Triptolide

Manufactured by Merck Group

Sourced in United States, Germany

Triptolide is a compound extracted from the Chinese herb Tripterygium wilfordii. It is commonly used as a research tool in laboratory settings to study various biological processes and pathways. Triptolide has been shown to have anti-inflammatory, anti-tumor, and immunosuppressive properties, making it a valuable tool for researchers investigating these areas. However, a detailed description of its core function and intended use cannot be provided while maintaining an unbiased and factual approach.

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Lab products found in correlation

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83 protocols using triptolide

Triptolide-Induced Chromatin Remodeling in K562 Cells

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Cell culture: K562 cells (ATCC, CCL-243) were cultured at 37 o C, 5% CO2 at a density between 0.3-1 x 10 6 cells/mL in RPMI medium (VWR 45000-396) topped up with 10% Fetal Bovine Serum (Genesee Scientific, cat: #25-514). Cells were split at a consistent interval of 3 days, when the cells reach 10 6 cells/mL.
Cell culture for Triptolide time course: 24h prior to Triptolide treatment, K562 cells were resuspended in fresh (RPMI) medium at a density of 0.6 x 10 6 cells/mL. On the day of the experiment, cells were recounted, aliquoted in equal cell numbers to 6 T-100 ThermoFisher Tissue Culture Flasks (each flask corresponding to one time point) and treated with Triptolide (Sigma-Aldrich, T3652-1MG) to a final concentration of 500 nM Triptolide. The Triptolide treatment was performed for 0 min, 15 min, 30 min, 1h, and respectively 4h.
Cells cross-linking for ChIP: After Triptolide treatment, K562 cells were cross-linked in 1% CH2O freshly prepared in 1x PBS on the day of the experiment to reach the final concentration of 0.1% CH2O in the media. Following a 5 min incubation at room temperature on a rocking platform, the cross-linker was quenched with 1M Glycine to reach a final concentration of 0.135 M Glycine. Lastly, cells were washed twice in 1x PBS, then harvested and snap frozen on dry ice.

Wang Z., Chivu A.G., Choate L.A., Rice E.J., Miller D.C., Chu T., Chou S., Kingsley N.B., Petersen J.L., Finno C.J., Bellone R.R., Antczak D.F., Lis J.T., & Danko C.G. (2021). Interdependence between histone marks and steps in Pol II transcription.

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Triptolide Mitigates Cerebral Ischemia-Reperfusion Injury

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Twenty-five 8-week-old healthy male SD rats, weighing 225 g to 245 g were selected as study subjects. The ambient temperature of the experiment was 22 ±1°C and the light time was 12 hours/ day. The above rats were randomly divided into 5 groups (n = 5): sham operation group (Sham group), ischemia-reperfusion model group (I/R group), low concentration of triptolide (Merck, Germany) group (12.5 mg/kg, TL-L group), medium concentration of triptolide group (25 mg/kg, TL-M group) and high concentration of triptolide group (50 mg/kg, TL-H group). In the I/R and triptolide treated groups the suture-occluded method was used to block the middle cerebral artery. In the Sham group, only internal and external cervical vessels were separated and no suture to block. In the TL-L, TL-M, TL-H groups, the rats were injected intraperitoneally with an appropriate amount of DMSO solvent mixed with triptolide at 24 hours before operation. The Sham group and the I/R group were also injected intraperitoneally with the same dose of DMSO solvent as in the triptolide treated groups before the operation.

Pan W, & Xu Z. (2020). Triptolide mediates Wnt/β-catenin signalling pathway to reduce cerebral ischemia-reperfusion injury in rats. Folia neuropathologica, 58(4).

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Triptolide Mitigates I/R Injury in Rats

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A total of 60 rats were randomly divided into four groups (n=15 in each group). 1) Sham group (sham), 2) I/R, 3) I/R + dimethyl sulfoxide [DMSO], and 4) I/R + Triptolide. Triptolide (5 mg/kg; Sigma-Aldrich Co., St Louis, MO, USA, dissolved in pure DMSO) was intraperitoneally injected at the onset of reperfusion in rats belonging to the Triptolide group; a corresponding volume of vehicle (pure DMSO) was administered to the rats in the I/R group. At the conclusion of the reperfusion period, the neurological deficit scores were evaluated. The rats were then sacrificed for the collection of tissue samples.

Zhang B., Song C., Feng B, & Fan W. (2016). Neuroprotection by triptolide against cerebral ischemia/reperfusion injury through the inhibition of NF-κB/PUMA signal in rats. Therapeutics and Clinical Risk Management, 12, 817-824.

