基于16S rDNA测序分析腹腔注射苦参碱的昆明小鼠肠道菌群结构

doi:10.11843/j.issn.0366-6964.2022.02.028

Abstract

Abstract: The study aimed to analyse the structure of the intestinal flora of Kunming mice injected with matrine by 16S rDNA sequencing. Twenty Kunming mice were randomly divided into 2 groups named as matrine group (MT group) and negative control group (NC group). The intraperitoneal administration was administered twice a day for 5 days, the feces and intestinal tissues of each group were collected for β diversity, Lefse and Metastats analysis. qPCR was used to detect the mRNA expression of different strains in each intestinal segment, and KEGG was used to analyze the differences in metabolic pathways caused by the changes in intestinal flora. The rarefaction curve results showed that the measured sample data was sufficient to reflect the species diversity in the samples. The results of β diversity analysis showed that matrine regulated the structure of the intestinal flora. Lefse and Metastats analysis results showed that matrine significantly increased the abundance of Bacteroidetes, Bacteroidales, Muribaculaceae and probiotic Lactobacillus acidophilus, and significantly reduced the abundance of Firmicutes, Ruminococcaceae and Desulfovibrio. The qPCR results showed that the expression of Lactobacillus acidophilus in the feces of mice in the matrine group was increased, which was consistent with the results of Lefse and Metastats analysis. It was also found that matrine enhanced the colonization of Lactobacillus acidophilus in various segments of the intestine. KEGG analysis found that there were significant differences in metabolic pathways such as glycan biosynthesis and metabolism, transport and catabolism between NC and MT groups. The study results showed that matrine significantly changed the structure of the intestinal flora in mice, increase the colonization of probiotic Lactobacillus acidophilus in the intestine and caused the differences in metabolic pathways such as glycan biosynthesis and metabolism, transport and catabolism. These results have accumulated data for further study of the mechanism of matrine exerting its pharmacological effects.

Key words: matrine, Kunming mice, intestinal flora, 16S rDNA sequencing, Lactobacillus acidophilus

CLC Number:

S852.6

CAO Zhigang, WANG Hong, ZHANG Hua, SUN Panpan, LI Hongquan, SUN Yaogui, YANG Huizhen, WANG Jianzhong, YIN Wei, FAN Kuohai, SUN Na. Influence of Matrine on Intestinal Flora of Kunming Mice through Intraperitoneal Injection Based on 16S rDNA Sequencing[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(2): 618-627.

