Accurate Assessment and Tracking the Process of Liver-Specific Injury by the Residual Tissue Activity of Carboxylesterase 1 and Dipeptidyl Peptidase 4

Accurately assessing and tracking the progression of liver-specific injury remains a major challenge in the field of biomarker research.Here,we took a retrospective validation approach built on the mutuality between serum and tissue biomarkers to characterize the liver-specific damage of bile duct cells caused by a-naphthyl isothiocyanate(ANIT).We found that carboxylesterase 1(CES1),as an intrahepatic marker,and dipeptidyl peptidase 4(DPP-IV),as an extrahepatic marker,can reflect the different pathophysiologies of liver injury.Levels of CES1 and DPP-IV can be used to identify liver damage itself and the inflammatory state,respectively.While the levels of the conventional serological biomarkers alkaline phosphatase(ALP),alanine aminotransferase(ALT),and aspartate aminotransferase(AST)were all concomitantly elevated in serum and tissues after ANIT-induced injury,the levels of bile acids decreased in bile,increased in serum,and ascended in intrahepatic tissue.Although the level of γ-glutamyl transpeptidase(γ-GT)changed in an opposite direction,the duration was much shorter than that of CES1 and was quickly restored to normal levels.Therefore,among the abovementioned biomarkers,only CES1 made it possible to specifically determine whether the liver cells were destroyed or damaged without interference from inflammation.CES1 also enabled accurate assessment of the anti-cholestasis effects of ursodeoxycholic acid(UDCA;single component)and Qing Fei Pai Du Decoction(QFPDD;multicomponent).We found that both QFPDD and UDCA attenuated ANIT-induced liver damage.UDCA was more potent in promoting bile excretion but showed relatively weaker anti-injury and antiinflammatory effects than QFPDD,whereas QFPDD was more effective in blocking liver inflammation and repairing liver damage.Our data highlights the potential of the combined use of CES1(as an intrahepatic marker of liver damage)and DPP-IV(as an extrahepatic marker of inflammation)for the accurate evaluation and tracking of liver-specific injury—an application that allows for the differentiation of liver damage and inflammatory liver injury.

Cisplatin increases carboxylesterases through increasing PXR mediated by the decrease of DEC1

cis-Diamminedichloroplatinum(CDDP)is widely used for the treatment of various solid cancers.Here we reported that CDDP increased the expression and enzymatic activities of carboxylesterase 1(CES1)and carboxylesterase 2(CES2),along with the upregulation of pregnane X receptor(PXR)and the downregulation of differentiated embryonic chondrocyte-expressed gene 1(DEC1)in human hepatoma cells,primary mouse hepatocytes,mouse liver and intestine.The overexpression or knockdown of PXR alone upregulated or downregulated the CES1 and CES2 expression,respectively.The increases in CES1 and CES2 expression levels induced by CDDP abolished or enhanced by PXR knockdown or overexpression,implying that CDDP induces carboxylesterases through the activation of PXR.Likewise,the overexpression or knockdown of DEC1 alone significantly decreased or increased PXR and its targets.Moreover,the increases of PXR and its targets induced by CDDP were abolished or alleviated by the overexpression or knockdown of DEC1.The overexpression or knockdown of DEC1 affected the response of PXR to CDDP,but not vice versa,suggesting that CDDP increases carboxylesterases by upregulating PXR mediated by the decrease of DEC1.In addition,CDDP did not increase DEC1 mRNA degradation but suppressed DEC1 promoter reporter activity,indicating that it suppresses DEC1 transcriptionally.The combined use of CDDP and irinotecan had a synergistic effect on two cell lines,especially when CDDP was used first.

