Ginsenoside Rh3 is a bacterial metabolite of Ginsenoside Rg5. Ginsenoside Rh3 treatment in human retinal cells induces Nrf2 activation.

Nrf2^[1]

Ginsenoside Rh3 inhibits UV-induced oxidative damages in retinal cells via activating nuclear-factor-E2-related factor 2 (Nrf2) signaling. Ginsenoside Rh3 treatment in retinal cells induces Nrf2 activation. The potential activity of Ginsenoside Rh3 is tested on Nrf2 signaling in the retinal pigment epithelium cells (RPEs). The qRT-PCR assay results demonstrate that treatment with Ginsenoside Rh3 dose-dependently increases mRNA transcription and expression of key Nrf2-regulated genes, including HO1, NQO1 and GCLC. Consequently, protein expressions of these Nrf2-dependent genes (HO1, NQO1 and GCLC) are also significantly increased in Ginsenoside Rh3 (3-10 μM)-treated RPEs. Notably, although Nrf2 mRNA level is unchanged after Ginsenoside Rh3 treatment, its protein level is significantly increased by Rh3^[1]. EZ-Cytox assay is used to assess the effect of ginsenoside-Rh3 on SP 1-keratinocytes viability. Ginsenoside Rh3 (0.01, 0.1, 1 and 10 μM) shows no cytotoxic effect at all concentrations^[2].

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

The potential effect of Ginsenoside Rh3 is examined on mouse retina, using the light-induced retinal damage model. Ginsenoside Rh3 intravitreal injection (5 mg/kg body weight, 30 min pre-treatment) significantly attenuates light-induced decrease of both a- and b-wave amplitude. The electroretinography (ERG)’s a-wave decreases to 46.03±1.62% % of control level after light exposure, which is back to 71.84±7.51% with Ginsenoside Rh3 administration. The b-wave is 40.19±3.34% of control level by light exposure, and Rh3 intravitreal injection brings back to 80.01±2.37% of control level^[1].

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

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C₃₆H₆₀O₇

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人参皂苷 Rh3；人参皂苷 Rh3

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• 方案 一

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

  Solubility: 0.67 mg/mL (1.11 mM); 悬浊液; 超声助溶

  此方案可获得 0.67 mg/mL的均匀悬浊液，悬浊液可用于口服和腹腔注射。

  以 1 mL 工作液为例，取 100 μL 6.7 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；再向上述体系中加入 50 μL Tween-80，混合均匀；然后再继续加入 450 μL 生理盐水 定容至 1 mL。

  生理盐水的配制：将 0.9 g 氯化钠，溶解于 ddH₂O 并定容至 100 mL，可以得到澄清透明的生理盐水溶液。

• 方案 二

  请依序添加每种溶剂： 10% DMSO    90% Corn Oil

  Solubility: ≥ 0.67 mg/mL (1.11 mM); 澄清溶液

  此方案可获得 ≥ 0.67 mg/mL（饱和度未知）的澄清溶液，此方案实验周期在半个月以上的动物实验酌情使用。

  以 1 mL 工作液为例，取 100 μL 6.7 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. Tang CZ, et al. Activation of Nrf2 by Ginsenoside Rh3 protects retinal pigment epithelium cells and retinal ganglion cells from UV. Free Radic Biol Med. 2018 Mar;117:238-246.  [Content Brief]

  [2]. Chung I, et al. Inhibitory mechanism of Korean Red Ginseng on GM-CSF expression in UVB-irradiated keratinocytes. J Ginseng Res. 2015 Oct;39(4):322-30.  [Content Brief]

SP-1 keratinocytes are seeded in 96 well plates (2×10⁴ cells/well). After 24 h, the media is replaced with media containing various concentrations of (A) SKRG, or (B) Ginsenoside Rh3 (0.01, 0.1, 1 and 10 μM). Control cells are treated with DMSO at a final concentration of 0.1%. After 24 h, the media containing the compounds or DMSO is replaced with media containing 10% EZ-Cytox. The cells are then incubated at 37°C for 1 h, and the absorbance is measured using a microplate reader at a wavelength of 450 nm. All assays are performed in triplicate^[2].

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

Mice^[1]
The BALB/c mice (Male, 5-6 week old, 17-18 g weight) are used. The pupillary dilation is performed before exposure to 5000 lx of white fluorescent light. Thirty min before light exposure, Ginsenoside Rh3 (at 5 mg/kg body weight) are injected intravitreally to the right eye. ERG recording after light exposure is also reported early. The b-wave amplitude is measured from the trough of the a-wave to the peak of the b-wave, and the amplitude of the a-wave is measured from the initial baseline.

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

• [1]. Tang CZ, et al. Activation of Nrf2 by Ginsenoside Rh3 protects retinal pigment epithelium cells and retinal ganglion cells from UV. Free Radic Biol Med. 2018 Mar;117:238-246.  [Content Brief]

  [2]. Chung I, et al. Inhibitory mechanism of Korean Red Ginseng on GM-CSF expression in UVB-irradiated keratinocytes. J Ginseng Res. 2015 Oct;39(4):322-30.  [Content Brief]