Gracillin
(Synonyms: 纤细薯蓣皂苷) 目录号 : GN10533
Gracillin 是西洋参的天然甾体皂苷成分，具有抗肿瘤、抗菌、抗氧化、促凋亡和抗炎等特性。
Cas No.:19083-00-2
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
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- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
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Cell lines |
A549 cells |
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Preparation Method |
A549 cells were cultured in 6-well plates at a density of 1.5 × 105/well and cultured for 12 h, and then treated with a series of concentrations of gracillin (0.25, 0.5, 1, 2, 4 µmol/L) for 24 h. The cells were stained with Hoechst 33,258 staining buffer and the morphology of apoptosis was observed by fluorescence microscopy. |
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Reaction Conditions |
0.25, 0.5, 1, 2, 4 µmol/L for 24 h |
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Applications |
It can be observed that gracillin induced morphological changes in apoptosis of A549 cells by fluorescence microscopy after staining with Hoechst33258. Compared with the blank group, the gracillin treatment group showed the typical morphological characteristics of apoptotic cells, specifically, the number of cells was relatively reduced, the nucleus was pyknotic, the chromatin condensed into lumps under the nuclear membrane, and showed bright blue fluorescence staining. |
| Animal experiment [2]: | |
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Animal models |
Five‐week‐old athymic female BALB/c nude mice |
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Preparation Method |
The mice were randomly divided into three groups, which were injected intraperitoneal every other day with gracillin (1 mg/kg or 2 mg/kg) or PBS (control) till the tumours' volume reached ~100 mm3. |
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Dosage form |
1 mg/kg or 2 mg/kg, i.p. |
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Applications |
Results clearly show that the volume and weight of the tumours were lower in the gracillin©\treated mice when compared to the control group. This inhibitory effect was in a clear dose©\dependent manner. |
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References: [1] Yang J, et al. Gracillin Isolated from Reineckia carnea Induces Apoptosis of A549 Cells via the Mitochondrial Pathway. Drug Des Devel Ther. 2021 Jan 20;15:233-243. |
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Gracillin, as a natural steroidal saponin component of D. quinqueloba, has antitumour, antimicrobial, antioxidant, pro‐apoptotic and anti‐inflammatory properties[1].
In vitro efficacy test it shown that gracillin markedly inhibits the viability of RKO, SW480 and HCT116 cells, with IC50 of 3.118, 2.671 and 5.473 μmol/L, respectively. Gracillin also stimulates apoptosis in human CRC cells in a dose-dependent manner (0, 2.5, 5.0 or 10.0 μmol/L)[1]. In vitro experiment it exhibited that gracillin obviously inhibited the proliferation of A549 cells with IC50 of 2.54 μmol/L in a concentration-dependent and time-dependent manner and induced morphological changes[2]. Furthermore, gracillin inhibited the gastric tumor BGC823 cell proliferation in a concentration-dependent manner with IC50 of 8.3 μM and gastric tumor SGC7901 cell proliferation with IC50 of 8.9 μM in the range of 0-15 μM. At concentrations of 3 μM, 6 μM, and 12 μM, gracillin has TIPE2 induction in concentration-dependent manner[3]. In vitro, gracillin has dose-dependent inhibitory effects on the mitochondrial function in various human cancer cell lines derived from lung, colorectum, prostate, pharynx, and liver with IC50 of ~1-5 μM[4].
In vivo, mice were administrated with 20 mg/kg gracillin significantly inhibited the growth of xenograft tumors[5]. In vivo efficacy test it demenstrated that fish treated with 1 mg/kg gracillin carried significantly fewer parasites than the control, and acute toxicity LD50 of gracillin for grass carp was 1.64 mg/kg[6].
References:
[1] Yang L, et al. Gracillin shows potent efficacy against colorectal cancer through inhibiting the STAT3 pathway. J Cell Mol Med. 2021 Jan;25(2):801-812.
