α-Hederin Regulates SREBP1/FASN Pathway and Inhibits Malignant Phenotype of Non-small Cell Lung Cancer / 肿瘤防治研究

Objective To explore the regulative effect of α-Hederin on the proliferation and invasion of NSCLC and investigate its related molecular mechanism. Methods After A549 and HCC-1833 cells were treated with a concentration gradient of α-Hederin for 24 and 48 h, the OD450nm was detected by using CCK8 assays, and the IC50 was calculated.The A549 and HCC-1833 cells were divided into the blank control and α-Hederin groups in accordance with IC50 values.Cell proliferation was detected by EdU assays, and cell cycle transformation and cell apoptosis were detected by flow cytometry.Cell mobility was detected by using Transwell and scratch assays.SREBP1 and FASN protein expression levels were detected through Western blot analysis, and cell lipid accumulation was detected via oil red O staining. Results The survival rate of lung cancer cells decreased significantly with the increase of α-Hederin concentration, and the IC50 values of A549 and HCC-1833 cells at 48 h were 15 and 25 μg/ml, respectively.Compared with the blank control group, cells proliferation and migration were significantly inhibited, cells were blocked in the G1/S phase, the apoptosis rate increased, and the protein expression and lipid accumulation of SREBP1/FASN significantly reduced after α-Hederin treatment. Conclusion α-Hederin can inhibit the proliferation and migration, G1/S phase transition and induce the apoptosis of NSCLC cells and hinder the malignant progression of NSCLC by downregulating the expression of SREBP1 and FASN and reducing the accumulation of cell lipids.

Anti-tumor Effect and Mechanisms of Hederin： A Review / 中国实验方剂学杂志

Hederin is a natural active component of triterpenoid saponins extracted from many medicinal herbs， such as Lithospermum erythrorhizon， Pulsatilla chinensis， and Clematis florida. It has attracted much attention from doctors for its anti-inflammatory， anti-convulsive， anti-oxidation and anti-leishmaniasis activities. Hederin has significant anti-tumor bioactivity and is expected to be a potential drug for the treatment of malignant tumors. The available studies have demonstrated that hederin can promote the apoptosis， inhibit the proliferation， metastasis， and invasion， and induce the autophagy of tumor cells， exhibiting a promising prospect in the treatment of breast cancer， lung cancer， liver cancer， and pancreatic cancer. Specifically， hederin can regulate the phosphoinositide 3-kinase/protein kinase B （PI3K/Akt） pathway， reactive oxygen species （ROS）， and microRNA （miRNA） to trigger tumor cell apoptosis. Its anti-proliferation activity is mainly reflected in the regulation of cyclin and cyclin-dependent kinase （CDK）. Hederin inhibits the metastasis and invasion of tumor cells by blocking epithelial-mesenchymal transformation （EMT）. In addition， hederin can influence metabolic reprogramming to induce tumor cell autophagy. Hederin is involved in a variety of pathways to exert its anti-tumor activity and may become a novel anti-tumor drug in the future， which give new sights into the study of hederin in the anti-tumor field. There are few studies about hederin and no systematic review of its anti-tumor mechanisms. Therefore， this study reviewed the studies about the anti-tumor mechanism of hederin， aiming to provide reference and information for researchers and clinical staff.

Alpha-hederin induces the apoptosis of oral cancer SCC-25 cells by regulating PI3K/Akt/mTOR signaling pathway

BACKGROUND: Oral cancer is one of the common malignant tumors of the head and neck. However, current treatments have numerous side effects, and drugs from natural sources may have better therapeutic potential. This research investigated the induction of apoptosis by α-hederin (α-HN), a constituent of Pulsatilla chinensis (Bunge) Regel, in the oral cancer cell line SCC-25 and its underlying mechanism. RESULTS: SCC-25 cells were treated with 50, 100, and 200 µmol/L α-HN. Cell proliferation; extent of apoptosis; activities of caspases-3, 8, and 9; and the expression of Bcl-2, Bax, phosphorylated (p)-phosphoinositide 3-kinase (PI3K), p-Akt, and p-mammalian target of rapamycin (mTOR) proteins were determined using the 3-(4,5)-2-thiazole-(2,5)-diphenyl tetrazolium bromide, flow cytometry, caspase activity detection kits, and western blot assays, respectively. The results showed that the proliferation of SCC-25 cells in the α-HN-treated groups decreased significantly, and the inhibitory effect was time and concentration dependent. Compared with cells in the control group, the extent of apoptosis increased significantly, caspase-3 and -9 activities were significantly enhanced, and the Bcl-2 level was lowered and the Bax level was elevated significantly in SCC-25 cells treated with α-HN for 48 h (P b 0.05). The expression of p-PI3K, p-Akt, and p-mTOR was also significantly lower in SCC-25 cells treated with α-HN than that in the control group (P b 0.05). CONCLUSION: These results indicate that α-HN can inhibit proliferation and induce apoptosis of SCC-25 cells and may exert these effects by inhibiting the PI3K/Akt/mTOR signaling pathway.

