中草药复方防治肉鸡骨病的研究

doi:10.16431/j.cnki.1671-7236.2021.01.042

摘要/Abstract

摘要： 为探明中草药复方对肉鸡骨病的防治效果，本研究选用160只1日龄健康科宝肉鸡，随机均分为8组，分别饲养至30和60日龄，由当归、枸杞、山药等8味中草药组成复方制剂，将中草药复方以高（0.6%）、中（0.4%）、低（0.2%）3个剂量组进行治疗试验，同时设置对照组饲喂基础饲粮，分别于30、60日龄时称重，采用生化分析仪检测骨代谢相关血液生化指标维生素D受体（VDR）、骨钙素（OT）、骨保护素（OPG）、碱性磷酸酶（ALP），通过万能试验机进行三点弯曲试验，以弹性模量、屈服强度和最大应力为指标检测胫骨的骨强度，运用实时荧光定量PCR方法检测胸椎、腿软骨组织中骨保护素（OPG）、骨形态发生蛋白-2（BMP-2）、成骨细胞转录因子2（Runx-2）、细胞核因子κB受体活化因子（RANK）、细胞核因子κB受体活化因子配体（RANKL）、肿瘤坏死因子α（TNF-α）基因mRNA表达量的差异。结果显示，与对照组相比添加复方中草药组试验鸡后体重略增加；60日龄时血清中VDR、OT、OPG、ALP含量显著或极显著提高（P<0.05；P<0.01）；肉鸡的胫骨骨强度指标（弹性模量、屈服强度和最大应力）显著增加（P<0.05），且弹性模量、屈服强度、最大应力3个指标均表现为30日龄高于60日龄，与血液生化指标相符；复方中草药可显著促进OPG、BMP-2、Runx-2基因表达（P<0.05），显著抑制TNF-α、RANK、RANKL基因表达（P<0.05）。综合试验结果，兼顾活重和骨强度相关指标，复方中草药高剂量组是改善科宝肉鸡骨骼特性最优配方，且中草药复方通过提高血清中VDR、OT、OPG、ALP含量，调控OPG/RANK/RANKL系统维持骨吸收和骨形成两者动态平衡，促进骨形成的重要通路BMP-2/Smads/Runx-2中BMP-2和Runx-2基因的表达，同时抑制TNF-α基因表达，以达到改善科宝肉鸡骨骼特性，最终有效防治肉鸡的骨病。

关键词: 中草药复方; 科宝肉鸡; 骨病; 相关基因

Abstract: In order to prove the prevention and treatment effect of Chinese herbal medicine compound on broiler bone disease,this experiment selected 160 1-day-old healthy Kosbao broilers and randomly divided into 8 groups to be raised to 30 and 60 days old,a compound of Chinese herbal medicine complex consistion of 8 Chinese herbal medicine such as Angelica sinensis,Fructus Lycii and Dioscorea oppositifolia L..In the experiment,high (0.6%),medium (0.4%),and low (0.2%) dose of Chinese herbal medicine were used,and control group was fed a basic diet.At the age of 30 and 60 days,weight were detected.Biochemical analyzer was used to detect the blood biochemical indexes bone metabolism related (VDR,OT,OPG,ALP),the bone strength (elastic modulus,yield strength and maximum stress) of the tibia was detected by the universal test machine three point bending test,and the Real-time PCR method was used to detect the difference of mRNA expression of OPG,Runx-2,RANK,RANKL,TNF-α and BMP-2 genes in the thoracic and leg cartilage tissue.The results showed that the Chinese herbal medicine compound could gain weight,increase the content of VDR,OT,OPG and ALP in serum (P<0.05) at 60 days old,30 days old was higher than that of 60 days old.It could enhance tibial bone strength (P<0.05),30 days old was higher than that of 60 days old.And it could also promote the expression of OPG,BMP-2 and Runx-2 genes and inhibit the expression of TNF-α,RANK and RANKL genes (P<0.05).From above mentioned results,considering the relative indexes of live weight and bone strength,compound Chinese herbal medicine high dose group was the best formula to improve the bone characteristics of the broilers.The compound Chinese herbal medicine could improve the content of VDR,OT,OPG and ALP in serum,regulate the OPG/RANK/RANKL system to maintain the dynamic balance of bone resorption and bone formation,promote the expression of BMP-2 and Runx-2 genes in the important pathway of bone formation BMP-2/Smads/Runx-2,and inhibit TNF-α gene expression,so as to improve the skeleton characteristics of Kosbao broilers,and effectively prevent bone disease of broilers.

