肾复康颗粒对人工诱发鸡肾肿病理模型的炎性细胞因子及IL-1R/NF-κB信号通路的影响

doi:10.11843/j.issn.0366-6964.2020.11.027

摘要/Abstract

摘要： 旨在探讨肾复康颗粒对鸡肾肿的抗炎机制。将90只健康雏鸡分为空白组（15只）和试验组（75只），试验组通过饲喂自制高钙高蛋白饲料人工复制鸡肾肿模型，造模成功后，随机分为模型组、阳性药物组、肾复康颗粒高、中、低剂量组，每组15只，肾复康颗粒高、中、低剂量组分别饮水给药肾复康颗粒2.0、1.0、0.5 g·L^-1，阳性药物组饮水给药苍蓝口服液1.0 mL·L^-1，模型组饮水给予生理盐水1.0 mL·L^-1，2次·d^-1，连用5 d。分别于造模前、后以及末次给药后2 d进行翅下静脉采血，分离血清，采用ELISA法检测血清IL-1、TNF-α、MCP-1、TGF-β1的含量变化；采用RT-PCR法检测肾IL-1R、IκBα、NF-κBp65 mRNA的转录。结果显示：与空白组相比，模型组雏鸡血清IL-1、TNF-α、MCP-1、TGF-β1含量显著升高（P<0.05），肾中IL-1R、NF-κBp65 mRNA转录量极显著升高（P<0.01），肾中IκBα mRNA转录量极显著降低（P<0.01）。饮水给药治疗后，肾复康颗粒高、低剂量组和阳性药物组雏鸡血清IL-1、TNF-α、MCP-1、TGF-β1含量显著降低（P<0.05）；肾复康颗粒中剂量组肾中IL-1R、NF-κBp65 mRNA转录量极显著降低（P<0.01），肾中IκBα mRNA转录量极显著增加（P<0.01）。肾复康颗粒通过降低肾肿鸡血清中IL-1、TNF-α、MCP-1、TGF-β1中含量，抑制IL-1R/NF-κB信号通路，从而发挥抗炎作用。

关键词: 肾复康颗粒, 肾肿, 抗炎, 转录, IL-1R/NF-κB信号通路

Abstract: The objective of the current study was to explore the anti-inflammatory mechanism of Shenfukang granule on chicken kidney swelling. Ninety healthy chickens were divided into blank group (n=15) and test group (n=75), the test group was replicated by the chicken kidney swelling model through feeding high-calcium and high-protein diet. After successful modeling, the test group was randomly divided into model group, positive drug group, Shenfukang high, medium and low dose group (n=15), Shenfukang granule 2.0, 1.0 and 0.5 g·L^-1, positive drug group, drinking water to Canglan oral liquid 1.0 mL·L^-1, The drinking water of the model group was given 1.0 mL·L^-1 of normal saline for 2 times·d^-1 and 5 days. Subpteral venous blood was collected before modeling, after modeling and 2 days after the last administration to isolate the serum. The content changes of serum IL-1, TNF-α, MCP-1 and TGF-β1 content were detected by ELISA. RT-PCR was used to detect renal IL-1R, IκBα, NF-κB mRNA transcription. Results were as follows: Compared with the blank group, the serum contents of IL-1, TNF-1, MCP-1 and TGF-β1 were significantly increased in the model group (P<0.05), the transcription of IL-1R, NF-κBp65 mRNA in kidney was significantly increased (P<0.01), and IκBα mRNA in kidney was significantly decreased (P<0.01). After drinking water treatment, the serum IL-1, TNF-α, MCP-1 and TGF-β1 contents of Shenfukang granule were significantly decreased in the high and low dose group and the positive drug group (P<0.01). The results of the current study suggested that Shenfukang granule played an anti-inflammatory role in inhibiting the IL-1R/NF-κB signaling pathway by reducing the contents of IL-1, TNF-α, MCP-1 and TGF-β1 in serum of kidney swelling chicken.

Key words: Shenfukang granule, renal swelling, anti-inflammatory, transcription, IL-1R/NF-κB signaling pathway

中图分类号:

S853.9

罗艺晨, 杨庆昌, 林春发, 叶芮伶, 尚杨斐, 胡宇, 刘娟. 肾复康颗粒对人工诱发鸡肾肿病理模型的炎性细胞因子及IL-1R/NF-κB信号通路的影响[J]. 畜牧兽医学报, 2020, 51(11): 2886-2894.

LUO Yichen, YANG Qingchang, LIN Chunfa, YE Ruiling, SHANG Yangfei, HU Yu, LIU Juan. The Effect of Shenfukang Granule on Content of Inflammatory Cytokines and the Signaling Pathway of IL-1R/NF-κB Induced by the Artificial Pathological Model of Chicken Kidney Swelling[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(11): 2886-2894.

