牛至香酚对LPS所致的免疫应激小鼠脾脏细胞因子及核转录因子mRNA表达的影响
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摘要
将80只小鼠随机分成5组(空白对照组、模型组及牛至香酚低、中、高剂量组),牛至香酚低、中、高剂量组小鼠分别按25、50、100 mg·kg~(-1)的剂量灌胃牛至香酚,空白对照组、模型组灌胃相同体积的橄榄油,每日1次,连续14 d.第15天时,牛至香酚试验组、模型组小鼠腹腔注射8 mg·kg~(-1)脂多糖(LPS),6 h后采集脾脏组织,用qRT-PCR检测小鼠脾脏细胞因子(TNF-α、IFN-γ、IL-6、IL-10、IL-1β)、NF-κB及核转录因子(T-bet、GATA-3)mRNA的表达量,研究牛至香酚对LPS所致的免疫应激小鼠脾脏细胞因子及核转录因子mRNA表达的影响.结果表明:与空白对照组相比,模型组小鼠TNF-α、IFN-γ、IL-6、IL-10、IL-1β、NF-κB、T-bet mRNA的表达量提高(P<0.01),GATA-3 mRNA的表达量降低(P<0.01);与模型组相比,牛至香酚低、高剂量组IL-1β mRNA的表达量降低(P<0.01),低、中、高剂量组IL-6、TNF-α mRNA的表达量降低(P<0.01、P<0.05),低、中、高剂量组IFN-γ mRNA的表达量极显著或显著降低(P<0.01或P<0.05),高剂量组NF-κB mRNA的表达量降低(P<0.05),低、中、高剂量组T-bet mRNA的表达量降低(P<0.05),中剂量组GATA-3 mRNA的表达量提高(P<0.05).由本试验结果可知,牛至香酚通过提高或降低小鼠脾脏细胞因子、核转录因子的表达,调节免疫应激小鼠的免疫功能.
To investigate whether oregano phenol promotes the expression of cytokines and positively regulate the nuclear factor mRNA in mice with immune stress arose from lipopolysaccharide(LPS), 80 mice were randomized into 5 groups, namely 3 experimental groups lavage with 25, 50, and 100 mg·kg~(-1) oregano phenol, respectively, 1 model group lavage with same amount of olive oil and last control group fed with water. After continuous gavage for 14 d, mice from 3 experimental groups and the model group were intraperitoneally injected 8 mg·kg~(-1) LPS, with the control group injected physiological saline solution. After 6 h, spleen were sampled and the expressions of spleen cytokines(TNF-α, IFN-γ, IL-6, IL-10, IL-1β), NF-κB and nuclear transcription factors(T-bet, GATA-3) mRNA were detected by qRT-PCR. The results showed that the expression levels of TNF-α, IFN-γ, IL-6, IL-10, IL-1β, NF-κB and T-bet mRNA were significantly elevated in the model group than the control(P<0.01), while the expression of GATA-3 mRNA was significantly decreased(P<0.01). Compared with the model group, the expression levels of IL-1β mRNA in low-and high-dose groups dropped significantly(P<0.01). The expressions of IL-6(P<0.01) and TNF-α mRNA(P<0.05) in the experimental groups went down. The expression of IFN-γ mRNA decreased or significantly decreased in the experiment groups(P<0.01 or P<0.05). The expression of NF-κB mRNA decreased only in the high-dose experimental group(P<0.05), while the expression of T-bet mRNA decreased in all experiment groups(P<0.05). And the expression of GATA-3 mRNA only increased in the medium-dose experiment group(P<0.05). To sum up, oregano phenol could regulate the immune function of mice with immune stress by adjusting the expression of cytokines and transcription factors in mouse spleen.
