术苦芩总多糖对湿热泄泻仔猪肠道菌群和免疫功能的影响

doi:10.11843/j.issn.0366-6964.2022.03.024

Abstract

Abstract: This study was carried out to investigate the effect of polysaccharides from ZhuKuQin (ZKQPs) on the intestinal flora and immune function of piglets with dampness-heat diarrhea. We analyzed the monosaccharide composition of ZKQPs by HPLC, and studied the direct effects of ZKQPs on common intestinal pathogens in vitro antibacterial experiments. At the same time, we establish a piglet diarrhea model, and treated with Pulsatilla powder and three doses of ZKQPs (25, 50 and 75 mg·kg^-1). After the treatment, the small intestine tissues and the contents of the cecum were collected. The cecal colony was identified and counted; the changes of intestinal flora were analyzed by 16S rDNA high-throughput sequencing, and the mRNA expression of small intestinal immune-related cytokines was determined. The results showed that the primary structure of ZKQPs is mainly composed of rhamnose, glucose, galactose and xylose. In vitro tests, E. coli, Salmonella, and Clostridium wischii were sensitive to ZKQPs and showed a concentration dependence within a certain range. In animal experiments, ZKQPs effectively promoted the growth of intestinal Lactobacilli in piglets with diarrhea, and had a strong inhibitory effect on the growth of pathogenic E. coli. Among them, 50 mg·kg^-1ZKQPs had the most significant effect. The results of 16S rDNA high-throughput sequencing showed that ZKQPs regulate the Alpha diversity of diarrhea piglets, and had a significant impact on the intestinal flora at the class and genera level. The relative abundance of Bacillus and Lactobacillus decreased significantly and the relative abundance of bacteria increased significantly in diarrhea piglets; after ZKQPs treatment, the relative abundances of Bacillus and Lactobacillus increased significantly and were higher than normal, and the relative abundance of Clostridia significantly decreased, which significantly improved the intestinal flora structure of diarrhea piglets. In piglets with diarrhea, the level of Th1 cytokine IL-2 mRNA in each segment of the small intestine increased (P<0.01), while the Th2 cytokine IL-4 and IL-5 mRNA levels decreased; after treatment with ZKQPs, the levels of IL-2 mRNA in the small intestine decreased, the levels of IL-4 and IL-5 mRNA generally increased, and the mRNA levels of different cytokines vary in different locations in the small intestine. ZKQPs 50 and 75 mg·kg^-1 have the most significant effects on them, comprehensive analysis ZKQPs 50 mg·kg^-1 treatment of diarrhea piglets has the greatest impact on the level of cytokine mRNA. This study shows that ZKQPs have a good inhibitory effect on the growth of intestinal pathogens in vitro, can change the abundance and diversity of the intestinal microbiota of diarrhea piglets, increase the relative abundance of Lactobacillus and improve the structure of the flora. At the same time, ZKQPs regulate the intestinal immune response and effectively enhance the anti-diarrhea ability of piglets.

Key words: polysaccharides from Zhukuqin, diarrhea piglets, intestinal flora, immune

CLC Number:

S853.74

TAO Weilai, LIU Jia, LIU Qiongdan, HAO Yongfeng, HU Yu, ZHU Zhaorong, LIU Juan. Effects of Total Polysaccharides from Zhukuqin on Intestinal Flora and Immune Function in Piglets with Dampness-heat Diarrhea[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(3): 913-924.