IL-33 Downregulates Hepatic Carboxylesterase 1 in Acute Liver Injury via Macrophage-derived Exosomal miR-27b-3p

Background and Aims:We previously reported that carboxylesterase 1(CES1)expression was suppressed following liver injury.The study aimed to explore the role of interleukin(IL)-33 in liver injury and examine the mechanism by which IL-33 regulates CES1.Methods:IL-33 and CES1 levels were determined in the livers of patients and lipopolysaccharide(LPS)-,acetaminophen(APAP)-treated mice.We constructed IL-33 and ST2 knockout(KO)mice.ST2-enriched immune cells in livers were screened to identify the responsible cells.Macrophage-derived exosome(MDE)activity was tested by adding exosome inhibitors.Micro-RNAs(miRs)were extracted from control and IL-33-stimulated MDEs(IL-33-MDEs)and subjected miR sequencing(miR-Seq).Candidate miR was tested in vitro and in vivo and its binding of a target gene was assessed by luciferase reporter assays.Lentivirus-vector cellular transfection and transcript silencing were used to examine pathways mediating IL-33 suppression of miR-27b-3p.Results:Patient liver IL-33 and CES1 expression levels were inversely correlated.CES1 downregulation in liver injury was rescued in both IL-33–deficient and ST2 KO mice.Macrophages were shown to be responsible for IL-33 effects.IL-33-MDEs reduced CES1 levels in hepatocytes.Exosomal miR-Seq and qRT-PCR demonstrated increased miR-27b-3p levels in IL-33-MDEs;miR-27b-3p was implicated in Nrf2 targeting.IL-33 inhibition of miR-27b-3p was found to be GATA3-dependent.Conclusions:IL-33–ST2–GATA3 pathway signaling increases miR-27b-3p content in MDEs,which upon being internalized by hepatocytes reduce CES1 expression by inhibiting Nrf2.The elucidation of this mechanism in this study contributes to a better understanding of CES1 dysregulation in liver injury.

Rapid bioluminescence assay for monitoring rat CES1 activity and its alteration by traditional Chinese medicines

In traditional Chinese medicine herbs(TCM),including Radix Salviae Miltiorrhizae(Danshen),Radix Puerariae Lobatae(Gegen),Radix Angelicae Sinensis(Danggui),and Rhizoma Chuanxiong(Chuanxiong)are widely used for the prevention and treatment of cardiovascular diseases and also often co-administered with Western drugs as a part of integrative medicine practice.Carboxylesterase 1(CES1)plays a pivotal role in the metabolisms of pro-drugs,Since(S)-2-(2-(6-dimethylamino)-benzothiazole)-4,5-dihydrothiazole-4-carboxylate(NLMe)has recently been identified by us as a selective CES1 bioluminescent sensor,we developed a rapid method using this substrate for the direct measurement of CES1 activity in rats.This bioluminescence assay was applied to determine CES1 activity in rat tissues after a two-week oral administration of each of the four herbs noted above.The results demonstrated the presence of CES1 enzyme in rat blood and all tested tissues with much higher enzyme activity in the blood,liver,kidney and heart than that in the small intestine,spleen,lung,pancreas,brain and stomach.In addition,the four herbs showed tissue-specific effects on rat CES1 expression.Based on the CES1 biodistribution and its changes after treatment in rats,the possibility that Danshen,Gegen and Danggui might alter CES1 activities in human blood and kidney should be considered.In summary,a selective and sensitive bioluminescence assay was developed to rapidly evaluate CES1 activity and the effects of orally administered TCMs in rats.

Bioactive neolignans and lignans from the roots of Paeonia lactiflora

Two new neolignans and one new lignan(1-3)were obtained from the roots of Paeonia lactiflora.Their structures were unambiguously elucidated based on extensive spectroscopic analysis,single-crystal X-ray crystallography,and the calculated and experimental electronic circular dichroism(ECD)spectra.Compound 1 was a racemic mixture and successfully resolved into the anticipated enantiomers via chiral-phase HPLC.Compound 3 demonstrated moderate inhibitory activity against human carboxylesterase 2A1(hCES2Al)with an IC_(50) value of 7.28±0.94 μmol·L^(-1).