Eupatilin, a lipophilic flavonoid isolated from Artemisia species, is a PPARα agonist, and possesses anti-apoptotic, anti-oxidative and anti-inflammatory activities.

Eupatilin is a PPARα agonist. Eupatilin (10, 30, 100 μM) suppresses IL-4 expression and degranulation in RBL-2H3 cells^[1]. Eupatilin (50-100 μM) slightly reduces cell viability of HaCaT cells. Eupatilin (10, 30, 50, 100 μM) increases PPARα transactivation and expression in HaCaT cells. Eupatilin (10, 30, 50 μM) also suppresses TNFα-induced MMP-2/-9 expression in HaCaT cells. Furthermore, Eupatilin inhibits TNFα-induced p65 translocation, IκBα Phosphorylation, AP-1 and MAPK signaling via PPARα^[2]. Eupatilin (10-50 μM) shows no cytotoxic effects on ARPE19 cells. Eupatilin (10, 25, 50 μM) elevates cell viability from oxidative stress, and inhibits H₂O₂-induced ROS production in ARPE19 cells. Moreover, Eupatilin (50 μM) inbibits H₂O₂-induced cells apoptosis and promotes the activation of PI3K/Akt pathway in RPE cells^[3].

Shanghai Jinpan Biotech Co Ltd has not independently confirmed the accuracy of these methods. They are for reference only.

Eupatilin (1.5% or 3.0%) restores PPARα mRNA expression, and improves atopic dermatitis (AD)-like symptoms in oxazolone-induced Balb/c mice. Eupatilin causes significant decrease in serum IgE, IL-4 levels, oxazolone-induced TNFα, IFNγ, IL-1β, TSLP, IL-33 and IL-25 mRNA expression in oxazolone-induced mice. Eupatilin also increases filaggrin and loricrin mRNA expression in oxazolone-induced mice^[1].

Shanghai Jinpan Biotech Co Ltd has not independently confirmed the accuracy of these methods. They are for reference only.

344.32

C₁₈H₁₆O₇

22368-21-4

异泽兰黄素

Room temperature in continental US; may vary elsewhere.

DMSO : 33.33 mg/mL (96.80 mM; Need ultrasonic)

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂： 10% DMSO    40% PEG300    5% Tween-80    45% saline

  Solubility: ≥ 2.5 mg/mL (7.26 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (7.26 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 2.

  请依序添加每种溶剂： 10% DMSO    90% (20% SBE-β-CD in saline)

  Solubility: ≥ 2.5 mg/mL (7.26 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (7.26 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明
• 3.

  请依序添加每种溶剂： 10% DMSO    90% corn oil

  Solubility: ≥ 2.5 mg/mL (7.26 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (7.26 mM，饱和度未知) 的澄清溶液，此方案不适用于实验周期在半个月以上的实验。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. Jung Y, et al. Eupatilin, an activator of PPARα, inhibits the development of oxazolone-induced atopic dermatitis symptoms in Balb/c mice. Biochem Biophys Res Commun. 2018 Feb 5;496(2):508-514.

  [2]. Jung Y, et al. Eupatilin with PPARα agonistic effects inhibits TNFα-induced MMP signaling in HaCaT cells. Biochem Biophys Res Commun. 2017 Nov 4;493(1):220-226.

  [3]. Du L, et al. Eupatilin prevents H2O2-induced oxidative stress and apoptosis in human retinal pigment epithelial cells. Biomed Pharmacother. 2017 Jan;85:136-140.

Cell viability is detected using a MTT assay. In brief, after treatment, the medium is replaced with fresh medium containing 0.5 mg/mL MTT for 4 h at 37°C. Then, the medium is gently aspirated and 150 μL of DMSO is added to each well to solubilize the formazan crystals. The absorbance is measured at 450 nm by a microplate reader. The relative cell viability is defined as the absorbance of treated wells divided by that of the control^[3].

Shanghai Jinpan Biotech Co Ltd has not independently confirmed the accuracy of these methods. They are for reference only.

Six-week-old female Balb/c mice are housed under conditions of controlled temperature (23  ±  2 °C), humidity (55  ±  5%), and 12 h light/dark cycles (06:00-18:00 h light, 18:00-06:00 dark). Briefly, Balb/c mice are sensitized on day −7 by a single application of 20 μL of 1.0% oxazolone in a mixture of acetone and olive oil (4:1) to the inner and outer surface of both ears. On day 0, the mouse ears are challenged with 20 μL of 0.1% oxazolone at 2-day intervals for 4 weeks post-sensitization. The mice are treated with the indicated concentrations of Eupatilin (1.5% or 3.0%) twice a day for 4 weeks. The control group is treated with vehicle alone (acetone and olive oil [4:1]). After 3 weeks, the mice are sacrificed and samples are collected. Ears are stored at −80 °C for RNA isolation and analysis or immediately fixed in 4% formalin for histological analysis^[1].

Shanghai Jinpan Biotech Co Ltd has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Jung Y, et al. Eupatilin, an activator of PPARα, inhibits the development of oxazolone-induced atopic dermatitis symptoms in Balb/c mice. Biochem Biophys Res Commun. 2018 Feb 5;496(2):508-514.

  [2]. Jung Y, et al. Eupatilin with PPARα agonistic effects inhibits TNFα-induced MMP signaling in HaCaT cells. Biochem Biophys Res Commun. 2017 Nov 4;493(1):220-226.

  [3]. Du L, et al. Eupatilin prevents H2O2-induced oxidative stress and apoptosis in human retinal pigment epithelial cells. Biomed Pharmacother. 2017 Jan;85:136-140.