VDR基因肠上皮特异性敲除小鼠的构建及其对炎症性肠病的影响
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摘要
采用Cre/loxP条件性基因敲除技术构建肠上皮敲除VDR(Vitamin D receptor)基因小鼠.利用Casp9方法构建的VDR~(flox/flox)小鼠与Villin-Cre转基因小鼠进行繁殖杂交获得VDR肠上皮特异敲除小鼠,其基因型鉴定为VDR~(flox/flox)Villin-Cre~+.通过建立小鼠TNBS诱导的炎症性肠病模型,比较肠上皮特异敲除VDR基因的小鼠与野生型小鼠的发病程度,结果显示VDR基因条件敲除小鼠,体重下降更加明显,肠损伤和病理评分更加严重,说明了肠上皮中的VDR缺失促进肠炎发生,预示其对肠上皮具有保护作用.本研究为进一步探讨肠上皮VDR的表达对炎症性肠病的作用机制提供坚实基础和强有力的研究平台.
To generate and identify the intestinal epithelium VDR(Vitamin D receptor)gene knockout mice through the Cre/loxP conditional gene knockout technology,VDR~(flox/flox) mice constructed by casp9 and the mice which expressed intestinal epithelium cell-specific Cre recombinase were bred. Then,the two kinds of mice were crossed,and the offspring mice which genotyping VDR~(flox/flox) Villin-Cre~+were the mice. By establishing a mouse model of inflammatory bowel disease(IBD)induced by TNBS,the incidence of intestinal epithelial-specific VDR knockdown mice and wild-type mice was compared. The results showed that intestinal epithelial-specific VDR knockdown mice had significant weight loss,intestinal injury score and the pathological score was more serious,indicating that the VDR loss in the intestinal epithelium promotes the occurrence of IBD,and that it had a protective effect on the intestinal epithelium. Furthermore,this study will provide a solid foundation and a powerful research platform for further exploration of the mechanism of action of intestinal epithelial VDR on inflammatory bowel disease.
To generate and identify the intestinal epithelium VDR(Vitamin D receptor)gene knockout mice through the Cre/loxP conditional gene knockout technology,VDR~(flox/flox) mice constructed by casp9 and the mice which expressed intestinal epithelium cell-specific Cre recombinase were bred. Then,the two kinds of mice were crossed,and the offspring mice which genotyping VDR~(flox/flox) Villin-Cre~+were the mice. By establishing a mouse model of inflammatory bowel disease(IBD)induced by TNBS,the incidence of intestinal epithelial-specific VDR knockdown mice and wild-type mice was compared. The results showed that intestinal epithelial-specific VDR knockdown mice had significant weight loss,intestinal injury score and the pathological score was more serious,indicating that the VDR loss in the intestinal epithelium promotes the occurrence of IBD,and that it had a protective effect on the intestinal epithelium. Furthermore,this study will provide a solid foundation and a powerful research platform for further exploration of the mechanism of action of intestinal epithelial VDR on inflammatory bowel disease.
引文
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[15] 李福兴,胡超,何薇等. 血液系统条件性DNMT3B基因敲除小鼠的构建及鉴定. 中国现代医学杂志,2018,28(6):23-27.(Li F X,Hu C,He W,et al. Generation of hematopoietic system-specific DNMT3B gene conditional knockout in mouse model. China Journal of Modern Medicine,2018,28(6):23-27.)
[16] Schmidt-Supprian M,Rajewsky K. Vagaries of conditional gene targeting. Nature Immunology,2007,8(7):665-668.
[17] Wu S P,Zhang YG,Lu R,et al. Intestinal epithelial vitamin D receptor deletion leads to defective autophagy in colitis. Gut,2015,64(7):1082-1094.
[2] Neurath M F. Cytokines in inflammatory bowel disease. Nature Reviews Immunology,2014,14(5):329-342.
[3] Lin R. Crosstalk between vitamin D metabolism,VDR signalling,and innate immunity. Bio Med Research International,2016,2016:1375858.
[4] Trochoutsou A I,Kloukina V,Samitas K,et al. Vitamin-D in the immune system:Genomic and non-genomic actions. Mini-Reviews in Medicinal Chemistry,2015,15(11):953-963.
[5] 王宽,黄任佳,吴焕淦等. 维生素D在炎症性肠病中的作用机制. 世界华人消化杂志,2015,23(35):5655-5661.(Wang K,Huang R J,Wu H G,et al. Mechanisms of action of vitamin D in inflammatory bowel disease. World Chinese Journal of Digestology,2015,23(35):5655-5661.)
[6] Shang M,Sun J.Vitamin D/VDR,probiotics,and gastrointestinal diseases. Current Medicinal Chemistry,2017,24(9):876-887.
[7] Christakos S,Ajibade D V,Dhawan P,et al. Vitamin D:Metabolism. Endocrinology and Metabolism Clinics of North America,2010,39(2):243-253.
[8] Deeb K K,Trump D L,Johnson C S. Vitamin D signalling pathways in cancer:Potential for anticancer therapeutics. Nature Reviews Cancer,2007,7(9):684-700.
[9] Dusso A S,Brown A J,Slatopolsky E. Vitamin D. American Journal of Physiology. Renal Physiology,2005,289(1):F8-F28.
[10] Holick M F. The vitamin D deficiency pandemic:Approaches for diagnosis,treatment and prevention. Reviews in Endocrine and Metabolic Disorders,2017,18(2):153-165.
[11] Del Pinto R,Ferri C,Cominelli F. Vitamin D axis in inflammatory bowel diseases:Role,current uses and future perspectives. International Journal of Molecular Sciences,2017,18(11):2360.
[12] Ulitsky A,Ananthakrishnan A N,Naik A,et al. Vitamin D deficiency in patients with inflammatory bowel disease:Association with disease activity and quality of life. Journal of Parenteral and Enteral Nutrition,2011,35(3):308-316.
[13] Liu W C,Chen Y Z,Golan M A,et al. Intestinal epithelial vitamin D receptor signaling inhibits experimental colitis. Journal of Clinical Investigation,2013,123(9):3983-3996.
[14] 孔维健,常宇鑫,昝春芳等. 基于Cre-loxP系统条件性基因敲除小鼠的构建及其应用进展. 中国实验诊断学,2017,21(12):2208-2211.
[15] 李福兴,胡超,何薇等. 血液系统条件性DNMT3B基因敲除小鼠的构建及鉴定. 中国现代医学杂志,2018,28(6):23-27.(Li F X,Hu C,He W,et al. Generation of hematopoietic system-specific DNMT3B gene conditional knockout in mouse model. China Journal of Modern Medicine,2018,28(6):23-27.)
[16] Schmidt-Supprian M,Rajewsky K. Vagaries of conditional gene targeting. Nature Immunology,2007,8(7):665-668.
[17] Wu S P,Zhang YG,Lu R,et al. Intestinal epithelial vitamin D receptor deletion leads to defective autophagy in colitis. Gut,2015,64(7):1082-1094.