References 42

[1]	孙盼盼.苦参碱抑制PAMs分泌IL-1β的机理及联合用药研究[D].晋中:山西农业大学,2019.SUN P P.Study on the mechanism of Matrine inhibiting PAMs secrete IL-1β and its combined effect with antibiotics[D].Jinzhong:Shanxi Agricultural University,2019.(in Chinese)
[2]	HU H G,WANG S Z,ZHANG C M,et al.Synthesis and in vitro inhibitory activity of matrine derivatives towards pro-inflammatory cytokines[J].Bioorg Med Chem Lett,2010,20(24):7537-7539.
[3]	王登科,田兴仓,王小萍.苦参碱对高脂血症大鼠血脂代谢和抗氧化能力的影响[J].青海医学院学报,2008,29(4):260-262.WANG D K,TIAN X C,WANG X P.Effects of matrine on the metabolization of serum lipid and anti-oxidation ability in hyperlipidemic rats[J].Journal of Qinghai Medical College,2008,29(4):260-262.(in Chinese)
[4]	ZHAO P,ZHOU R,ZHU X Y,et al.Matrine attenuates focal cerebral ischemic injury by improving antioxidant activity and inhibiting apoptosis in mice[J].Int J Mol Med,2015,36(3):633-644.
[5]	金光虎,高吉,毛小强,等.苦参碱诱导人前列腺癌pc-3 m细胞凋亡及对骨桥蛋白表达的影响[J].中国老年学杂志,2009,29(21):2758-2759.JIN G H,GAO J,MAO X Q,et al.Matrine induces apoptosis of human prostate cancer pc-3 m cells and its effect on the expression of osteopontin[J].Chinese Journal of Gerontology,2009,29(21):2758-2759.(in Chinese)
[6]	WANG W,YOU R L,QIN W J,et al.Anti-tumor activities of active ingredients in Compound Kushen Injection[J].Acta Pharmacol Sin,2015,36(6):676-679.
[7]	WANG H Y,LI Y X,DUN L L,et al.Antinociceptive effects of matrine on neuropathic pain induced by chronic constriction injury[J].Pharm Biol,2013,51(7):844-850.
[8]	杨明炜,陈锋,朱定俊,等.苦参碱氯化钠注射液治疗40例新型冠状病毒肺炎的临床疗效分析[J].中国中药杂志,2020,45(10):2221-2231.YANG M W,CHEN F,ZHU D J,et al.Clinical efficacy of matrine and sodium chloride injection in treatment of 40 cases of COVID-19[J].China Journal of Chinese Materia Medica,2020,45(10):2221-2231.(in Chinese)
[9]	丁园园,张荣生,张冬华,等.基于网络药理学和分子对接探讨苦参碱抗新冠病毒机制研究[J].中药药理与临床,2020,36(4):18-23.DING Y Y,ZHANG R S,ZHANG D H,et al.Mechanism study of matrine against SARS-CoV-2 based on network pharmacology and molecular docking[J].Pharmacology and Clinics of Chinese Materia Medica,2020,36(4):18-23.(in Chinese)
[10]	ZHENG J G,XU Y L,KHAN A,et al.In vitro screening of traditional chinese medicines compounds derived with anti-encephalomyocarditis virus activities[J].Biotechnol Bioproc Eng,2020,25(2):181-189.
[11]	SUN N,YU T,ZHAO J X,et al.Antiviral activities of natural compounds derived from traditional Chinese medicines against porcine circovirus type 2(PCV2)[J].Biotechnol Bioproc Eng,2015,20(1):180-187.
[12]	SUN N,WANG Z W,WU C H,et al.Antiviral activity and underlying molecular mechanisms of matrine against porcine reproductive and respiratory syndrome virus in vitro[J].Res Vet Sci,2014,96(2):323-327.
[13]	SUN N,ZHANG H,SUN P P,et al.Matrine exhibits antiviral activity in a PRRSV/PCV2 co-infected mouse model[J].Phytomedicine,2020,77:153289.
[14]	HORIE M,MIURA T,HIRAKATA S,et al.Comparative analysis of the intestinal flora in type 2 diabetes and nondiabetic mice[J].Exp Anim,2017,66(4):405-416.
[15]	DING Q,HU Y,FU Y,et al.Systematic review and meta-analysis of the correlation between intestinal flora and gestational diabetes mellitus[J].Ann Palliat Med,2021,10(9):9752-9764.
[16]	YANG Y Z,MISRA B B,LIANG L,et al.Integrated microbiome and metabolome analysis reveals a novel interplay between commensal bacteria and metabolites in colorectal cancer[J].Theranostics,2019,9(14):4101-4114.
[17]	ZHANG Y X,LI T,YUAN H Q,et al.Correlations of inflammatory factors with intestinal flora and gastrointestinal incommensurate symptoms in children with asthma[J].Med Sci Monit,2018,24:7975-7979.
[18]	SUN B M,MENG L,LIU H,et al.Changes in intestinal flora in preeclampsia rats and effects of probiotics on their inflammation and blood pressure[J].Eur Rev Med Pharmacol Sci,2020,24(19):10155-10161.
[19]	WANG Y L,WANG Z Y,WANG Y,et al.The gut-microglia connection:implications for central nervous system diseases[J].Front Immunol,2018,9:2325.
[20]	朱琳琳,武子函,邹战明,等.猪圆环病毒2型感染的仔猪空肠菌群变化[J].微生物学通报,2017,44(12):2951-2957.ZHU L L,WU Z H,ZOU Z M,et al.Change of the jejunum fora from PCV2-infected piglets[J].Microbiology China,2017,44(12):2951-2957.(in Chinese)
[21]	NIEDERWERDER M C,JAING C J,THISSEN J B,et al.Microbiome associations in pigs with the best and worst clinical outcomes following co-infection with porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2(PCV2)[J].Vet Microbiol,2016,188:1-11.
