苦参苍术颗粒对大肠杆菌感染肉鸡肝组织胆固醇代谢的调节作用

doi:10.11843/j.issn.0366-6964.2023.01.036

摘要/Abstract

摘要： 旨在探究苦参苍术颗粒是否通过调节肝组织胆固醇代谢，改善致病性大肠杆菌诱导的肉鸡肝组织胆固醇合成异常。将40只21日龄白羽肉鸡随机分为空白对照组、模型对照组、苦参苍术低剂量组、中剂量组和高剂量组，每组8只。模型对照组和苦参苍术组分别胸部肌肉注射0.7mL致病性大肠杆菌菌液(8.37×10⁸ CFU·mL^-1)，苦参苍术组在接种菌液24h后，在饮水中分别添加不同剂量的苦参苍术颗粒(低剂量组：3.6g·L^-1，中剂量组：5.5g·L^-1，高剂量组：7.3g·L^-1)，自由饮水，连用7d。试验结束，所有试验肉鸡处死，收集血液，采集组织样品，称体重和各器官重；采用石蜡切片技术观察肝组织形态变化；检测血清和肝组织中总胆固醇(total cholesterol,TCH)、甘油三酯(hepatic triglyceride,TG)、高密度脂蛋白胆固醇(high-density lipoprotein-cholesterol,HDL-C)、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)含量的变化；采用RT-qPCR检测肝组织中胆固醇代谢相关基因的变化。结果显示，与空白对照组相比，模型对照组肉鸡体重显著降低，肝脏指数和血清TG含量显著增加(P＜0.05)，肝小叶中央静脉周围和肝血窦内有炎性细胞浸润，肝细胞排列紊乱，同时肝组织中TG含量以及SREBP裂解激活蛋白(SREBP cleavage activating protein，SCAP) mRNA相对表达量也显著降低(P＜0.05)；和模型对照组相比，低剂量组显著增加了肉鸡体重和肝中LDL-C含量(P＜0.05)，降低了肝脏指数和血清中TG含量(P＜0.05)，且肝组织形态结构得到改善；和模型对照组相比，中剂量组显著升高了肝HDL-C、LDL-C含量及LDL-C/HDL-C的比值(P＜0.05)，3羟基3甲基戊二酸单酰辅酶A还原酶(3-hydroxy-3-methylglutaryl coenzyme A reductase,HMGCR)、调节元件结合蛋白2(sterol regulator element-binding protein-2,SREBP2)、SCAP mRNA相对表达量也显著提高(P＜0.05)。和模型对照组相比，高剂量组显著降低了血清中TG含量(P＜0.05)，显著增加肝中LDL-C含量(P＜0.05)，同时显著增加了LDL-C/HDL-C比值(P＜0.05)。综上所述，苦参苍术颗粒可通过调节肉鸡肝组织胆固醇代谢，缓解大肠杆菌感染所致的肝组织胆固醇合成异常。

关键词: 苦参苍术颗粒剂, 大肠杆菌, 白羽肉鸡, 肝, 胆固醇代谢

Abstract: The aim of this experiment was to investigate whether kushen-cangzhu (Sophora flavescens Ait. and Atractylodes Lancea) granules could improve the abnormal cholesterol synthesis induced by Escherichia coli (E. coli) through regulating liver cholesterol metabolism in broilers. Forty 21 days old white-feathered broilers were randomly divided into 5 groups (8 in each group): Blank control group, model control group, low-, medium-and high-dose of kushen-cangzhu groups. The model control group and different doses of kushen-cangzhu groups were intramuscular thoracic injection treated with pathogenic E. coli bacterial solution for 0.7 mL (8.37×10⁸ CFU·mL^-1). After inoculating pathogenic E. coli bacterial solution for 24 h, different doses of kushen-cangzhu granules (low-dose group: 3.6 g·L^-1, medium-dose group: 5.5 g·L^-1, high-dose group: 7.3 g·L^-1) were supplemented in drinking water for chickens in low-, medium-and high-dose groups, respectively, and the trail lasted for 7 days. At the end of the experiment, all the broilers were slaughtered, body and organs were weighed, blood and tissue samples were collected. The morphological changes of liver were observed by hematoxylin-eosin (HE) staining. The levels of serum and liver total cholesterol (TCH), hepatic triglyceride (TG), high-density lipoprotein-cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) were detected, RT-qPCR was used to detect the expression of cholesterol metabolism-related genes in liver. The results showed that, compared with the blank control group, the broilers in the model control group had significantly lower body weight, higher liver index and serum TG content (P＜0.05), while infiltrated inflammatory cells in the central vein and the hepatic sinusoid and disordered hepatocyte was observed in model control group. TG and SCAP mRNA expression was significantly lower (P＜0.05) in liver of model control group compared with blank control group. Compared with the model control group, the body weight and liver LDL-C content was significantly increased in the low-dose group (P＜0.05), while the liver index and serum TG content were significantly decreased (P＜0.05), and the hepatic morphological structure was at a better state. The HDL-C, LDL-C and the LDL-C/HDL-C ratio in liver were significantly increased in the medium-dose group compared with model control group. Meanwhile, the mRNA expression of HMGCR, SREBP2 and SCAP was significantly enhanced (P＜0.05) in medium-dose group than model control group. In the high-dose group, the serum TG concentration was significantly reduced (P＜0.05) but hepatic LDL-C was significantly increased (P＜0.05), and the ratio of LDL-C/HDL-C was significantly raised (P＜0.05) compared with the model control group. In conclusion, the liver abnormal synthesis of cholesterol in broilers induced by E. coli was alleviated by kushen-cangzhu granules through regulating cholesterol metabolic pathway.

