khellactone and Disease-Models--Animal

Psoriasis is a chronic inflammatory skin disease with unresolved pathogenesis. Studies on the pathogenesis of psoriasis have been extensively carried out, but treatments are still not satisfactory. In this study, we found improvement after treatment with cis-khellactone, a small molecular natural product, in imiquimod-challenged C57BL/6 mice. cis-Khellactone clearly reduced the level of cytokines in psoriatic skin, including IL-23, TNF-α, IL-1β, and IL-6, while limiting the inhibition of IL-17A, which is produced by T helper type 17 cells. cis-Khellactone treatment specifically decreased dermal macrophage infiltration in psoriatic skin but not in neutrophils or T cells. Additionally, compared with the control group, cis-khellactone significantly decreased the activation of NF-κB p65 in these infiltrated macrophages. Further study showed that cis-khellactone suppressed proinflammatory phenotypic macrophages by promoting autophagy. Blocking autophagy by silencing Beclin1 or Atg7 abrogated the effect of cis-khellactone on macrophages. The autophagy-dependent improvement in psoriasis from cis-khellactone treatment was further manifested by its limited effects on skin lesions in chloroquine-treated mice. Moreover, cis-khellactone showed lower toxicity levels than methotrexate in macrophages and primary hepatocytes. Taken together, cis-khellactone selectively modulated macrophage function and phenotype by inducing autophagy to ameliorate imiquimod-induced psoriasis in mice. Our research provides an effective strategy for the treatment of psoriasis.

Topics: Adjuvants, Immunologic; Animals; Autophagy; Autophagy-Related Protein 7; Beclin-1; Cells, Cultured; Coumarins; Disease Models, Animal; Female; Gene Knockdown Techniques; Hepatocytes; Humans; Imiquimod; Macrophages; Male; Methotrexate; Mice; Mice, Inbred C57BL; Primary Cell Culture; Psoriasis; Toxicity Tests

Many chiral drugs are used as the racemic mixtures in clinical practice. The occurrence of enantioselectively pharmacological activities calls for the development of enantiospecific analytical approaches during pharmacokinetic studies of enantiomers. Sample preparation plays a key role during quantitative analysis of biological samples. In current study, a rapid and reliable online solid phase extraction-chiral high performance liquid chromatography-tandem mass spectrometry (online SPE-chiral LC-MS/MS) method was developed for the simultaneously enantiospecific quantitation of (+)-trans-khellactone (dTK), (+/-)-cis-khellactone (d/lCK), (+/-)-praeruptorin A (d/lPA), (+/-)-praeruptorin B (d/lPB) and (+)-praeruptorin E (dPE), the main active angular-type pyranocoumarins (APs) in Peucedani Radix (Chinese name: Qian-hu) or the major metabolites of those APs, in rat plasma. The validation assay results described here show good selectivity and enantiospecificity, extraction efficiency, accuracy and precision with quantification limits (LOQs) of 2.57, 1.28, 1.28, 1.88, 4.16, 4.16 and 4.18ngmL(-1) for dTK, lCK, dCK, dPA, dPB, lPB and dPE, respectively, while lPA was not detected in rat plasma due to the carboxylesterase(s)-mediated hydrolysis. In addition, the validated system was satisfactorily applied to characterize the pharmacokinetic properties of those components in normal and chronic obstructive pulmonary disease (COPD) rats following oral administration of Qian-hu extract. dCK and lCK were observed as the main herb-related compounds in plasma. Enantioselectively pharmacokinetic profiles occurred for dCK vs lCK, dPA vs lPA, and dPB vs lPB in either normal or COPD rats. The proposed whole system is expected to be a preferable analytical tool for in vivo study of chiral drugs, in particular for the characterization of enantioselectively pharmacokinetic profiles.

Topics: Administration, Oral; Animals; Calibration; Chromatography, High Pressure Liquid; Coumarins; Disease Models, Animal; Hydrolysis; Linear Models; Male; Pulmonary Disease, Chronic Obstructive; Rats; Rats, Wistar; Reproducibility of Results; Stereoisomerism; Tandem Mass Spectrometry