Texts of Kolima dialect of Yukaghir

<p>Clinical chemistry data of monkeys fed on diets containing GM rice or non-GM rice.</p

Primer sequences for PCR amplification.

<p>Primer sequences for PCR amplification.</p

Effect of triptolide and SFN on activities of antioxidant enzymes in liver of BALB/C mice.

<p>Mice were given vehicle control, SFN (25 mg/kg), TP (1.0 mg/kg), SFN (25 mg/kg)+TP (1.0 mg/kg) by intraperitoneal injection. Mice were sacrificed under anesthesia 24 h after triptolide treatment. Values are expressed as mean ± SD. n = 6.</p>a<p><i>P<0.05</i> compared to the control group.</p>b<p><i>P<0.01</i> compared to the control group.</p>c<p><i>P<0.05</i> compared to the triptolide-treated group.</p>d<p><i>P<0.01</i> compared to the triptolide-treated group.</p

siRNA-Nrf2 enhances triptolide-induced oxidative stress in HepG2 cells.

<p>HepG2 cells transfected with siRNA-control or siRNA-Nrf2 were treated with triptolide at various concentrations for 6 h. (A) Levels of intracellular GSH were measured. (B) The intracellular ROS levels were measured using the fluorescent probe DCFH-DA.The data are represented as the mean ± SD from three independent experiments. *<i>P</i><0.05, **<i>P</i><0.01 versus vehicle control; ^#<i>P</i><0.05, ^##<i>P</i><0.01 versus si-control cells treated with the same concentration of triptolide. Con: control (0.1% DMSO).</p

Activation of Nrf2 Protects against Triptolide-Induced Hepatotoxicity

<div><p>Triptolide, the major active component of <i>Tripterygium wilfordii</i> Hook f. (TWHF), has a wide range of pharmacological activities. However, the toxicities of triptolide, particularly the hepatotoxicity, limit its clinical application. The hepatotoxicity of triptolide has not been well characterized yet. The aim of this study was to investigate the role of NF-E2-related factor 2 (Nrf2) in triptolide-induced toxicity and whether activation of Nrf2 could protect against triptolide-induced hepatotoxicity. The results showed that triptolide caused oxidative stress and cell damage in HepG2 cells, and these toxic effects could be aggravated by Nrf2 knockdown or be counteracted by overexpression of Nrf2. Treatment with a typical Nrf2 agonist, sulforaphane (SFN), attenuated triptolide-induced liver dysfunction, structural damage, glutathione depletion and decrease in antioxidant enzymes in BALB/C mice. Moreover, the hepatoprotective effect of SFN on triptolide-induced liver injury was associated with the activation of Nrf2 and its downstream targets. Collectively, these results indicate that Nrf2 activation protects against triptolide-induced hepatotoxicity.</p></div

Effects of triptolide and SFN on Nrf2-related downstream targets in livers of BALB/C mice.

<p>(A) The mRNA levels of NQO1, GCLC and HO-1 were analyzed by real-time PCR. GAPDH was used as an internal control. The data are represented as the mean ± SD; n = 6; *<i>P<0.05</i>, **<i>P<0.01</i> vs Con group. (B) The protein levels of NQO1, GCLC and HO-1 were analyzed by Western blot. β-actin was used as an internal control. A representative blot from three independent experiments is shown. The density of the immunoreactive bands was analyzed, and the data are represented as the means ± SD; n = 6; *<i>P</i><0.05, **<i>P</i><0.01 versus Con group.</p

The role of Nrf2 in the protection against triptolide-induced cytotoxicity in HepG2 cells.

<p>Overexpression experiments (A, B): HepG2 cells were transiently transfected with PEF or PEF_NRF2 plasmids. The cells were treated 24 h later with triptolide at various concentrations for 24 h. (A) Western blot analysis of Nrf2 expression 24 h after transfection. Expression of β-actin was used as an internal control. Representative blots from three independent experiments are shown. The density of the immunoreactive bands was analyzed, and the data are represented as the means ± SD. (B) Cytotoxicity was determined by an MTT assay. The data are represented as the mean ± SD from three independent experiments. Knockdown experiments (C, D, E): HepG2 cells were transiently transfected with control siRNA or siRNA targeting Nrf2. The cells were treated 48 h later with triptolide at various concentrations for 24 h. (C) Western blot analysis of Nrf2 expression 48 h after transfection. Expression of β-actin was used as an internal control. A representative blot from three independent experiments is shown. The density of the immunoreactive bands was analyzed, and the data are represented as the means ± SD. (D) Cytotoxicity was determined by an MTT assay. The data are represented as the mean ± SD from three independent experiments. (E) Morphological changes in normal and Nrf2 knockdown HepG2 cells treated with triptolide (20 nM) were observed under an inverted phase contrast microscope (100×, Olympus, Japan). The representative results from three independent experiments are shown. *<i>P</i><0.05, **<i>P</i><0.01 versus control cells treated with the same concentration of triptolide.</p

Effect of triptolide and SFN on ALT, AST, ALP and LDH in BALB/C mice.

<p>Mice were given vehicle control, SFN (25 mg/kg), TP (1.0 mg/kg), SFN (25 mg/kg)+TP (1.0 mg/kg) by intraperitoneal injection. Mice were sacrificed under anesthesia 24 h after triptolide treatment. Values are expressed as mean ± SD. n = 6.</p>a<p><i>P<0.05</i> compared to the control group.</p>b<p><i>P<0.01</i> compared to the control group.</p>c<p><i>P<0.01</i> compared to the triptolide-treated group.</p

Triptolide induces oxidative stress in HepG2 cells.

<p>HepG2 cells were treated with triptolide at various concentrations for 6(A) and GSH (B) were measured respectively using the fluorescent probe DCFH-DA and a GSH Detection Kit. The data are represented as the mean ± SD from three independent experiments. <i>*P</i><0.05, <i>**P</i><0.01 versus vehicle control. Con: control (0.1% DMSO).</p

Triptolide induces cytotoxicity in HepG2 cells.

<p>(A) HepG2 cells were treated with triptolide at various concentrations for 6, 12 and 24 h. Cell viability was determined by an MTT assay. The viability of the cells without triptolide treatment is defined as 100%. (B) HepG2 cells were treated with triptolide at various concentrations for 24 h. LDH leakage was measured. The data are represented as the mean ± SD from three independent experiments. <i>*P</i><0.05, <i>**P</i><0.01 versus vehicle control (0.1% DMSO).</p