神经肽Y对猪小肠上皮细胞NaPi-Ⅱb蛋白表达及无机磷吸收的影响

doi:10.11843/j.issn.0366-6964.2014.10.009

Abstract

Abstract:

This experiment was conducted to investigate the effect of different concentration of neuropeptide Y on expression of NaPi-Ⅱb protein and absorption of inorganic phosphorus through culturing porcine small intestinal epithelial cells in vitro.To interpret the regulation mechanism of inorganic phosphorus absorption in small intestine affected by neuropeptide Y，and provide experiment basis for further study on molecular mechanism of animal phosphorus absorption and regulation pathways.Six treatments were designed according to different concentrations of neuropeptide Y，and the control group without neuropeptide Y，other treatments of adding different concentrations of neuropeptide Y (10^-11，10^-10，10^-9，10^-8 and 10^-7 mol•L^-1)，respectively，with 6 or 8 repetition for each treatment.Proliferation of porcine small intestinal epithelial cells was determined by MTT assay.Phosphorus content in the cell supernatant fluid was detected by phosphorus detection kits.The relative expression quantity of NaPi-Ⅱb mRNA in porcine small intestinal epithelial cells was detected by RT-PCR.The relative expression levels of NaPi-Ⅱb protein was detected by Western blotting.The results showed that there was no significant difference (P>0.05) in cell proliferation in each treatment group，stimulating intestinal epithelial cells with different concentration of neuropeptide Y after 24 h；Phosphorus content in the cell supernatant fluid declined on the form of significant linear (P<0.01) and quadratic curve(P<0.01) over time.Neuropeptide Y and phosphorus content in IEC were very significantly correlated (P<0.01) after stimulated 24 h by NPY.Compared with the control group，the treatment group of 10^-10 ，10^-9 ，10^-8 and 10^-7 mol•L^-1 very significantly (P<0.01) decreased phosphorus content，and the amount of phosphorus uptake in small intestinal epithelial cells increased by 161%，171%，168% and 139%，respectively than that of the control group.There were significant quadratic correlations between relative expression quantity of NaPi-Ⅱb mRNA and NaPi-Ⅱb protein and neuropeptide Y adding concentration (P<0.01，P<0.05，respectively).We concluded that the appropriate concentration of neuropeptide Y can stimulate NaPi-Ⅱb gene transcription，increase expression levels of NaPi-Ⅱb protein，and then induce inorganic phosphorus in the extracellular fluid to transport across the membrane into the intracellular fluid，so as to promote porcine small intestinal epithelial cells to absorb inorganic phosphorus，and the optimum concentration of neuropeptide Y in the cell culture fluid between 10^-10 and 10^-8 mol•L^-1.

CLC Number:

S828
S816.2

HU Long-chang，FANG Re-jun. Effect of Neuropeptide Y on Expression of NaPi-Ⅱb Protein and Absorption of Inorganic Phosphorus in Porcine Small Intestinal Epithelial Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(10): 1640-1647.

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Effect of Neuropeptide Y on Expression of NaPi-Ⅱb Protein and Absorption of Inorganic Phosphorus in Porcine Small Intestinal Epithelial Cells

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