Key words: Chinese herbal medicine; Kosbao broiler; bone disease; related gene

中图分类号:

S853.74

段小花, 徐志强, 葛长荣, 贾俊静, 豆腾飞. 中草药复方防治肉鸡骨病的研究[J]. 中国畜牧兽医, 2021, 48(1): 385-394.

DUAN Xiaohua, XU Zhiqiang, GE Changrong, JIA Junjing, DOU Tengfei. Prevention and Treatment of a Chinese Herbal Medicine Compound for Bone Disease in Chickens[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(1): 385-394.

参考文献

[1] 郭晓宇.维生素A过量对肉鸡钙、磷沉积及碱性磷酸酶和骨钙素表达的影响[D].呼和浩特:内蒙古农业大学,2010. GUO X Y.Effects of excessive vitamin A on deposition of calcium and phosphorus and expression of alkaline phosphatase and bone Gla-protein in broiler chickens[D].Hohhot:Inner Mongolia Agricultural University,2010.
[2] WANG S J,YUE W,RAHMAN K,et al.Mechanism of treatment of kidney deficiency and osteoporosis is similar by traditional Chinese medicine[J].Current Pharmaceutical Design,2016,22(3):312-320.
[3] XIE Q F,XIE J H,DONG T X,et al.Effect of a derived herbal recipe from an ancient Chinese formula,Danggui Buxue Tang,on ovariectomized rats[J].Journal of Ethnopharmacology,2012,144(2):567-575.
[4] 贾朝娟,鞠大宏,刘梅洁,等.山药对卵巢切除大鼠骨质疏松症的治疗作用及其机理探讨[J].中国中医基础医学杂志,2009,15(4):268-271. JIA C J,JU D H,LIU M J,et al.Therapeutic effect and mechanism of Chinese yam on osteoporosis in ovariectomized rats[J].Journal of Basic Medicine in Traditional Chinese Medicine,2009,15(4):268-271.(in Chinese)
[5] HE X F,SHEN Q. Salvianolic acid B promotes bone formation by increasing activity of alkaline phosphatase in a rat tibia fracture model:A pilot study[J].BMC Complementary and Alternative Medicine,2014,14:493.
[6] KIM E J,LEE H,KIM M H,et al.Inhibition of RANKL-stimulated osteoclast differentiation by Schisandra chinensis through down-regulation of NFATc1 and c-fos expression[J].BMC Complementary and Alternative Medicine, 2018,18(1):270-278.
[7] ZHANG Z R,LEUNG W N,LI G,et al.Osthole enhances osteogenesis in osteoblasts by elevating transcription factor osterix via cAMP/CREB signaling in vitro and in vivo[J].Nutrients,2017,9(6):588.
[8] LEE Y K,JUNG S K,CHANG Y H,et al.Highly bioavailable nanocalcium from oyster shell for preventing osteoporosis in rats[J].International Journal of Food Sciences and Nutrition,2017,68(8):931-940.
[9] 冯海鹏,辛蕊华,张凯,等.中兽药复方组合与抗生素防治肉鸡呼吸道疾病的协同效应[J].中国畜牧兽医,2020,47(3):940-947. FENG H P,XIN R H,ZHANG K,et al.Synergistic effect of combination of Chinese veterinary drugs and antibiotics on the prevention and treatment of respiratory diseases in broilers[J].China Animal Husbandry & Veterinary Medicine,2020,47(3):940-947.(in Chinese)
[10] YANG R,CHEN J,ZHANG J,et al.1,25-dihydroxyvitamin D protects against age-related osteoporosis by a novel VDR-Ezh2-p16 signal axis[J].Aging Cell, 2020,19(2):13095-13115.
[11] 郑敏,刘强.维生素D及维生素D受体的研究进展[J].医学综述,2013,19(21):3965-3967. ZHENG M,LIU Q.The progress in the studies of vitamin D and vitamin D receptor[J].Medical Recapitulate,2013,19(21):3965-3967.(in Chinese)
[12] 刘红,廖二元,伍贤平.骨钙素与代谢性骨病[J].国外医学:内分泌学分册,2004,24(4):239-240. LIU H,LIAO E Y,WU X P.Osteocalcin and metabolic bone disease[J].Foreign Medicine:Endocrino-logy Branch, 2004,24(4):239-240.(in Chinese)
[13] 邓宏明,李世生.年龄、性别及某些内分泌疾病对血清骨钙素水平的影响[J].广西医学,2001,23(6):1341-1343. DENG H M,LI S S.Study on effect of sex,age and common endocrine disease on serum BGP level[J].Guangxi Medical Journal,2001,23(6):1341-1343.(in Chinese)
