Multiple microRNAs regulate tacrolimus metabolism through CYP3A5.

The CYP3A5 gene polymorphism accounts for the majority of inter-individual variability in tacrolimus pharmacokinetics. We found that the basal expression of CYP3A5 in donor grafts also played a significant role in tacrolimus metabolism under the same genetic conditions after pediatric liver transplantation. Thus, we hypothesized that some potential epigenetic factors could affect CYP3A5 expression and contributed to the variability. We used a high-throughput functional screening for miRNAs to identify miRNAs that had the most abundant expression in normal human liver and could regulate tacrolimus metabolism in HepaRG cells and HepLPCs. Four of these miRNAs (miR-29a-3p, miR-99a-5p, miR-532-5p, and miR-26-5p) were selected for testing. We found that these miRNAs inhibited tacrolimus metabolism that was dependent on CYP3A5. Putative miRNAs targeting key drug-metabolizing enzymes and transporters (DMETs) were selected using an in silico prediction algorithm. Luciferase reporter assays and functional studies showed that miR-26b-5p inhibited tacrolimus metabolism by directly regulating CYP3A5, while miR-29a-5p, miR-99a-5p, and miR-532-5p targeted HNF4α, NR1I3, and NR1I2, respectively, in turn regulating the downstream expression of CYP3A5; the corresponding target gene siRNAs markedly abolished the effects caused by miRNA inhibitors. Also, the expression of miR-29a-3p, miR-99a-5p, miR-532-5p, and miR-26b-5p in donor grafts were negatively correlated with tacrolimus C/D following pediatric liver transplantation. Taken together, our findings identify these miRNAs as novel regulators of tacrolimus metabolism.

Deceleration of Liver Regeneration by Knockdown of Heme Oxygenase-1 is Associated With Impairment of Liver Injury Recovery After Reduced-Size Liver Transplantation in Rats.

AIM: It has been reported that heme oxygenase-1 (HO-1) is upregulated during hepatocyte proliferation. Herein, we used a half-size liver transplantation (HSLT) model to study the impact of HO-1 on liver grafts proliferation. To the best of our knowledge, this is the first time that HO-1 has been characterized as a regulator of liver graft regeneration. MATERIALS AND METHODS: Saline and tin protoporphyrin (SnPP, a HO-1 competitive inhibitor) were separately administered in vehicle and SnPP group before rats HSLT. Plasma samples were collected at 0, 1, 3, and 5 days after HSLT for liver function analysis. Liver tissues were obtained at 0, 1, 3, and 5 days after HSLT for analyses of histologic, apoptosis, and proliferation index by immunohistochemical, enzyme-linked immunosorbent assay, quantitative real-time polymerase chain reaction, and Western blotting. RESULTS: HO-1 level was upregulated by the treatment of HSLT along with accelerated liver proliferation, which was reversed by SnPP. The reduced regeneration by SnPP lead to higher Suzuki's scores, alanine aminotransferase, and aspartate aminotransferase levels. The interleukin-6 levels, p-Stat3/t-Stat3, c-myc, and c-jun were decreased in the SnPP group than the vehicle group. CONCLUSIONS: Our findings suggest that inhibition of HO-1 mitigates liver regeneration in part by downregulation of an interleukin-6/Stat3 axis. Targeted specific pharmacologic induction of HO-1 may be applicable in clinical practice.

Inducible nitric oxide synthase and vein graft performance in patients undergoing coronary artery bypass surgery: physiological or pathophysiological role?

Coronary artery disease is the major cause of mortalilty in the West with coronary artery bypass surgery (CABG) being a means of restoring blood supply to ischaemic myocardium. The long saphenous vein is the most commonly used bypass conduit but its patency is inferior to the internal thoracic artery, the 'gold standard' graft. In conventional procedures the saphenous vein is harvested in such a manner that considerable vascular damage is inflicted. The structures mainly affected by this vascular trauma are the endothelium, autonomic nerves and vascular smooth muscle all containing cells with the potential to release nitric oxide (NO). While the majority of studies into the potential role of NO in vein graft performance have focussed on the involvement of endothelial nitric oxide synthase (eNOS) less information is available regarding the role of the inducible isoform of nitric oxide synthase (iNOS). While the effects of eNOS-derived NO are principally beneficial, iNOS is generally associated with pathological conditions. While potential pathophysiological roles of iNOS are discussed in this review we also outline many studies suggesting that this isoenzyme plays an important role in maintaing vein graft patency in patients undergoing CABG, particularly when the saphenous vein is harvested with minimal surgical trauma.