Ginsenoside Ro (Polysciasaponin P3; Chikusetsusaponin 5; Chikusetsusaponin V) exhibits a Ca²⁺-antagonistic antiplatelet effect with an IC₅₀ of 155  μM. Ginsenoside Ro reduces the production of TXA₂ more than it reduces the activities of COX-1 and TXAS.

Ginsenoside Ro in Panax ginseng is a beneficial novel Ca²⁺-antagonistic compound and may prevent platelet aggregation-mediated thrombotic disease. Ginsenoside Ro dose-dependently reduces thrombin-stimulated platelet aggregation, and IC₅₀ is approximately 155 μM^[1]. Ginsenoside Ro inhibits TXA₂ production to abolish thrombin-induced platelet aggregation. Thromboxane A₂ (TXA₂) induces platelet aggregation and promotes thrombus formation. Ginsenoside Ro dose-dependently (50-300 μM) reduces the TXB₂ level that is induced by thrombin; Ginsenoside Ro (300 μM) inhibits the thrombin-mediated elevation in TXB₂ level by 94.9%. COX-1 activity in the absence of Ginsenoside Ro (negative control) is 2.3±0.1 nmol/mg protein. However, Ginsenoside Ro dose-dependently (50-300 μM) reduces its activity; at 300 μM, COX-1 activity is reduced by 26.4% of that of the negative control. TXA₂ synthase (TXAS) activity in the absence of Ginsenoside Ro (negative control) is 220.8±1.8 ng/mg protein/min. However, Ginsenoside Ro dose-dependently (50-300 μM) reduces its activity; at 300 μM, TXAS activity is reduced by 22.9% of that of the negative control. The inhibitory effect of Ginsenoside Ro (300 μM) on TXB₂ production (94.9%) is significantly higher than those on COX-1 (26.4%) and TXAS (22.9%) activities^[2]. To assess the toxicity of Ginsenoside Ro in Raw 264.7 cells, they are first treated with various concentrations (10 μM, 50 μM, 100 μM, and 200 μM) of Ginsenoside Ro for 24 h. Ginsenoside Ro exhibits no significant dose dependent toxicity. The effect of Ginsenoside Ro is next determined on cell viability and ROS levels, a marker of oxidative stress, following treatment with 1 μg/mL LPS. LPS reduces cell viability by ∼70% compared with nontreated controls. Pretreatment with 100 μM and 200 μM Ginsenoside Ro for 1 h prior to 1 μg/mL LPS incubation for 24 h leads to a significant increase in cell viability. The changes in ROS levels and NO production are consistent with the effects of Ginsenoside Ro on viability^[3].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Ginsenoside Ro dissolved in water is administrated by gavage to mice at doses of 25 and 250 mg/kg/day for 4 days before i.v. injection of HT29 in order to keep blood concentrations of Ginsenoside Ro above a certain level before HT29 i.v. injection followed by 40 days of oral administration of Ginsenoside Ro to the mice. After 38 days of treatment, the animals are euthanized, and the number of pulmonary metastatic nodules is counted in addition to evaluation of toxicity of Ginsenoside Ro and mouse pathology by HT29. Ginsenoside Ro (250 mg/kg/day) produces a significant decrease in the number of tumor nodules on the lung surface, yielding inhibition rates of 88% (P < 0.01)^[4].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

957.11

C₄₈H₇₆O₁₉

34367-04-9

人参皂苷 Ro；人参皂苷 Ro

Room temperature in continental US; may vary elsewhere.

DMSO : 100 mg/mL (104.48 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 (2.61 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (2.61 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 (2.61 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (2.61 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 (2.61 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (2.61 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]. Kwon HW, et al. Inhibitory Effects of Cytosolic Ca²⁺ Concentration by Ginsenoside Ro Are Dependent on Phosphorylation of IP3RI and Dephosphorylation of ERK in Human Platelets. Evid Based Complement Alternat Med. 2015;2015:764906.

