Construction of Pnpla3 I148M and Tm6sf2 E167K double mutant mouse model

Mengke WANG; Shousheng LIU; Xueru CHU; Yifen WANG; Yongning XIN

doi:10.3969/j.issn.1001-5256.2022.08.013

Volume 38 Issue 8

Aug. 2022

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Article Navigation > Journal of Clinical Hepatology > 2022 > 38(8): 1784-1789

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Construction of Pnpla3 I148M and Tm6sf2 E167K double mutant mouse model

DOI: 10.3969/j.issn.1001-5256.2022.08.013

1a.
Department of Infectious Diseases, The Affiliated Qingdao Municipal Hospital of Qingdao University, Qingdao, Shandong 266071, China
1b.
Clinical Research Center, The Affiliated Qingdao Municipal Hospital of Qingdao University, Qingdao, Shandong 266071, China
2.
School of Medicine and Pharmacy, Ocean University of China, Qingdao, Shandong 266071, China

Research funding:

National Natural Science Foundation of China (31770837)

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Corresponding author: XIN Yongning, xinyongning@163.com(ORCID: 0000-0002-3692-7655)
Received Date: 2022-01-21
Accepted Date: 2022-02-25
Published Date: 2022-08-20

Abstract

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.
- Non-alcoholic Fatty Liver Disease,
- Point Mutation,
- Models, Animal,
- Mice

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References(18)

References

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Construction of Pnpla3 I148M and Tm6sf2 E167K double mutant mouse model

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Construction of Pnpla3 I148M and Tm6sf2 E167K double mutant mouse model

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