Interleukin-6 Protein Expression Is More Important Than Interleukin-6 mRNA Levels in Assessing Surgical Invasiveness
ABSTRACT Interleukin-6 (IL-6) protein has been recognized as a sensitive marker of surgical stress response. However, little is known about the clinical significance of IL-6 mRNA levels as a marker of surgical stress. This study aims to examine the role of IL-6 mRNA expression in comparing the tissue invasiveness of microendoscopic discectomy (MED) and open discectomy (OD).
Twenty-three consecutive patients were randomly selected to undergo either MED or OD. The total RNA was extracted from the peripheral whole blood of patients at pre-op and at 1, 2, 4, 8, 12 h post-op. The real-time reverse transcription polymerase chain reaction (RT-PCR) using the SYBR Green I fluorescence dye and the 2(-DeltaDeltaCt) method was adopted to measure the IL-6 gene expression.
The quantitative changes of IL-6 mRNA expression in MED and OD patients at different times post-op differed significantly, P = 0.04. Experimental results indicate that the changes in IL-6 mRNA expression in OD and MED groups varied significantly at 1 h, 12 h post-op, 10.26-fold versus 4.42-fold and 52.15-fold versus 26.78-fold increase, respectively. Although IL-6 mRNA expression demonstrated an earlier difference than protein levels at 1 h post-op, IL-6 mRNA levels were found to be significantly affected after surgical procedures. Furthermore, compared with our enzyme-linked immunosorbent assay data, no significant correlation existed between IL-6 mRNA and protein levels at any post-op time interval.
We conclude that IL-6 mRNA expression using RT-PCR to extract the total RNA from a patient's peripheral whole blood is more sensitive than protein levels but can be significantly affected by surgical procedures. The enzyme-linked immunosorbent assay data on IL-6 protein expression are more consistent and significant than IL-6 mRNA levels in comparing tissue invasiveness between MED and OD.
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ABSTRACT: Endothelial dysfunction is associated with cardiovascular diseases. The Ca2+ influx occurring via activation of plasmalemma Ca2+ channels was shown to be critical in signaling the increase in endothelial permeability in response to a variety of permeability-increasing mediators. It has been reported that angiotensin II (AngII) could induce Ca2+ signaling in some cells, and transient receptor potential canonical 1 (TRPC1) had an important role in this process. The objective of this study was to examine the mechanism of AngII-induced Ca2+ entry and vascular endothelial hyperpermeability. Human umbilical vein endothelial cells (HUVECs) exposed to AngII exhibited dose-dependent increase in [Ca2+]i and endothelial permeability. Quantitative real-time RT-PCR and Western blotting showed that the level of TRPC1 expression had increased significantly at 12h and at 24h after treatment of HUEVCs with AngII. The expression of p65 was suppressed using an RNAi strategy. The results showed that the NF-κB signaling pathway and type-1 receptor of AngII was involved in AngII-induced TRPC1 upregulation. Moreover, knockdown of TRPC1 and NF-κB expression attenuates AngII-induced [Ca2+]i and endothelial permeability. NF-κB and TRPC1 have critical roles in AngII-induced Ca2+ entry and endothelial permeability.Peptides 07/2009; 30(7):1368-1373. DOI:10.1016/j.peptides.2009.04.007 · 2.62 Impact Factor
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