Immunohistochemical and mRNA expression of RANK, RANKL, OPG, TLR2 and MyD88 during apical periodontitis progression in mice

Abstract Objective To evaluate and correlate, in the same research, the mRNA expression and the staining of RANK, RANKL, OPG, TLR2 and MyD88 by immunohistochemistry in the apical periodontitis (AP) progression in mice. Material and Methods AP was induced in the lower first molars of thirty-five C57BL/6 mice. They were assigned to four groups according to their euthanasia periods (G0, G7, G21 and G42). The jaws were removed and subjected to histotechnical processing, immunohistochemistry and real-time reverse transcription-PCR (qRT-PCR). Data were analyzed with parametric and nonparametric tests (α=0.05). Results An increase of positive immunoreactivity for RANK, RANKL, OPG, TLR2 and MyD88 was observed over time (p<0.05). The RANKL expression was different between the groups G0 and G42, G21 and G42 (p=0.006), with G42 presenting the higher expression in both comparations. The OPG expression was statistically different between the groups G0 and G7, G7 and G21 and G7 and G42 (p<0.001), with G7 presenting higher expression in all the time points. The TLR2 expression was different between the groups G0 and G42 (p=0.03), with G42 showing the higher expression. The MyD88 expression presented a statistical significant difference between groups G7, G21 and G42 compared with G0 (p=0.01), with G0 presenting the smallest expression in all the comparisons. The Tnfrsf11/Tnfrsf11b (RANKL/OPG) ratio increased with the AP progression (p=0.002). A moderate positive correlation between MyD88 and RANKL (r=0.42; p=0.03) and between MyD88 and TLR2 (r=0.48; p<0.0001) was observed. Conclusion The expression of the RANK, RANKL, OPG, MyD88 and TLR2 proteins as well as the ratio Tnfrsf11/Tnfrsf11b (RANKL/OPG) increased with AP progression. There was also a moderate positive correlation between the expression Myd88-Tnfrsf11 and Tlr2-Myd88, suggesting the relevance of Tlr2-Myd88 in bone loss due to bacterial infection.

Study on Effect of TLR4/MyD8 8 Protein on Chronic Renal Failure / 现代检验医学杂志

Objective To explore the expression of TLR4 and MyD88 in chronic renal failure and its role in the development of chronic renal failure.Methods 40 cases of patients diagnosed chronic renal failure during 2011.07~2014.05 were select-ed as observation subjects,and renal tissues without invasion of 40 cases patients with renal carcinoma resection were chosen as control.The expression of TLR4 and MyD88 in chronic renal failure was detected by IMH.The mice model was stabled establish through gavage of adenine (200 mg/kg).And the TLR4 and MyD88 expression was detected by RT-PCR and Western blot.The models was divided into three groups:TLR4 blocking group,TLR4 non-blocking and control group.And the BUN and CRE were detected by biochemical analyzer in three groups.Results The expression of TLR4 and MyD88 were higher in chronic renal failure than in normals.The TLR4 and MyD88 were also higher in chronic renal failure model mice.The levels of BUN (15.65±3.97 mmol/L)in TLR4 blocking group were lower than which in TLR4 non-blocking group (23.33±7.62 mmol/L)and which in IgG group (26.33±6.77mmol/L)(t=2.887,P=0.045).The CRE levels were the lowest in TLR4-blocking group (523.89 ± 52.67μmol/L)compared with the TLR4 non-blocking group (789.51 ± 98.17μmol/L)and the IgG group (809.51±94.19μmol/L)(t=4.125,P=0.015).Conclusion The increased expression of TLR4 and MyD88 in chronic renal failure significantly would promote the development of chronic renal failure.