Involvement of miR-148a-5p in ApoE-/- mouse hepatocyte apoptosis induced by high methionine diet

doi:10.3969/j.issn.2095-4344.2349

Abstract

Abstract:

BACKGROUND: High methionine diet can cause liver damage in ApoE^-/-mice, and microRNAs (miRNAs) are involved in various cell processes such as cell survival, differentiation, and apoptosis, which is of great significance.

OBJECTIVE: To investigate the effect of miR-148a-5p on ApoE^-/- mouse hepatocyte apoptosis induced by high methionine diet.

METHODS: Twelve ApoE^-/- mice aged 5 weeks were randomly divided into two groups with six ApoE^-/- mice in each group: ApoE^-/- control group was fed with a normal diet, and ApoE^-/- high methionine diet group (ApoE^-/-+HMD) was fed with a high methionine diet. Hematoxylin-eosin staining was used to observe the morphological changes of liver tissues in the two groups. TUNEL staining was used to observe the apoptosis of hepatocytes. The expressions of Bax and Bcl-2 were detected by western blot. Fluorescence quantitative PCR was used to detect the expression of miR-148a-5p. Target Scan prediction software was used to predict the target gene of miR-148a-5p, and double luciferase activity assay was used to verify the targeted relationship.

RESULTS AND CONCLUSION: Compared with the control group, the hepatic lobular structure of the ApoE^-/- mice with high methionine diet was obviously disturbed, and some cells showed cytoplasmic osteoporosis; hepatocyte apoptosis increased; the expression of Bax significantly increased and the expression of Bcl-2 significantly decreased (P < 0.01). Fluorescence quantitative PCR results showed that the expression of miR-148a-5p increased in the ApoE^-/- mice with high methionine diet (P < 0.05). The target gene prediction software indicated that Bcl-2 was a potential target gene of miR-148a-5p, and the targeted relationship between miR-148a-5p and Bcl-2 was confirmed by the double luciferase activity assay (P < 0.05). After miR-148a-5p was overexpressed in hepatocytes, Bax expression increased and Bcl-2 expression decreased significantly (P < 0.01). In conclusion, miR-148a-5p promoted hepatocyte apoptosis through negative regulation of Bcl-2 in ApoE^-/- mouse hepatocytes induced by high methionine diet, which may be one of the mechanisms causing liver injury.

Key words: miR-148a-5p, Bcl-2, high-methionine diet, hepatocyte, apoptosis, animal

CLC Number:

Ma Fang, Zhang Hui, Li Guizhong, Ma Shengchao, Shen Jiangyong, Sun Lei, Hao Yinju, Ma Shengxian, Jiang Yideng. Involvement of miR-148a-5p in ApoE-/- mouse hepatocyte apoptosis induced by high methionine diet[J]. Chinese Journal of Tissue Engineering Research, 2020, 24(35): 5632-5637.

Figures/Tables 5

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