参考文献 35

[1]	程金科, 范国雄. 高钙、低磷饲料致鸡痛风的病理学研究[J]. 畜牧兽医学报, 1992, 23(1):80-86.CHENG J K, FAN G X. The pathology of chicken gout induced by high dietary calcium and low dietary phosphorus[J]. Acta Veterinaria et Zootechnica Sinica, 1992, 23(1):80-86. (in Chinese)
[2]	骆建兵. 高钙高蛋白日粮对鸡肝肾功能与酸碱平衡的影响及痛风形成机理研究[D]. 南京:南京农业大学, 2006.LUO J B. Effects of diet with high calcium and/or protein on function of liver and kidney and acid-base balance of pullets and study of mechanism of avian gout[D]. Nanjing:Nanjing Agricultural University, 2006. (in Chinese)
[3]	汪海. 肉鸡肾肿的病因、症状及防治措施[J]. 现代畜牧科技, 2016(9):90.WANG H. Etiology, symptoms and prevention and treatment of kidney swelling in broilers[J]. Modern Animal Husbandry Science & Technology, 2016(9):90. (in Chinese)
[4]	李志鹏. 鸡肾肿症的防治措施[J]. 江西畜牧兽医杂志, 2009(5):43-44.LI Z P. Prevention and treatment of chicken kidney disease[J]. Jiangxi Journal of Animal Husbandry and Veterinary Medicine, 2009(5):43-44. (in Chinese)
[5]	STANHEWICZ A E, KENNEY W L. Determinants of water and sodium intake and output[J]. Nutr Rev, 2015, 73(S2):73-82.
[6]	BOCKENHAUER D, BICHET D G. Pathophysiology, diagnosis and management of nephrogenic diabetes insipidus[J]. Nat Rev Nephrol, 2015, 11(10):576-588.
[7]	RANGAN G, WANG Y P, HARRIS D. NF-κB signalling in chronic kidney disease[J]. Front Biosci (Landmark Ed), 2009, 14:3496-3522.
[8]	张爱华, 陈荣华. 核因子NF-κB与肾脏纤维化[J]. 国外医学·泌尿系统分册, 2001, 20(S1):188-190.ZHANG A H, CHEN R H. Nuclear factor NF-κB and renal fibrosis[J]. Foreign Medical Sciences, 2001, 20(S1):188-190. (in Chinese)
[9]	MORRISSEY J, KLAHR S. Transcription factor NF-κB regulation of renal fibrosis during ureteral obstruction[J]. Semin Nephrol, 1998, 18(6):603-611.
[10]	BAUERLE K T, SCHWEPPE R E, HAUGEN B R. Inhibition of nuclear factor-kappa B differentially affects thyroid cancer cell growth, apoptosis, and invasion[J]. Mol Cancer, 2010, 9:117.
[11]	HABELHAH H. Emerging complexity of protein ubiquitination in the NF-κB pathway[J]. Genes Cancer, 2010, 1(7):735-747.
[12]	HUTTON H L, OOI J D, HOLDSWORTH S R, et al. The NLRP3 inflammasome in kidney disease and autoimmunity[J]. Nephrology (Carlton), 2016, 21(9):736-744.
[13]	SANZ A B, SANCHEZ-NIÑO M D, RAMOS A M, et al. NF-κB in renal inflammation[J]. J Am Soc Nephrol, 2010, 21(8):1254-1262.
[14]	FU Z J, LIAO W T, MA H K, et al. Inhibition of neddylation plays protective role in lipopolysaccharide-induced kidney damage through CRL-mediated NF-κB pathways[J]. Am J Transl Res, 2019, 11(5):2830-2842.
[15]	苏友新, 陈伟宏, 王和鸣, 等. 高尿酸血症及痛风性关节炎的动物模型研究概况[J]. 中国骨伤, 2004, 17(7):444-445.SU Y X, CHEN W H, WANG H M, et al. Advance in the animal model of hyperuricemia and gouty arthritis[J]. China Journal of Orthopaedics and Traumatology, 2004, 17(7):444-445. (in Chinese)
[16]	杨佐君, 王晓霞, 苏玲, 等. 锌和高钙日粮对肉用仔鸡肾脏组织影响的观察[J]. 北京农学院学报, 1998, 13(4):36-41.YANG Z J, WANG X X, SU L, et al. Effects of dietary zinc and excessive calcium on renal histology of broilers[J]. Journal of Beijing Agricultural College, 1998, 13(4):36-41. (in Chinese)
[17]	DAS A, CHAKRABARTI A, BASAK D K. Visceral gout in Japanese quail[J]. J Indian Vet, 1992, 69:167-168.
[18]	BLAXLAND J D, BORLAND E D, SILLER W G, et al. An investigation of urolithiasis in two flocks of laying fowls[J]. Avian Pathol, 1980, 9(1):5-19.
[19]	WAKEFIELD A P, HOUSE J D, OGBORN M R, et al. A diet with 35% of energy from protein leads to kidney damage in female Sprague-Dawley rats[J]. Br J Nutr, 2011, 106(5):656-663.
