Effect of Lipopolysaccharide and 4-(Methylnitro-samino)-1- (3-pyridyl)-1-butanone on the Proliferation of Mouse Bone Marrow Stem Cells

Liu Xiaoping

doi:10.18483/ijsci.925

Abstract

BMSCs were isolated, cultured and proliferated in vitro. The proliferation of mouse BMSCs treated with different concentrations of LPS and NNK were detected by MTT assay, the expression of NF-κB and CUGBP1 in the BMSCs were analyzed by Immunocytochemistry assay and Western Blot. MTT results shown that different concentrations of LPS and NNK could promote the proliferation of BMSCs and the promotion effect of LPS12.5μg/ml, NNK10μg/ml and LPS12.5μg/ml combined NNK 10μg/ml was much more significant than others. Immunocytochemistry and Western Blot revealed that the location and the expression level of NF-κB and CUGBP1 in BMSCs treated with LPS and NNK were changed with different incubation time. The highest expression of NF-κB was detected at the LPS48h, NNK24h and LPS combined NNK24h and significant difference can be seen in-group compared. The highest expression of CUGBP1 was LPS96h, NNK48h and LPS combined NNK24h and significant difference can be seen in-group compared. The irritation of LPS and NNK on the proliferation of mouse BMSCs may mediate by up-regulating the expression of NF-κB and CUGBP1.

Highlights

Stem cells is the most primitive, pluripotent and self-renewing cells in tissue and has the ability to differentiate into osteoblast, chondroblast, adipocyte, neurocyte, cardiomyocytes, hepatocyte, with different inductions, this can be applied to therapy of various organs damage and become the hot research of life science and medicine [1]
1.4 The expression features of NF-κB and CUGBP1 in Bone Marrow Stem Cells (BMSCs) detected by Immunocytochemistry assay The concentration of LPS, NNK and LPS combined NNK we used were based on the result of MTT assay
BMSCs were incubated with the medium containing LPS12.5μg/ml, NNK10μg/ml, LPS12.5μg/ml combined NNK10μg/ml respectively for 24h, 48h, 96h, the cells were fixed by cold acetone for 30min, followed by permeabilized with 0.5% TritonX-100 for 20min at room temperature, rinsed several times with PBS, treated with 3% H2O2 for 30min to inactivate the endogenous enzyme, incubated with primary antibody (Rabbit anti Mouse NF-κB p65 polyclonal antibody, Rabbit anti Mouse CUGBP1 polyclonal antibody) in a humidified chamber overnight at 4°C, incubated with secondary antibody (Goat anti Rabbit HRP conjugated) for 2h at 37°C, colorated with DAB in dark for 6min

Summary

Introduction

Stem cells is the most primitive, pluripotent and self-renewing cells in tissue and has the ability to differentiate into osteoblast, chondroblast, adipocyte, neurocyte, cardiomyocytes, hepatocyte, with different inductions, this can be applied to therapy of various organs damage and become the hot research of life science and medicine [1]. Effect of Lipopolysaccharide and 4-(Methylnitro-samino)-1-(3-pyridyl)-1-butanone on the Proliferation of Mouse Bone Marrow Stem Cells one of the most harmful components of cigarette smoke and can cause a variety abnormal change of immune function. It can result in the production of toxins that bind DNA to form adducts that causes the mutation of oncogene and anti-oncogenewhen it is metabolized in body. In this study the effect of different concentrations of LPS, NNK and LPS combined NNK on the proliferation and the expression of NF-κB and CUGBP1 of mouse BMSCs were detected to reveal the possible mechanism of BMSCs proliferation stimulated by inflammatory

Material and Method

Statistical Analyses

Results