National Center for International Research on Animal Gut Nutrition, Nanjing Agricultural University, No.1 WeiGang, Xuanwu region, Nanjing, 210095, Jiangsu, China.
Laboratory of Gastrointestinal Microbiology, College of Animal Science and Technology, Nanjing Agricultural University, No.1 WeiGang, Xuanwu region, Nanjing, 210095, Jiangsu, China.
Environ Sci Pollut Res Int. 2022 Nov;29(53):80952-80967. doi: 10.1007/s11356-022-20737-6. Epub 2022 Jun 21.
Heat stress (HS) has detrimental effects on intestinal health by altering digestive and immune responses in animals. Dietary Moringa oleifera leaf powder (MOLP) has been implicated in ameliorating the impact of HS, but its effects in terms of intestinal function improvement under HS remain poorly characterized. Therefore, the current study investigated the impact of HS and MOLP supplementation on tight junction barriers, intestinal microbiota (jejunal digesta), and differentially expressed genes (DEGs) in jejunal mucosa of heat-stressed rabbits by using the next-generation sequencing techniques. A total of 21 male New Zealand White rabbits (32 weeks old mean body weight of 3318 ± 171 g) were divided into three groups (n = 7/group) as control (CON, 25 °C), heat stress (HS, 35 °C for 7 h daily), and HS with MOLP supplementation (HSM, 35 °C for 7 h daily) gavage at 200 mg/kg body weight per day for 4 weeks. The results indicated that MOLP supplementation increased mRNA expression of tight junction proteins and glutathione transferase activity, while the malonaldehyde concentration was decreased in the jejunal mucosa compared to HS group (P < 0.05). Furthermore, MOLP decreased the concentrations of lipopolysaccharide, pro-inflammatory cytokines, and myeloperoxidase compared with HS group (P < 0.05). Intestinal microbiota analysis revealed that at phyla level, the relative abundance of Bacteroidetes was higher in HSM group compared to CON and HS groups. MOLP supplementation also resulted in higher abundance of putatively health-associated genera such as Christensenellaceae R-7 gut group, Ruminococcaceae NK4A214 group, Ruminococcus 2, Lachnospiraceae NK4A136 group, and Lachnospiraceae unclassified along with higher butyrate levels in HSM group as compared to HS group. The analysis of DEGs revealed that MOLP reversed inflammatory response by downregulation of genes, such as TNFRSF13C, LBP, and COX2 in enriched KEGG pathway of NF-kβ pathway. MOLP supplementation also significantly upregulated the expression of genes in protein digestion and absorption pathway, including PRSS2, LOC100349163, CPA1, CPB1, SLC9A3, SLC1A1, and SLC7A9 in HSM group. Three genes of fibrillar collagens, i.e., COL3A1, COL5A3, and COL12A1 in protein digestion were also down-regulated in HSM group. In conclusion, MOLP supplementation could improve jejunal permeability and digestive function, positively modulate microbiota composition and mucosal immunity in heat-stressed rabbits.
热应激(HS)通过改变动物的消化和免疫反应对肠道健康产生有害影响。饲料辣木叶粉(MOLP)已被证明可以改善 HS 的影响,但它在 HS 下改善肠道功能的作用仍知之甚少。因此,本研究通过下一代测序技术研究了 HS 和 MOLP 补充对热应激兔的紧密连接屏障、肠道微生物群(空肠内容物)和空肠黏膜中差异表达基因(DEGs)的影响。总共 21 只雄性新西兰白兔(32 周龄,平均体重 3318±171g)分为三组(每组 7 只):对照组(CON,25°C)、热应激组(HS,35°C 每天 7 小时)和热应激+MOLP 补充组(HSM,35°C 每天 7 小时,每天 200mg/kg 体重),补充 4 周。结果表明,与 HS 组相比,MOLP 补充组空肠黏膜中紧密连接蛋白的 mRNA 表达增加,谷胱甘肽转移酶活性增加,而丙二醛浓度降低(P<0.05)。此外,与 HS 组相比,MOLP 降低了脂多糖、促炎细胞因子和髓过氧化物酶的浓度(P<0.05)。肠道微生物群分析表明,在门水平上,HSM 组厚壁菌门的相对丰度高于 CON 和 HS 组。MOLP 补充还导致假定与健康相关的属的丰度增加,如 Christensenellaceae R-7 肠道群、Ruminococcaceae NK4A214 群、Ruminococcus 2、Lachnospiraceae NK4A136 群和 Lachnospiraceae 未分类群,以及 HSM 组丁酸水平升高。与 HS 组相比,DEGs 分析表明,MOLP 通过下调 TNFRSF13C、LBP 和 COX2 等基因,逆转了 NF-kβ 途径中富集的关键基因途径的炎症反应。MOLP 补充还显著上调了 HSM 组中蛋白消化吸收途径的基因表达,包括 PRSS2、LOC100349163、CPA1、CPB1、SLC9A3、SLC1A1 和 SLC7A9。在蛋白消化中,三个原纤维胶原基因,即 COL3A1、COL5A3 和 COL12A1,在 HSM 组中也下调。
总之,MOLP 补充可改善热应激兔的空肠通透性和消化功能,正向调节微生物群组成和黏膜免疫。