Key words: kushen-cangzhu granules, E. coli, white-feathered broilers, liver, cholesterol metabolism

中图分类号:

S853.74

未张怡, 韩飞, 范婷婷, 陈子贤, 李寒梅, 袁非凡, 丛日华, 李贤. 苦参苍术颗粒对大肠杆菌感染肉鸡肝组织胆固醇代谢的调节作用[J]. 畜牧兽医学报, 2023, 54(1): 392-402.

WEI Zhangyi, HAN Fei, FAN Tingting, CHEN Zixian, LI Hanmei, YUAN Feifan, CONG Rihua, LI Xian. Regulation of Kushen-cangzhu Granules on Hepatic Cholesterol Metabolism in Broilers Infected with Escherichia coli[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 392-402.

参考文献 38

[1]	王亚芳,崔小七,贾召锋,等.鸡大肠杆菌病的模型建立与中兽医辨证论治研究进展[J].中国畜牧兽医,2014,41(9):269-272.WANG Y F,CUI X Q,JIA Z F,et al.Research advance on disease model and Chinese traditional veterinary diagnosis and treatment of chicken Escherichia coli disease[J].China Animal Husbandry & Veterinary Medicine,2014,41(9):269-272.(in Chinese)
[2]	DA ROSA G,ALBA D F,SILVA A D,et al.Impact of Escherichia coli infection in broiler breeder chicks:The effect of oxidative stress on weight gain[J].Microb Pathog,2020,139:103861.
[3]	ZHAO P F,PIAO X S,PAN L,et al.Forsythia suspensa extract attenuates lipopolysaccharide-induced inflammatory liver injury in rats via promoting antioxidant defense mechanisms[J].Anim Sci J,2017,88(6):873-881.
[4]	SONG H T,ZUO Z C,YANG Z Z,et al.Hepatic histopathology and apoptosis in diet-induced-obese mice under Escherichia coli pneumonia[J].Aging (Albany NY),2019,11(9):2836-2851.
[5]	CARPINO G,DEL BEN M,PASTORI D,et al.Increased liver localization of lipopolysaccharides in human and experimental NAFLD[J].Hepatology,2020,72(2):470-485.
[6]	彭梦玲.胚胎期肉鸡脂肪代谢的蛋白质组学和代谢组学分析及(-)-羟基柠檬酸调控机制研究[D].南京:南京农业大学,2018.PENG M L.Proteomics and Metabolomics Analyze the Lipids Metabolism during Embryonic Chicken and the Mechanisms of Lipids Metabolism-regulation of (-)-Hydroxylcitrate[D].Nanjing:Nanjing Agricultural University,2018.(in Chinese)
[7]	HOPKINS E G D,ROUMELIOTIS T I,MULLINEAUX-SANDERS C,et al.Intestinal epithelial cells and the microbiome undergo swift reprogramming at the inception of colonic Citrobacter rodentium infection[J].mBio,2019,10(2):e00062-19.
[8]	ZHANG C J,ZHU N,LIU Z,et al.Wnt5a/Ror2 pathway contributes to the regulation of cholesterol homeostasis and inflammatory response in atherosclerosis[J].Biochim Biophys Acta (BBA)-Mol Cell Biol Lipids,2020,1865(2):158547.
[9]	苏丽丽.苦参的化学成分及药理作用研究进展[J].化学工程师,2021,35(3):58-61.SU L L.Advances in the study of chemical constituents and pharmacological effects of Sophora flavescens Ait.[J].Chemical Engineer,2021,35(3):58-61.(in Chinese)
[10]	KIM H Y,JEONG D M,JUNG H J,et al.Hypolipidemic effects of Sophora flavescens and its constituents in poloxamer 407-induced hyperlipidemic and cholesterol-fed rats[J].Biol Pharm Bull,2008,31(1):73-78.
[11]	唐彬,艾正琳,姚树坤.苦参碱对高脂-脂肪肝模型大鼠肝脏COX-2、iNOS表达的影响[J].中华临床医师杂志:电子版,2013,7(5):2011-2015.TANG B,AI Z L,YAO S K.Effects of matrine on the expression of COX-2 and iNOS in rats with non-alcoholic fatty liver disease[J].Chinese Journal of Clinicians:Electronic Edition,2013,7(5):2011-2015.(in Chinese)
