中药十味配伍复方通过 MAPK/ERK/JNK 通路缓解鸡毒支原体感染对鸡免疫功能和生产性能的影响

doi:10.1016/j.jia.2023.11.043

摘要/Abstract

摘要：

鸡毒支原体（Mycoplasma gallisepticum，MG）是养禽业中一种常见且易引起严重问题的病原体。MG感染可导致多种疾病，包括鸡慢性呼吸道疾病，气管炎和结膜炎等。其典型症状表现为呼吸急促、打喷嚏和眼部分泌物增多。此外，MG感染不仅会破坏免疫器官引起免疫抑制，还会导致鸡的生产性能下降，包括体重增长减缓、蛋产量减少，甚至死亡，严重威胁了养禽业的可持续发展。

目前，防治 MG 感染的有效措施是使用抗生素。然而，随着抗生素的使用和滥用，MG对抗生素的耐药性也在增加，同时引发食品中的药物残留问题，对人类食品安全构成潜在威胁。为了寻找可持续的防治方法，本研究采用了葛根、虎杖、黄芩、甘草等九味中草药配伍的复方，结合体内和体外实验，探究了其对MG的防治效果及作用机制。

研究结果显示，中药复方在体内外均能有效抑制MG的增殖。在生产性能方面，中药复方显著降低了MG感染引起的死亡率，提高了饲料转化率，并显著改善了体重增长减缓的问题。此外，与未治疗组相比，中药复方显著提高了免疫器官指数，增强了鸡体的免疫力，缓解了气管和肺组织的损伤，并抑制了组织和细胞的炎症反应。更重要的是，研究结果显示，中药复方对MG感染具有很好的预防效果。

为深入揭示中药复方抵抗MG感染的分子机制，我们进行了网络药理学分析，并通过体内外实验验证了MAPK/ERK/JNK通路是中药复方抑制MG感染的关键调节通路。本研究不仅结合体内外的实验为解析MG的致病机制提供了理论支持，同时也强有力地表明，中药复方可以有效替代抗生素，缓解MG感染对养禽业的危害。这一发现对于养禽业的可持续发展具有重要的实际意义，并为未来的研究提供了有力支持。

Abstract:

Mycoplasma gallisepticum (MG) is a common avian pathogen that mainly infects poultry, causing significant reductions in body weight gain and egg production, along with damage to immune organs and immunosuppression. MG is susceptible to co-infections with other pathogens, leading to increased mortality rates and significant economic losses in the global poultry industry. While antibiotics have been extensively applied worldwide to treat MG infections in poultry production, concerns regarding antibiotic resistance and residue remain prevalent. Traditional Chinese medicine (TCM), renowned for its natural, safe, and non-toxic properties, has shown significant anti-inflammatory and immune-enhancing effects. This study aimed to investigate the protective effect of TCM on production performance and its impact on MG-induced immunosuppression through the MAPK/ERK/JNK signaling pathway in chickens. Our results showed that TCM alleviated the negative effects of MG infection on production performance, as evidenced by improvements in body weight gain, feed conversion rate, survival rate, and immune organ index. TCM exhibited direct inhibition of the MG proliferation in vitro and in vivo. Furthermore, TCM treatment promoted the normalization of trachea and lung tissue structure in MG-infected chickens, leading to a significant reduction in inflammatory damage. Moreover, following the treatment with the TCM, the production of pro-inflammatory cytokines Interleukin-1β (IL-1β) and Tumor Necrosis Factor-α (TNF-α) decreased significantly, accompanied by the downregulation of pro-apoptotic genes caspase3, caspase9, and BAX, both in vitro and in vivo. A mechanism-based study showed that in vitro and in vivo treatment with the TCM significantly reduced the expression of key proteins, including early growth response gene1 (EGR1), p-ERK, p-JNK, and p-JUN. Altogether, TCM improved body weight gain, inhibited pro-inflammation responses, and alleviated tissue damage by inhibiting the MAPK/ERK/JNK signaling pathway to protect the performance and immune system of MG-infected chickens.

Key words: Mycoplasma gallisepticum (MG) , , Traditional Chinese medicine (TCM) , , production performance , , inflammation , , MAPK/ERK/JNK , , chicken

. 中药十味配伍复方通过 MAPK/ERK/JNK 通路缓解鸡毒支原体感染对鸡免疫功能和生产性能的影响[J]. Journal of Integrative Agriculture, DOI: 10.1016/j.jia.2023.11.043.

WANG Teng-fei, FAN Chang-yong, XIAO Yu-fei, LV Shan, JIANG Guang-yang, ZOU Meng-yun, WANG Ying-jie, GUO Qiao, CHE Zheng-hao, PENG Xiu-li. [J]. Journal of Integrative Agriculture, DOI: 10.1016/j.jia.2023.11.043.

