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1.
Article in Chinese | WPRIM | ID: wpr-921673

ABSTRACT

Triptolide(TP), the main active and toxic component of Tripterygium wilfordii, has the limitations of low bioavailability, poor absorption, low concentration in plasma, and small lethal dose. Microneedle(MN), the hybrid of hypodermic needle and transdermal patch, is a physical penetration-enhancing system. Dissolving microneedles(DMNs) can be tailored to specific needs of degradation rate. In this study, the TP-loaded DMNs(DMNs-TP) were prepared with the two-step centrifugation method. The optimal ratio of PVA to PVP K30, water content in matrix solution, demoulding method, and plasticizer for preparing DMNs were investigated with the indexes of formability and mechanical strength. The drug loading capacity was determined by HPLC and morphological characteristics were observed under an optical microscope. The mechanical properties were investigated by H&E staining and Franz diffusion cell was used to detect the in vitro skin permeation characteristics. Through the experiment, we confirmed that the optimal backing material should be PVA and PVP K30(3∶1) and the optimal ratio of matrix material to water should be 3∶4. The prepared DMNs-TP were pyramidal with smooth surface and length of approximately 550 μm. Each patch(2.75 cm~2) had the drug loading capacity of(153.41±2.29) μg, and TP was located in the upper part of the needle. The results of in vitro skin permeation assay demonstrated that the cumulative penetration of TP in DMNs-TP reached 80% in 24 h, while little TP solution penetrated the skin, which proved that DMNs promoted the transdermal delivery of TP.


Subject(s)
Administration, Cutaneous , Diterpenes , Drug Delivery Systems , Epoxy Compounds , Needles , Phenanthrenes , Skin
2.
Article in Chinese | WPRIM | ID: wpr-887974

ABSTRACT

Overtaking lung cancer,breast cancer is now the most commonly diagnosed cancer seriously threatening people's health and life. As the main effective component of Tripterygium wilfordii,triptolide( TP) has attracted increasing attention due to its multitarget and multi-pathway anti-tumor activity. Recent studies have revealed that breast cancer-sensitive TP enables the inactivation of breast cancer cells by inducing tumor cell apoptosis and autophagy,interfering in tumor cell metastasis,resisting drug resistance,arresting tumor cell cycle,and influencing tumor microenvironment. It has been recognized as a promising clinical antitumor agent by virtue of its widely accepted therapeutic efficacy. This paper reviewed the anti-breast cancer action and its molecular mechanisms of TP on the basis of the relevant literature in the past ten years,and proposed application strategies in view of the inadequacy of TP to provide a reference for further research on the application of TP in the treatment of breast cancer.


Subject(s)
Breast Neoplasms/genetics , Diterpenes/pharmacology , Epoxy Compounds , Female , Humans , Phenanthrenes , Tumor Microenvironment
3.
Chinese Journal of Biotechnology ; (12): 2039-2049, 2021.
Article in Chinese | WPRIM | ID: wpr-887780

ABSTRACT

Triptolide has wide clinical applications due to its anti-inflammatory, anti-tumor and immunosuppressive activities. In this study, we investigated the effect of blocking isopentenyl pyrophosphate (IPP) translocation on the biosynthesis of triptolide by exogenously adding D,L-glyceraldehyde (DLG) to the suspension cells of Ttripterygium wilfordii at different stages (7 d, 14 d). Subsequently, the cell viability, biomass accumulation, triptolide contents, as well as the profiles of the key enzyme genes involved in the upstream pathway of triptolide biosynthesis, were analyzed. The results showed that IPP translocation is involved in the biosynthesis of triptolide. IPP is mainly translocated from the plastid (containing the MEP pathway) to the cytoplasm (containing the MVA pathway) in the early stage of the culture, but reversed in the late stage. Blocking the translocation of IPP affected the expression of key enzyme genes involved in the upstream pathway of triptolide, which in turn affected the accumulation of triptolide. Understanding the characteristics and mechanism of IPP translocation provides a theoretical basis for further promoting triptolide biosynthesis through synthetic biology.


Subject(s)
Diterpenes , Epoxy Compounds , Hemiterpenes , Organophosphorus Compounds , Phenanthrenes
4.
Article in Chinese | WPRIM | ID: wpr-880776

ABSTRACT

OBJECTIVE@#To investigate the protective effect of arctiin with anti-inflammatory bioactivity against triptolide-induced nephrotoxicity in rats and explore the underlying mechanism.@*METHODS@#Forty SD rats were divided into 4 groups for gastric lavage of normal saline, arctiin (500 mg/kg), triptolide (500 μg/kg), or both arctiin (500 mg/kg) and triptolide (500 μg/kg). Blood samples were collected for analysis of biochemical renal parameters, and the renal tissues were harvested for determining the kidney index and for pathological evaluation with HE staining. In the @*RESULTS@#In SD rats, arctiin significantly antagonized triptolide-induced elevation of BUN, Scr and kidney index (@*CONCLUSIONS@#Arctiin can protect the kidney from triptolide-induced damages in rats possibly through the anti-inflammatory pathway.