[12]	张明发,沈雅琴.苍术及其有效成分对心血管和代谢系统药理作用的研究进展[J].抗感染药学,2017,14(2):244-249.ZHANG M F,SHEN Y Q.Research advances in pharmacologic actions of Atractylodis rhizoma and its effective components on cardiovascular and metabolic systems[J].Anti-Infection Pharmacy,2017,14(2):244-249.(in Chinese)
[13]	KIM C K,KIM M,OH S D,et al.Effects of Atractylodes macrocephala koidzumi rhizome on 3T3-L1 adipogenesis and an animal model of obesity[J].J Ethnopharmacol,2011,137(1):396-402.
[14]	SONG M Y,LIM S K,WANG J H,et al.The Root of Atractylodes macrocephala koidzumi prevents obesity and glucose intolerance and increases energy metabolism in mice[J].Int J Mol Sci,2018,19(1):278.
[15]	高艳艳,张志伟,康慧.苦参苍术口服液的抗炎与止泻作用研究[J].中国兽药杂志,2013,47(4):26-28.GAO Y Y,ZHANG Z W,KANG H.Study on the anti-inflammatory and anti-diarrhea effects of Kushencangzhu oral liquid in mice[J].Chinese Journal of Veterinary Drug,2013,47(4):26-28.(in Chinese)
[16]	周德刚,高艳艳,张晓会.苦参苍术口服液对鸡耐药大肠杆菌的疗效试验[J].中国动物保健,2012,14(12):25-27.ZHOU D G,GAO Y Y,ZHANG X H.Effect of Sophora flavescens and Atractylodes lancea oral liquid on drug-resistant Escherichia coli in chickens[J].China Animal Health,2012,14(12):25-27.(in Chinese)
[17]	丛日华.品种和母猪日粮蛋白水平对仔猪肝脏胆固醇代谢的影响及其机制[D].南京:南京农业大学,2011.CONG R H.Effect of breed and maternal dietary protein level on hepatic cholesterol metabolism in piglets and mechanisms involved[D].Nanjing:Nanjing Agricultural University,2011.(in Chinese)
[18]	程林,谌俊,韩景河,等.母猪妊娠和哺乳饲粮中硒源和维生素E水平对后代仔猪器官指数和血清激素水平的影响[J].动物营养学报,2018,30(3):918-928.CHENG L,CHEN J,HAN J H,et al.Effects of dietary selenium sources and vitamin E levels during sows' gestation and lactation on organ indices and serum hormone levels of their progeny[J].Chinese Journal of Animal Nutrition,2018,30(3):918-928.(in Chinese)
[19]	YAN C Y,OUYANG S H,WANG X,et al.Celastrol ameliorates Propionibacterium acnes/LPS-induced liver damage and MSU-induced gouty arthritis via inhibiting K63 deubiquitination of NLRP3[J].Phytomedicine,2021,80:153398.
[20]	POHLMANN S,SCHEU S,ZIEGLER V,et al.Hepatic rac1 GTPase contributes to liver-mediated basal immune homeostasis and LPS-induced endotoxemia[J].Biochim Biophys Acta (BBA)-Mol Cell Res,2018,1865(9):1277-1292.
[21]	张志翔,曹颖光,徐恋祎.大肠杆菌脂多糖对小鼠骨髓间充质干细胞炎症因子分泌的影响[J].临床口腔医学杂志,2020,36(12):707-711.ZHANG Z X,CAO Y G,XU L Y.Effects of Escherichia coli lipopolysaccharide on the secretion of inflammatory factors in mouse bone marrow mesenchymal stem cells[J].Journal of Clinical Stomatology,2020,36(12):707-711.(in Chinese)
[22]	ZHANG Z J,TIAN L,JIANG K.Propofol attenuates inflammatory response and apoptosis to protect d-galactosamine/lipopolysaccharide induced acute liver injury via regulating TLR4/NF-κB/NLRP3 pathway[J].Int Immunopharmacol,2019,77:105974.
[23]	ANDERS L C,LANG A L,ANWAR-MOHAMED A,et al.Vinyl chloride metabolites potentiate inflammatory liver Injury caused by LPS in mice[J].Toxicol Sci,2016,151(2):312-323.
[24]	HUNG Y L,FANG S H,WANG S C,et al.Corylin protects LPS-induced sepsis and attenuates LPS-induced inflammatory response[J].Sci Rep,2017,7(1):46299.