References 36

[1]	ZHANG H H, XU Y P, ZHANG Z J, et al. Protective immunity of a multivalent vaccine candidate against piglet diarrhea caused by enterotoxigenic Escherichia coli (ETEC) in a pig model[J]. Vaccine, 2018, 36(5):723-728.
[2]	WANG W W, LIU Y, TANG H, et al. ITGB5 plays a key role in Escherichia coli F4ac-induced diarrhea in piglets[J]. Front Immunol, 2019, 10:2834.
[3]	刘钟杰,许剑琴.中兽医学[M]. 4版.北京:中国农业出版社, 2011:49-50.LIU Z J, XU J Q. Traditional Chinese veterinary medicine[M]. 4th ed. Beijing:China Agriculture Press, 2005:353-354.(in Chinese)
[4]	WANG X M, TANG Y Q, LIU C X, et al. Effects of restrictive prescribing on antibiotic consumption in primary care in China, 2012-17:an interrupted time-series analysis[J]. Lancet, 2019, 394(S1):S97.
[5]	YANG J H,BHARGAVA P,MCCLOSKEY D,et al. Antibiotic-induced changes to the host metabolic environment inhibit drug efficacy and alter immune function[J].Cell Host Microbe,2017,22(6):757-765.
[6]	O'HARA A M, SHANAHAN F. The gut flora as a forgotten organ[J]. EMBO Rep, 2006, 7(7):688-693.
[7]	PICKARD J M, ZENG M Y, CARUSO R, et al. Gut microbiota:role in pathogen colonization, immune responses, and inflammatory disease[J]. Immunol Rev, 2017, 279(1):70-89.
[8]	BÄUMLER A J, SPERANDIO V. Interactions between the microbiota and pathogenic bacteria in the gut[J]. Nature, 2016, 535(7610):85-93.
[9]	YANG Y, CHEN G, YANG Q, et al. Gut microbiota drives the attenuation of dextran sulphate sodium-induced colitis by Huangqin decoction[J]. Oncotarget, 2017, 8(30):48863-48874.
[10]	WU T R, LIN C S, CHANG C J, et al. Gut commensal Parabacteroides goldsteinii plays a predominant role in the anti-obesity effects of polysaccharides isolated from Hirsutella sinensis[J]. Gut, 2019, 68(2):248-262.
[11]	BOULANGÉ C L, NEVES A L, CHILLOUX J, et al. Impact of the gut microbiota on inflammation, obesity, and metabolic disease[J]. Genome Med, 2016, 8(1):42.
[12]	ALLAIRE J M, CROWLEY S M, LAW H T, et al. The intestinal epithelium:central coordinator of mucosal immunity[J]. Trends Immunol, 2018, 39(9):677-696.
[13]	TILG H, ZMORA N, ADOLPH T E, et al. The intestinal microbiota fuelling metabolic inflammation[J]. Nat Rev Immunol, 2020, 20(1):40-54.
[14]	BEL S, PENDSE M, WANG Y H, et al. Paneth cells secrete lysozyme via secretory autophagy during bacterial infection of the intestine[J]. Science, 2017, 357(6355):1047-1052.
[15]	FUJIMOTO K, KAWAGUCHI Y, SHIMOHIGOSHI M, et al. Antigen-specific mucosal immunity regulates development of intestinal bacteria-mediated diseases[J]. Gastroenterology, 2019, 157(6):1530-1543.
[16]	SHEN J, CHEN J J, ZHANG B M, et al. Baicalin is curative against rotavirus damp heat diarrhea by tuning colonic mucosal barrier and lung immune function[J]. Dig Dis Sci, 2020, 65(8):2234-2245.
[17]	ZENG P J, LI J, CHEN Y L, et al. The structures and biological functions of polysaccharides from traditional Chinese herbs[J]. Prog Mol Biol Transl Sci, 2019, 163:423-444.
[18]	林春发,郝永峰,刘娟.术苦芩总多糖对湿热泄泻仔猪小肠杯状细胞数量以及MUC-2和ITF-3 mRNA转录的影响[J].畜牧兽医学报, 2019, 50(6):1301-1311.LIN C F, HAO Y F, LIU J. Effect of polysaccharides from ZhuKuQin on the number of intestinal goblet cell and the transcription of MUC-2 and ITF-3 mRNA in the small intestine of damp-heat diarrhea piglets[J]. Acta Veterinaria et Zootechnica Sinica, 2019, 50(6):1301-1311.(in Chinese)
[19]	CUI L, WANG W, LUO Y, et al. Polysaccharide from Scutellaria baicalensis Georgi ameliorates colitis via suppressing NF-κB signaling and NLRP3 inflammasome activation[J]. Int J Biol Macromol, 2019, 132:393-405.
