HSP70在牦牛肠道中的分布与变化规律

doi:10.11843/j.issn.0366-6964.2018.08.019

摘要/Abstract

摘要：

为了探索HSP70在牦牛肠道组织中的分布与变化规律，选择6头份健康成年（3~6岁）牦牛的肠道组织，采用免疫组织化学（IHC）、蛋白免疫印迹（WB）和实时荧光定量PCR（RT-qPCR）检测HSP70在牦牛十二指肠、空肠、回肠、盲肠、结肠和直肠组织中的分布与变化规律。免疫组织化学结果显示，HSP70在牦牛小肠的黏膜层、黏膜下层和肌层中均有表达，其中小肠黏膜上皮的柱状细胞呈强阳性表达，杯状细胞未见表达；小肠腺的肠腺细胞和肌层的平滑肌细胞也呈阳性表达，阳性反应主要分布于阳性细胞的细胞质和细胞核。HSP70在大肠中的定位表达与小肠相似，在大肠壁黏膜层、黏膜下层和肌层三层结构中也均有表达，其阳性反应主要位于黏膜上皮和固有层内散在分布的结缔组织细胞中，而固有层内大肠腺细胞仅在细胞核有表达，细胞质呈阴性。蛋白免疫印迹和免疫组化积分光密度值分析得出，从小肠的前端到大肠末端，HSP70的表达呈先增高后降低的趋势，其中在盲肠中的表达量达到峰值，且盲肠和结肠中HSP70的相对蛋白表达量极显著高于其他肠段（P<0.01）。实时荧光定量PCR结果显示，HSP70 mRNA在盲肠中的相对转录量最高，分别是十二指肠、空肠、回肠、结肠和直肠的2、1.68、1.06、1.43和1.52倍。本试验显示牦牛肠道中HSP70呈广泛性阳性反应，HSP70基因和HSP70蛋白在牦牛小肠和大肠中具有不同的表达模式，揭示了HSP70在牦牛小肠和大肠中所发挥生物学作用的异同，为HSP70在反刍动物组织中的表达和功能研究提供了资料。

Abstract:

In order to explore the distribution and change rule of HSP70 in yak intestine, six healthy adult yaks (3-6 years old) were selected, and the distribution and change rule of HSP70 in yak intestine were observed and detected by immunohistochemistry(IHC), Western blotting (WB) and real-time fluorescence quantitative PCR (RT-qPCR) methods. Immunohistochemical results showed that HSP70 was expressed in the mucosa, submucosa and muscular layer of yak small intestine, among which the columnar cells of small intestinal mucosa epithelium were strongly positive, but the goblet cells were negative. The intestinal glandular cells of the intestinal glands and the smooth muscle cells of the muscular layer also showed positive expression, and the positive particles were mainly distributed in the cytoplasm and nucleus of the above cells. Similar to the small intestine, HSP70 was also expressed in the mucosa, submucosa and muscular layer of yak large intestine. The positive reaction was mainly located in scattered connective tissue cells of the mucosal epithelium and lamina propria,while positive reaction of colonic gland cells in the lamina propria was found only in the nucleus, but not in the cytoplasm. The statistical analysis of Western blotting and integrated optical density value of immunohistochemistry showed that, from the anterior segment of small intestine to the distal segment of large intestine, the expression of HSP70 increased first and then decreased, and peaked in the cecum. In addition, the relative protein expression of HSP70 in the cecum and colon was significantly higher than that of other bowel segments (P<0.01). Real-time fluorescent quantitative PCR showed that HSP70 mRNA had the highest relative expression in the cecum, which was twice as much as that of the duodenum, 1.68 times of the jejunum, 1.06 times of the ileum, 1.43 times of the colon and 1.52 times of that of the rectum. This study revealed that HSP70 showed a broad positive reaction in yak intestine, and HSP70 gene, HSP70 protein had different expression patterns in the small intestine and the large intestine of yaks. The results revealed the similarities and differences in biological functions played by HSP70 protein between the small intestine and the large intestine and provided information for the study of the expression and function analysis of HSP70 in ruminant tissues.

中图分类号:

S852.16

廖博, 崔燕, 杨雪, 何俊峰, 宋亮丽, 杨晓晴, 刘鹏刚. HSP70在牦牛肠道中的分布与变化规律[J]. 畜牧兽医学报, 2018, 49(8): 1735-1742.

LIAO Bo, CUI Yan, YANG Xue, HE Jun-feng, SONG Liang-li, YANG Xiao-qing, LIU Peng-gang. Distribution and Change Rule of HSP70 in Yak Intestine[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(8): 1735-1742.

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