摘要
目的利用Pnpla3^(148M/M)纯合小鼠与Tm6sf2^(167K/K)纯合小鼠杂交方法构建Pnpla3^(148M/M)Tm6sf2^(167K/K)双突变小鼠模型。方法利用Pnpla3Ⅰ148M和Tm6sf2 E 167K单突变纯合小鼠交配出Pnpla3^(148M/M)Tm6sf2^(167K/K)双突变杂合小鼠,再通过自交得到Pnpla3^(148M/M)Tm6sf2^(167K/K)双突变纯合小鼠。选取同窝的Pnpla3^(148M/M)Tm6sf2^(167K/K)(n=6)、Pnpla3^(148M/M)Tm6sf2^(167E)/E(n=6)、Pnpla3^(148I/I) Tm6sf2^(167K/K)(n=6)和野生(Wt)(n=6)雄性小鼠普通饮食喂养8周,在第8周检测小鼠葡萄糖及脂质代谢等指标。多组间比较采用方差分析,进一步两两比较采用LSD-t检验。结果琼脂糖凝胶电泳和核酸测序结果表明Pnpla3^(148M/M)Tm6sf2^(167K/K)双突变小鼠模型构建成功。Pnpla3^(148M/M)Tm6sf2^(167K/K)小鼠体质量与其他3种基因型小鼠比较,差异无统计学意义(P值均>0.05),Pnpla3^(148M/M)Tm6sf2^(167K/K)小鼠肝湿重高于Wt小鼠(P<0.05)。Pnpla3^(148M/M)Tm6sf2^(167K/K)小鼠的空腹血糖低于Tm6sf2^(167K/K)单突变小鼠和Wt小鼠(P值均<0.05),Pnpla3^(148M/M)Tm6sf2^(167K/K)小鼠葡萄糖耐受能力较Tm6sf2^(167K/K)单突变小鼠有所下降(P<0.05),四组基因型小鼠的胰岛素水平无明显差异(P值均>0.05)。Pnpla3^(148M/M)Tm6sf2^(167K/K)双突变小鼠的血浆生化指标与其他三种基因型小鼠比较,差异均无统计学意义(P值均>0.05)。肝脏油红O切片染色结果显示Pnpla3^(148M/M)Tm6sf2^(167K/K)双突变小鼠的肝脏较Pnpla3^(148M/M)单突变小鼠和Wt小鼠更容易发生脂质积累。结论Pnpla3^(148M/M)Tm6sf2^(167K/K)双突变小鼠模型构建成功,Pnpla3Ⅰ148M和Tm6sf2 E 167K双突变可引起小鼠葡萄糖代谢异常。
Objective To construct a Pnpla3^(148M/M)Tm6sf2^(167K/K)double mutant mouse model by crossbreeding Pnpla3^(148M/M)homozygous mice and Tm6sf2^(167K/K)homozygous mice.Methods Pnpla3148I/M Tm6sf2167E/K heterozygous mice were bred by hybridization of Pnpla3^(148M/M)Tm6sf2167E/E and Pnpla3148I/I Tm6sf2^(167K/K)homozygous mice,and the Pnpla3^(148M/M)Tm6sf2^(167K/K)mice were obtained by the self-crossbreeding of Pnpla3^(148I/M) Tm6sf2167E/K mice.Male mice of Pnpla3^(148M/M)Tm6sf2^(167K/K)(n=6),Pnpla3^(148M/M)Tm6sf2167E/E(n=6),and Pnpla3148I/I Tm6sf2^(167K/K)(n=6)genotypes and Wt mice(n=6)were fed with normal diet for 8 weeks,and then the glucose and lipid metabolism indices were measured.A one-way analysis of variance was used for comparison between multiple groups,and the least significant difference t-test was used for further comparison bewteen two groups.Results Agarose gel electrophoresis and nucleic acid sequencing results showed that the Pnpla3^(148M/M)Tm6sf2^(167K/K)double mutant mouse model was successfully constructed.There were no significant difference in body weight between the Pnpla3^(148M/M)Tm6sf2^(167K/K)mice and the Pnpla3^(148M/M)Tm6sf2167E/E,Pnpla3148I/I Tm6sf2^(167K/K),and Wt mice(all P>0.05).The Pnpla3^(148M/M)Tm6sf2^(167K/K)mice had a significantly higher liver wet weight than the Wt mice(P<0.05).The fasting blood glucose of Pnpla3^(148M/M)Tm6sf2^(167K/K)mice was significantly lower than that of Pnpla3148I/I Tm6sf2^(167K/K)mice and Wt mice(both P<0.05).The glucose tolerance of Pnpla3^(148M/M)Tm6sf2^(167K/K)mice was significantly reduced compared with the Pnpla3148I/I Tm6sf2^(167K/K)mice(P<0.05).There were no significant differences in insulin level between the four groups of mice(all P>0.05).Also,there were no significant differences in the serum levels of biochemical indices between the Pnpla3^(148M/M)Tm6sf2^(167K/K)mice and the Pnpla3^(148M/M)Tm6sf2167E/E,Pnpla3148I/I Tm6sf2^(167K/K),and Wt mice(all P>0.05).Oil red O staining of the liver showed that more lipid accumulation was observed in the Pnpla3^(148M/M)Tm6sf2^(167K/K)mice than in the Pnpla3^(148M/M)Tm6sf2167E/E and Wt mice.Conclusion The Pnpla3^(148M/M)Tm6sf2^(167K/K)double mutant mouse model was successfully constructed.Pnpla3Ⅰ148M and Tm6sf2 E167K double mutations can cause abnormal glucose metabolism in mice.
作者
王孟轲
刘守胜
褚雪汝
王艺奋
辛永宁
WANG Mengke;LIU Shousheng;CHU Xueru;WANG Yifen;XIN Yongning(Department of Infectious Diseases,The Affiliated Qingdao Municipal Hospital of Qingdao University,Qingdao,Shandong 266071,China;Clinical Research Center,The Affiliated Qingdao Municipal Hospital of Qingdao University,Qingdao,Shandong 266071,China;School of Medicine and Pharmacy,Ocean University of China,Qingdao,Shandong 266071,China)
出处
《临床肝胆病杂志》
CAS
北大核心
2022年第8期1784-1789,共6页
Journal of Clinical Hepatology
基金
国家自然科学基金面上项目(31770837)。
关键词
非酒精性脂肪性肝病
点突变
模型
动物
小鼠
Non-alcoholic Fatty Liver Disease
Point Mutation
Models,Animal
Mice