[20]	叶文初,袁富威,班贇,等.苦豆草提取物对奶牛乳房炎致病菌的体外抑菌作用[J].中国兽医学报, 2016, 36(9):1568-1571.YE W C, YUAN F W, BAN Y, et al. The antibacterial effect of sophora alopecuroides extract on mastitis pathogens in vitro[J]. Chinese Journal of Veterinary Science, 2016, 36(9):1568-1571.(in Chinese)
[21]	DIAO H, ZHENG P, YU B, et al. Effects of dietary supplementation with benzoic acid on intestinal morphological structure and microflora in weaned piglets[J]. Livestock Sci, 2014, 167:249-256.
[22]	GUILLIAMS M, GINHOUX F, JAKUBZICK C, et al. Dendritic cells, monocytes and macrophages:a unified nomenclature based on ontogeny[J]. Nat Rev Immunol, 2014, 14(8):571-578.
[23]	FERREIRA I C F R, HELENO S A, REIS F S, et al. Chemical features of Ganoderma polysaccharides with antioxidant, antitumor and antimicrobial activities[J]. Phytochemistry, 2015, 114:38-55.
[24]	GRESSE R, CHAUCHEYRAS-DURAND F, FLEURY M A, et al. Gut microbiota dysbiosis in postweaning piglets:understanding the keys to health[J]. Trends Microbiol, 2017, 25(10):851-873.
[25]	CÖR D, KNEZŽ, KNEZ HRN AČU I AČU M. Antitumour, antimicrobial, antioxidant and antiacetylcholinesterase effect of Ganoderma lucidum terpenoids and polysaccharides:a review[J]. Molecules, 2018, 23(3):649.
[26]	杨莉,杨茂生,史开志,等.猪细菌性腹泻的病原分离及血清型鉴定[J].黑龙江畜牧兽医, 2017(8):100-102.YANG L, YANG M S, SHI K Z, et al. Pathogen isolation and serotype identification of swine bacterial diarrhea[J].Heilongjiang Animal Science and Veterinary Medicine, 2017(8):100-102.(in Chinese)
[27]	DONG L N, WANG M, GUO J, et al. Role of intestinal microbiota and metabolites in inflammatory bowel disease[J]. Chin Med J (Engl), 2019, 132(13):1610-1614.
[28]	PIEWNGAM P, ZHENG Y, NGUYEN T H, et al. Pathogen elimination by probiotic Bacillus via signalling interference[J]. Nature, 2018, 562(7728):532-537.
[29]	MEDZHITOV R. Origin and physiological roles of inflammation[J]. Nature, 2008, 454(7203):428-435.
[30]	HU R Z, HE Z Y, LIU M, et al. Dietary protocatechuic acid ameliorates inflammation and up-regulates intestinal tight junction proteins by modulating gut microbiota in LPS-challenged piglets[J]. J Anim Sci Biotechnol, 2020, 11(1):92.
[31]	YUE S S, LI Z T, HU F L, et al. Curing piglets from diarrhea and preparation of a healthy microbiome with Bacillus treatment for industrial animal breeding[J]. Sci Rep, 2020, 10(1):19476.
[32]	JARRET A, JACKSON R, DUIZER C, et al. Enteric nervous system-derived IL-18 orchestrates mucosal barrier immunity[J]. Cell, 2020, 180(1):50-63.
[33]	ZHOU B L, YUAN Y T, ZHANG S S, et al. Intestinal flora and disease mutually shape the regional immune system in the intestinal tract[J]. Front Immunol, 2020, 11:575.
[34]	ZHOU L, CHU C, TENG F, et al. Innate lymphoid cells support regulatory T cells in the intestine through interleukin-2[J]. Nature, 2019, 568(7752):405-409.
[35]	WYNN T A. Type 2 cytokines:mechanisms and therapeutic strategies[J]. Nat Rev Immunol, 2015, 15(5):271-282.
[36]	FENG J S, WANG L, XIE Y S, et al. Effects of antimicrobial peptide cathelicidin-BF on diarrhea controlling, immune responses, intestinal inflammation and intestinal barrier function in piglets with postweaning diarrhea[J]. Int Immunopharmacol, 2020, 85:106658.

Effects of Total Polysaccharides from Zhukuqin on Intestinal Flora and Immune Function in Piglets with Dampness-heat Diarrhea

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