[2] Yang J, et al. Gracillin Isolated from Reineckia carnea Induces Apoptosis of A549 Cells via the Mitochondrial Pathway. Drug Des Devel Ther. 2021 Jan 20;15:233-243.
[3] Liu W, et al. Beneficial Effects of Gracillin From Rhizoma Paridis Against Gastric Carcinoma via the Potential TIPE2-Mediated Induction of Endogenous Apoptosis and Inhibition of Migration in BGC823 Cells. Front Pharmacol. 2021 Sep 24;12:669199.
[4] Min HY, et al. The natural compound gracillin exerts potent antitumor activity by targeting mitochondrial complex II. Cell Death Dis. 2019 Oct 24;10(11):810.
[5] Min HY, et al. Potent Anticancer Effect of the Natural Steroidal Saponin Gracillin Is Produced by Inhibiting Glycolysis and Oxidative Phosphorylation-Mediated Bioenergetics. Cancers (Basel). 2020 Apr 8;12(4):913.
[6] Zheng W, et al. Antiparasitic efficacy of Gracillin and Zingibernsis newsaponin from Costus speciosus (Koen ex. Retz) Sm. against Ichthyophthirius multifiliis. Parasitology. 2015 Mar;142(3):473-9.
Gracillin 是西洋参的天然甾体皂苷成分，具有抗肿瘤、抗菌、抗氧化、促凋亡和抗炎等特性[1]。
体外药效试验表明，gracillin显着抑制RKO、SW480和HCT116细胞的活力，IC50分别为3.118、2.671和5.473 μmol/L。 Gracillin 还以剂量依赖性方式（0、2.5、5.0 或 10.0 μmol/L）刺激人 CRC 细胞凋亡[1]。体外实验表明，gracillin明显抑制A549细胞增殖，IC50为2.54 μmol/L，呈浓度依赖性和时间依赖性，并诱导细胞形态学改变[2]< /sup>。此外，gracillin 以浓度依赖性方式抑制胃肿瘤 BGC823 细胞增殖，IC50 为 8.3 μM，抑制胃肿瘤 SGC7901 细胞增殖，IC50 为 8.9 μM。范围为 0-15 μM。在 3 μM、6 μM 和 12 μM 浓度下，gracillin 以浓度依赖性方式诱导 TIPE2[3]。在体外，gracillin 对来自肺癌、结直肠癌、前列腺癌、咽癌和肝脏的多种人类癌细胞系的线粒体功能具有剂量依赖性抑制作用，IC50 约为 1-5 μM<sup >[4].
在体内，给予小鼠 20 mg/kg gracillin 显着抑制异种移植肿瘤的生长[5]。体内药效试验表明，用1 mg/kg gracillin处理的鱼携带的寄生虫明显少于对照，gracillin对草鱼的急性毒性LD50为1.64 mg/kg[6]。
| Cas No. | 19083-00-2 | SDF | |
| 别名 | 纤细薯蓣皂苷 | ||
| Canonical SMILES | CC1CCC2(C(C3C(O2)CC4C3(CCC5C4CC=C6C5(CCC(C6)OC7C(C(C(C(O7)CO)O)OC8C(C(C(C(O8)CO)O)O)O)OC9C(C(C(C(O9)C)O)O)O)C)C)C)OC1 | ||
| 分子式 | C45H72O17 | 分子量 | 884.48 |
| 溶解度 | ≥ 174.4mg/mL in DMSO, Limited solubility in EtOH | 储存条件 | 4°C, protect from light |
| General tips | 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 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.1306 mL | 5.653 mL | 11.3061 mL |
| 5 mM | 0.2261 mL | 1.1306 mL | 2.2612 mL |
| 10 mM | 0.1131 mL | 0.5653 mL | 1.1306 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
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| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
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Gracillin shows potent efficacy against colorectal cancer through inhibiting the STAT3 pathway
Colorectal cancer (CRC) accounts for about 10% of all annually diagnosed cancers and cancer-related deaths worldwide. STAT3 plays a vital role in the occurrence and development of tumours. Gracillin has shown a significant antitumour activity in tumours, but its mechanism remains unknown. The human CRC cell lines HCT116, RKO, and SW480 and immunodeficient mice were used as models to study the effects of gracillin on cell proliferation, migration and apoptosis. These were evaluated by cell viability, colony formation, wound-healing migration and cell apoptosis assays. Luciferase reporter assay, and immunostaining and western blot analyses were used to explore the specific mechanism through which gracillin exerts its effects. Gracillin significantly reduces viability and migration and stimulates apoptosis in human CRC cells. It also significantly inhibits tumour growth with no apparent physiological toxicity in animal model experiments. Moreover, gracillin is found to inhibit STAT3 phosphorylation and STAT3 target gene products. In addition, gracillin inhibits IL6-induced nuclear translocation of P-STAT3. Gracillin shows potent efficacy against CRC by inhibiting the STAT3 pathway. It should be further explored as a unique STAT3 inhibitor for the treatment of CRC.
