柴术抗激颗粒对LPS诱导大鼠神经-内分泌-免疫相关因子调节机制研究

doi:10.3864/j.issn.0578-1752.2014.23.016

摘要/Abstract

摘要： 【目的】“仔猪早期断奶应激综合征”给养猪业带来了巨大的经济损失。中药制剂柴术抗激散(CZKJS)通过现代制药技术制成柴术抗激颗粒(CZKJKL)，已被证实其对早期断奶仔猪应激及早期断奶仔猪脂多糖（LPS）免疫应激都具有明显的缓减作用。现进一步探究CZKJKL对LPS免疫应激断奶大鼠应激相关神经递质、细胞因子和HPA轴相关激素的影响，从机理上证实其抗应激作用。【方法】选取体重相近的18日龄断奶的Wistar大鼠72只，随机分为3个处理组，即Ⅰ组（空白对照组）、Ⅱ组（LPS免疫应激模型组）、Ⅲ组（LPS+CZKJKL组）。每个处理设6个重复，每个重复4只。在18—24日龄期间，Ⅲ组的大鼠每天灌服5 000 mg·kg^-1BW柴术抗激颗粒，Ⅰ组、Ⅱ组的大鼠每天灌服等剂量生理盐水；21日龄时，Ⅱ组、Ⅲ组的大鼠腹腔注射4 mg·kg^-1BW的LPS，Ⅰ组的大鼠腹腔注射等剂量生理盐水，注射后6 h采血并取样。测定指标有：①海马：去甲肾上腺素(NE)和5-羟色胺(5-HT)；②下丘脑：β-内啡肽（β-EP）和促肾上腺皮质激素释放因子（CRF）；③血清：β-内啡肽(β-EP)、促肾上腺皮质激素(ACTH)、皮质醇(COR)、白介素-1β (IL-1β)、白介素-6 (IL-6)、肿瘤坏死因子-α (TNF-α)、一氧化氮(NO)、诱导性一氧化氮合酶(iNOS)。【结果】与Ⅰ组相比，Ⅱ组大鼠海马NE和5-HT的含量均极显著升高（P＜0.01），下丘脑β-EP的含量极显著升高（P＜0.01），血清β-EP、IL-1β、IL-6、TNF-α、NO、ACTH、COR的含量均极显著升高（P＜0.01），血清iNOS的活性极显著增强（P＜0.01）。与Ⅱ组相比，Ⅲ组大鼠海马NE和5-HT的含量均极显著降低（P＜0.01），下丘脑β-EP的含量极显著降低（P＜0.01），血清β-EP、IL-1β、IL-6、TNF-α、NO、ACTH、COR的含量均极显著降低（P＜0.01），血清iNOS的活性极显著减弱（P＜0.01）。【结论】CZKJKL可通过有效调节LPS免疫应激状态下机体神经-内分泌-免疫系统相关神经递质、细胞因子和HPA轴激素的紊乱而发挥其抗免疫应激作用。

关键词: 柴术抗激颗粒, 脂多糖, 免疫应激, 断奶大鼠

Abstract:

【Objective】“Piglet early weaning stress syndrome” brought huge economic losses to the pig industry, immunological stress is one of the important factors. Chaizhukangji particles (CZKJKL) were prepared through modern pharmaceutical technology, total saikosaponins (TSS, the composition of the main therapeutic effects of Bupleuri Radix), polysaccharide of Atractyles macroceohala koidz (PAM, the composition of the main therapeutic effects of Atractylodis Macrocephalae Rhizoma), total glycosides of paeony (TGP, the composition of the main therapeutic effects of Paeoniae Radix Alba)were also selected, and proved that the preparation has a significant mitigation effect on stress of early weaned piglets and stress of early weaning piglets after immunological stress by lipopolysaccharide. The experiment was conducted to investigate the effects of CZKJKL on stress-related neurotransmitters, cytokines and hormones in HPA axis of nerve-endocrine-immune system of early weanling rats after LPS immunological stress, to further explore the anti-stress effects of CZKJKL.【Method】Total 72 early weanling wistar rats at the age of 18 days were randomly divided into 3 treatments of 24 rats each, with 6 replicates in each group and 4 rats in each replicate. The 3 treatments were group Ⅰ(control group), group Ⅱ (LPS group) and group Ⅲ (LPS+CZKJKL group). In the period of 18-24 days of age, the rats in group Ⅲ were fed with 5 000 mg·kg^-1BW CZKJKL daily by gavage, the rats in group Ⅰ and group Ⅱ were fed with equivalent amount of sterile saline. At the age of 21 days, the rats in group Ⅱ and group Ⅲ were injected intraperitoneally with 4 mg·kg^-1BW of LPS, the rats in group Ⅰ were injected with equivalent amount of sterile saline. Blood and tissue samples were obtained at the time of 6 h after injecting LPS. The following indicators were determinated: The contents of NE,5-HT of Hippocampus; the contents of β-EP, CRF of Hypothalamus; the contents or activities of β-EP, ACTH, COR, IL-1β, IL-6, TNF-α, NO, and iNOS of serum.【Result】Compared with groupⅠ, the contents of NE and 5-HT in hippocampi of groupⅡ were significantly increased (P＜0.01), the levels of β-EP in hypothalamus were significantly increased (P＜0.01), the contents of β-EP, IL-1β, IL-6, TNF-α, NO, ACTH, and COR in serum were significantly increased (P＜0.01), and the activity of iNOS in serum was significantly enhanced (P＜0.01). Compared with groupⅡ, the contents of NE and 5-HT in hippocampi of group Ⅲ were significantly decreased (P＜0.01), the levels of β-EP in hypothalamus were significantly decreased (P＜0.01), the contents of β-EP, IL-1β, IL-6, TNF-α, NO, ACTH, and COR in serum were significantly decreased (P＜0.01), and the activity of iNOS in serum was significantly reduced (P＜0.01).【Conclusion】The results indicate that CZKJKL can play a role of anti-immunological stress by efficiently alleviating the disorder of stress-related neurotransmitters, cytokines and hormones in HPA axis of nerve-endocrine-immune system of early weanling rats after LPS immunological stress.

Key words: CZKJKL, lipopolysaccharide, immunological stress, weanling rats

孙耀贵，程佳，李宏全，王俊东. 柴术抗激颗粒对LPS诱导大鼠神经-内分泌-免疫相关因子调节机制研究[J]. 中国农业科学, 2014, 47(23): 4718-4725.

SUN Yao-gui, CHENG Jia, LI Hong-quan, WANG Jun-dong. Effects of CZKJKL on Stress-Related Factors of Nerve - Endocrine - Immune System of Rats After Immunological Induction[J]. Scientia Agricultura Sinica, 2014, 47(23): 4718-4725.

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https://www.chinaagrisci.com/CN/Y2014/V47/I23/4718

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