Inhibition of Paeoniflorin on TNF-α-induced TNF-α Receptor Type I /Nuclear Factor-κB Signal Transduction in Endothelial Cells / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the inhibitory effect of paeoniflorin (PAE) on TNF-α-induced TNF receptor type I (TNFR1)-mediated signaling pathway in mouse renal arterial endothelial cells (AECs) and to explore its underlying molecular mechanisms.</p><p><b>METHODS</b>Mouse AECs were cultured in vitro and then they were treated by different concentrations PAE or TNF-α for various time periods. Expression levels of intercellular cell adhesion molecule-1 (ICAM-1) were detected in the normal group (cultured by serum-free culture media), the TNF-α group (cultured by 2-h serum-free culture media plus 6-h TNF-α 30 ng/mL), the low dose PAE group (cultured by 2-h PAE 0.8 μmo/L plus 6-h TNF-α 30 ng/mL), the middle dose PAE group (cultured by 2-h PAE 8 μmol/L plus 6-h TNF-α 30 ng/mL), the high dose PAE group (cultured by 2-h PAE 80 μmol/L plus 6-h TNF-α 30 ng/mL) with Western blot analysis. Nuclear translocation of transcription factor NF-κB (NE-κB) was detected in the normal group (cultured by serum-free culture media), the TNF-α group (cultured by 2-h serum-free culture media plus 45-mm TNF-α 30 ng/mL), and the high dose PAE group (cultured by 2-h PAE 80 μmol/L plus 45-min TNF-α 30 ng/mL) by immunofluorescent staining. Expression levels of the phosphorylation of extracellular signal-regulated (protein) kinase (ph-ERK) and p38 (ph- p38) were detected in the normal group (cultured by serum-free culture media) and the high dose PAE group (2-h PAE 80 μmol/L culture) by Western blot. NF-κB inhibitor-α (IκBα) protein expressions were detected in the normal group (cultured by serum-free culture media), the TNF-α group (cultured by 2-h serum-free culture media plus 30-min TNF-α 30 ng/mL), the high dose PAE group (cultured by 2-h PAE 80 μmol/L plus 30-min TNF-α 30 ng/mL), the p38 inhibitor group (SB group, pretreatment with SB238025 25 μmol/L for 30 min, then treated by PAE 80 μmol/L for 2 h, and finally treated by TNF-α 30 ng/mL for 30 min), the ERK inhibitor group (PD group, treated by PD98059 50 μmol/L for 30 min, then treated by PAE 80 μmol/L for 2 h, and finally treated by TNF-α 30 ng/mL for 30 min) by Western blot.</p><p><b>RESULTS</b>Compared with the normal group, ICAM-1 protein expression levels obviously increased (P < 0.01). Compared with the TNFα group, ICAM-1 protein expression levels were obviously inhibited in the high dose PAE group (P < 0.05). Protein expression levels of ph-p38 and ph-ERK were obviously higher in the hIgh dose PAE group (P < 0.05). Compared with the normal group, IκBα protein expression levels obviously decreased in the TNF-α group (P < 0.01). Compared with the TNFα group, TNF-α-induced IκBα degradation could be significantly inhibited in the high dose PAE group (P < 0.01); the inhibition of PAE on IκBα degradation could be significantly inhibited in the SB group (P < 0.05). NF-κB/p65 signal was mainly located in cytoplasm in the normal group. NF-κB/p65 was translocated from cytoplasm to nucleus after stimulated by 45 min TNF-α in the TNF-α group, while it could be significantly inhibited in the high dose PAE group.</p><p><b>CONCLUSIONS</b>PAE inhibited TNF-α-induced expression of lCAM-1. Its action might be associated with inhibiting TNFR1/NF-κB signaling pathway. p38 participated and mediated these actions.</p>

Expression of Oct2 and its significance in lymphoma diagnosis / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the specificity and sensitivity of Oct2 protein expression in lymphoma cells and its significance in diagnosis and classification of lymphoma.</p><p><b>METHODS</b>Formalin-fixed and paraffin-embedded materials from 129 cases of lymphoma and 10 cases of reactive lymphoid hyperplasia (RLH) were studied by EnVision immunohistochemistry for Oct2 protein.</p><p><b>RESULTS</b>Oct2 was mainly expressed in germinal center cells of RLH. It was diffusely expressed in B-cell lymphoma cells. 97.7% cases (85/87) of B-cell lymphoma and 3.8% cases (1/26) of T-cell lymphoma were positive for Oct2 protein. In comparison, the expression rates for CD20 and CD79alpha in B-cell lymphomas were 90.8% (79/87) and 84.7% (61/72) respectively. The difference in expression rates between Oct2 protein and CD20 was not statistically significant (P > 0.05) There was, however, significant difference in expression rates between Oct2 protein and CD79alpha (P < 0.05). The expression rates of Oct2 protein in nodular lymphocyte-predominant Hodgkin lymphoma and classic Hodgkin lymphoma were 3/3 and 46.2% (6/13) respectively. The difference in expression rates of Oct2 protein in these two groups showed no statistical significance (P > 0.05).</p><p><b>CONCLUSION</b>As a relatively sensitive and specific marker for B cells, Oct2 can serve as a useful antibody for the diagnosis and differential diagnosis of lymphoma.</p>