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Objective To investigate the effect and the underlying molecular mechanism of zinc finger protein Zpr1 on differentiation and the insulin signaling pathway of pre-adipocytes. Methods A high-fat diet-induced insulin resistance mouse model (IR group, n=7) was established by feeding 8-week-old C57BL/6J male mice using high-fat feed for 16 weeks, with conventional diet feeding mice as control group (n=7). Glucose tolerance and insulin tolerance tests were used to observe the blood glucose values of mice in each group and verify the insulin resistance phenotype of mice in the induction group. Western blotting and RT-PCR were performed to detect the protein and mRNA expression of Zpr1 in subcutaneous and visceral adipose tissue from high-fat diet induced insulin resistance mice and regular diet mice. Bioinformatic prediction combined with dual-luciferase reporter gene system were employed to detect the influence of single nucleotide polymorphism (SNP) rs964184 GG genotype and CC genotype in human ZPR1 against miR-4286 binding to the 3'UTR of ZPR1. After mouse preadipocyte 3T3-L1 transfected with miR-4286 simulant to observe the effect of miR-4286 regulating Zpr1 expression on adipocyte differentiation and insulin signaling pathway. Results The protein and mRNA expression levels of Zpr1 decreased obviously in the subcutaneous and especially in the visceral adipose tissues of high-fat diet induced insulin resistant (IR) mice compared with the mice in control group (P<0.05, P<0.01).The rs964184 SNP locus of ZPR1 made it difficult that ZPR1 3'UTR forms a multi-hairpin structure, but may promote miR-4286 to recognize and bind to ZPR1 3'UTR. The miR-4286 level increased obviously in subcutaneous and especially in visceral adipose tissues of IR model group mice (P<0.05); meanwhile, the protein expression level decreased of Zpr1 in mouse pre-adipose cells 3T3-L1 regulated by miR-4286 simulacrum (P<0.05). At the same time, after transfection of miR-4286 simulacrum, the expression levels of fat cell differentiation and insulin signaling pathway-related proteins PPAR-γ, Perilipin A, pAkt, and IRS-1 in 3T3-L1 cells were reduced markedly (P<0.05). Conclusions The GG genotype rs964184 locus can promote the post-transcriptional regulation of Zpr1 by miR-4286. The decreased expression of Zpr1 can inhibit the differentiation of 3T3-L1 preadipocytes and interfere the insulin signaling pathway.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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