OBJECTIVES: To investigate the protective effect of catalpol against triptolide-induced liver injury and explore its mechanism to test the Fuzheng Zhidu theory for detoxification. METHODS: C57BL/6J mice were randomized into blank control group, catalpol group, triptolide group and triptolide+catalpol group. After 13 days of treatment with the agents by gavage, the mice were examined for liver tissue pathology, liver function, hepatocyte subcellular structure, lipid peroxidation, ferrous ion deposition and expressions of ferroptosis-related proteins in the liver. In a liver cell line HL7702, the effect of catalpol or the ferroptosis inhibitor Fer-1 on triptolide-induced cytotoxicity was tested by examining cell functions, Fe²⁺ concentration, lipid peroxidation, ROS level and the ferroptosis-related proteins. RESULTS: In C57BL/6J mice, catalpol significantly alleviated triptolide-induced hepatic injury, lowered the levels of ALT, AST and LDH, and reversed the elevation of Fe²⁺ concentration and MDA level and the reduction of GP_X level. In HL7702 cells, inhibition of ferroptosis by Fer-1 significantly reversed triptolide-induced elevation of ALT, AST and LDH levels. Western blotting and qRT-PCR demonstrated that catalpol reversed abnormalities in expressions of SLC7A11, FTH1 and GPX4 at both the mRNA and protein levels in triptolide-treated HL7702 cells. CONCLUSIONS The combined use of catalpol can reduce the hepatotoxicity of triptolide in mice by inhibiting excessive hepatocyte ferroptosis through the SLC7A11/GPX4 pathway.
Animals ; Phenanthrenes/toxicity* ; Ferroptosis/drug effects* ; Diterpenes/toxicity* ; Epoxy Compounds/toxicity* ; Mice, Inbred C57BL ; Hepatocytes/metabolism* ; Mice ; Phospholipid Hydroperoxide Glutathione Peroxidase ; Iridoid Glucosides/pharmacology* ; Liver/metabolism* ; Chemical and Drug Induced Liver Injury/prevention & control* ; Male ; Amino Acid Transport System y+/metabolism*

Thirteen novel diterpenoids, comprising seven tiglianes (walliglianes G-M, 1-7), four rhamnofolanes (wallinofolanes A-D, 8-11), and two daphnanes (wallaphnanes A and B, 12 and 13), together with two known rhamnofolane diterpenoids (euphorwallside H and euphorwallside I, 14 and 15), were isolated and characterized from Euphorbia wallichii(E. wallichii). The chemical structures of these compounds were elucidated through nuclear magnetic resonance (NMR), mass spectrometry (MS), and quantum chemical calculations. Compounds 9 and 11 demonstrated protective effects against H₂O₂-induced BV-2 microglial cell damage. Molecular docking analyses indicated that compound 9 exhibited binding affinity to the anti-oxidant-related targets HMGCR, GSTP1, and SHBG.
Euphorbia/chemistry* ; Antioxidants/isolation & purification* ; Diterpenes/isolation & purification* ; Molecular Structure ; Mice ; Molecular Docking Simulation ; Animals ; Hydrogen Peroxide/toxicity* ; Cell Line ; Microglia/drug effects*

OBJECTIVE: The transformations that occur in diterpenoid alkaloids during the process of sand frying for Chinese herbal medicine preparation have yet to be clarified. This study investigated the structural changes that take place in 3-acetylaconitine during a simulation of heat-processing and evaluated the toxicity and biological activity of the pyrolysis products. METHODS: The diterpenoid alkaloid 3-acetylaconitine was heated at 180 °C for 15 min to simulate the process of sand frying. The pyrolysis products were separated using column chromatography, and their structures were investigated using high-resolution electrospray ionization mass spectroscopy and nuclear magnetic resonance spectroscopy. Further, in vivo cardiotoxicity and acute toxicity of 3-acetylaconitine and its pyrolysis products were compared, and the aconitine-induced arrhythmia model was employed to evaluate the antiarrhythmic effect of the pyrolysis products. RESULTS: Two new diterpenoid alkaloids, pyroacetylaconitine and 16-epi-pyroacetylaconitine, a pair of epimers at C-16, were isolated. After comparing the structures of these compounds, possible transformation pathways were proposed. Compared with the prototype compound, 3-acetylaconitine, the cardiotoxicity and acute toxicity of the heat-transformed products were significantly decreased. In the biological activity assay, the two pyrolysis products exhibited an effective increase in ventricular premature beat latency, a reduction in the occurrence of ventricular tachycardia, as well as an increase in the rate of arrhythmia inhibition, implying strong antiarrhythmic activity. CONCLUSION Compared with 3-acetylaconitine, its pyrolysis products displayed lower toxicity and good antiarrhythmic effects; thus, they have potential for being developed into antiarrhythmic medicines. Please cite this article as: Wang YJ, Wang Y, Tao P. Structural characterization, in vivo toxicity and biological activity of two new pyro-type diterpenoid alkaloids derived from 3-acetylaconitine. J Integr Med. 2023; 21(3): 302-314.
Humans ; Aconitine/chemistry* ; Cardiotoxicity ; Sand ; Alkaloids/toxicity* ; Arrhythmias, Cardiac/drug therapy* ; Diterpenes/toxicity*