[14] 文娱,陈雪梅.骨保护素在临床中的应用进展[J].中国当代医药,2013,20(3):12-13. WEN Y,CHEN X M.Research progress of osteoprotegerin in clinic[J].China Modern Medicine,2013,20(3):12-13.(in Chinese)
[15] HASS M,LEKOO-MOHR Z,ROSCHGER P,et al.Osteoprotegerin and parathyroid hormone as markers of high-tumover osteodystrophy and decreased bone mineralization in hemodialysis patients[J].American Journal of Kidney Diseases:The Official Journal of the National Kidney Foundation,2002,39(3):580-586.
[16] OSAMA M A,MARIAN Y W,RAHMA K A,et al.Potential effectiveness of exenatide in experimentally-induced osteoporosis[J].The Egyptian Rheumatologist,2020,42(1):57-62.
[17] 张美莉,魏忠义.艾维菌肉种母鸡骨骼、肌肉、脂肪组织生长发育规律研究[J].安徽农业科学,2007,35(24):7480-7481. ZHANG M L,WEI Z Y.Study on the growth and development laws of skeleton,muscle and adipose tissue of hen[J].Anhui Agricultural Sciences, 2007,35(24):7480-7481.(in Chinese)
[18] 陶振阳,张耀文,曹忠洋,等.粪肠球菌对产蛋后期蛋鸡生产性能、蛋品质及肠道组织形态的影响[J].中国畜牧兽医,2020,47(5):1413-1420. TAO Z Y,ZHANG Y W,CAO Z Y,et al.Effects of Enterococcus faecalis on production performance,egg quality and intestinal morphology of laying hens during the late laying period[J].China Animal Husbandry & Veterinary Medicine,2020,47(5):1413-1420.(in Chinese)
[19] 王丽赛,廖秀冬,冯艳忠,等.日粮硒水平对22~42日龄肉仔鸡生长性能及肉品质的影响[J].中国畜牧兽医,2020,47(7):2063-2070. WANG L S,LIAO X D,FENG Y Z,et al.Effect of dietary selenium level on growth performance and meat quality of broilers from 22 to 42 days of age[J].China Animal Husbandry & Veterinary Medicine,2020,47(7):2063-2070.(in Chinese)
[20] COUSMINER D L,MITCHELL J A,CHESI A,et al.Genetically determined later puberty impacts lowered bone mineral density in childhood and adulthood[J].Journal of Bone and Mineral Research:The Official Journal of the American Society for Bone and Mineral Research,2018;33(3):430-436.
[21] SANCHEZ-SILES J M,TAMIMI-MARINO I,CORTES A R,et al.Age related changes in the bone microstructure in patients with femoral neck fractures[J].Injury,2020,51(1):S12-S18.
[22] ZHAO Y,WANG H L,LI T T,et al.Baicalin ameliorates dexamethasone-induced osteoporosis by regulation of the RANK/RANKL/OPG signaling pathway[J].Drug Design,Development and Therapy,2020,14:195-206.
[23] TOBEIHA M,MOGHADASIAN M H,AMIN N,et al.RANKL/RANK/OPG pathway:A mechanism involved in exercise-induced bone remodeling[J].BioMed Research International,2020,69(10):3-12.
[24] SIEKLUCKA B,DOMANIEWSKI T,ZIEMINSKA M,et al.P0690 correlations between OPG/RANKL/RANK axis,vitamin D satus,PTH and vascular calcification in an adenine-induced model of chronic kidney disease[J].Nephrology Dialysis Transplantation,2020,35(3):690-695.
[25] INJAMURI S,RAHAMAN M N,SHEN Y,et al.Relaxin enhances bone regeneration with BMP-2-loaded hydroxyapatite microspheres[J].Journal of Biomedical Materials Research Part A,2020,108(5):1231-1242.
[26] QI S,HE J,ZHENG H,et al.Zinc supplementation increased bone mineral density,improves bone histomorphology,and prevents bone loss in diabetic rat[J].Biological Trace Element Research,2020,194(2):493-501.
[27] OHORI F,KITAURA H,OGAWA S,et al.IL-33 inhibits TNF-α-induced osteoclastogenesis and bone resorption[J].International Journal of Molecular Sciences,2020,21(3):1130-1137.
[28] YANG B,SUN H,XU X,et al.YAP1 inhibits the induction of TNF-α stimulated bone resorbing mediators by suppressing the NF-κB signaling pathway in MC3T3-E1 cells[J].Journal of Cellular Physiology,2020,235(5):4698-4708.