Subject(s)
Animals , Anti-Inflammatory Agents , Diterpenes/toxicity , Epoxy Compounds/toxicity , Furans , Glucosides , Kidney/drug effects , Phenanthrenes/toxicity , Rats , Rats, Sprague-Dawley
5.
Article in English | WPRIM | ID: wpr-776622

ABSTRACT

As a major active component extracted from traditional Chinese herb Tripterygium wilfordii Hook F, triptolide exhibits multiple pharmacological effects. Autophagy is an evolutionary conserved intracellular catabolic process involved in cytoplasmic materials degradation. Autophagic dysfunction contributes to the pathologies of many human diseases, which makes it a promising therapeutic target. Recent studies have shown that triptolide exerts neuroprotection, anti-tumor activities, organ toxicity, and podocyte protection by modulating autophagy. This article highlights the current information on triptolide-modulated autophagy, analyzes the possible pathways involved, and describes the crosstalk between autophagy and apoptosis modulated by triptolide, in hope of providing implications for the roles of autophagy in pharmacological effects of triptolide and expanding its novel usage as an autophagy modulator.


Subject(s)
Animals , Apoptosis , Autophagy , Diterpenes , Pharmacology , Epoxy Compounds , Pharmacology , Humans , Neoplasms , Drug Therapy , Pathology , Neuroprotective Agents , Pharmacology , Phenanthrenes , Pharmacology , Podocytes
6.
Journal of Experimental Hematology ; (6): 1088-1093, 2019.
Article in Chinese | WPRIM | ID: wpr-775759

ABSTRACT

OBJECTIVE@#To investigate the effects of inhibiting proliferation and inducing apoptosis of low-dose triptolide and sorafenib alone or in combination on FLT3-ITD acute myeloid leukemia cell line MV4-11 and STAT5 pathway.@*METHODS@#The MV4-11 cells were treated with low dose triptolide(IC) and sorafenib(IC) alone or in combination for 48 hours. The cell proliferation and inhibition were detected by using CCK-8 kit, the cell apoptosis was detected by flow cytometry, the expression of FLT3,STAT5 in mRNA and protein levels was detected by RT-PCR and Western blot respectively.@*RESULTS@#The treatment of MV4-11 cells with low dose triptolide and sorafenib alone and in combination for 48 hours could inhibit cell proliferation and induce cell apoptosis, moreover the inhibitory rate and apoptotic rate of MV4-11 cells in drug-combination group both were higher than those in single drug group. The mRNA expression and protein expression of FLT3,STAT5 signaling pathway in drug combination group were significantly lower than those in single drug group.@*CONCLUSION@#Low-dose triptolide combined with sorafenib can synergistically inhibit the proliferation and induce the apoptosis of MV4-11 cells, which may be related with the inhibition of FLT3 and STAT5 pathway.


Subject(s)
Apoptosis , Cell Line, Tumor , Diterpenes , Epoxy Compounds , Humans , Leukemia, Myeloid, Acute , Phenanthrenes , STAT5 Transcription Factor , Sorafenib , fms-Like Tyrosine Kinase 3
7.
Journal of Experimental Hematology ; (6): 1691-1695, 2019.
Article in Chinese | WPRIM | ID: wpr-775664

ABSTRACT

OBJECTIVE@#To investigate the effect of triptolide on the excursion of Tc and Th cells in peripheral blood of systemic lupus erythematosus (SLE) BALB/c-un nude mice induced by pristane.@*METHODS@#Eighteen female BALB/c-un nude mice were randomly divided into blank, SLE and triptolide group, each with 6 mice by random table method. Group SLE and group triptolide were established by single intraperitoneal injection of pristane, and blank group was used as blank control group. SLE model was established by single intraperitoneal injection. Triptolide group was fed with triptolide at the dose of 5 mg/(kg·d), and the blank group and SLE group were fed normally. Blood samples were collected from the caudal vein before treatment and 1, 3 and 6 months after treatment respectively. Fluorescence labeled flow cytometry was used to delect Tc and Th lymphocyte subsets at different stages of treatment.@*RESULTS@#After treatment for 3 and 6 moths, the percentages of Tcl, Thl cells and CD8, Tcl/Tc2, Thl/Th2 and CD4/CD8 all decreased in the group of triptolide, and the percentage of CD4, Tc2 and Th2 cells increased (P<0.05).@*CONCLUSION@#The mechanism of triptolide in the treatment of SLE may be related with the excursion of Tc and Th cells to Tcl and Tc2 to maintain the relative homeostasis of Tc and Th cells at different stage, thus affecting the immune response and the inflammatory reaction.