Gracillin Isolated from Reineckia carnea Induces Apoptosis of A549 Cells via the Mitochondrial Pathway
Background: Reineckia carnea is commonly used to treat cough, pneumonia and other diseases in China. In our previous study, it was found that the ethanol extracts of Reineckia carnea have a strong inhibitory effect on the proliferation of human lung cancer A549 cells. Here, we isolated gracillin from ethanol extracts for the first time.
Purpose: Clarify the antiproliferation effect of gracillin on A549 cells and further explore its mechanisms via the mitochondrial pathway.
Methods: Gracillin was isolated and purified by silica gel, D-101 macroporous resin and preparative RP-HPLC, then identified by NMR and HR-MS. The inhibitory effects of gracillin on the proliferation of A549 cells were detected by the MTS method. Its mechanisms were further explored by flow cytometry and Western blot.
Results: A steroid saponin, gracillin, was isolated and identified from Reineckia carnea for the first time. In a concentration-dependent and time-dependent manner, gracillin significantly inhibited the proliferation of A549 cells with an IC50 value at 2.54 μmol/L and induced morphological changes. The results of flow cytometry analysis showed that the apoptosis rate of A549 cells was significantly increased (p < 0.05), and the cells proportion was obviously arrested in S phase. The concentration of intracellular calcium was raised (p < 0.01), and the mitochondrial membrane potential was greatly decreased (p < 0.01). In addition, the expression levels of Bax, caspase-3, cleaved caspase-3, and cytochrome C were dramatically up-regulated while Bcl-2 was down-regulated (p < 0.05) in A549 cells.
Conclusion: This study confirmed that gracillin has a significant antiproliferative effect on A549 cells. Gracillin could induce the apoptosis of A549 cells through the mitochondrial pathway, which might be associated with regulation of the concentration of intracellular calcium, the mitochondrial membrane potential and the expression levels of Bax, Bcl-2, caspase-3, cleaved caspase-3, and cytochrome C.