参考文献

Abd El-Hamid M I, Ibrahim S M, Eldemery F, El-Mandrawy S A M, Metwally A S, Khalifa E, Elnahriry S S, Ibrahim D. 2021. Dietary cinnamaldehyde nanoemulsion boosts growth and transcriptomes of antioxidant and immune related genes to fight Streptococcus agalactiae infection in Nile tilapia (Oreochromis niloticus). Fish Shellfish Immunol, 113, 96-105.

Aljazzar A, Abd El-Hamid M I, El-Malt R M S, El-Gharreb W R, Abdel-Raheem S M, Ibrahim A M, Abdelaziz A M, Ibrahim D. 2022. Prevalence and Antimicrobial Susceptibility of Campylobacter Species with Particular Focus on the Growth Promoting, Immunostimulant and Anti-Campylobacter jejuni Activities of Eugenol and Trans-Cinnamaldehyde Mixture in Broiler Chickens. Animals (Basel), 12.

An X, Zhang Y, Duan L, Jin D, Zhao S, Zhou R, Duan Y, Lian F, Tong X. 2021. The direct evidence and mechanism of traditional Chinese medicine treatment of COVID-19. Biomed Pharmacother, 137, 111267.

Cakarova L, Marsh L M, Wilhelm J, Mayer K, Grimminger F, Seeger W, Lohmeyer J, Herold S. 2009. Macrophage tumor necrosis factor-alpha induces epithelial expression of granulocyte-macrophage colony-stimulating factor: impact on alveolar epithelial repair. Am J Respir Crit Care Med, 180, 521-532.

de Jong A, Youala M, Klein U, El Garch F, Simjee S, Moyaert H, Rose M, Gautier-Bouchardon A V, Catania S, Ganapathy K, Gyuranecz M, Moller Palau-Ribes F, Pridmore A, Ayling R D. 2021. Minimal inhibitory concentration of seven antimicrobials to Mycoplasma gallisepticum and Mycoplasma synoviae isolates from six European countries. Avian Pathol, 50, 161-173.

Dhondt A A, Altizer S, Cooch E G, Davis A K, Dobson A, Driscoll M J, Hartup B K, Hawley D M, Hochachka W M, Hosseini P R, Jennelle C S, Kollias G V, Ley D H, Swarthout E C, Sydenstricker K V. 2005. Dynamics of a novel pathogen in an avian host: Mycoplasmal conjunctivitis in house finches. Acta Trop, 94, 77-93.

Feberwee A, de Wit S, Dijkman R. 2022. Clinical expression, epidemiology, and monitoring of Mycoplasma gallisepticum and Mycoplasma synoviae: an update. Avian Pathol, 51, 2-18.

Ge H, Wang Y F, Xu J, Gu Q, Liu H B, Xiao P G, Zhou J, Liu Y, Yang Z, Su H. 2010. Anti-influenza agents from Traditional Chinese Medicine. Nat Prod Rep, 27, 1758-1780.

Hashem Y M, Abd El-Hamid M I, Awad N F S, Ibrahim D, Elshater N S, El-Malt R M S, Hassan W H, Abo-Shama U H, Nassan M A, El-Bahy S M, Samy O M, El Sharkawy R B, Algabri N, Elnahriry S S. 2022. Insights into growth-promoting, anti-inflammatory, immunostimulant, and antibacterial activities of Toldin CRD as a novel phytobiotic in broiler chickens experimentally infected with Mycoplasma gallisepticum. Poult Sci, 101, 102154.

Hu F, Zhao C, Bi D, Tian W, Chen J, Sun J, Peng X. 2016. Mycoplasma gallisepticum (HS strain) surface lipoprotein pMGA interacts with host apolipoprotein A-I during infection in chicken. Appl Microbiol Biotechnol, 100, 1343-1354.

Huang Q, Niu Z, Han J, Liu X, Lv Z, Li H, Yuan L, Li X, Sun S, Wang H, Huang X. 2017. HTLV-1 Tax upregulates early growth response protein 1 through nuclear factor-kappaB signaling. Oncotarget, 8, 51123-51133.

Ibrahim D, Ismail T A, Khalifa E, Abd El-Kader S A, Mohamed D I, Mohamed D T, Shahin S E, Abd El-Hamid M I. 2021. Supplementing Garlic Nanohydrogel Optimized Growth, Gastrointestinal Integrity and Economics and Ameliorated Necrotic Enteritis in Broiler Chickens Using a Clostridium perfringens Challenge Model. Animals (Basel),11.

Ishfaq M, Hu W, Khan M Z, Ahmad I, Guo W, Li J. 2020. Current status of vaccine research, development, and challenges of vaccines for Mycoplasma gallisepticum. Poult Sci, 99, 4195-4202.