Subject(s)
Animals , Diterpenes , Epoxy Compounds , Female , Lupus Erythematosus, Systemic , Mice , Mice, Inbred BALB C , Mice, Nude , Phenanthrenes , T-Lymphocytes, Helper-Inducer
8.
Article in Chinese | WPRIM | ID: wpr-773714

ABSTRACT

Triptolide( TP) is isolated from the traditional Chinese medicine Tripterygium wilfordii,which exhibits notable immuneregulative effect. Th17 cells involve in inflammatory response and Treg cells contribute to immune tolerance. They both play an important role in immune response. Previous studies have investigated that TP induced hepatic Th17/Treg imbalance. However,the effect of TP on spleen Th17/Treg cells remains unclear. Therefore,the aim of present study was to investigate the effect of TP on Th17/Treg cells in spleen. In this study,the effect of TP on the proliferation of splenic lymphocyte was detected by cytotoxicity test in vitro. After different concentrations of TP( 2. 5,5,20,40 nmol·L~(-1)) were given to splenic lymphocyte,cytokines secreted from the supernatant of splenic lymphocyte were detected by cytometric bead array,and the expression of suppressor of cytokine signaling( SOCS) mRNA was detected by qRT-PCR. Female C57 BL/6 mice were continuously observed for 24 h after treatment of 500 μg·kg-1 TP. The effects of TP on the splenic tissue structure and the percentage of Th17/Treg cells were examined. The results showed that the IC50 of TP was19. 6 nmol·L~(-1) in spleen lymphocytes. TP inhibited the secretion of IL-2 and IL-10 and induced the expression of SOCS-1/3 mRNA in spleen lymphocytes at the dosage of 2. 5 and 5 nmol·L~(-1) after 24 h in vitro. Administration of TP at dosage of 500 μg·kg-1 had no significant spleen toxicity in vivo. TP treatment increased the percentage of Th17 cells after 12 h and inhibited the proportion of Treg cells after 12 and 24 h. In conclusion,TP reduced the secretion of IL-2 and IL-10 through SOCS-1/3 signaling pathway,thereby induced the percentage of Th17 cells and inhibited the percentage of Treg cells.


Subject(s)
Animals , Cytokines , Metabolism , Diterpenes , Pharmacology , Epoxy Compounds , Pharmacology , Female , Mice , Mice, Inbred C57BL , Phenanthrenes , Pharmacology , Signal Transduction , Spleen , Cell Biology , Suppressor of Cytokine Signaling 1 Protein , Metabolism , Suppressor of Cytokine Signaling 3 Protein , Metabolism , T-Lymphocytes, Regulatory , Cell Biology , Th17 Cells , Cell Biology
9.
Article in Chinese | WPRIM | ID: wpr-773705

ABSTRACT

Tumors are major chronic diseases and seriously threaten human health all over the world. How to effectively control and cure tumors is one of the most pivotal problems in the medical field. At present,surgery,radiotherapy and chemotherapy are still the main treatment methods. However,the side effects of radiotherapy and chemotherapy cannot be underestimated. Therefore,it is of great practical significance to find new anti-cancer drugs with low toxicity,high efficiency and targeting to cancer cells. With the increasing incidence of tumor,the anti-tumor effect of traditional Chinese medicine has increasingly become a research hotspot. Triptolide,which is a natural diterpenoid active ingredient derived from of Tripterygium wilfordii,as one of the highly active components,has anti-inflammatory,immunosuppressive,anti-tumor and other multiple effects. A large number of studies have confirmed that it has good anti-tumor activity against various tumors in vivo and in vitro. It can play an anti-tumor role by inhibiting the proliferation of cancer cells,inducing apoptosis of cancer cells,inducing autophagy of cancer cells,blocking the cell cycle,inhibiting the migration,invasion and metastasis of cancer cells,reversing multidrug resistance,mediating tumor immunity and inhibiting angiogenesis. On the basis of literatures,this paper reviews the anti-tumor effect and mechanism of triptolide,and analyzes the current situation of triptolide combined with other chemotherapy drugs,in order to promote deep research and better clinical application about triptolide.