[25]	HUYNH D T N,BAEK N,SIM S,et al.Minor ginsenoside Rg2 and Rh1 attenuates LPS-Induced acute liver and kidney damages via downregulating activation of TLR4-STAT1 and inflammatory cytokine production in macrophages[J].Int J Mol Sci,2020,21(18):6656.
[26]	XU Q M,XU J,ZHANG K F,et al.Study on the protective effect and mechanism of Dicliptera chinensis (L.) Juss (Acanthaceae) polysaccharide on immune liver injury induced by LPS[J].Biomed Pharmacother,2021,134:111159.
[27]	张明发,沈雅琴.氧化苦参碱抗慢性肝损伤的药理作用及其机制研究进展[J].药物评价研究,2019,42(7):1466-1473.ZHANG M F,SHEN Y Q.Research advances on effects of oxymatrine against chronic liver injuries[J].Drug Evaluation Research,2019,42(7):1466-1473.(in Chinese)
[28]	沙多依,杭永付,宋菲,等.北苍术炮制前后挥发油部位保肝作用比较研究[J].现代中药研究与实践,2010,24(4):41-43.SHA D Y,HANG Y F,SONG F,et al.Study on the protecting-liver effect of volatile oil of pre and post processing Cangzhu[J].Research and Practice on Chinese Medicines,2010,24(4):41-43.(in Chinese)
[29]	GONG Z W,TAS E,YAKAR S,et al.Hepatic lipid metabolism and non-alcoholic fatty liver disease in aging[J].Mol Cell Endocrinol,2017,455:115-130.
[30]	于茹,赵立岭,李娥娥,等.细菌中I型毒素蛋白-细胞膜作用机制的研究进展[J/OL].生物化学与生物物理进展,2022:1-16.(2022-02-23).http://kns.cnki.net/kcms/detail/11.2161.Q.20220222.1805.004.html. YU R,ZHAO L L,LI E E,et al.Research Progress on mechanisms of Type Ⅰ toxin protein-membrane in bacteria[J/OL].Progress in Biochemistry and Biophysics,2022:1-16.(2022-02-23).http://kns.cnki.net/kcms/detail/11.2161.Q.20220222.1805.004.html. (in Chinese)
[31]	SONG Y O,KIM M,WOO M,et al.Chondroitin sulfate-rich extract of skate cartilage attenuates lipopolysaccharide-induced liver damage in mice[J].Mar Drugs,2017,15(6):178.
[32]	路筱涛,祝晨蔯.苦参碱降血脂作用的实验研究[J].广东药学,2003,13(6):32-34.LU X T,ZHU C C.Experimental research on antilipidic effect of matrine[J].Guangdong Pharmaceutical Journal,2003,13(6):32-34.(in Chinese)
[33]	ZHOU Y F,MAO S G,ZHOU M X.Effect of the flavonoid baicalein as a feed additive on the growth performance,immunity,and antioxidant capacity of broiler chickens[J].J Poult Sci,2019,98(7):2790-2799.
[34]	LI L C,VARGHESE Z,MOORHEAD J F,et al.Cross-talk between TLR4-MyD88-NF-κB and SCAP-SREBP2 pathways mediates macrophage foam cell formation[J].Amer J Physiol Heart Circ Physiol,2013,304(6):H874-H884.
[35]	HE M Y,ZHANG W H,DONG Y Y,et al.Pro-inflammation NF-κB signaling triggers a positive feedback via enhancing cholesterol accumulation in liver cancer cells[J].J Exp Clin Cancer Res,2017,36(1):15.
[36]	NA S F,DUAN X J,WANG R Y,et al.Chronic neuroinflammation induced by lipopolysaccharide injection into the third ventricle induces behavioral changes[J].J Mol Neurosci,2021,71(6):1306-1319.
[37]	MOTAMED M,ZHANG Y X,WANG M L,et al.Identification of luminal Loop 1 of SCAP protein as the sterol sensor that maintains cholesterol homeostasis[J].J Biol Chem,2011,286(20):18002-18012.
[38]	林莹,杨军红,司萍,等.米油对内毒素导致大鼠急性肝损伤的保护作用[J].营养学报,2019,41(1):102-104.LIN Y,YANG J H,SI P,et al.Protective effect of rice oil on endotoxin-induced liver injury in rats[J].Acta Nutrimenta Sinica,2019,41(1):102-104.(in Chinese).