Gracillin Shows Potential Efficacy Against Non-Small Cell Lung Cancer Through Inhibiting the mTOR Pathway
The leading cause of cancer deaths is lung cancer, non-small cell lung cancer (NSCLC), the most common type of lung cancers, remains a difficult cancer to treat and cure. It is urgent to develop new products to treat NSCLS. Gracillin, extracted from Reineckia carnea, Dioscorea villosa, and other medicinal plants, has anti-tumor potential with toxic effect on a variety of tumor cells such as NSCLC. However, the anti-NSCLC mechanism of gracillin is not completely clear. In this study, A549 cells and athymic nude mice were used as models to evaluate the anti-NSCLC effects of gracillin. The antiproliferative activity of gracillin on A549 cells was conducted by CCK-8, and obvious autophagy was observed in gracillin-treated A549 through transmission electron microscopy. Furthermore, the expressions of Beclin-1, LC3-II, and WIPI1 were upregulated, while the expression of p62 was downregulated in gracillin-treated A549. The further mechanism study found that the mTOR signaling pathway was significantly inhibited by gracillin. Accordingly, the PI3K/Akt pathway positively regulating mTOR was inhibited, and AMPK negatively regulating mTOR was activated. Meanwhile, LC3-II transformation was found to be significantly reduced after WIPI1 was silenced in A549 cells but increased after gracillin treatment. It also proves that WIPI is involved in the process of gracillin regulating A549 autophagy. At last, the anti-tumor growth activity of gracillin in vivo was validated in A549-bearing athymic nude mice. In conclusion, gracillin has anti-NSCLC activity by inducing autophagy. The mechanism maybe that gracillin inhibited the mTOR signaling pathway. Gracillin has the potential to be a candidate product for the treatment of NSCLC in the future.
Beneficial Effects of Gracillin From Rhizoma Paridis Against Gastric Carcinoma via the Potential TIPE2-Mediated Induction of Endogenous Apoptosis and Inhibition of Migration in BGC823 Cells
Tumor necrosis factor-α inducible protein-8 (TIPE2), initially recognized as a negative immune regulator, exerts an important role in suppressing the progression of numerous cancers. In our previous investigation, we found that TIPE2 expression displayed a decrease or absence in gastric tumor tissue, and the overexpression of TIPE2 suppressed the growth of gastric cancer tumors and cells, demonstrating that TIPE2 could be a potential medicinal target for gastric cancer treatment. However, it's seldomly reported that several medicinal agents or candidates targeted TIPE2 for treating diseases, including gastric cancer. To identify the candidate targeting TIPE2 to fight against gastric cancer, several extractions from traditional natural medicinal plants with anti-tumor functions were employed to screen the active compounds according to bioassay-guided isolation. Interestingly, gracillin, a component from the ethyl acetate extraction of Rhizoma Paridis, was identified to induce the expression of TIPE2 and inhibit the cell proliferation in gastric cancer BGC-823 cells. Furthermore, the underlying mechanisms that restrain gastric cancer were evaluated by clone formation, EdU staining, flow cytometry, and other assays. Meanwhile, the role of TIPE2 in the anti-tumor effect of gracillin was elucidated via the use of siTIPE2 RNA. It was determined that gracillin could fight against gastric cancer cells by inhibiting the cell proliferation participated by the PI3K/AKT pathway and cell cycle arrest, suppressing the EMT pathway-regulating cell migration, and inducing bcl2-associated mitochondrial apoptosis. Additionally, TIPE2 maybe contribute to the benefits of gracillin. These results of the present study are an important step toward the medicinal development of gracillin, and are also of use in understanding the effect of TIPE2 as a potential tumor target.
Anti-Atopic Properties of Gracillin Isolated from Dioscorea quinqueloba on 2,4-Dinitrochlorobenzene-Induced Skin Lesions in Mice
Naturally occurring saponins have been reported to have anti-inflammatory and immunomodulatory effects. However, the effects of gracillin, a main saponin component of Dioscorea quinqueloba (D. quinqueloba), on atopic dermatitis (AD), have not been previously studied. The aim of this study was to determine whether gracillin isolated from D. quinqueloba has an anti-AD effect on 2,4-dinitrochlorobenzene (DNCB)-induced AD-like skin lesions in SKH-1 hairless mice. Topical co-treatment of gracillin and DNCB for two weeks markedly reduced symptoms typical of AD (redness, itching, swelling and skin lichenification), decreased transepidermal water loss (TEWL) and increased skin hydration. In addition, gracillin strongly inhibited PI-induced IL-4 expression in RBL-2H3 cells and in the skins of AD mice. Our results suggest gracillin is a potential candidate for the prevention and treatment of AD and other inflammatory skin disorders.