Subject(s)
Antineoplastic Agents, Phytogenic , Pharmacology , Apoptosis , Autophagy , Cell Cycle Checkpoints , Diterpenes , Pharmacology , Epoxy Compounds , Pharmacology , Humans , Neoplasms , Drug Therapy , Phenanthrenes , Pharmacology , Tripterygium , Chemistry
10.
Article in Chinese | WPRIM | ID: wpr-773701

ABSTRACT

To investigate the effect of triptolide on cognitive dysfunction in vascular dementia rats and its effect on SIRT1/NF-κB pathway,fifty healthy male Sprague-Dawley rats were randomly divided into 5 groups: Sham operation group( Sham group),vascular dementia model group( 2 VO group),triptolide intraperitoneal injection group( TR group),triptolide intraperitoneal injection + EX527 intracerebroventricular administration group( T+E group),EX527 intracerebroventricular administration group( EX527 group). After 4 weeks of modeling,Morris water maze test and object recognition test were used to evaluate the learning and memory ability of rats. The morphological changes of hippocampus in each group were observed in brain tissue. The chemical colorimetry was used to detect the activities of SOD and MDA in hippocampus. IL-6 and TNF-α levels were detected by ELISA. Western blot was used to detect the expression of SIRT1,NF-κB,IκBα and caspase 3 in hippocampus. The results showed that compared with the Sham group,the learning and memory ability of the vascular dementia model rats was reduced,the SOD activity in the hippocampus was decreased,the MDA activity and IL-6 level were increased,the neuronal degeneration changed significantly,the expression of SIRT1 and IκBα was decreased and the expression of caspase 3 and NF-κB was significantly increased. After intervention by triptolide,the level of oxidative stress and the degenerative changes in hippocampus were significantly slowed down. The expression of SIRT1 and IκBα protein was increased and the expression of caspase 3 and NF-κB was significantly decreased. While,after intervention by triptolide and EX527,the expression of SIRT1 was decreased,the levels of oxidative stress and neuronal degeneration in the hippocampus were aggravated,and the learning and memory ability was reduced. The results showed that triptolide could improve cognitive impairment in vascular dementia rats and its mechanism may be related to SIRT1/NF-κB signaling pathway.


Subject(s)
Animals , Cognitive Dysfunction , Drug Therapy , Dementia, Vascular , Drug Therapy , Diterpenes , Pharmacology , Epoxy Compounds , Pharmacology , Hippocampus , Male , NF-kappa B , Metabolism , Oxidative Stress , Phenanthrenes , Pharmacology , Random Allocation , Rats , Rats, Sprague-Dawley , Signal Transduction , Sirtuin 1 , Metabolism
11.
Article in Chinese | WPRIM | ID: wpr-773700

ABSTRACT

The aim of this paper was to observe the concentration,time and mechanism of autophagy induced by triptolide( TP) in ovarian granulosa cells( OGCs). CCK-8 method was used to compare the inhibitory effects of TP at different concentrations on primary cultured rat OGCs and IC50 was calculated. The effects of TP at different concentrations and time points on the expression of OGCs autophagy factor protein and the cascade of PI3 K/AKT/m TOR pathway were detected by Western blot. The effects of TP,autophagy inducer( brefeldin A) and PI3 K/m TOR inhibitor( NVP-BEZ235) on the expression of PI3 K/AKT/m TOR cascade and autophagy related factor protein were detected by Western blot. The results show that the IC50 of different concentrations of TP on OGCs of rat ovary was14. 65 μmol·L-1,and the minimum inhibitory concentration of TP was 0. 1 μmol·L-1( 100 nmol·L-1). Compared with the control group,the expression levels of beclin1 and LC3Ⅱ in each group were significantly higher than those in the control group( P<0. 05 or P<0. 01). After 12 hours of treatment with TP,brefeldin A and NVP-BEZ235,respectively,compared with the control group,TP could significantly promote the expression level of downstream autophagy effect or molecule beclin1,LC3Ⅱ and inhibit the expression level of LC3Ⅰ,p62 protein( P<0. 05 or P< 0. 01). Moreover,the expression of beclin1 and LC3Ⅱ/LC3Ⅰ in TP group was higher than that in brefeldin A group( P<0. 05 or P<0. 01),and the expression of p62 in TP group was lower than that in brefeldin A group( P<0. 05 or P<0. 01). At the same time,TP could significantly inhibit the expression of p-PI3 K,p-AKT,p-mTOR protein,and the inhibitory effect of TP was better than that of NVP-BEZ235 group. This study suggests that 100 nmol·L-1 TP could induce OGCs autophagy successfully in cultured rat ovary for 12 h; TP may induce OGCs autophagy by inhibiting PI3 k/Akt/m TOR signaling pathway.


