| Objective:In order to explore the roles of TLR2 and TLR4 in HepG2cells, and to discover whether LPS can affect hepatocytes without immuno-mechanism, we detected the changes of TLR2 and TLR4 expressions inhuman hepatoma cell lines: HepG2. Methods: HepG2 is cells are most similar to normal humanhepatocytes. We selected this cell line as model of hepatocyte to study thedifferences of TLR2 and TLR4 expression between hepatocytes before andafter LPS stimulation. In this research, all of the HepG2 cells werestimulated with 0 ug /ml, 1ug /ml , 10 ug /ml , 100ug /ml , 1 mg/ml and10mg/ml LPS. Then the expression of protein of TLR2 and TLR4 wereexamined by immuno- histochemistry (IHC), the mRNA examined byreversal transcription- polymerase chain reaction (RT-PCR). Results: IHC and RT-PCR analysis revealed that TLR2 and TLR4expressions were detected in all of the HepG2 cells. Moreover, withoutimmune activation, TLR2 and TLR4 expressions were higher when theconcentrations groups of LPS were higher. Conclusion: TLR2 and TLR4 can be expressed in HepG2 cells ,and the expressions intensity of TLR2 and TLR4 in HepG2 cell lines canbe enhanced by increasing concentration of LPS. Objective: In order to explore the roles of TLR2 and TLR4 inhepatocyte damage after hepatitis B virus infection, and to discoverwhether LPS can affect hepatocytes without immuno-mechanism pre- andpost- HBV infection, we detected the changes of TLR2 and TLR4expressions in human hepatocyte lines HepG2 cells and 2.2.15 cells. Methods: HepG2 is cells are most similar to normal humanhepatocytes and 2.2.15 cells are HepG2 cells infected with HBV. Weselected these two cell lines to study the differences of TLR2 and TLR4expression between HepG2 cells before and after HBV infection. In thisresearch, both HepG2 and 2.2.15 cells were stimulated with 0 ug /ml ,1ug/ml , 10 ug /ml , 100ug /ml , 1 mg/ml and 10mg/ml LPS. Then theexpression of protein of TLR2 and TLR4 were examined by immuno-histochemistry (IHC). The mRNA of HepG2 and 2.2.15 cells stimulatedwith 0 ug /ml and 10mg/ml LPS were examined by reversaltranscription-polymerase chain reaction (RT-PCR). The apoptosis ofHepG2 and HepG2.2.15 cells were examined by flow cytometry (FC), andthe expressions of HBsAg and HBeAg of HepG2.2.15 cells tested withAbbott kits. Results: IHC and RT-PCR analysis revealed that TLR2 and TLR4expressions were detected in both HepG2 and 2.2.15 cells. Moreover,without immune activation, TLR2 and TLR4 expressions were higher whenthe concentrations of LPS were higher. FC analysis revealed that noapoptosis detected in HepG2 cells stimulated with LPS in this research, butapoptosis have detected in 2.2.15 cells when treated with the same factors.Furthermore, the apoptosis ratios increased with the increase inconcentrations of LPS. When concentrations of LPS were 1ug /ml , 10 ug/ml , 100ug /ml , 1 mg/ml and 10mg/ml, the apoptosis ratios were 1.94%,3.03%, 3.50%, 3.72%, 5.30%, respectively. Abbott analysis revealed thatexpressions of HBsAg and HBeAg of 2.2.15 cells stimulated with LPSwere lower than those not stimulated with LPS. Conclusion: HBV can affect expressions of TLR2 and TLR4 inHepG2 cell lines. LPS can lead 2.2.15 cells to apoptosis but not HepG2cells. Although LPS cannot damage normal hepatocytes, it might aggravatehepatocytes damage when their microenvironment was changed by HBVinfection. Therefore, we feel the mechanism involved in this phenomenaneeds to be identified through more research. Objective: To explore the roles of TLR2 and TLR4 in HepG2 cellstransfected with HBV X gene, and whether HBV X gene or HBx proteinplays an important role in the mechanism of that hepatocytes with HBVinfection damaged by LPS. Methods: In this research, PECP4-X-HepG2 cells, one HepG2 cellline transfected with HBV X gene, were stimulated with 0ug/ml,1ug/ml,10ug/ml, 100ug/ml,1mg/ml and 10mg/ml LPS. Then the expressionof protein of TLR2 and TLR4 were examined by immuno-histochemistry(IHC). The mRNA of HepG2 cells, HepG2.2.15 cells andPECP4-X-HepG2 cells were exam... |
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