复方术苦芩提取液对腹泻小鼠iIELs及十二指肠INF-γ、IL-4 mRNA转录的影响

doi:10.11843/j.issn.0366-6964.2020.03.023

摘要/Abstract

摘要： 旨在研究复方术苦芩提取液对腹泻小鼠小肠上皮内淋巴细胞（intestinal intraepithelial lymphocyte，iIELs）及十二指肠INF-γ、IL-4 mRNA转录的影响。将72只小鼠随机分为6个组，分别为空白组、感染组、阳性药物组（黄芪多糖口服液，浓度为1.2 mg·mL^-1）和药物Ⅰ、Ⅱ、Ⅲ组（复方术苦芩提取液，生药浓度分别为0.1、1和10 mg·mL^-1）。除空白组外，小鼠连续给药7 d[灌胃0.5 mL·（次·d）^-1]后腹腔注射5×10⁷ CFU大肠杆菌，并将感染组小鼠出现腹泻时设定为试验0 h。于试验6 h和4 d时各处死6只小鼠，制作小肠病理切片并计数iIELs数量，qRT-PCR分析十二指肠段INF-γ、IL-4 mRNA水平。结果显示：阳性药物组和药物Ⅰ、Ⅱ、Ⅲ组对腹泻小鼠的保护率分别为66.7%、50.0%、66.7%、75.0%。试验6 h时，与空白组相比，感染组小鼠iIELs数量增加，十二指肠INF-γ mRNA水平降低、IL-4 mRNA水平增高，差异均极显著（P<0.01）；与感染组相比，阳性药物组和药物Ⅰ、Ⅱ、Ⅲ组小鼠iIELs数量减少，阳性药物组和药物Ⅲ组小鼠十二指肠INF-γ mRNA水平增高、IL-4 mRNA水平降低。试验4 d时，与空白组相比，感染组小鼠十二指肠和回肠的iIELs数量增加，空肠iIELs数量减少，其十二指肠INF-γ、IL-4 mRNA水平增高，差异均极显著（P<0.01）；与感染组相比，阳性药物组和药物Ⅰ、Ⅱ、Ⅲ组小鼠十二指肠和回肠的iIELs数量（Ⅰ组十二指肠除外）均减少，空肠iIELs数量增加，阳性药物组和药物Ⅲ组十二指肠段INF-γ、IL-4 mRNA水平均降低。结果表明，复方术苦芩提取液通过调控小鼠小肠iIELs数量，平衡十二指肠INF-γ/IL-4 mRNA转录，达到对小鼠腹泻的保护作用。

关键词: 复方术苦芩, 肠黏膜屏障, iIELs, INF-γ, IL-4

Abstract: To study the effects of Zhukuqin compound extractive fluid on intestinal intraepithelial lymphocyte (iIELs) and the transcription of INF-γ and IL-4 mRNA in the duodenum of diarrhea mice. A total of 72 mice were randomly divided into six groups equally:control group, infection group, positive drug group (Astragalus polysaccharide oral liquid, the concentration was 1.2 mg·mL^-1), and drug group Ⅰ, Ⅱ, Ⅲ (Zhukuqin extractive fluid, the concentrations were 0.1, 1 and 10 mg·mL^-1, respectively). After continuously pre-administration for 7 days (0.5 mL, once daily dosing), except the control group, the mice in each group were intraperitoneally injected with E. coli (5×10⁷CFU). The time point was selected as 0 h when mice appeared diarrhea in the infection group. Six mice were killed for preparing pathological sections, the number of iIELs in the small intestine was counted, and the mRNA level of INF-γ and IL-4 in the duodenum were analyzed by quantitative real-time PCR at 6 h and 4 d, respectively. The diarrhea protective rate of positive drug group and drug group Ⅰ, Ⅱ, Ⅲ were 66.7%, 50.0%, 66.7%, and 75.0%, respectively. At 6 h, compared with control group, the number of iIELs in infection group was significantly increased, and the mRNA level of INF-γ and IL-4 were observably reduced and strengthened, respectively (P<0.01); Compared with infection group, the number of iIELs in positive drug group and drug group Ⅰ, Ⅱ, Ⅲ were decreased, and the mRNA level of INF-γ and IL-4 in positive drug group and drug group Ⅲ were augmented and lessened, respectively. At 4 d, compared with control group, the quantity of iIELs in the duodenum and ileum in infection group were markedly heightened, while that in the jejunum was dropped, and the mRNA level of INF-γ and IL-4 were dramatically enhanced (P<0.01); Compared with infection group, the number of iIELs in the duodenum and ileum in positive drug group and drug group Ⅰ, Ⅱ, Ⅲ were significantly diminished (except that in duodenum of group Ⅰ), and that in the jejunum was aggrandized, and the mRNA level of INF-γ and IL-4 in positive drug group and drug Ⅲ group were decreased. The results of the current study show that Zhukuqin compound extractive fluid can protect the diarrhea mice, via regulating the number of iIELs in the small intestine and balancing the transcription of INF-γ and IL-4 mRNA in the duodenum of mice.

Key words: Zhukuqin compound, intestinal mucosa barrier, iIELs, INF-γ, IL-4

中图分类号:

S853.74

王忠清, 吴秋月, 胡巧, 周梅芳, 刘娟. 复方术苦芩提取液对腹泻小鼠iIELs及十二指肠INF-γ、IL-4 mRNA转录的影响[J]. 畜牧兽医学报, 2020, 51(3): 630-638.

WANG Zhongqing, WU Qiuyue, HU Qiao, ZHOU Meifang, LIU Juan. Effects of Zhukuqin Compound Extractive Fluid on iIELs and the Transcription of INF-γ and IL-4 mRNA in the Duodenum of Diarrhea Mice[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(3): 630-638.

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