To investigate whether oregano phenol promotes the expression of cytokines and positively regulate the nuclear factor mRNA in mice with immune stress arose from lipopolysaccharide(LPS), 80 mice were randomized into 5 groups, namely 3 experimental groups lavage with 25, 50, and 100 mg·kg~(-1) oregano phenol, respectively, 1 model group lavage with same amount of olive oil and last control group fed with water. After continuous gavage for 14 d, mice from 3 experimental groups and the model group were intraperitoneally injected 8 mg·kg~(-1) LPS, with the control group injected physiological saline solution. After 6 h, spleen were sampled and the expressions of spleen cytokines(TNF-α, IFN-γ, IL-6, IL-10, IL-1β), NF-κB and nuclear transcription factors(T-bet, GATA-3) mRNA were detected by qRT-PCR. The results showed that the expression levels of TNF-α, IFN-γ, IL-6, IL-10, IL-1β, NF-κB and T-bet mRNA were significantly elevated in the model group than the control(P<0.01), while the expression of GATA-3 mRNA was significantly decreased(P<0.01). Compared with the model group, the expression levels of IL-1β mRNA in low-and high-dose groups dropped significantly(P<0.01). The expressions of IL-6(P<0.01) and TNF-α mRNA(P<0.05) in the experimental groups went down. The expression of IFN-γ mRNA decreased or significantly decreased in the experiment groups(P<0.01 or P<0.05). The expression of NF-κB mRNA decreased only in the high-dose experimental group(P<0.05), while the expression of T-bet mRNA decreased in all experiment groups(P<0.05). And the expression of GATA-3 mRNA only increased in the medium-dose experiment group(P<0.05). To sum up, oregano phenol could regulate the immune function of mice with immune stress by adjusting the expression of cytokines and transcription factors in mouse spleen.
引文
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[15] 惠永岗,杨宇焦,陈昌林,等.极化液对内毒素血症小鼠脾脏的保护作用[J].川北医学院学报,2016,31(5):659-662.
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[18] CHENG Y,ZHANG J,HOU W,et al.Immunoregulatory effects of sinomenine on the T-bet/GATA-3 ratio and Th1/Th2 cytokine balance in the treatment of mesangial proliferative nephritis [J].International Immunopharmacology,2009,9(7/8):894-899.
[19] 孙骆生.动物免疫学[M].北京:高等教育出版社,2003.
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[21] 黄丽群,薛峰,沈小雁,等.系统性红斑狼疮患者转录因子T-bet和GATA-3基因的表达[J].中华皮肤科杂志,2004,37(12):724-726.
[22] PI C C,CHU L,LU C Y,et al.Polysaccharides from Ganoderma formosanum function as a Th1 adjuvant and stimulate cytotoxic T cell response in vivo [J].Vaccine,2014,32(3):401-408.
[23] 马玉芳,李萍,潘晓丽,等.金线莲多糖对环磷酰胺所致免疫抑制小鼠脾脏细胞因子及转录因子mRNA表达的影响[J].中国兽医学报,2017,37(7):1 322-1 326.
[24] 刘汉斌,彭娇娇,魏文康,等.强鸡散对免疫抑制小鼠脾转录因子GATA-3和T-bet表达量的影响[J].广东农业科学,2014,41(19):122-125.
[25] 李鹏,汪中兴,吴梦郡,等.复合植物精油对脂多糖刺激仔猪肝脏的保护作用[J].中国畜牧杂志,2018,54(3):87-91.
[2] 余勋信,林伯全,杨慧.牛至香酚替代抗生素对育肥土黑猪生长性能和肉质的影响[J].黑龙江畜牧兽医,2016(3):129-131.
[3] 郝丹丹,张旭,陈嘉,等.牛至油对成年蛋鸡生长性能和免疫功能的影响[J].中国兽医学报,2017,37(6):1 121-1 127.
[4] KINTZIOS S,PAPAGEORGIOU K,YIAKOUMETTIS I,et al.Evaluation of the antioxidants activities of four Slovene medicinal plant species by traditional and novel biosensory assays [J].Journal of Pharmaceutical & Biomedical Analysis,2010,53(3):773-776.
[5] 王宇,王琳,刘蕾,等.牛至油体外抗肿瘤活性研究[J].中国微生态学杂志,2010,22(12):1 101-1 102.
[6] 蔡旭滨.牛至油对断奶仔猪的保健作用及机理研究[D].福州:福建农林大学,2017.