Effects of drying temperature, growing area and plucking time on two saponins from leaves of Hedera helix / 中成药

AIM To investigate the effects of drying temperature,growing area and plucking time on hederacoside C,α-hederin from leaves of Hedera helix L..METHODS Three batches of H.helix leaves plucked in different time from two growing areas were dried in a vacuum oven to the constant weight at 60 ℃,70 ℃,80 ℃,90 ℃ and 105 ℃,respectively.Two saponins in the processed leaves were determined by HPLC.The powders of the processed H.helix leaves of different batches were mixed with proper ratios,which were determined by least squares optimization method with constraints.RESULTS The content of hederacoside C in the processed H.helix leaves of the three batches increased while that of α-hederin decreased with increasing temperature.The relative error between measured value and desired contents of hederacoside C and α-hederin in the mixed H.helix leaves was less than 5.5％.CONCLUSION The effects of three factors on the content of two saponins in the H.helix leaves are in the order of drying temperature,growing area and plucking time.Mixing processed H.helix leaves of different quality statues reasonably can control the contents of two saponins in a certain range.

Dynamic accumulation of saponins in Hedera helix / 中草药

Objective: This study detected the contents of four saponins (hederacoside C, hederacoside D, hederacoside D, α-hederin) from different parts of annual cutting Hedera helix in one year growth cycle and analyzed the dynamic accumulation, in order to provide theoretical basis to harvest Hedera helix L. as medicinal raw materials reasonably. Methods: This study designed a reasonable method of collecting materials and detected the contents of saponins by HPLC. Results: The contents of saponins in different parts were as following order: mature leaf > new leaf > stem > root, and four saponins in different parts had different dynamic accumulations in one year growth cycle. Conclusion: The leaf and stem are the reasonable collecting parts of raw materials which have higher medicinal quality. The reasonable collecting time of leaf is the growth stage in March and stem is from March to November before the beginning of dormancy.

Pharmacokinetics and tissue distribution of α-hederin sodium salt in rats / 中国中药杂志

To study the pharmacokinetics and tissue distribution characteristics of α-hederin sodium salt in rats. 100 mg•kg⁻¹ α-hederin sodium salt was given to the rats by intragastric administration, and LC-MS/MS method was used to determine its concentration at different time in plasma and tissues. Plasma and tissue samples were treated with methanol protein deposition method. Main pharmacokinetic parameters were as follows： tmax (0.97±1.23) h, Cmax (222.53±57.28) μg•L⁻¹, AUC0-t (1 262±788.9) h•μg•L⁻¹, T1/2 (17.94±9.50) h. α-hederin can be detected in heart, liver, spleen, lung, kidney, brain, muscle and adipose. The results showed that α-hederin sodium salt was absorbed fast and eliminated slowly in rats after oral administration. It was widely distributed in body tissues and livers kept the highest concentrations among various tissues at different time, so it can be speculated that α-hederin may have certain targeting property on livers.

In vivo intestinal absorption characteristics of α-hederin in rats / 中草药

Objective: To investigate the intestinal absorption characteristics of α-hederin and to explore the causes of poor bioavailability. Methods: In vivo single-pass perfusion model was used and the concentration of α-hederin was determined by HPLC. The effects of intestinal segment, drug concentration, pH value, gut microflora, and P-gp inhibitor on the intestinal absorption of the drug were investigated. Results: The absorption rate constant (Ka) of α-hederin decreased following the sequence of ileum > colon > jejunum > duodenum. Absorption parameters of α-hederin had no significant difference at different concentration of 75, 150, and 300 μg/mL and those increased with the increase of pH value. The intestinal flora which were disrupted may affect the absorption of α-hederin. There was no significant difference in Ka and Peff values between P-gp inhibitor and no P-gp inhibitor groups. Conclusion: α-Hederin can be absorbed in whole intestine, but better in lower intestine. The saturate phenomena was not observed under the test range of drug concentration, and the absorption mechanism may be the passive diffusion transport. The absorption can be better under basic condition. The absorption is significantly affected by the intestinal flora and α-hederin is not the substrate of P-gp.

Analysis on in vivo metabolites of α-hederin in rats by UPLC-MS/MS / 中草药

Objective: To investigate the metabolites of α-hederin in feces of rats after ig administration. Methods: An UPLC-Q/Trap-MS method was used for the identification of metabolites of α-hederin in feces of rats after ig administration of α-hederin (150 mg/kg). Results: Seven metabolites were detected in rat feces, including demethylation of α-hederin (M1), 3-O-α-L-arabinopyranoside hederagenin (M2), hederagenin (M3), glucuronideconjugate of α-hederin (M4), and double bond bonus of α-hederin (M5, M6-1, and M6-2). Conclusion: α-Hederin experiences a variety of metabolic reactions in rat gastrointestinal tract, mainly including deglycosylation, glucuronidation, demethylation, etc.