Subject(s)
Animals , Apoptosis , Autophagy , Cell Proliferation , Cells, Cultured , Diterpenes , Pharmacology , Epoxy Compounds , Pharmacology , Female , Granulosa Cells , Phenanthrenes , Pharmacology , Phosphatidylinositol 3-Kinases , Metabolism , Proto-Oncogene Proteins c-akt , Metabolism , Rats , Signal Transduction , TOR Serine-Threonine Kinases , Metabolism
12.
Article in Chinese | WPRIM | ID: wpr-773697

ABSTRACT

The aim of this paper was to study the influence of triptolide in the immune response pathways of acquired immune deficiency syndrome( AIDS). Target proteins of triptolide and related genes of AIDS were searched in PubChem and Gene databases on line. Molecular networks and canonical pathways comparison analyses were performed by bioinformatics software( IPA). There were 15 targets proteins of triptolide and 258 related genes of AIDS. Close biological relationships of molecules of triptolide and AIDS were established by networks analysis. There were 21 common immune response pathways of triptolide and AIDS,including neuroinflammation signaling pathway,Th1 and Th2 activation pathway and role of pattern recognition receptors in recognition of bacteria and viruses. Triptolide stimulated immune response pathways by the main molecules of IFNγ,JAK2,NOD1,PTGS2,RORC. IFNγ is the focus nodes of triptolide and AIDS,and regulates genes of AIDS directly or indirectly. Triptolide may against AIDS by regulating molecules IFNγ in immune response pathways.


Subject(s)
Acquired Immunodeficiency Syndrome , Drug Therapy , Allergy and Immunology , Computational Biology , Diterpenes , Pharmacology , Epoxy Compounds , Pharmacology , Gene Regulatory Networks , Humans , Interferon-gamma , Genetics , Phenanthrenes , Pharmacology , Receptors, Pattern Recognition , Allergy and Immunology , Signal Transduction , T-Lymphocytes , Allergy and Immunology
13.
Article in Chinese | WPRIM | ID: wpr-773694

ABSTRACT

Tripterygium wilfordii multiglycoside( GTW),an extract derived from T. wilfordii,has been used for rheumatoid arthritis and other immune diseases in China. However its potential hepatotoxicity has not been investigated completely. Firstly,the content of triptolid( TP) in GTW was 0. 008% confirmed by a LC method. Then after oral administration of GTW( 100,150 mg·kg-1) and TP( 12 μg·kg-1) in female Wistar rats for 24 h,it was found that 150 mg·kg-1 GTW showed more serious acute liver injury than 12 μg·kg-1 TP,with the significantly increased lever of serum ALT,AST,TBA,TBi L,TG and bile duct hyperplasia even hepatocyte apoptosis. The expression of mRNA and proteins of liver bile acid transporters such as BSEP,MRP2,NTCP and OATP were down-regulated significantly by GTW to inhibit bile acid excretion and absorption,resulting in cholestatic liver injury. Moreover,GTW was considered to be involved in hepatic oxidative stress injury,although it down-regulated SOD1 and GPX-1 mRNA expression without significant difference in MDA and GSH levels. In vitro,we found that TP was the main toxic component in GTW,which could inhibit cell viability up to 80% in Hep G2 and LO2 cells at the dose of 0. 1 μmol·L-1. Next a LC-MS/MS method was used to detect the concentration of triptolid in plasma from rats,interestingly,we found that the content of TP in GTW was always higher than in the same amount of TP,suggesting the other components in GTW may affect the TP metabolism. Finally,we screened the substrate of p-glycoprotein( p-gp) in Caco-2 cells treated with components except TP extrated from GTW,finding that wilforgine,wilforine and wilfordine was the substrate of p-gp. Thus,we speculated that wilforgine,wilforine and wilfordine may competitively inhibit the excretion of TP to bile through p-gp,leading to the enhanced hepatotoxity caused by GTW than the same amount of TP.


Subject(s)
Animals , Caco-2 Cells , Chemical and Drug Induced Liver Injury , Pathology , Chromatography, Liquid , Diterpenes , Toxicity , Drugs, Chinese Herbal , Toxicity , Epoxy Compounds , Toxicity , Female , Glycosides , Toxicity , Humans , Liver , Phenanthrenes , Toxicity , Plant Extracts , Toxicity , Rats , Rats, Wistar , Tandem Mass Spectrometry , Tripterygium , Toxicity
14.
Article in Chinese | WPRIM | ID: wpr-773688

ABSTRACT

The effect of triptolide( TP) on VEGFA,SDF-1,CXCR4 pathway were investigated in vitro to explore the mechanism in improving platelet activation in patients with ankylosing spondylitis( AS). Peripheral blood mononuclear cells( PBMC) were used for the experiment and divided into 4 groups: normal group( NC),model group( MC),triptolide group( TP),and AMD3100 group. The optimal concentration of TP was measured by the MTT method. The expressions of TNF-α,IL-1β,IL-4,IL-10,VEGFA and VEGFR were detected by ELISA. The expressions of SDF-1,CXCR4 and VEGFA were detected by real-time quantitative PCR( RT-qPCR).The expressions of SDF-1,CXCR4,VEGFA and VEGFR were detected by Western blot. The expression levels of CD62 p,CD40 L and PDGFA were detected by immunofluorescence. MTT results showed that medium-dose TP had the strongest inhibitory effect on cells at24 h. The results of ELISA and PCR showed that TP inhibited mRNA expressions of IL-1β,TNF-α,VEGFA,VEGFR and SDF-1,CXCR4 and VEGFA. The results of Western blot indicated that TP inhibited SDF-1,CXCR4 and VEGFA,VEGFR protein expressions; immunofluorescence results indicate that TP can inhibit the expressions of CD62 p,CD40 L,PDGFA. TP may regulate platelet activation by down-regulating SDF-1,CXCR4,VEGFA and VEGFR mRNA expressions,thereby down-regulating IL-1β and TNF-αexpressions,and up-regulating the expressions of IL-4 and IL-10 cytokines.


