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目的饲养、繁殖并鉴定CXC趋化因子配体4[chemokine(C-X-C motif)ligand 4,CXCL4]基因敲除小鼠,为深入研究CXCL4的生物学功能奠定基础。方法将引进的杂合子小鼠进行饲养并繁殖,剪取繁殖成功的子代小鼠耳朵,提取其基因组DNA,采用PCR法鉴定小鼠的基因型;比较野生型小鼠和CXCL4敲除的纯合子小鼠的体重变化及结直肠的长度,通过HE染色观察野生型小鼠和CXCL4敲除的纯合子小鼠的结直肠形态结构;采用ELISA法验证CXCL4野生型和纯合子小鼠血清中CXCL4的表达。结果繁殖成功的子代小鼠有3种基因型:野生型、杂合子及纯合子;4、10周龄雌性纯合子小鼠体重明显重于同龄的野生型小鼠(P<0.05),6、10周龄雄性纯合子小鼠体重明显重于同龄的野生型小鼠(P<0.05);6周龄雌性纯合子小鼠结直肠长度明显长于同周龄的野生型小鼠(P<0.05),但结直肠的形态结构无显著改变;纯合子小鼠血清中CXCL4的表达量明显低于WT小鼠(P<0.05)。结论成功获得了CXCL4基因敲除纯合子小鼠,为深入研究CXCL4的生物学功能奠定了基础。
Objective To raise, breed and identify CXC chemokine ligand 4 (CXCL4) knockout mice and lay a foundation for further study on the biological function of CXCL4. Methods Heterotrophic mice were bred and propagated. The ears of the offspring mice were cut out and their genomic DNA was extracted. The genotypes of the mice were identified by PCR. The wild-type mice and CXCL4-knockout pure Zygotic mice weight changes and colorectal length by HE staining observed wild-type mice and CXCL4 knockout homozygous mice colorectal morphology; ELISA CXCL4 wild-type and homozygous mice serum CXCL4 expression. Results There were 3 genotypes of wild-type, heterozygous and homozygous reproductively successful offspring mice; 4 and 10-week-old female homozygous mice were significantly heavier than their wild-type mice at the same age (P <0.05); 6 The body weight of male homozygous mice at 10 weeks of age was significantly higher than that of wild-type mice at the same age (P <0.05). The length of colorectal tissue of 6-week-old female homozygous mice was significantly longer than that of wild-type mice at the same age ), But there was no significant change in the morphology of colorectal tissue. The expression of CXCL4 in serum of homozygous mice was significantly lower than that of WT mice (P <0.05). Conclusion CXCL4 knockout homozygous mice were successfully obtained, which laid the foundation for further study on the biological function of CXCL4.