RESUMO
Pegylated liposomal doxorubicin (PLD) has excellent therapeutic efficacy in the treatment of cancers, but can cause serious adverse reactions such as hand-foot syndrome (HFS). Our previous research suggests that both PLD-induced HFS may be associated with injury to tight junctions (TJs) in the skin and that calcium dobesilate (CaD) can alleviate HFS. However, the underlying molecular mechanism is not well understood. Here, we created an in vitro PLD-treated model using Human Microvascular Endothelial Cell line-1 (HMEC-1) and an in vivo HFS rat model to investigate the underlying pathways. Treatment with PLD increased the expression of HYAL-1, CD44, and hyaluronic acid (HA) concentration, while reducing ZO-1 and Claudin-5 expression. Moreover, PLD treatment induced the degradation of higher molecular weight HA to its lower molecular weight counterpart, elevating the permeability of both HEMC-1 cell membranes and rat paw skin capillaries. AD-01 (CD44 inhibitor) inhibited the effect of PLD on the expression of ZO-1 and Claudin-5. Furthermore, CaD treatment suppressed the expression of HYAL-1 and CD44, mitigated HA degradation, and enhanced the expression of ZO-1 and Claudin-5. This resulted in decreased permeability in HEMC-1 cells and rat skin capillaries. In summary, our data suggest that PLD may promote the destruction of TJs via the HA/CD44 pathway, thereby leading to HFS through increased skin permeability and exacerbated doxorubicin extravasation. Moreover, CaD can inhibit this pathway, offering a potential therapeutic avenue to alleviate HFS.
RESUMO
OBJECTIVE: To develop and validate a normal phase HPLC-MS/MS method for the determination of donepezil enantiomer in human plasma. METHODS: Donepezil was extracted from plasma by n-hexane:isopropanol (98:2, V/V) with lidocaine serving as an internal standard. The analytes went through the column of CHIRALCEL OJ-H (250 mm x 4.6 mm, 5 microm) with mobile phase n-hexane:n-propanol:diethylamine (60:40: 0.1, V/V/V). Donepezil enantiomer was determined by API 3000 in MRM mode. RESULTS: The retention time of S-DN and R-DN were 15.56 min and 18.41 min, respectively. The calibration curves were linear in a range from 0.051 to 7.596 ng/mL for S-DN, and from 0.049 to 7.404 ng/mL for R-DN, respectively, both with more than 0.99 correlation coefficients. The relative recovery were 95.10%-103.70% for S-DN and 93.58%-98.00% for R-DN, respectively; the pretreatment recovery were 58.42%-61.08% for S-DN and 53.24%-61.87% for R-DN, respectively; the within-day RSD ranged from 8.35% to 11.28% for S-DN and from 6.78% to 11.58% for R-DN, respectively; the between-day RSD ranged from 5.82% to 9.02% for S-DN and from 6.87% to 9.19% for R-DN, respectively. CONCLUSION: This normal phase HPLC-MS/MS method is simple, rapid, sensitive and accurate for the determination of donepezil enantiomer in human plasma and is suitable for pharmacokinetic studies of donepezil enantiomers.