Subject(s)
Cells, Cultured , Chemokine CXCL12 , Metabolism , Cytokines , Metabolism , Diterpenes , Pharmacology , Epoxy Compounds , Pharmacology , Heterocyclic Compounds , Pharmacology , Humans , Leukocytes, Mononuclear , Phenanthrenes , Pharmacology , Platelet Activation , Receptors, CXCR4 , Metabolism , Spondylitis, Ankylosing , Vascular Endothelial Growth Factor A , Metabolism
15.
Article in Chinese | WPRIM | ID: wpr-773680

ABSTRACT

To detect the concentration of triptolide in skin and joint after percutaneous administration,an HPLC-MS/MS method and skin and joint micro-dialysis( MD) method of triptolide were established in this study. The separation was achieved on triple quadrupole( AB QTRAP4500) and phenomenex-C18( 4. 6 mm×150 mm,5 μm,luna) column with acetonitrile-water with 0. 1% formic acid( 65 ∶35) as the mobile phase at a flow rate of 0. 7 m L·min-1. An electrospray ionization( ESI) source was applied and operated in the positive multiple reaction monitoring( MRM) mode. The fragment ion for triptolide was m/z 361. 1→145. 0. The effects of different perfusion [Ringer's,PBS( p H 7. 4),30% ethanol saline]drug concentrations and flow rates on the recovery rate,as well as the relationship between the recovery rate and the loss rate were determined by incremental( dialysis) and reduction( retrodialysis) methods.The reduction method was applied in the in vivo study to investigate and determine the stability of the probe recovery rate in 10 h. The results of HPLC-MS/MS detection method conformed to the requirements of biological samples. The perfusion fluid was 30% ethanol saline. The recovery rate of skin and joint probes in vitro of triptolide increased within the flow rate of 0. 5-2. 5 μL·min-1. In order to increase the timeliness of data and the accuracy,the flow rate was determined to be 1 μL·min-1,and the sample interval was determined to be 0. 5 h. The recovery rate of triptolide in skin and joint probes in vitro and the loss rate were stable and equal despite of change of triptolide concentration within 10-200 μg·L-1. This indicated that the effect of drug concentration on the MD probe recovery rate was small,and the recovery rate could be replaced by the loss rate. The loss rate in vivo using MD method was measured at 10 h,indicating that the transfer rate of triptolide was stable within 10 h. The established method of triptolide in MD and HPLC-MS/MS can be applied to investigate the kinetic in skin and joint after percutaneous administration of triptolide.


Subject(s)
Chromatography, High Pressure Liquid , Diterpenes , Pharmacokinetics , Epoxy Compounds , Pharmacokinetics , Humans , Joints , Metabolism , Phenanthrenes , Pharmacokinetics , Reproducibility of Results , Skin , Metabolism , Tandem Mass Spectrometry
16.
Rev. biol. trop ; 66(2): 826-835, abr.-jun. 2018. tab, graf
Article in Spanish | LILACS, SaludCR | ID: biblio-977348

ABSTRACT

Resumen La leishmaniasis es una enfermedad con una alta incidencia en el ser humano, que puede ser controlada, pero como los tratamientos tienen efectos secundarios importantes se han realizado estudios de diversas plantas con el fin de encontrar compuestos con actividad antileishmaniásica que presenten pocos efectos nocivos para el ser humano. El presente estudio consistió en realizar un tamizaje fitoquímico de la planta, para identificar la presencia de cumarinas, terpenos, triterpenos y azúcares reductores. El objetivo fue encontrar componentes químicos puros con actividad contra el parásito Leishmania sp. Por tal razón se purificaron los compuestos: trans-Z-alfabisaboleno y el Safrol, a los que se les realizaron pruebas del efecto anti parasitario que presentaron un CI50 de 50.0 µg/mL y 0.0 µg/mL, respectivamente. Además, se discute la importancia de estos nuevos hallazgos. El compuesto mayoritario presente en los aceites esenciales (Safrol) no es el componente que presentó la actividad. Es importante realizar estudios sobre su proyección en el tratamiento de la leishmaniasis.