[22]	IDDIR M,BRITO A,DINGEO G,et al.Strengthening the immune system and reducing inflammation and oxidative stress through diet and nutrition:considerations during the COVID-19 crisis[J].Nutrients,2020,12(6):1562.
[23]	徐凯进,蔡洪流,沈毅弘,等.2019冠状病毒病(COVID-19)诊疗浙江经验[J].浙江大学学报(医学版),2020,49(2):147-157.XU K J,CAI H L,SHEN Y H,et al.Management of COVID-19:the Zhejiang experience[J].Journal of Zhejiang University(Medical Sciences),2020,49(2):147-157.(in Chinese)
[24]	MJÖSBERG J,RAO A.Lung inflammation originating in the gut[J].Science,2018,359(6371):36-37.
[25]	SEQUEIRA R P,MCDONALD J A K,MARCHESI J R,et al.Commensal bacteroidetes protect against Klebsiella pneumoniae colonization and transmission through IL-36 signalling[J].Nat Microbiol,2020,5(2):304-313.
[26]	STEFAN K L,KIM M V,IWASAKI A,et al.Commensal microbiota modulation of natural resistance to virus infection[J].Cell,2020,183(5):1312-1324.
[27]	KUHN K A,SCHULZ H M,REGNER E H,et al.Bacteroidales recruit IL-6-producing intraepithelial lymphocytes in the colon to promote barrier integrity[J].Mucosal Immunol,2018,11(2):357-368.
[28]	PEREIRA F C,WASMUND K,COBANKOVIC I,et al.Rational design of a microbial consortium of mucosal sugar utilizers reduces Clostridiodes difficile colonization[J].Nat Commun,2020,11(1):5104.
[29]	ZAGATO E,POZZI C,BERTOCCHI A,et al.Endogenous murine microbiota member Faecalibaculum rodentium and its human homologue protect from intestinal tumour growth[J].Nat Microbiol,2020,5(3):511-524.
[30]	CHEN K,CHEN H,FAAS M M,et al.Specific inulin-type fructan fibers protect against autoimmune diabetes by modulating gut immunity,barrier function,and microbiota homeostasis[J].Mol Nutr Food Res,2017,61(8):1601006.
[31]	ASTBURY S,ATALLAH E,VIJAY A,et al.Lower gut microbiome diversity and higher abundance of proinflammatory genus Collinsella are associated with biopsy-proven nonalcoholic steatohepatitis[J].Gut Microbes,2020,11(3):569-580.
[32]	丁俊荣.不同人群脱硫弧菌数量及肠道菌群多样性分析[D].无锡:江南大学,2012.DING J R.Quantity of desulfovibrio and analysis of intestinal microbiota diversity in different people[D].Wuxi:Jiangnan University,2012.(in Chinese)
[33]	陈德龙,朱宏亮,许光勇,等.乳酸菌对IPEC-J2细胞黏着斑激酶磷酸化及紧密连接蛋白Occludin表达的影响[J].畜牧兽医学报,2013,44(2):283-288.CHEN D L,ZHU H L,XU G Y,et al.Study of lactobacillus influence the FAK phosphorylation and tight junction protein occludin expression of IPEC-J2 cells[J].Acta Veterinaria et Zootechnica Sinica,2013,44(2):283-288.(in Chinese)
[34]	KAWASE M,HE F,KUBOTA A,et al.Oral administration of lactobacilli from human intestinal tract protects mice against influenza virus infection[J].Lett Appl Microbiol,2010,51(1):6-10.
[35]	GOTO H,SAGITANI A,ASHIDA N,et al.Anti-influenza virus effects of both live and non-live Lactobacillus acidophilus L-92 accompanied by the activation of innate immunity[J].Br J Nutr,2013,110(10):1810-1818.
[36]	TAKEDA S,TAKESHITA M,KIKUCHI Y,et al.Efficacy of oral administration of heat-killed probiotics from Mongolian dairy products against influenza infection in mice:alleviation of influenza infection by its immunomodulatory activity through intestinal immunity[J].Int Immunopharmacol,2011,11(12):1976-1983.
[37]	FUJIMURA K E,DEMOOR T,RAUCH M,et al.House dust exposure mediates gut microbiome Lactobacillus enrichment and airway immune defense against allergens and virus infection[J].Proc Natl Acad Sci U S A,2014,111(2):805-810.
[38]	GUILLEMARD E,TONDU F,LACOIN F,et al.Consumption of a fermented dairy product containing the probiotic Lactobacillus casei DN-114001 reduces the duration of respiratory infections in the elderly in a randomised controlled trial[J].Br J Nutr,2010,103(1):58-68.
[39]	KONISHI H,FUJIYA M,TANAKA H,et al.Probiotic-derived ferrichrome inhibits colon cancer progression via JNK-mediated apoptosis[J].Nat Commun,2016,7:12365.
[40]	AINDELIS G,TIPTIRI-KOURPETI A,LAMPRI E,et al.Immune responses raised in an experimental colon carcinoma model following oral administration of Lactobacillus casei[J].Cancers,2020,12(2):368.
[41]	GIRI S S,YUN S,JUN J W,et al.Therapeutic effect of intestinal autochthonous P16 against waterborne lead toxicity in Cyprinus carpio[J].Front Immunol,2018,9(9):2208.
[42]	SAEEDI B J,LIU K H,OWENS J A,et al.Gut-resident Lactobacilli activate hepatic Nrf2 and protect against oxidative liver injury[J].Cell Metab,2020,31(5):956-968.e5.

Influence of Matrine on Intestinal Flora of Kunming Mice through Intraperitoneal Injection Based on 16S rDNA Sequencing

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