Construction of Pnpla3 I148M and Tm6sf2 E167K double mutant mouse model
10.3969/j.issn.1001-5256.2022.08.013
- VernacularTitle:Pnpla3 I148M和Tm6sf2 E167K双突变纯合小鼠模型的构建
- Author:
Mengke WANG
1
;
Shousheng LIU
2
;
Xueru CHU
3
;
Yifen WANG
1
;
Yongning XIN
1
Author Information
1. Department of Infectious Diseases, The Affiliated Qingdao Municipal Hospital of Qingdao University, Qingdao, Shandong 266071, China
2. Clinical Research Center, The Affiliated Qingdao Municipal Hospital of Qingdao University, Qingdao, Shandong 266071, China
3. School of Medicine and Pharmacy, Ocean University of China, Qingdao, Shandong 266071, China
- Publication Type:Original Articles_Fatty Liver Diseases
- Keywords:
Non-alcoholic Fatty Liver Disease;
Point Mutation;
Models, Animal;
Mice
- From:
Journal of Clinical Hepatology
2022;38(8):1784-1789
- CountryChina
- Language:Chinese
-
Abstract:
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 Pnpla3 148I/M Tm6sf2 167E/K heterozygous mice were bred by hybridization of Pnpla3 148M/M Tm6sf2 167E/E and Pnpla3 148I/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 Tm6sf2 167E/K mice. Male mice of Pnpla3 148M/M Tm6sf2 167K/K ( n =6), Pnpla3 148M/M Tm6sf2 167E/E ( n =6), and Pnpla3 148I/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 Tm6sf2 167E/E , Pnpla3 148I/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 Pnpla3 148I/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 Pnpla3 148I/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 Tm6sf2 167E/E , Pnpla3 148I/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 Tm6sf2 167E/E and Wt mice. Conclusion The Pnpla3 148M/M Tm6sf2 167K/K double mutant mouse model was successfully constructed. Pnpla3 Ⅰ 148M and Tm6sf2 E 167K double mutations can cause abnormal glucose metabolism in mice.
