Based on LC-MS technology explored the metabolomics of Agrimonia pilosa intervening in non-small cell lung cancer A549 cells / 药学学报

The objective of this study was to analyze the effects on cell viability, apoptosis, and cell cycle of non-small cell lung cancer (NSCLC) A549 cells after intervention with Agrimonia pilosa (AP) and investigate Agrimonia pilosa anti-tumor activity in vitro. Meanwhile, liquid chromatography mass spectrometry (LC-MS) metabolomics technology was used to analyze the changes of cellular metabolites and metabolic pathways. The results of this study will provide a theoretical and experimental basis for investigating the mechanism of the effect of Agrimonia pilosa on non-small cell lung cancer A549 cells. The results showed that the cell nucleus of A549 cells crumpled and apoptosis occurred with the increase of drug concentration. The survival rate of the cells decreased, and the inhibition rate reached 21.5% and 91.74% under the low and high dose conditions, respectively. Lactate dehydrogenase (LDH) content increased (P < 0.05). Metabolomics results showed significant differences in metabolism between groups, thirty-three distinct metabolites including LysoPC(24:0/0:0), LysoPC(17:0/0:0) and PC(O-40:5) were deduced. The pathway enrichment showed that the Agrimonia pilosa plays an anti-tumor role mainly by regulating the metabolism of glycerophosphate and purine in A549 cells, in which the effect on glycerophosphate metabolism pathway was most significant. The results of combined pharmacodynamics suggested that Agrimonia pilosa might induce apoptosis and inhibit the growth of A549 cells by regulating LysoPC(24:0/0:0), LysoPC(17:0/0:0) and PC(O-40:5) metabolites in A549 cells.

Study on the different extraction fractions of Agrimonia pilosa against hepatic fibrosis / 中国药房

OBJECTIVE To study different extraction fractions of Agrimonia pilosa against h epatic fibrosis. METHODS Using hepatic stellate cells HSC-T 6 of rats as objects ，the effects of different extraction fractions （total extract ，ethyl acetate fraction ， petroleum ether fraction and n-butanol fraction ）with different concentrations （0.5，5，50，500，5 000 μg/mL，calculated by raw drug）of A. pilosa on the proliferation of HSC-T 6 cells were detected （after treated for 24，48，72 h）；median inhibition concentration（IC50）was also caculated. Platelet-derived growth factor （PDGF-BB）was used to induce the activation of HSC-T 6 cells to establish hepatic fibrosis cell model. Flow cytometry was used to detect the effects of different extraction fractions of A. pilosa on apoptosis of HSC-T 6 cells. The expression of collagen Ⅰ（Col-Ⅰ）in the supernatant was detected by enzyme linked immunosorbent assay. The expressions of α-smooth muscle actin （α-SMA），Col-Ⅰ，B-cell lymphoma- 2（Bcl-2），Bcl-2-associated X protein （Bax）and caspase- 3 were detected by Western blot assay. RESULTS Total extract ，ethyl acetate fraction ，petroleum ether fraction and n-butanol fraction of A. pilosa could significantly increase the apoptotic rate of HSC-T 6 cells（P＜0.01）. After treated for 24 h，IC50 of above fractions were 50.17，20.75，5.82，4.09 μg/mL，respectively. After intervened with PDGF-BB ，the expression of Col- Ⅰ in supernatant of HSC-T 6 cells as well as protein expression of Col- Ⅰ，α-SMA，Bcl-2，Bax and caspase- 3 in HSC-T6 cells were increased significantly （P＜0.01）. After intervened with different extraction fractions of A. pilosa ，most of the expressions of above proteins in HSC-T 6 cell culture supernatant or cells were significantly reversed compared with PDGF-BB group （P＜0.05 or P＜0.01）， and the intervention effect of n-butanol fraction of A. pilosa was the most significant. CONCLUSIONS Different extraction fractions of A. pilosa can inhibite the proliferation of HSC-T 6 cells and induce their apoptosis；n-butanol fraction from A. pilosa may be an effective fraction to exert the effect of anti-hepatic fibrosis.

Review on Anti-tumor Effects Mechanism and Clinical Application of Agrimonia pilosa / 世界科学技术-中医药现代化

Agrimonia pilosa is one of the traditional Chinese herbal medicines for 2000 years history. Due to its various pharmacological effects, it is widely used in the clinical treatment of tumors and other diseases. Presently malignancy has become an important factor threatening human health that the anti-tumor effect of Agrimonia pilosa has attracted great attention. The author reviewed its anti-tumor effect mechanism by articles arrangement of experimental study, clinical study and experience introduction. A series of study revealed 3 mechanisms: blocking cell cycle, inducing apoptosis and strengthening the cellular immunity. As a kind of traditional Chinese herbal medicine which has anti-tumor effect, the Agrimonia pilosa should be reasonably applied according to the related research and previous experience.