[20]	EL-SAWI N M, BACKER W, ALY M M, et al. Assessment of therapeutic value of black costus (Saussurea lappa) using several parameters[J]. J Int Environ Appl Sci, 2010, 5(5):832-841.
[21]	CONG F, LIU X L, HAN Z X, et al. Transcriptome analysis of chicken kidney tissues following coronavirus avian infectious bronchitis virus infection[J]. BMC Genomics, 2013, 14:743.
[22]	RANDALL C J, BLANDFORD T B, BORLAND E D, et al. A survey of mortality in 51 caged laying flocks[J]. Avian Pathol, 1977, 6(2):149-170.
[23]	BROWN T P, GLISSOM J R, ROSALES G, et al. Studies of avian urolithiasis associated with an infectious bronchitis virus[J]. Avian Dis, 1987, 31(3):629-636.
[24]	程金科, 范国雄. 禽痛风研究进展[J]. 中国兽医杂志, 1989, 15(6):43-45, 52.CHENG J K, FAN G X. Advances in the study of avian gout[J]. Chinese Journal of Veterinary Medicine, 1989, 15(6):43-45, 52. (in Chinese)
[25]	臧长江. 热应激条件下日粮阴阳离子平衡对奶牛生产性能及血液生理生化指标的影响[D]. 乌鲁木齐:新疆农业大学, 2008.ZANG C J. Effect of dietary cation-anion balance on production performance and blood biochemistry indicators of dairy cows in the condition of heatstres[D]. Urumqi:Xinjiang Agricultural University, 2008. (in Chinese)
[26]	丁厚康, 马卫明, 刘莲莲, 等. 大肠杆菌对兔血液生理生化指标和实质器官组织结构的影响[J]. 江西农业大学学报, 2013, 35(3):556-562, 567.DING H K, MA W M, LIU L L, et al. Effect of colibacillosis on blood physiobiochemical indices and histological structure of parenchymal organs in rabbits[J]. Acta Agriculturae Universitatis Jiangxiensis, 2013, 35(3):556-562, 567. (in Chinese)
[27]	丁瑞, 洪权, 耿晓东, 等. 土茯苓治疗小鼠高尿酸血症的机制研究[J]. 中国中西医结合肾病杂志, 2019, 20(2):97-100.DING R, HONG Q, GENG X D, et al. Mechanism of rhizoma smilacis glabrae in the treatment of hyperuricemia mouse model[J]. Chinese Journal of Integrated Traditional and Western Nephrology, 2019, 20(2):97-100. (in Chinese)
[28]	黄意婷, 赵艺蔓, 庾雪鹰, 等. 基于网络药理学的土茯苓治疗痛风性肾病的作用机制研究[J]. 中国医药, 2020, 15(7):1063-1066.HUANG Y T, ZHAO Y M, YU X Y, et al. Study on the mechanism of smilacis glabrae rhizoma treating gouty nephropathy based on network pharmacology[J]. China Medicine, 2020, 15(7):1063-1066. (in Chinese)
[29]	DE OLIVEIRA D C, HASTREITER A A, MELLO A S, et al. The effects of protein malnutrition on the TNF-RI and NF-κB expression via the TNF-α signaling pathway[J]. Cytokine, 2014, 69(2):218-225.
[30]	BRENNER E, BLASER H, MAK T W. Regulation of tumour necrosis factor signalling:live or let die[J]. Nat Rev Immunol, 2015, 15(6):362-374.
[31]	SUN S C. The non-canonical NF-κB pathway in immunity and inflammation[J]. Nat Rev Immunol, 2017, 17(9):545-558.
[32]	ZHANG Q, LENARDO M J, BALTIMORE D. 30 years of NF-κB:A blossoming of relevance to human pathobiology[J]. Cell, 2017, 168(1-2):37-57.
[33]	BAUERLE K T, SCHWEPPE R E, HAUGEN B R. Inhibition of nuclear factor-kappa B differentially affects thyroid cancer cell growth, apoptosis, and invasion[J]. Mol Cancer, 2010, 9:117.
[34]	HSU L C, ENZLER T, SEITA J, et al. IL-1β-driven neutrophilia preserves antibacterial defense in the absence of the kinase IKKβ[J]. Nat Immunol, 2011, 12(2):144-150.
[35]	陈继红, 周栋, 高坤, 等. NF-κB、IκB在阿霉素肾病大鼠中的表达及大黄蛰虫丸的干预作用[J]. 中国临床药理学与治疗学, 2011, 16(1):22-26.CHEN J H, ZHOU D, GAO K, et al. Expression of NF-κB and IκB in rats with adriamycin nephropathy and the role of Dahuangzhechong Wan[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2011, 16(1):22-26. (in Chinese)