Abstract：Vitamin E plays a very important role in improving the reproductive function of livestock and poultry, improving immunity and protecting the liver, and its storage and metabolism mainly occurs in the liver. α-tocopherol transfer protein (α-TTP) is mainly expressed in the liver, which can specifically bind α-tocopherol and promote its transport from the liver to peripheral tissues for absorption and utilization. However, some studies have shown that α-TTP may also require other interacting factors in the process of transporting α-tocopherol. In this study, using co-immunoprecipitation (Co-IP) and Pull-down screening, the α-TTP-interacting protein-myosin 1d (MYO1D) was obtained. Immunofluorescence observation showed that MYO1D and α-TTP co-localized significantly. The Myo1d-shRNA lentivirus plasmid was used to construct the interference vector of the Myo1d gene, and the hepatocytes line stably infected with the lentiviral interference vector was obtained by screening. The results of qRT-PCR showed that the Myo1d gene expression in the interference group decreased significantly (P<0.01), and the interference efficiency was 46.83%, while the expression of α-ttp gene in the interference group had no significant change. The results of the kit test showed that the content of α-tocopherol in the Myo1d interference group increased significantly (P<0.05). The above results suggest that MYO1D could interact with α-TTP to mediate α-tocopherol transport in hepatocytes, and might provide power in the process of transporting α-tocopherol-TTP complexes. These findings provide experimental bases for screening the interaction factors in the process of α-TTP transporting α-tocopherol, and provide a reference basis for further research on the mechanism of α-TTP transporting α-tocopherol in the liver.
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