紫茎泽兰对大鼠肠道结构和肠道黏膜免疫屏障的影响

doi:10.11686/cyxb2021171

摘要/Abstract

摘要：

为探究紫茎泽兰对动物肠道损伤及致毒机制，本试验选取16只7周龄雄性SD大鼠经7 d适应期后随机分为对照组（饲喂不含紫茎泽兰饲料，10 g·100 g^-1 BW）与试验组（饲喂含30%紫茎泽兰饲料，10 g·100 g^-1 BW），试验周期为14 d，试验结束后取各肠段（取十二指肠、空肠、回肠、盲肠、结肠和直肠）组织样品，观察结构损伤，计数免疫细胞，测定sIgA（secretory immunoglobulin A，sIgA）和炎性细胞因子分泌量。结果表明：与对照组相比，紫茎泽兰可使十二指肠绒毛出血及顶端轻度坏死脱落，肠绒毛高度、隐窝深度及其比值极显著增加（P<0.01）；空肠充血并伴有绒毛顶端糜烂性坏死脱落，绒毛高度及绒毛高度与隐窝深度比值极显著增加（P<0.01）；回肠大量肠绒毛顶端凝固性坏死，并伴有出血与炎性浸润，绒毛高度、隐窝深度及其比值极显著增加（P<0.01）；盲肠水肿及充血；结肠淋巴细胞增多、直肠大量淋巴细胞浸润及淋巴细胞增生，其中以回肠和直肠机械损伤最为严重，回肠评分增加722.36%，直肠评分增加976.00%；试验组各肠段上皮内淋巴细胞（intestinal intraepithelial lymphocytes， IELs）、固有层淋巴细胞（lamina propria lymphocytes， LPLs）和杯状细胞（goblet cells， GCs）数量极显著增加（P<0.01），同时还可极显著增加sIgA的分泌量（P<0.01），极显著增加促炎因子IL-1β、IL-2、TNF-α和IFN-γ的表达量（P<0.01），极显著降低抑炎因子IL-4、IL-10的表达（P<0.01），激活肠道炎症反应，从而破坏肠道黏膜免疫屏障，造成肠道损伤。

关键词: 紫茎泽兰, 肠道结构, 肠道黏膜免疫

Abstract:

In order to analyze the toxicity mechanisms of Ageratina adenophora on the intestine of animals， sixteen 7-week-old male Sprague Dawley rats were randomly divided into two groups， a control group （fed 10 g·100 g^-1 body weight （BW） normal diet without A. adenophora powder supplementation） and a treatment group （fed 10 g·100 g^-1 BW of experimental diet prepared by mixing the normal diet and A. adenophora powder in a ratio of 7∶3）， after a 7-day period of adaptation to the normal diet. The test cycle was 14 days. After the experiment， we observed the structural damage， counted the immune cells， and determined the levels of sIgA and inflammatory cytokines of each intestinal segment （duodenum， jejunum， ileum， cecum， colon and rectum） tissue samples. It was found that A. adenophora caused duodenal villi bleeding， mild necrosis， abscission at the top of the duodenum and increased the villi height， crypt depth and and their ratio compared with the control group （P<0.01）. The jejunum was congested with erosive necrosis and abscission at the top of the villi， and also showed a significant increase in the villi height and villi height∶crypt depth ratio （P<0.01）. A large number of intestinal villi at the top of ileum showed coagulated necrosis， accompanied by bleeding， inflammatory infiltration， and a significant increase in villi height， crypt depth and their ratio （P<0.01）. In addition， cecum edema and congestion， colonic lymphocytosis， large numbers of lymphocytes infiltrating and lymphocyte proliferation in the rectum were also observed. Among the intestinal sections the ileum and rectum had the most serious loss of structural integrity with the ileum injury score increasing 722% and rectum score increasing 976%. A. adenophora also significantly increased the number of intestinal intraepithelial lymphocytes， lamina propria lymphocytes and goblet cells （GC_S）（P<0.01） in each intestinal segment. It also significantly increased the secretion of sIgA （P<0.01） and expression of pro-inflammatory factor （IL-1β， IL-2、TNF-α and IFN-γ）， however， the expression of IL-4 and IL-10 was significantly decreased （P<0.01）. In summary， A. adenophora induced an extensive intestinal inflammatory response， destroyed the intestinal mucosal immune barrier and caused intestinal injury.