[7] 马玉芳,王冠淞,闵思明,等.牛至油对断奶仔猪生长性能、血清激素指标及小肠组织胰岛素样生长因子-Ⅰ含量的影响[J].动物营养学报,2018,30(6):2 303-2 309.
[8] XIAO K,ZOU W H,YANG Z,et al.The role of visfatin on the regulation of inflammation and apoptosis in the spleen of LPS-treated rats [J].Cell & Tissue Research,2015,359(2):605-618.
[9] 陈语嫣.太子参茎叶多糖对DSS诱导溃疡性结肠炎小鼠的治疗与干预作用研究[D].福州:福建农林大学,2017.
[10] 檀新珠,陈赛红,陈俊宇,等.太子参茎叶多糖对小鼠脾淋巴细胞因子含量及对细胞因子和转录因子mRNA表达量的影响[J].中国兽医科学,2018,48(1):124-129.
[11] HOTCHKISS R S,TINSLEY K W,SWANSON P E,et al.Depletion of dendritic cells,but not macrophages,in patients with sepsis [J].Journal of Immunology,2002,168(5):2 493-2 500.
[12] AYALA A,HERDON C D,LEHMAN D L,et al.The induction of accelerated thymic programmed cell death during polymicrobial sepsis:control by corticosteroids but not tumor necrosis factor [J].Shock,1995,3(4):259-267.
[13] GRIVENNIKOV S I,KARIN M.Dangerous liaisons:STAT3 and NF-κB collaboration and crosstalk in cancer [J].Cytokine & Growth Factor Reviews,2010,21(1):11-19.
[14] 梁德洁.百里香酚对LPS诱发的乳腺炎小鼠的保护作用及机制研究[D].长春:吉林大学,2014.
[15] 惠永岗,杨宇焦,陈昌林,等.极化液对内毒素血症小鼠脾脏的保护作用[J].川北医学院学报,2016,31(5):659-662.
[16] 韩琳,王洪新,鲁美丽.黄芪多糖通过抑制NF-κB和JNK信号通路减轻LPS诱导的小鼠心肌细胞凋亡[J].中国药理学通报,2018,34(2):243-249.
[17] CHAKIR H,WANG H,LEFEBVRE D E,et al.T-bet/GATA-3 ratio as a measure of the Th1/Th2 cytokine profile in mixed cell populations:predominant role of GATA-3 [J].Journal of Immunological Methods,2003,278(1):157-169.
[18] CHENG Y,ZHANG J,HOU W,et al.Immunoregulatory effects of sinomenine on the T-bet/GATA-3 ratio and Th1/Th2 cytokine balance in the treatment of mesangial proliferative nephritis [J].International Immunopharmacology,2009,9(7/8):894-899.
[19] 孙骆生.动物免疫学[M].北京:高等教育出版社,2003.
[20] 袁颖,金素安,何世民.中药对Th1/Th2细胞因子平衡调节作用的研究进展[J].上海中医药杂志,2011,45(5):90-92.
[21] 黄丽群,薛峰,沈小雁,等.系统性红斑狼疮患者转录因子T-bet和GATA-3基因的表达[J].中华皮肤科杂志,2004,37(12):724-726.
[22] PI C C,CHU L,LU C Y,et al.Polysaccharides from Ganoderma formosanum function as a Th1 adjuvant and stimulate cytotoxic T cell response in vivo [J].Vaccine,2014,32(3):401-408.
[23] 马玉芳,李萍,潘晓丽,等.金线莲多糖对环磷酰胺所致免疫抑制小鼠脾脏细胞因子及转录因子mRNA表达的影响[J].中国兽医学报,2017,37(7):1 322-1 326.
[24] 刘汉斌,彭娇娇,魏文康,等.强鸡散对免疫抑制小鼠脾转录因子GATA-3和T-bet表达量的影响[J].广东农业科学,2014,41(19):122-125.
[25] 李鹏,汪中兴,吴梦郡,等.复合植物精油对脂多糖刺激仔猪肝脏的保护作用[J].中国畜牧杂志,2018,54(3):87-91.