Effects of dietary inclusion of Radix Bupleuri extract on the growth performance, and ultrastructural changes and apoptosis of lung epithelial cells in broilers exposed to atmospheric ammonia

Abstract

Abstract To explore whether Radix Bupleuri extract (RBE) could protect lung injury of broilers under ammonia (NH3) exposure, 360 one-d-old male broilers were randomly allocated to four groups of six replicates each in a 2 × 2 factorial design with two diets (the basal diet [control; CON] and the basal diet supplemented with RBE [RB]) and two air conditions (normal condition [<2 ppm of NH3; NOR] and NH3 exposure [70 ppm of NH3; NH70]). The RB diet contained 80 mg saikosaponins/kg diet. On day 7, the lung tissues were collected and the lung epithelial cells (LEC) were isolated. Our experimental results showed that the NH3 exposure decreased body weight gain and feed intake irrespective of dietary treatments during days 1 to 7. However, the RBE addition decreased feed consumption to body weight gain ratio in broilers under NH70 conditions. In the LEC of CON-fed broilers under NH70 conditions, Golgi stacks showed the dilation of cisternaes and reduced secretory vesicles, mitochondria enlarged, the inner membrane of mitochondria became obscure, and the cristae of mitochondria ruptured, whereas only a mild enlargement of Golgi cisternaes and the part rupture of mitochondrial cristaes occurred in the LEC of RB-fed broilers under NH70 conditions. The NH3 exposure increased malondialdehyde (MDA) level, but decreased total antioxidant capacity (T-AOC) in the lungs of CON-fed broilers. However, the RBE addition decreased MDA level and increased T-AOC in the lungs of broilers under NH70 conditions. Simultaneously, the NH3 exposure increased apoptotic rate (AR), mitochondrial membrane potential (MMP), and reactive oxygen species (ROS) level in the isolated LEC of CON-fed broilers. The RBE addition decreased AR, MMP, and ROS in the isolated LEC of broilers under NH70 condition. Besides, the NH3 exposure increased mRNA expression of B-cell lymphoma-2 associated X protein (BAX), caspase-3, and tumor necrosis factor α (TNF-α), but increased interferon γ (IFN-γ) mRNA abundance in the lungs of CON-fed broilers. The RBE supplement decreased mRNA levels of BAX, caspase-3, and TNF-α, but increased IFN-γ, interleukin (IL)-4, and IL-17 mRNA levels in the lungs of broilers under NH70 conditions. These results indicated that dietary RBE addition alleviated NH3 exposure-induced intercellular ultrastructural damage via mitochondrial apoptotic pathway, possibly due to RBE-induced increase of antioxidant capacity and immunomodulatory function in the lungs of broilers under NH3 exposure.