The Effect Of Progesterone And Oestaradiol On The Oviductin And Osteopontin Produced By Oviduct Epithelial Cells In Yak

The aim of this study is to verify whether steroid hormones involved in mammalian embryonic development by Somatic cell.This paper discusses the effects of Progesterone?P₄?or Estradiol?E₂?on oviductin?Ovn?and Osteopontin?OPN?by oviduct epithelial cells.The yak oviduct epithelial cells as the research object,tissue culture method and two times of trypsin? for yak oviduct epithelial cells,we cloned OPN c DNA by RT-PCR and nucleic acid sequencing.The fourth generation of yak oviduct epithelial cells were cultured in vitro supplemented with different concentrations of P₄ and E₂.The relative expression level of Ovn and OPN gene or protein in different groups were detected by q RT-PCR and immunofluorescence.The content of OPN in the culture supernatant with different concentrations of P₄ and E₂ was detected by enzyme linked immunosorbent assay.These results suggestted that:?1?The range of P₄?5²5 ng/m L?,expression of Ovn increased gradually with the increase of P₄ concentration;The Ovn value was highest when the P₄ concentration was 25 ng/m L;However m RNA and protein expression was decreased when P₄ concentration over 25 ng/m L. ?2?E₂ improved the relative levels of Ovn m RNA and protein.These improvements were the highest in the 25 ng/m L E₂ group,followed by 50,10,100 and 200 ng/m L groups;The relative levels of Ovn were the lowest in control group ?3?The yak OPN was closely related to its cattle counterpart,reached 99.2% with Bos taurus,followed by 93.8% with Capra hircas,93.4% with Ovis aries,73.6% with Sus scrofa.The 1135 bp length OPN gene in yak?Gen Bank accession No: KY402045?,which contained the full open reading frame and determined to be 837 nucleotides encoding for a deduced protein of 278 amino acids. ?4?The OPN m RNA and protein levels were increased,when the in vitro culture system with different concentrations of P₄.The OPN value was highest when the P₄ concentration was 25 ng / m L;OPN m RNA and protein levels were decreased,when the in vitro culture system with different concentrations of E₂.The OPN value was lowest when the E₂ concentration was 100 ng / m L.?5?The effective range of P₄?10²5 ng/m L?,and the highest protein concentration numbers at 25 ng/m L,and reduced protein concentration level during all others.However,the OPN protein content was decreased when P₄ concentration over 50 ng/m L.The relative levels of OPN protein content were the lowest in control group?P₄=0 ng/m L?;Range of E₂?10¹00 ng/m L?,the content of OPN decreased gradually with the increase of E₂ concentration.However,OPN protein content was increased when E₂ concentration over 100 ng/m L.The OPN content was the lowest,When the E₂ concentration of 100ng/m L.