Anti-tumor effects of Agrimonia Pilosa Ledeb .on SMMC-7721 hepatocellular carcinoma cells and its mechanisms / 重庆医学

Objective To observe the anti-tumor effects of Agrimonia Pilosa Ledeb(APL) ,a Chinese herbal medicine ,on hepato-cellular carcinoma cells in vitro and investigate the underlined mechanisms preliminarily .Methods APL water extracts were pre-pared .SMMC-7721 cells were cultured with the medium containing different concentrations of APL water extracts ,and at different time points ,cell viabilities were measured by the MTT assay and inhibitory rates (IR) were calculated ;cell morphologic changes were observed under a light microscope ;apoptotic ratios were measured by flow cytometry ;and the expressions of Bcl-2 and P53 proteins were examined by immunocytochemistry .Results After the cells were cultured with the medium containing APL water ex-tracts for 24 h ,48 h and 72 h ,no obvious effects were found on the cell proliferation in 5 mg/mL group and 10 mg/mL group ,but IR were 0 .5% ,23 .9% and 27 .5% in 20 mg/mL group and 23 .3% ,51 .7% and 71 .6% in the 40 mg/mL group ,respectively .In the groups with effects on the cells proliferation ,morphological characteristics of apoptosis were obvious ,and the cell apoptotic ratios were 19 .5% and 23 .0% in 20 mg/mL group and 33 .4% and 42 .7% in 40 mg/mL group at 48 h and 72 h .The expressions of Bcl-2 protein were 71 .9% and 58 .5% in 20 mg/mL group and 47 .9% and 26 .5% in 40 mg/mL group at 48 h and 72 h ,and the ex-pressions of P53 protein were 22 .9% and 50 .6% in 20 mg/mL group and 48 .7% and 83 .7% in 40 mg/mL group at 48 h and 72 h . Conclusion The water extracts of APL are able to inhibit proliferation and induce apoptosis of SMMC-7721 cells dose-time depend-ently in vitro ,which might be associated with the expression changes of Bcl-2 and P53 protein .

Effect of Agrimonia pilosa Ledeb Extract on the Antinociception and Mechanisms in Mouse

In the present study, the antinociceptive profiles of Agrimonia pilosa Ledeb extract were examined in ICR mice. Agrimonia pilosa Ledeb extract administered orally (200 mg/kg) showed an antinociceptive effect as measured by the tail-flick and hot-plate tests. In addition, Agrimonia pilosa Ledeb extract attenuated the writhing numbers in the acetic acid-induced writhing test. Furthermore, the cumulative nociceptive response time for intrathecal (i.t.) injection of substance P (0.7 microg) was diminished by Agrimonia pilosa Ledeb extract. Intraperitoneal (i.p.) pretreatment with yohimbine (alpha2-adrenergic receptor antagonist) attenuated antinociceptive effect induced by Agrimonia pilosa Ledeb extract in the writhing test. However, naloxone (opioid receptor antagonist) or methysergide (5-HT serotonergic receptor antagonist) did not affect antinociception induced by Agrimonia pilosa Ledeb extract in the writhing test. Our results suggest that Agrimonia pilosa Ledeb extract shows an antinociceptive property in various pain models. Furthermore, this antinociceptive effect of Agrimonia pilosa Ledeb extract may be mediated by alpha2-adrenergic receptor, but not opioidergic and serotonergic receptors.

Genetic toxicity of Agrimonia Pilosa Ledeb in male mouse genital cells / 吉林大学学报(医学版)

0.05),indicating that Agrimonia Pilosa Ledeb could not induce any UDS in mouse sperms.Conclusion Agrimonia Pilosa Ledeb has no genetic toxicity in male mouse reproductive cells at the range of doses that are applied under the experimental conditions.

Effect of Agrimonia Pilosa Ledeb. Extract on the Growth of Food-Borne Pathogens

This study was performed to investigate the antimicrobial effect of the Agrimonia pilosa Ledeb. extracts against food-borne pathogens. First, the Agrimonia pilosa Ledeb. was extracted with methanol at room temperatures, and fractionation of the methanol extracts from Agrimonia pilosa Ledeb. was carried out by using petroleum ether, chloroform, and ethyl acetate, and methanol respectively. The antimicrobial activity of the Agrimonia pilosa Ledeb. extracts was determined using a paper disc method against food-borne pathogens and food spoilage bacteria. The petroleum ether extracts of Agrimonia pilosa Ledeb. showed the highest antimicrobial activity against Pseudomonas aeruginosa. The synergistic effect has been found in combined extracts of Agrimonia pilosa Ledeb. and Perillae folium as compared to each extracts alone. Finally, the growth inhibition curve was determined using ethyl acetate extracts of Agrimonia pilosa Ledeb. against Bacillus Cereus and Salmonella Enteritidis. The petroleum ether extract of Agrimonia pilosa Ledeb. showed strong antimicrobial activity against Bacillus Cereus at the concentration of 4,000 ppm. The 4,000 ppm of petroleum ether extract from Agrimonia pilosa Ledeb. retarded the growth of Bacillus Cereus more than 24 hours and Salmonella Enteritidis up to 36 hours. The petroleum ether extracts of Agrimonia pilosa Ledeb. has been shown the antimicrobial effect against Bacillus Cereus and Salmonella Enteritidis.