目的繁殖和鉴定CXC趋化因子配体5(CXCchemokineligand5,CxcL5)基因敲除小鼠,以获得CXCL5-/-纯合子小鼠并与野生型小鼠进行比较。方法将CXCL5+/-杂合子小鼠繁殖后,剪取子代小鼠耳组织,提取基因组DNA,用PCR法鉴定子代小鼠的基因型。采用独立样本t检验比较同周龄同性别CXCL5基因敲除纯合子小鼠与野生型小鼠的体重和结直肠长度,并通过苏木素-伊红染色观察两种基因型小鼠的结肠形态结构。结果经繁殖,共获得60只CXCL5-/-纯合子小鼠和55只野生型小鼠。4、6、8、10周龄雌性CXCL5-/-小鼠的体重分别为(10.08±0.78)、(14.55±0.92)、(18.01±0.68)和(19.75±1.23)g,明显轻于同龄雌性野生型小鼠[(11.43±0.85)、(18.42±0.76)、(20.42±1.63)和(21.27±1.66)g](t值分别为-2.622、-7.253、-3.042、-1.644,P值均〈0.05);6、8周龄雄性CXCL5-/-小鼠的体重分别为(17.46±0.74)和(20.62±1.62)g,也明显轻于同龄雄性野生型小鼠[(20.47±1.66)和(23.33±1.67)g](t值分别为-3.688、-2.885,P值均〈0.05)。6周龄雌性CXCL5-/-小鼠的结直肠长度为(6.86±0.17)cm,与同龄雌性野生型小鼠(7.66±0.11)cm的长度相比,明显较短(t=-8.835,P〈0.05),但结肠的形态结构无明显改变。结论成功获得了CXCL5基因敲除纯合子小鼠,为深入研究CXCL5的生物学功能奠定了基础。
Objective To breed and identify the genotype of CXC chemokine ligand 5 (CXCLS) gene knockout mice to acquire CXCL5-/- homozygote mice and compare them with wild type mice. Methods CXCL5+/- heterozygote mice were bred. Genomic DNA was extracted from ear tissue of offspring mice and subjected to PCR for genotype identification. The body weight and colorectal length were compared between CXCL5-/- and wild type mice using independent-sample t-test. Morphological structure of colon of the two genotype mice was observed by hernatoxylin-eosin staining. Resdts Sixty CXCL5-/- homozygote mice and 55 wild type mice were obtained after breeding. The weight of CXCL5-/- female mice was significantly lighter than that of wild type female mice at the age of 4, 6, 8, 10 weeks [(10.08±0.78) gvs(11. 43±0.85) g, (14. 55±0.92) g vs (18. 42±0.76) g,(18.01 ±0.68) g vs (20.42±1.63) g, (19.75±1.23) gvs (21.27±1.66) g; t=-2. 622, -7.253, -3.042, - 1.644, respectively,all P 〈0. 05]. CXCL5-/- male mice were also significantly lighter compared with wild type male mice at the age of 6, 8 weeks[(17. 46 ±0. 74) g vs (20. 47± 1.66) g, (20. 62± 1.62) g vs (23.33 ±1.67) g; t=- 3. 688, -2. 885, respectively, all P 〈0. 05]. The colorectal length of CXCL5-/- female mice was significantly shorter than that of wild type female mice at the age of 6 weeks [(6. 86 ± 0. 17) cm vs (7. 66 ± 0. 11) cm; t=- 8. 835, P 〈0. 05], but the morphological structure of colon had no obvious change. Conclusion The CXCL5 gene knockout mice are obtained successfully, which lays a foundation for further research on CXCL5 biological function.