Abstract The leishmaniosis disease incidence is high in tropical regions, and its current treatment has shown severe secondary effects. Considering this problem, many studies have focused on plants, looking for chemical components that have anti-leishmanial activity, and are free of adverse effects for human beings. The purpose of this work was to find a chemical component with this kind of activity in Piper auritum. In a phytochemical screening of this plant, we found some cumarins, terpens, triterpens and reducing sugars; and later, we identified the components trans-Z-α-bisabolene epoxide and Safrol. The first component presented a CI50 of 50.0 µg/mL of anti-Leishmania activity. The Safrol, which is the major component of the essential oils of this plant, did not show antiparasitic activity. These results are discussed considering treatment of leishmaniasis. Rev. Biol. Trop. 66(2): 826-835. Epub 2018 June 01.


Subject(s)
Safrole/therapeutic use , Leishmaniasis , Trypanosomatina , Piperaceae/parasitology , Epoxy Compounds , Phytochemicals/administration & dosage , Monocyclic Sesquiterpenes/therapeutic use , Leishmania
17.
Article in Chinese | WPRIM | ID: wpr-690985

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect and possible mechanism of low concentration of triptolide (TPL) combined with homoharringtonine (HHT) on the proliferation and apoptosis of KG-1α cells.</p><p><b>METHODS</b>CCK-8 method was used to detect the antiproliferating effects of different concentrations of TPL and HHT single-use and combined use on KG-1α cells, and the combined index (CI) was calculated. The colony formation ability was also determined by methylcellulose colony formation assay, cell surface molecules, apoptosis rate and cell cycle changes were detected by flow cytometry. Westerrn blot was used to detect the expression of Akt signaling pathway related proteins before and after low dose TPL combined with HHT using.</p><p><b>RESULTS</b>High expression of CD34 and CD123 were on KG-1a cells, which being lack expression of CD38. TPL and HHT dose-dependently inhibited the proliferation of KG-1α cells. Compared with low dosage TPL and HHT single-use groups, the cell proliferation and colony formation efficiency were lower, and the cell apoptosis rate was higher in the combined group. CI values also indicated that low concentration TPL combined with HHT possessed highly synergistic effect. After the combination of the 2 drugs, the expressions of P-Akt, P-Akt, BCL-2, PARP and survivin protein were down-regulated and the cleavage of PARP protein was increased.</p><p><b>CONCLUSION</b>Low concentration of TPL combined with HHT can synergistically inhibit KG-1α cell proliferation and induce its apoptosis through the PI3K/Akt signaling pathway and downstream protein.</p>


Subject(s)
Apoptosis , Cell Line, Tumor , Cell Proliferation , Diterpenes , Epoxy Compounds , Harringtonines , Homoharringtonine , Humans , Phenanthrenes , Phosphatidylinositol 3-Kinases
18.
Article in English | WPRIM | ID: wpr-812363

ABSTRACT

Triptolide (TP) induces severe liver injury, but its hepatotoxicity mechanisms are still unclear. Inflammatory responses may be involved in the pathophysiology. Neutrophils are the first-line immune effectors for sterile and non-sterile inflammatory responses. Thus, the aim of the present study was to investigate the neutrophilic inflammatory response in TP-induced liver injury in C57BL/6 mice. Our results showed that neutrophils were recruited and accumulated in the liver, which was parallel to or slightly after the development of liver injury. Neutrophils induced release of myeloperoxidase and up-regulation of CD11b, which caused cytotoxicity and hepatocyte death. Hepatic expressions of CXL1, TNF-α, IL-6, and MCP1 were increased significantly to regulate neutrophils recruitment and activation. Up-regulation of toll like receptors 4 and 9 also facilitated neutrophils infiltration. Moreover, neutrophils depletion using an anti-Gr1 antibody showed mild protection against TP overdose. These results indicated that neutrophils accumulation might be the secondary response, not the cause of TP-induced liver injury. In conclusion, the inflammatory response including neutrophil infiltration may play a role in TP-induced hepatotoxicity, but may not be severe enough to cause additional liver injury.