  [2]. Jung-HaeShin, et al. Inhibitory effects of thromboxane A2 generation by ginsenoside Ro due to attenuation of cytosolic phospholipase A2 phosphorylation and arachidonic acid release. J Ginseng Res. 9 Jan 2018.

  [3]. Kim S, et al. Upregulation of heme oxygenase-1 by ginsenoside Ro attenuates lipopolysaccharide-induced inflammation in macrophage cells. J Ginseng Res. 2015 Oct;39(4):365-70.

  [4]. Jiang Z, et al. The traditional Chinese medicine Achyranthes bidentata and our de novo conception of its metastatic chemoprevention: from phytochemistry to pharmacology. Sci Rep. 2017 Jun 20;7(1):3888.

The microsomal fraction of platelets is preincubated with Ozagrel (11 nM, IC₅₀), a positive control, or with various concentrations of Ginsenoside Ro and other reagents at 37°C for 5 min. The reaction is initiated by adding prostaglandin H₂, and the samples are incubated at 37°C for 1 min; the reaction is terminated by adding citric acid (1 M). After neutralization with 1 N NaOH, the amount of TXB2 is determined using a TXB₂ EIA kit^[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Cell viability is determined with an MTT assay kit. Briefly, Raw 264.7 cells are plated in 48-well plates at a density of 2.0×10⁴ cells per well, incubated for 24 h, and treated with various concentrations of Ginsenoside Ro for 24 h. How 1 h of pretreatment with Ginsenoside Ro (50 μM, 100 μM, and 200 μM) affects the viability of Raw 264.7 cells is then investigated treated with 1 μg/mL LPS for 24 h. After the incubation period, 10 μL of MTT reagent is added to each well and incubated for 3 h at 37°C in 5% CO₂. The resulting formazan crystals are subsequently dissolved in MTT solubilization solution. The absorbance is determined at 540 nm using a microplate reader^[3].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Mice^[4]
Female BALB/c mice (20-25 g, 6-8 weeks old) are used. The experimental model of lung metastasis is established by tail vein injection of HT29 cells to mimic the dissemination of CTCs. HT29 cells in the number of 2×10⁶ cells in 0.2 mL PBS are infected into the tail vein of six-week-old female Balb/c mice. Before the HT29 inoculation, oral gavage pretreatment of PBS-suspended B (Ginsenoside Ro) is given daily for 4 days, followed by a 40-day treatment. Treatment groups (N = 10) include: 0 mg/kg, 25 mg/kg and 250 mg/kg Ginsenoside Ro. Body weight is measured and recorded every four days. Mice are sacrificed after 40 days of tumor metastasis and growth and 44 days of treatment with B. The number of surface lung metastasis nodules is evaluated in each treatment group. Slides with 4-5 μm thick lung section are prepared, paraffin embedded and then stained with hematoxylin and eosin^[4].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Kwon HW, et al. Inhibitory Effects of Cytosolic Ca²⁺ Concentration by Ginsenoside Ro Are Dependent on Phosphorylation of IP3RI and Dephosphorylation of ERK in Human Platelets. Evid Based Complement Alternat Med. 2015;2015:764906.

  [2]. Jung-HaeShin, et al. Inhibitory effects of thromboxane A2 generation by ginsenoside Ro due to attenuation of cytosolic phospholipase A2 phosphorylation and arachidonic acid release. J Ginseng Res. 9 Jan 2018.

  [3]. Kim S, et al. Upregulation of heme oxygenase-1 by ginsenoside Ro attenuates lipopolysaccharide-induced inflammation in macrophage cells. J Ginseng Res. 2015 Oct;39(4):365-70.

  [4]. Jiang Z, et al. The traditional Chinese medicine Achyranthes bidentata and our de novo conception of its metastatic chemoprevention: from phytochemistry to pharmacology. Sci Rep. 2017 Jun 20;7(1):3888.