Ishfaq M, Zhang W, Liu Y, Wang J, Wu Z, Shah S W, Li R, Miao Y, Chen C, Li J. 2021. Baicalin attenuated Mycoplasma gallisepticum-induced immune impairment in chicken bursa of fabricius through modulation of autophagy and inhibited inflammation and apoptosis. J Sci Food Agric, 101, 880-890.

Karp B E, Tate H, Plumblee J R, Dessai U, Whichard J M, Thacker E L, Hale K R, Wilson W, Friedman C R, Griffin P M, McDermott P F. 2017. National Antimicrobial Resistance Monitoring System: Two Decades of Advancing Public Health Through Integrated Surveillance of Antimicrobial Resistance. Foodborne Pathog Dis, 14, 545-557.

Kunnumakkara A B, Rana V, Parama D, Banik K, Girisa S, Henamayee S, Thakur K K, Dutta U, Garodia P, Gupta S C, Aggarwal B B. 2021. COVID-19, cytokines, inflammation, and spices: How are they related? Life Sci, 284, 119201.

Li H, Yang L, Liu F F, Ma X N, He P L, Tang W, Tong X K, Zuo J P. 2020. Overview of therapeutic drug research for COVID-19 in China. Acta Pharmacol Sin, 41, 1133-1140.

Li Z Y, Hao E W, Cao R, Lin S, Chen S Y, Huang X T, Xu W R, Hou X T, Deng J G. 2022. [Medication law and mechanism of traditional Chinese medicine in prevention and treatment of epidemic diseases: based on traditional Chinese medicine theory of cold pestilence]. Zhongguo Zhong Yao Za Zhi, 47, 4765-4777.

Luo H, Tang Q L, Shang Y X, Liang S B, Yang M, Robinson N, Liu J P. 2020. Can Chinese Medicine Be Used for Prevention of Corona Virus Disease 2019 (COVID-19)? A Review of Historical Classics, Research Evidence and Current Prevention Programs. Chin J Integr Med, 26, 243-250.

Luo R, Wang Y, Guo Q, Fan C, Jiang G, Wang L, Zou M, Wang T, Sun Y, Peng X. 2022. Andrographolide attenuates Mycoplasma gallisepticum-induced inflammation and apoptosis by the JAK/PI3K/AKT signal pathway in the chicken lungs and primary alveolar type II epithelial cells. Int Immunopharmacol, 109, 108819.

Machado L D, Santos F F, Togashi C K, Abreu D L, Pimentel J C, Sesti L, Pereira V L, Nascimento E R. 2017. Egg quality in laying hens exposed to Mycoplasma gallisepticum F-strain attenuated vaccine. Poult Sci, 96, 976-979.

Mancuso G, Midiri A, Gerace E, Biondo C. 2021. Bacterial Antibiotic Resistance: The Most Critical Pathogens. Pathogens, 10.

Md.Sh B, M A M, Md.M R, M N. 2021. Isolation and molecular detection of Mycoplasma gallisepticum in commercial layer chickens in Sylhet, Bangladesh. . World Vet. J., 614–620.

Miao Y, Niu D, Wang Z, Wang J, Wu Z, Bao J, Jin X, Li R, Ishfaq M, Li J. 2022. Methylsulfonylmethane ameliorates inflammation via NF-kappaB and ERK/JNK-MAPK signaling pathway in chicken trachea and HD11 cells during Mycoplasma gallisepticum infection. Poult Sci, 101, 101706.

Much P, Winner F, Stipkovits L, Rosengarten R, Citti C. 2002. Mycoplasma gallisepticum: Influence of cell invasiveness on the outcome of experimental infection in chickens. FEMS Immunol Med Microbiol, 34, 181-186.

Mugunthan S P, Kannan G, Chandra H M, Paital B. 2023. Infection, Transmission, Pathogenesis and Vaccine Development against Mycoplasma gallisepticum. Vaccines (Basel), 11.

Musarra-Pizzo M, Pennisi R, Ben-Amor I, Mandalari G, Sciortino M T. 2021. Antiviral Activity Exerted by Natural Products against Human Viruses. Viruses, 13.

Natalicchio A, De Stefano F, Orlando M R, Melchiorre M, Leonardini A, Cignarelli A, Labarbuta R, Marchetti P, Perrini S, Laviola L, Giorgino F. 2010. Exendin-4 prevents c-Jun N-terminal protein kinase activation by tumor necrosis factor-alpha (TNFalpha) and inhibits TNFalpha-induced apoptosis in insulin-secreting cells. Endocrinology, 151, 2019-2029.

Niu L, Luo R, Zou M, Sun Y, Fu Y, Wang Y, Peng X. 2020. Puerarin inhibits Mycoplasma gallisepticum (MG-HS)-induced inflammation and apoptosis via suppressing the TLR6/MyD88/NF-kappaB signal pathway in chicken. Int Immunopharmacol, 88, 106993.