Key words: Ageratina adenophora, intestinal structure, intestinal mucosal immunity

崔玉晶, 高佩, 文娟, OKYERE Kumi Samuel, 胡延春. 紫茎泽兰对大鼠肠道结构和肠道黏膜免疫屏障的影响[J]. 草业学报, 2022, 31(6): 211-220.

Yu-jing CUI, Pei GAO, Juan WEN, Kumi Samuel OKYERE, Yan-chun HU. The effect of Ageratina adenophora on the structure of various intestinal segments and on intestinal mucosal immune barrier function in rats[J]. Acta Prataculturae Sinica, 2022, 31(6): 211-220.

图/表 8

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表现Appearance	评分Score
水肿Edema	0无NO，1有Yes
出血Hemorrhage	0无NO，1有Yes
肌层增厚Muscle thickening	0无NO，1轻度Light，2中度Moderate，3重度Heavy
细胞浸润Cellular infiltration	0无NO，1轻度Light，2中度Moderate，3重度Heavy
黏膜结构破坏Loss of mucosal architecture	0无NO，1轻度Light，2中度Moderate，3重度Heavy

表现Appearance	评分Score
水肿Edema	0无NO，1有Yes
出血Hemorrhage	0无NO，1有Yes
肌层增厚Muscle thickening	0无NO，1轻度Light，2中度Moderate，3重度Heavy
细胞浸润Cellular infiltration	0无NO，1轻度Light，2中度Moderate，3重度Heavy
黏膜结构破坏Loss of mucosal architecture	0无NO，1轻度Light，2中度Moderate，3重度Heavy

基因名称Gene	引物Primers	引物序列Sequence（5'-3'）
IL-1β	Forward	GACTTCACCATGGAACCCGT
IL-1β	Reverse	CAGGGAGGGAAACACACGTT
IL-2	Forward	GCAGCGTGTGTTGGATTTGA
IL-2	Reverse	GGCTCATCATCGAATTGGCAC
IL-4	Forward	AAGGAACACCACGGAGAACG
IL-4	Reverse	CAGACCGCTGACACCTCTAC
IL-10	Forward	TTGAACCACCCGGCATCTAC
IL-10	Reverse	CCAAGGAGTTGCTCCCGTTA
TNF-α	Forward	AACTCAGCGAGGACACCAAG
TNF-α	Reverse	GCCAGTGTATGAGAGGGACG
IFN-γ	Forward	GGCAAAAGGACGGTAACACG
IFN-γ	Reverse	TCTGTGGGTTGTTCACCTCG
β-actin	Forward	ACGGTCAGGTCATCACTATCG
β-actin	Reverse	GGCATAGAGGTCTTTACGGATG

基因名称Gene	引物Primers	引物序列Sequence（5'-3'）
IL-1β	Forward	GACTTCACCATGGAACCCGT
IL-1β	Reverse	CAGGGAGGGAAACACACGTT
IL-2	Forward	GCAGCGTGTGTTGGATTTGA
IL-2	Reverse	GGCTCATCATCGAATTGGCAC
IL-4	Forward	AAGGAACACCACGGAGAACG
IL-4	Reverse	CAGACCGCTGACACCTCTAC
IL-10	Forward	TTGAACCACCCGGCATCTAC
IL-10	Reverse	CCAAGGAGTTGCTCCCGTTA
TNF-α	Forward	AACTCAGCGAGGACACCAAG
TNF-α	Reverse	GCCAGTGTATGAGAGGGACG
IFN-γ	Forward	GGCAAAAGGACGGTAACACG
IFN-γ	Reverse	TCTGTGGGTTGTTCACCTCG
β-actin	Forward	ACGGTCAGGTCATCACTATCG
β-actin	Reverse	GGCATAGAGGTCTTTACGGATG

肠段Part	对照组评分Control group score	试验组评分Treatment group score	P值P-value
十二指肠Duodenum	0.63±0.48	3.75±0.83	<0.01
空肠Jejunum	0.88±0.78	5.38±0.70	<0.01
回肠Ileum	0.76±0.66	6.25±0.66	<0.01
盲肠Cecum	0.75±0.66	3.50±0.50	<0.01
结肠Colon	0.63±0.48	3.38±0.48	<0.01
直肠Rectum	0.50±0.50	5.38±0.70	<0.01