Asperuloside Promotes Apoptosis of Cervical Cancer Cells through Endoplasmic Reticulum Stress-Mitochondrial Pathway / 中国结合医学杂志

OBJECTIVE@#To investigate the effects of asperuloside on cervical cancer based on endoplasmic reticulum (ER) stress and mitochondrial pathway.@*METHODS@#Different doses (12.5-800 µg/mL) of asperuloside were used to treat cervical cancer cell lines Hela and CaSki to calculate the half maximal inhibitory concentration (IC50) of asperuloside. The cell proliferation was analyzed by clone formation assay. Cell apoptosis, intracellular reactive oxygen species (ROS) and mitochondrial membrane potential were determined by flow cytometry. The protein expressions of cleaved-caspase-3, Bcl-2, Bax, Cyt-c, cleaved-caspase-4 and glucose-regulated protein 78 (GRP78) were analyzed by Western blot. And the inhibitor of ER stress, 4-phenyl butyric acid (4-PBA) was used to treat cervical cancer cells to further verify the role of ER stress in the apoptosis of cervical cancer cells induced by asperuloside.@*RESULTS@#Asperuloside of 325, 650, and 1300 µg/mL significantly inhibited the proliferation and promoted apoptosis of Hela and CaSki cells (P<0.01). All doses of asperuloside significantly increased intracellular ROS levels, reduced mitochondrial membrane potential, significantly reduced Bcl-2 protein expression level, and increased Bax, Cyt-c, GRP78 and cleaved-caspase-4 expressions (P<0.01). In addition, 10 mmol/L 4-PBA treatment significantly promoted cell proliferation and reduced apoptosis (P<0.05), and 650 µg/mL asperuloside could reverse 4-PBA-induced increased cell proliferation, decreased apoptosis and cleaved-caspase-3, -4 and GRP78 protein expressions (P<0.05).@*CONCLUSION@#Our study revealed the role of asperuloside in cervical cancer, suggesting that asperuloside promotes apoptosis of cervical cancer cells through ER stress-mitochondrial pathway.

Establishment of UPLC fingerprints of Zhidong Granules and determination of three constituents / 中草药

Objective To establish the UPLC fingerprints of Zhidong Granules (ZG) and determine the content of three constituents. Methods With baicalin as a reference peak, the UPLC fingerprints of 11 batches of samples were established. The analysis of ZG was performed on a 40 ℃ thermostatic Acquity HSS T3 column (150 mm × 2.1 mm, 1.8 μm), with the mobile phase comprising of acetonitrile-0.1% formic acid flowing at 0.3 mL/min in a gradient elution manner, and the detection wavelength was set at 254 nm. Results The UPLC fingerprint was established and 16 components were identified (gallic acid, asperuloside, methyl deacetylcyrrhizin, chlorogenic acid, genipin 1-gentiobioside, geniposide, paeoniflorin, chrysin-6-C-β-D-glucopyranosyl-8-C-α-L- arabinopyranoside, chrysin-6-C-α-L-arabopylanose-8-C-β-D-glucoside or isomer, baicalin, baicalin isomer, oroxylin A-7-O- glucuronide, wogonoside, wogonin, and schizandrin). The similarity of 11 batches of preparations were at least 0.98. Baicalin, paeoniflorin, and geniposide showed good linear relationships within the ranges of 12.15-388.80 μg/mL, 6.52-209.00 μg/mL, and 8.38-268.00 μg/mL, whose average recoveries (RSD) were 100.98% (3.04%), 100.49% (0.60%), and 100.55% (2.73%), respectively. The content of baicalin in the 11 batches of preparations was between 7.46 and 12.40 mg/g, the content of geniposide was between 7.01 and 13.27 mg/g, and the content of paeoniflorin was between 7.68 and 12.76 mg/g. Conclusion The method is accurate, simple, stable, and reliable, and can be used for quality control of ZG.

Anti-angiogenic activity of iridoids from Galium tunetanum

ABSTRACT The phytochemical study of Galium tunetanum Lam., Rubiaceae, leaves led to the isolation of 13 compounds from the chloroform-methanol and the methanol extracts, including six iridoid glycosides, one non-glycoside iridoid, two p-coumaroyl iridoid glycosides, two phenolic acids, and two flavonoid glycosides. The structural determination of the isolated compounds was performed by mono- and bidimensional NMR spectroscopic data, as well as ESI-MS experiments. All compounds were isolated from this species for the first time. The anti-angiogenic effects of the isolated iridoids were also reported on new blood vessels formation using the chick embryo chorioallantoic membrane as in vivo model. Results showed that among the isolated iridoids tested at the dose of 2 µg/egg, asperuloside (1), geniposidic acid (2), and iridoid V1 (3) reduced microvessel formation of the chorioallantoic membrane on morphological observations using a stereomicroscope. The anti-angiogenic effects of the active compounds, expressed as percentages of inhibition versus control, were 67% (1), 59% (2), and 54% (3), respectively. In addition, the active compounds were able to inhibit angiogenesis in the chorioallantoic membrane assay, in a dose-dependent manner (0.5-2 µg/egg) as compared to the standard retinoic acid.