Park J I. 2023. MAPK-ERK Pathway. Int J Mol Sci, 24.

Seidavi A, Tavakoli M, Slozhenkina M, Gorlov I, Hashem N M, Asroosh F, Taha A E, Abd El-Hack M E, Swelum A A. 2021. The use of some plant-derived products as effective alternatives to antibiotic growth promoters in organic poultry production: a review. Environ Sci Pollut Res Int, 28, 47856-47868.

Sun Y, Wang Y, Zou M, Wang T, Wang L, Peng X. 2022. Lnc90386 Sponges miR-33-5p to Mediate Mycoplasma gallisepticum-Induced Inflammation and Apoptosis in Chickens via the JNK Pathway. Front Immunol, 13, 887602.

Tian W, Zhao C, Hu Q, Sun J, Peng X. 2016. Roles of Toll-like receptors 2 and 6 in the inflammatory response to Mycoplasma gallisepticum infection in DF-1 cells and in chicken embryos. Dev Comp Immunol, 59, 39-47.

Trizzino M, Zucco A, Deliard S, Wang F, Barbieri E, Veglia F, Gabrilovich D, Gardini A. 2021. EGR1 is a gatekeeper of inflammatory enhancers in human macrophages. Sci Adv, 7.

Van Belleghem J D, Dabrowska K, Vaneechoutte M, Barr J J, Bollyky P L. 2018. Interactions between Bacteriophage, Bacteria, and the Mammalian Immune System. Viruses, 11.

Wang T, Xiao Y, Luo R, Wang Y, Zou M, Sun Y, Wang L, Guo Q, Peng X. 2022. Host resistance to Mycoplasma gallisepticum infection is enhanced by inhibiting PI3K/Akt pathway in Andrographolide-treating chickens. Int Immunopharmacol, 113, 109419.

Wang Y, Sun H, Zhao W, Wang T, Zou M, Han Y, Sun Y, Peng X. 2023. Low let-7d microRNA levels in chick embryos enhance innate immunity against Mycoplasma gallisepticum by suppressing the mitogen-activated protein kinase pathway. Vet Res, 54, 50.

Yang Y, Wang Y, Zou M, Deng G, Peng X. 2021. gga-miR-142-3p negatively regulates Mycoplasma gallisepticum (HS strain)-induced inflammatory cytokine production via the NF-kappaB and MAPK signaling by targeting TAB2. Inflamm Res, 70, 1217-1231.

Yoon H S, Kim H A. 2004. Prologation of c-Jun N-terminal kinase is associated with cell death induced by tumor necrosis factor alpha in human chondrocytes. J Korean Med Sci, 19, 567-573.

Yu J, Chen Y, Zhai L, Zhang L, Xu Y, Wang S, Hu S. 2015. Antioxidative effect of ginseng stem-leaf saponins on oxidative stress induced by cyclophosphamide in chickens. Poult Sci, 94, 927-933.

Zhang D H, Wu K L, Zhang X, Deng S Q, Peng B. 2020. In silico screening of Chinese herbal medicines with the potential to directly inhibit 2019 novel coronavirus. J Integr Med, 18, 152-158.

Zhang J, Li Q, Chang S. 2019. The effects of particle density in moxa smoke on the ultrastructure of knee cartilage and expressions of TNF-alpha, IL-1b, BAX, and Bcl-2 mRNA in a rat model for osteoarthritis. J Cell Biochem, 120, 6589-6595.

Zhao Y, Fu Y, Zou M, Sun Y, Yin X, Niu L, Gong Y, Peng X. 2020. Analysis of deep sequencing exosome-microRNA expression profile derived from CP-II reveals potential role of gga-miRNA-451 in inflammation. J Cell Mol Med, 24, 6178-6190.

Zhou H, Dai C, Cui X, Zhang T, Che Y, Duan K, Yi L, Nguyen A D, Li N, De Souza C, Wan X, Wu Y, Li K, Liu Y, Wu Y. 2022. Immunomodulatory and antioxidant effects of Glycyrrhiza uralensis polysaccharide in Lohmann Brown chickens. Front Vet Sci, 9, 959449.

Zou M, Yang W, Niu L, Sun Y, Luo R, Wang Y, Peng X. 2020. Polydatin attenuates Mycoplasma gallisepticum (HS strain)-induced inflammation injury via inhibiting the TLR6/ MyD88/NF-kappaB pathway. Microb Pathog, 149, 104552.

Zou M, Yang L, Niu L, Zhao Y, Sun Y, Fu Y, Peng X. 2021. Baicalin ameliorates Mycoplasma gallisepticum-induced lung inflammation in chicken by inhibiting TLR6-mediated NF-kappaB signalling. Br Poult Sci, 62, 199-210.