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.
Animals ; Anti-Inflammatory Agents ; Diterpenes/toxicity* ; Epoxy Compounds/toxicity* ; Furans ; Glucosides ; Kidney/drug effects* ; Phenanthrenes/toxicity* ; Rats ; Rats, Sprague-Dawley

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.
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 ; drug effects ; Phenanthrenes ; toxicity ; Plant Extracts ; toxicity ; Rats ; Rats, Wistar ; Tandem Mass Spectrometry ; Tripterygium ; toxicity

Jatrogricaine A (1), a new diterpenoid possessing a 5/6/6/4 carbon ring system, together with eight known diterpenoids (2-9) were isolated from the stems of Jatropha podagrica. Their structures were elucidated by extensive spectroscopic methods and the absolute configuration of 1 was determined by single crystal X-ray diffraction analysis. All compounds were evaluated for their anti-inflammatory activities in vitro, and compound 3 showed significant inhibitory effects against nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW264.7 macrophage cells with an IC of 13.44 ± 0.28 μmol·L, being comparable to the positive control, quercetin (IC 17.00 ± 2.10 μmol·L).
Animals ; Anti-Inflammatory Agents ; chemistry ; pharmacology ; Carbon ; analysis ; Diterpenes ; chemistry ; pharmacology ; Inhibitory Concentration 50 ; Jatropha ; chemistry ; Lipopolysaccharides ; toxicity ; Macrophages ; drug effects ; metabolism ; Mice ; Molecular Structure ; Nitric Oxide ; metabolism ; Plant Extracts ; chemistry ; pharmacology ; Plant Stems ; chemistry ; RAW 264.7 Cells

A new lignan, tirpitzin A (17) together with 20 known compounds (1-16, and 18-21) were isolated from the ethyl acetate soluble fraction of ethanol extract of the aerial parts of Tirpitzia ovoidea. The structure of new compound was elucidated by means of spectroscopic analysis. Of the known compounds, 7-21 were isolated from Linaceae family for the first time. The pharmacological activity of the crude extracts was tested using a mouse inflammation model induced by dimethyl benzene. The results demonstrated that the ethyl acetate soluble fraction had anti-inflammatory activity. Moreover, the cytotoxic and anti-inflammatory activities of some compounds were studied. The new compound 17 showed moderate cytotoxic effect against BxPC-3 cell line (IC = 19.51μmol·L) and Compound 10 showed significant cytotoxicity against HepG2, HL-60, U87 and BxPC-3 cell lines with IC values in the range 4.2-8.3μmol·L. Additionally, Compounds 2, 10, 11, and 13 exhibited potent inhibitory effects on LPS-induced nitric oxide production in RAW 264.7 macrophages at the concentration of 50μmol·L.
Animals ; Anti-Inflammatory Agents ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Survival ; drug effects ; Disease Models, Animal ; Diterpenes ; chemistry ; pharmacology ; toxicity ; HL-60 Cells ; Hep G2 Cells ; Humans ; Inhibitory Concentration 50 ; Lignans ; chemistry ; pharmacology ; toxicity ; Linaceae ; chemistry ; Macrophages ; drug effects ; Mice ; Nitric Oxide ; metabolism ; Plant Components, Aerial ; chemistry ; Plant Extracts ; chemistry ; pharmacology ; toxicity ; RAW 264.7 Cells