Subject(s)
Animals , Chemical and Drug Induced Liver Injury , Allergy and Immunology , Chemokine CCL2 , Genetics , Allergy and Immunology , Diterpenes , Drugs, Chinese Herbal , Epoxy Compounds , Female , Humans , Interleukin-6 , Genetics , Allergy and Immunology , Intracellular Signaling Peptides and Proteins , Genetics , Allergy and Immunology , Liver , Allergy and Immunology , Mice , Mice, Inbred C57BL , Neutrophil Infiltration , Neutrophils , Allergy and Immunology , Phenanthrenes , Tripterygium , Chemistry , Tumor Necrosis Factor-alpha , Genetics , Allergy and Immunology
19.
Article in Chinese | WPRIM | ID: wpr-776411

ABSTRACT

To explore the effects and molecular mechanisms of triptolide(TP)on improving podocyte epithelial-mesenchymal transition(EMT)induced by high dose of D-glucose(HG), the immortalized podocytes of mice were divided into the normal group(N), the high dose of D-glucose group(HG), the low dose of TP group(L-TP), the high dose of TP group(H-TP)and the mannitol group(MNT), and treated by the different measures respectively. More specifically, the podocytes in each group were separately treated by D-glucose(DG, 5 mmol·L⁻¹)or HG(25 mmol·L⁻¹)or HG(25 mmol·L⁻¹)+ TP(3 μg·L⁻¹)or HG(25 mmol·L⁻¹)+ TP(10 μg·L⁻¹)or DG(5 mmol·L⁻¹)+ MNT(24.5 mmol·L⁻¹). After the intervention for 24, 48 and 72 hours, firstly, the activation of podocyte proliferation was investigated. Secondly, the protein expression levels of the epithelial markers in podocytes such as nephrin and podocin, the mesenchymal markers such as desmin and collagen Ⅰ and the EMT-related mediators such as snail were detected respectively. Finally, the protein expression levels of Wnt3α and β-catenin as the key signaling molecules in Wnt3α/β-catenin pathway were examined severally. The results indicated that, HG could cause the low protein expression levels of nephrin and podocin and the high protein expression levels of desmin, collagen Ⅰ and snail in podocytes, and inducing podocyte EMT. On the other hand, HG could cause the high protein expression levels of Wnt3α and β-catenin in podocytes, and activating Wnt3α/β-catenin signaling pathway. In addition, L-TP had no effect on the activation of podocyte proliferation, the co-treatment of L-TP and HG could significantly recover the protein expression levels of nephrin and podocin, inhibit the protein expression levels of desmin, collagen I and snail in podocytes, thus, further improving podocyte EMT. And that, the co-treatment of L-TP and HG could obviously decrease the high protein expression levels of Wnt3α and β-catenin induced by HG in podocytes, and inhibit Wnt3α/β-catenin signaling pathway activation. On the whole, HG can induce podocyte EMT by activating Wnt3α/β-catenin signaling pathway; L-TP can ameliorate podocyte EMT through inhibiting Wnt3α/β-catenin signaling pathway activation, which may be one of the effects and molecular mechanisms .


Subject(s)
Animals , Cells, Cultured , Diterpenes , Pharmacology , Epithelial-Mesenchymal Transition , Epoxy Compounds , Pharmacology , Glucose , Mice , Phenanthrenes , Pharmacology , Podocytes , Wnt Signaling Pathway , Wnt3A Protein , Metabolism , beta Catenin , Metabolism
20.
Article in English | WPRIM | ID: wpr-773574

ABSTRACT

Triptolide (TP) induces severe liver injury, but its hepatotoxicity mechanisms are still unclear. Inflammatory responses may be involved in the pathophysiology. Neutrophils are the first-line immune effectors for sterile and non-sterile inflammatory responses. Thus, the aim of the present study was to investigate the neutrophilic inflammatory response in TP-induced liver injury in C57BL/6 mice. Our results showed that neutrophils were recruited and accumulated in the liver, which was parallel to or slightly after the development of liver injury. Neutrophils induced release of myeloperoxidase and up-regulation of CD11b, which caused cytotoxicity and hepatocyte death. Hepatic expressions of CXL1, TNF-α, IL-6, and MCP1 were increased significantly to regulate neutrophils recruitment and activation. Up-regulation of toll like receptors 4 and 9 also facilitated neutrophils infiltration. Moreover, neutrophils depletion using an anti-Gr1 antibody showed mild protection against TP overdose. These results indicated that neutrophils accumulation might be the secondary response, not the cause of TP-induced liver injury. In conclusion, the inflammatory response including neutrophil infiltration may play a role in TP-induced hepatotoxicity, but may not be severe enough to cause additional liver injury.


Subject(s)
Animals , Chemical and Drug Induced Liver Injury , Allergy and Immunology , Chemokine CCL2 , Genetics , Allergy and Immunology , Diterpenes , Drugs, Chinese Herbal , Epoxy Compounds , Female , Humans , Interleukin-6 , Genetics , Allergy and Immunology , Intracellular Signaling Peptides and Proteins , Genetics , Allergy and Immunology , Liver , Allergy and Immunology , Mice , Mice, Inbred C57BL , Neutrophil Infiltration , Neutrophils , Allergy and Immunology , Phenanthrenes , Tripterygium , Chemistry , Tumor Necrosis Factor-alpha , Genetics , Allergy and Immunology
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