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Zscan4 Silencing and DNA Damage Analysis in Mouse Embryos

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The siRNA diced pools against Zscan4 and luciferase (control) were generated using recombinant Giardia lamblia Dicer. Experimental setup was done as previously described (39 (link)). Briefly, 4 μM siRNAs against Zscan4 or luciferase control siRNA were coinjected with dextran-tetramethyl-rhodamine (Invitrogen, 3000 molecular weight 100 μg/ml) in late G₁ stage (4 to 5 hpf) zygotes derived from in vitro–fertilized mouse oocytes. After incubation at 37°C in a humidified atmosphere of 5% CO₂ and 95% air, Zscan4 or control siRNA–injected 2C embryos at 30 hpf were briefly washed in M2 medium, treated with acidic tyrodes, and fixed in 3.7% paraformaldehyde in PBS at 4°C. After permeabilization with 0.2% Triton X-100 in PBS for 10 min at RT, embryos were blocked overnight at 4°C in 1% BSA and 0.1% Triton X-100 in PBS. For DNase/MNase (micrococcal nuclease) treatment, fixed and permeabilized embryos were incubated with TURBO DNase 1 (2 U/μl, Ambion) and MNase (80 gel units, NEB) for 2 hours at 37°C under mineral oil. Embryos were analyzed for Zscan4 γH2A.X by immunostaining. For triptolide experiments, 2C embryos at 29 hpf were incubated with triptolide (10 μM; Sigma-Aldrich) or dimethyl sulfoxide (DMSO) for 2 hours in the presence of 5-ethynyl-uridine (5 mM; EU, Thermo Fisher Scientific) before fixation at 31 hpf and analyzed for Zscan4, γH2A.X-foci, and EU signals.

Srinivasan R., Nady N., Arora N., Hsieh L.J., Swigut T., Narlikar G.J., Wossidlo M, & Wysocka J. (2020). Zscan4 binds nucleosomal microsatellite DNA and protects mouse two-cell embryos from DNA damage. Science Advances, 6(12), eaaz9115.

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Cytotoxicity Assay of Small Molecules

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5 × 10³ cells/well (A2780 and A2780cis) were seeded in 96-well plates overnight and afterwards incubated with different concentrations of SB216763 (12.5 µM, 25 µM, 50 µM, 100 µM) (Sigma-Aldrich, Taufkirchen, Germany), XAV939 (6.25 µM, 12.5 µM, 25 µM, 50 µM) (Sigma-Aldrich) and triptolide (6.25 nM, 12.5 nM, 25 nM, 50 nM) (Sigma-Aldrich).
S-phase-dependent synthesis of DNA during the cell cycle and, therefore, cellular proliferation was analyzed with thymidine analog 5-bromo-2′-deoxyuridine (BrdU) enzyme-linked immunosorbent assay (ELISA) after 72 h. Cell viability was determined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) (Sigma-Aldrich) colorimetric assay after 48 h and 72 h. The procedures of both techniques were already reported by our colleagues (Geiger et al. 2016 (link); Zhu et al. 2018 ).
Controls of untreated cells (for incubation with triptolide), cells treated with 2‰ (for incubation with SB216763) and 5‰ (for incubation with XAV939) dimethyl sulfoxide (DMSO) were carried out. For the evaluation the control without DMSO was set 100%.

Kaltofen T., Preinfalk V., Schwertler S., Fraungruber P., Heidegger H., Vilsmaier T., Vattai A., Czogalla B., Mayr D., Mahner S., Jeschke U, & Trillsch F. (2020). Potential of platinum-resensitization by Wnt signaling modulators as treatment approach for epithelial ovarian cancer. Journal of Cancer Research and Clinical Oncology, 146(10), 2559-2574.

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Transcardial Perfusion and Tissue Dissociation

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Once anesthetized the animals were transcardially perfused with ice-cold Choline solution (above) containing the following small molecule cocktail for 5 minutes: 1 uM TTX (Sigma), 100 uM AP-V (Thermo Fisher Scientific), 5 ug/ml Actinomycin D (Sigma) and 10 uM Triptolide (Sigma). V1 was then microdissected, cut into 300um slices, and incubated on ice for 30 minutes in Dissociation Solution containing 1uM TTX, 100 uM AP-V (Thermo Fisher Scientific), 5 ug/ml Actinomycin D (Sigma), 10 uM Triptolide (Sigma) and 10 ug/ml Anicomycin (Sigma). Papain was added to a final concentration of 20 U/ml and the tissue dissociated for 1h at 37°C with gentle agitation in a total volume of 3.2ml. The remaining procedures were performed per manufacturer’s instructions without the small molecule cocktail. Following gradient centrifugation the cells were washed in Dissociation Solution containing 0.04% BSA and resuspended in Dissociation Solution containing 0.04% BSA and 15% Optiprep (Sigma) for single cell RNA-Seq.

