (8). extract (MCE) has been reported to have a broad spectrum of biological activities, such as anti-viral, anti-inflammatory, antimicrobial, and detoxifying effects (9). This is mainly attributed to its active principles including benzophenanthridine alkaloids (sanguinarine and chelerythrine) and isoquinoline alkaloids (protopine and allocryptopine) (10). Sanguinarine and chelerythrine from MCE have been exploited as natural feed additives in the European Union, and are widely used as substitutes for AGP in poultry and livestock Cyclopamine production (11, 12). A previous study also demonstrated that protopine and allocryptopine have the beneficial biological activities of anti-inflammatory, anti-bacterial, and hepatoprotective effects (13). However, there is little literature evaluating the effects of dietary supplementation with MCE containing protopine and allocryptopine on the growth performance and health status of broiler chickens. Therefore, this study was conducted to explore Gja5 the effects of dietary supplementation with MCE containing protopine and allocryptopine on the growth performance and liver health in broiler chickens and to highlight the application of MCE containing isoquinoline alkaloids as an effective alternative to AGP. Materials and Methods Experimental Design and Management A total of 486 newly hatched male Arbor Acres broiler chickens with average body weight (BW) of 48.76 0.25 g were obtained from a local hatchery and randomly assigned to three dietary treatments according to their BW with 6 replicates per treatment in a 42-d study. Each replicate contained 27 chickens housed in a three-level cage. The three treatment groups were designed as follows: (1) control (CON) group, broiler chickens fed a basal diet; (2) AGP group (positive control), broiler chickens fed a basal diet Cyclopamine supplemented with 50 mg/kg aureomycin (14); (3) MCE group, broiler chickens fed a basal diet supplemented with 0.6 mg/kg MCE including 0.4 mg/kg protopine and 0.2 mg/kg allotypotopine. The MCE was provided by the Micolta Bioresource Company Limited (Changsha 410331, PR China) and premixed with starch before supplementation. The 42-day trial spanned two phases including the starter period (days 0C21) and the grower period (days 21C42). The basal diets (Table 1) were formulated to respect the nutritional requirements of broiler chickens recommended by the Ministry of Agriculture of P.R. China (2004). Feed and water were offered during the experiment. All broiler chickens were housed in a temperature- and light-controlled room with continuous light at the Shandong Agricultural University. Cyclopamine The room temperature was kept at 32C34C for the first week and then reduced by 3C per week until it reached 21C. In addition, all broiler chickens were vaccinated with inactivated Newcastle disease vaccine at 7 days of age and inactivated infectious bursal disease vaccine at 14 days of age. Feed intake per replicate was recorded daily to calculate average daily feed intake (ADFI), and all broilers were weighed by replicates at 21 and 42 days to calculate average daily gain (ADG). Then the feed-to-gain ratio (F/G) was obtained based on feed.