Impact of normalization strategy on cardiac expression of pro-inflammatory cytokines: Evaluation of reference genes in different human myocardial regions after Left Ventricular Assist Device support

IL-33/ST2 Pathway and Classical Cytokines in End-Stage Heart Failure Patients Submitted to Left Ventricular Assist Device Support: A Paradoxic Role for Inflammatory Mediators?

“…Five myocardial samples from post-LVAD and HT groups were collected at the time of transplantation from the similar areas of left (LV) and right ventricle (RV), as previously reported [18]. Sample collection resulted in n = 22 myocardial samples in the pre-LVAD group, n = 30 in the post-LVAD group, and n = 35 in the HT group.…”

“…In order to normalize the inflammatory gene expression, genes previously selected [18] and confirmed in these set of samples were used as reference. The reaction conditions of all primer pairs were assessed determining the optimal annealing temperature, conducting a gradient PCR, and to verify efficiency, generating a standard curve obtained by scalar dilution of a cDNA pool.…”

Effect of siRNA-induced inhibition of IL-6 expression in rat cerebral gliocytes on cerebral edema following traumatic brain injury

“…However, under pathological stress, the secretion of cerebral astrocytes and microglia increases, thus IL-6 expression increases (2,3,19,20), which mediates a series of pathophysiological reactions. Such IL-6-induced pathophysiological reactions include: (i) Changes in cerebral microcirculation, resulting in increases in vascular permeability and damage to the BBB, thus promoting the formation and progression of cerebral edema; (ii) increases in leukocyte adhesion and endothelial cells, promoting extravascular infiltration and activation of inflammatory cells (4,19); (iii) increases in the proliferation and reparation of cerebral glial cells in damaged brain regions (5,22); (iv) induction of complications associated with TBI, as well as multiple organ failure, for example, increases in IL-6 have a negative muscle strength effect and cytotoxic effect, which causes damage to the structure and function of the left ventricle, promoting deterioration of patient cardiac function and hemodynamics (7,8,23,24); and (v) problems associated with fever and local metabolism, affecting patient prognosis and rehabilitation. As an inflammatory cytokine, IL-6 expression has been found to increase significantly in the CNS following TBI and the levels of IL-6 in the CSF has been negatively correlated with Glasgow Coma Scale and Glasgow Outcome Scale scores following injury (9,25).…”

Myocardial interleukin-6 in the setting of left ventricular mechanical assistance: relation with outcome and C-reactive protein

“…Total RNA was extracted from myocardial specimens and RNA concentration and purity were evaluated [13] . Starting from total RNA, fi rst-strand cDNA was synthesized by iScript cDNA Synthesis kit (Bio-Rad, Hercules, CA, USA).…”

“…Real-time PCR was performed in duplicate using the Bio-Rad C1000TM thermal cycler (CFX-96 real-time PCR detection systems; Bio-Rad, Hercules, CA, USA). IL-6 relative quantifi cation was obtained by Δ Δ Ct method using previously selected reference genes [13] .…”