肝细胞特异性HuR基因敲除小鼠模型的表型研究

目的 探讨肝细胞特异性敲除HuR基因对小鼠肝脏功能的影响。方法 从美国Jackson实验室引进LoxP标记的人抗原R（human antigen R，HuR）基因小鼠（HuR^fl/fl）和Alb-Cre转基因小鼠。杂交后，经数代选育配种，建立肝细胞特异性HuR基因敲除小鼠（HuR^LKO）模型。PCR技术鉴别小鼠基因型，蛋白免疫印迹和免疫荧光实验检验小鼠肝脏的HuR表达水平，同时HE染色观察肝脏结构变化。高脂喂养HuR^fl/fl/Alb-Cre和HuR^fl/fl两组小鼠，观察小鼠体重、肝重和肝体比变化，提取血清检测肝脏损伤和脂肪代谢相关指标。结果 PCR成功检测筛选子代小鼠基因型，包括HuR^fl/fl/Alb-Cre和HuR^fl/fl小鼠。蛋白免疫印迹实验和免疫荧光实验证明小鼠肝细胞特异性HuR基因敲除成功。HE染色结果提示HuR^LKO小鼠的肝细胞出现变性水肿、局部坏死。高脂喂养实验提示两组小鼠在肝重、肝体比、AST、ALT、HDL-C、血糖上的差异具有统计学意义（P＜0.05），但在体重、TG、LDL-C的差异没有统计学意义（P＞0.05）。结论 HuR基因是维持小鼠肝细胞功能的必要基因，Cre-LoxP技术可成功构建HuR^LKO小鼠模型，且特异性高，表型明显，为研究HuR在肝脏各项疾病的发生发展中所扮演的角色，提供了一个很好的疾病模型。

Phenotype study of hepatocyte-specific HuR gene knockout in mice model

Objective To investigate the effect of hepatocyte-specific HuR gene knockout on liver function in mice model. Methods LoxP-labeled human antigen R (HuR) mice (HuR^fl/fl) and Alb-Cre transgenic mice were imported from the Jackson Laboratory in the United States. After several generations of crossing and breeding,hepatocyte-specific HuR gene knockout mice (HuR^LKO) model was established. PCR technology was used to identify the genotypes of mice. Western blotting and immunofluorescence experiments were used to examine the expression of HuR in mice liver. HE staining was used to observe the structural changes in mice liver. HuR^fl/fl/Alb-Cre and HuR^fl/fl mice were fed with high-fat diet, and the changes of body weight and liver weight were recorded. Serum was extracted to detect hepatic impairment and lipid metabolism related indexes. Results PCR detected the genotypes of the progeny mice successfully, including HuR^fl/fl/Alb-Cre mice and HuR^fl/flmice. Western blotting and immunofluorescence experiments proved that the hepatocyte-specific HuR gene knockout in mice was successful. The results of HE staining suggested that the hepatocytes of HuR^LKOmice had degeneration and local necrosis. The high-fat feeding experiment showed that the differences in liver weight, AST, ALT, HDL-C and blood glucose between the two groups of mice were statistically significant (P<0.05), but the difference in body weight, TG and LDL-C was not statistically significant (P>0.05). Conclusion HuR gene is an essential gene to maintain the function of hepatocytes in mice model. Cre-LoxP technology can successfully construct the HuR^LKOmice model with high specificity and obvious phenotype, which provides a good disease model for the study of the role of HuR in the occurrence and development of various liver diseases.