The present study was designed to explore the influence of water extracts of Atractylodes lancea rhizomes on the toxicity and anti-inflammatory effects of triptolide (TP). A water extract was prepared from A. lancea rhizomes and co-administered with TP in C57BL/6 mice. The toxicity was assayed by determining serum biochemical parameters and visceral indexes and by liver histopathological analysis. The hepatic CYP3A expression levels were detected using Western blotting and RT-PCR methods. The data showed that the water extract of A. lancea rhizomes reduced triptolide-induced toxicity, probably by inducing the hepatic expression of CYP3A. The anti-inflammatory effects of TP were evaluated in mice using a xylene-induced ear edema test. By comparing ear edema inhibition rates, we found that the water extract could also increase the anti-inflammatory effects of TP. In conclusion, our results suggested that the water extract of A. lancea rhizomes, used in combination with TP, has a potential in reducing TP-induced toxicity and enhancing its anti-inflammatory effects.
Animals ; Anti-Inflammatory Agents ; isolation & purification ; pharmacology ; Atractylodes ; chemistry ; Cytochrome P-450 Enzyme System ; genetics ; Diterpenes ; toxicity ; Edema ; chemically induced ; pathology ; Enzyme Induction ; drug effects ; Epoxy Compounds ; toxicity ; Gene Expression Regulation ; drug effects ; Herb-Drug Interactions ; Liver ; drug effects ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Phenanthrenes ; toxicity ; Plant Extracts ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry ; Water ; chemistry

Triptolide (TP) from Tripterygium wilfordii has been demonstrated to possess anti-inflammatory, immunosuppressive, and anticancer activities. TP is specially used for the treatment of awkward rheumatoid arthritis, but its clinical application is confined by intense side effects. It is reported that licorice can obviously reduce the toxicity of TP, but the detailed mechanisms involved have not been comprehensively investigated. The current study aimed to explore metabolomics characteristics of the toxic reaction induced by TP and the intervention effect of licorice water extraction (LWE) against such toxicity. Obtained urine samples from control, TP and TP + LWE treated rats were analyzed by UPLC/ESI-QTOF-MS. The metabolic profiles of the control and the TP group were well differentiated by the principal component analysis and orthogonal partial least squares-discriminant analysis. The toxicity of TP was demonstrated to be evolving along with the exposure time of TP. Eight potential biomarkers related to TP toxicity were successfully identified in urine samples. Furthermore, LWE treatment could attenuate the change in six of the eight identified biomarkers. Functional pathway analysis revealed that the alterations in these metabolites were associated with tryptophan, pantothenic acid, and porphyrin metabolism. Therefore, it was concluded that LWE demonstrated interventional effects on TP toxicity through regulation of tryptophan, pantothenic acid, and porphyrin metabolism pathways, which provided novel insights into the possible mechanisms of TP toxicity as well as the potential therapeutic effects of LWE against such toxicity.
Animals ; Biomarkers ; Chromatography, High Pressure Liquid ; methods ; Diterpenes ; toxicity ; Epoxy Compounds ; toxicity ; Glycyrrhiza ; Male ; Metabolomics ; Phenanthrenes ; toxicity ; Plant Extracts ; therapeutic use ; Principal Component Analysis ; Rats ; Rats, Sprague-Dawley ; Spectrometry, Mass, Electrospray Ionization ; methods

The arrenotokous toxicity of triptolide was evaluated, and the rate of sperm abnormality, the changes of the lipid peroxide, the enzyme activity and the hormone in male rats were observed. With the negative and positive control group, the healthy rats were respectively given by gavage triptolide suspension at the dose of 0.025, 0.05, 0.1 mg x kg(-1) for 30 days. Then the rats were killed for the measurement of the indicators in testis and serum, as well as the study on the sperm abnormality. The results showed that the positive control group had significant difference, compared with the negative control group. The content of SOD, LDH, G-6-PD, Na+ -K+ -ATPase, Ca+ -Mg+ -ATPase decreased significantly in 0.05 mg x kg(-1) group, and reduced more obviously with exposure to the dose of 0.1 mg x kg(-1). The levels of GSH-Px and beta-G showed a significant decrease in the testis of rats only at the dose of 0.1 mg x kg(-1). Nevertheless, the MDA levels, the FSH levels and the LH levels showed no significant difference. The deformity rate of sperm increased significantly in 0.05 mg x kg(-1) group and 0.1 mg x kg(-1) group. The results indicated the triptolide had the effect of the lipid peroxidation to damage Spermatogenic cells, Sertolis cells and Leydig cells. At the same time, the triptolide interfered not only with the energy supply process of aerobic and anaerobic glycolysis,but also with the energy utilization in testis by affecting the activities of testis marker enzymes, and produced a damage chain of the male reproductive system
Animals ; Diterpenes ; toxicity ; Drugs, Chinese Herbal ; toxicity ; Epoxy Compounds ; toxicity ; Lipid Peroxidation ; drug effects ; Male ; Organ Size ; drug effects ; Phenanthrenes ; toxicity ; Rats ; Rats, Wistar ; Reproduction ; drug effects ; Spermatozoa ; abnormalities ; drug effects ; metabolism ; Testis ; drug effects ; growth & development ; metabolism ; Tripterygium ; chemistry ; toxicity