Hrvatin S., Hochbaum D.R., Nagy M.A., Cicconet M., Robertson K., Cheadle L., Zilionis R., Ratner A., Borges-Monroy R., Klein A.M., Sabatini B.L, & Greenberg M.E. (2017). Single-Cell Analysis of Experience-Dependent Transcriptomic States in Mouse Visual Cortex. Nature neuroscience, 21(1), 120-129.

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Transcriptional Regulation in Symbiodiniaceae

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For α-amanitin treatment, B. minutum cells at a density of ∼1 × 10⁶ cells per ml were treated with α-amanitin (Sigma-Aldrich, catalog no. A2263) at concentrations of 1 µg ml⁻¹ (‘normal’ dose) and 4 µg ml⁻¹ (‘high’ dose).
Samples were collected at 0, 24 and 48 h after treatment.
For triptolide treatment, B. minutum cells at a density of ∼1 × 10⁶ cells per ml were treated with triptolide (Sigma-Aldrich, catalog no. T3652) at concentrations of 10 µM (‘normal’ dose) and 40 µM (‘high’) dose. Samples were collected at 0, 8, 24 and 48 h after treatment.

Marinov G.K., Trevino A.E., Xiang T., Kundaje A., Grossman A.R, & Greenleaf W.J. (2021). Transcription-dependent domain-scale three-dimensional genome organization in the dinoflagellate Breviolum minutum. Nature Genetics, 53(5), 613-617.

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Transcardial Perfusion and Tissue Dissociation

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Monitoring mRNA Stability in T Cells

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To monitor mRNA stability, 3 million CD4+ T cells were incubated in the presence of 5,6-Dichloro-1-β-D-ribofuranosylbenzimidazole (DRB, Sigma-Aldrich, Cat: D1916) at a final concentration of 65µM or Triptolide (Sigma-Aldrich, Cat: T3652) at a final concentration of 25µM for 15min, 1h or 3h. At each time point (0, 15min, 1h and 3h), cells were collected, counted and total RNA was extracted from the same amount of cells (3 million) at each time point using trizol in the presence of 1µl of a 1/10 dilution of ERCC Spike-In RNA (Thermo Fisher Scientific, Cat: 4456740). To monitor mRNA stability in conditions where mRNA translation is impaired, cells were incubated in the presence of either DRB or Triptolide and Cycloheximide (final concentration of 100µg.ml-1, Sigma-Aldrich, Cat: 01810) or Harringtonine (final concentration of 2µg.ml-1, Interchim, Cat: H0169) for 1 or 3h and total RNA extracted as described above. Total RNA was depleted from ribosomal RNA using Ribo-Zero rRNA Removal Kit (Human/Mouse/Rat, Illumina) followed by cDNA library preparation as described below.

Mercier B.C., Labaronne E., Cluet D., Bicknell A.A., Corbin A., Guiguettaz L., Aubé F., Modolo L., Auboeuf D., Moore M.J., & Ricci E.P. (2020). Translation-dependent and independent mRNA decay occur through mutually exclusive pathways that are defined by ribosome density during T Cell activation.

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Triptolide-Induced Apoptosis in Thyroid Cancer

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The thyroid cancer cell line TPC-1 was selected for this study. TPC-1 cells were purchased from Bena Culture Collection (Henan, China). RPMI 1640 medium (Corning, 10-040-cv) was supplemented with 10% fetal bovine serum (FBS) (Gibco, 10099141), Penicillin-Streptomycin solution (Corning, 30-001-CI). TPC-1 cells were maintained under humidified conditions with 5% CO2 at 37°C. Triptolide was purchased from Merck (T3652). Cell apoptosis kits containing propidium iodide (PI) and Annexin V- Fluorescein isothiocyanate (FITC) were purchased from Abcam (ab14085). The anti- cyclin-dependent kinase inhibitor 1A (CDKN1A) primary antibody was purchased from Sigma-Aldrich (SAB5700742). Anti-phospho-c-JUN (CST, 3270s), anti-phospho-p53 (CST, 2521s), anti-phospho-NF-κB p65 (CST, 3031s), and anti- glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (CST, 5174s) antibodies were purchased from Cell Signaling Technology. Goat anti-rabbit secondary antibody was purchased from Abcam (ab6721).

Wang F., An S.J., Yin Y., Li J.J., Sun C.H., Lan J., Zhao W.J, & Li C.Q. (2021). Triptolide is a Promising Therapeutic Approach in Treating Thyroid Cancer Based on in silico and in vitro Experiment. Drug Design, Development and Therapy, 15, 4275-4287.

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