Hunan Fangsheng Pharmaceutical Co., Ltd.

We previously reported that activation of the cell cycle in human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) enhances their remuscularization capacity after human cardiac muscle patch transplantation in infarcted mouse hearts. Herein, we sought to identify the effect of magnesium lithospermate B (MLB) on hiPSC-CMs during myocardial repair using a myocardial infarction (MI) mouse model.

In C57BL/6 mice, MI was surgically induced by ligating the left anterior descending coronary artery. The mice were randomly divided into five groups (n = 10 per group); a MI group (treated with phosphate-buffered saline only), a hiPSC-CMs group, a MLB group, a hiPSC-CMs + MLB group, and a Sham operation group. Cardiac function and MLB therapeutic efficacy were evaluated by echocardiography and histochemical staining 4 weeks after surgery. To identify the associated mechanism, nuclear factor (NF)-κB p65 and intercellular cell adhesion molecule-1 (ICAM1) signals, cell adhesion ability, generation of reactive oxygen species, and rates of apoptosis were detected in human umbilical vein endothelial cells (HUVECs) and hiPSC-CMs.

After 4 weeks of transplantation, the number of cells that engrafted in the hiPSC-CMs + MLB group was about five times higher than those in the hiPSC-CMs group. Additionally, MLB treatment significantly reduced tohoku hospital pediatrics-1 (THP-1) cell adhesion, ICAM1 expression, NF-κB nuclear translocation, reactive oxygen species production, NF-κB p65 phosphorylation, and cell apoptosis in HUVECs cultured under hypoxia. Similarly, treatment with MLB significantly inhibited the apoptosis of hiPSC-CMs via enhancing signal transducer and activator of transcription 3 (STAT3) phosphorylation and B-cell lymphoma-2 (BCL2) expression, promoting STAT3 nuclear translocation, and downregulating BCL2-Associated X, dual specificity phosphatase 2 (DUSP2), and cleaved-caspase-3 expression under hypoxia. Furthermore, MLB significantly suppressed the production of malondialdehyde and lactate dehydrogenase and the reduction in glutathione content induced by hypoxia in both HUVECs and hiPSC-CMs in vitro.

MLB significantly enhanced the potential of hiPSC-CMs in repairing injured myocardium by improving endothelial cell function via the NF-κB/ICAM1 pathway and inhibiting hiPSC-CMs apoptosis via the DUSP2/STAT3 pathway.

Seven hexahydrodispiro[indole-3,3′-indolizine-2′,3″-piperidine]-2(1 H),4″-dione compounds were synthesized by a catalyst-free 1,3-dipolar cycloaddition reaction in a one-pot three-component system containing 3,5-diarylidene-1-benzylpiperidin-4-one, isatin and l-pipecolic acid. The structures of these compounds were characterized by nuclear magnetic resonance, high-resolution mass spectrometry and X-ray data analysis. These chemical entities were evaluated for their cytotoxicity (brine shrimp assay), and five compounds were found potential in this bioassay. Compound 2e was the most prominent target against Artemia salina with an LC50 value of 7.27 μg/mL.

OBJECTIVE To establish an LC-MS/MS method for simultaneous determination of irbesartan(IBST) and hydrochlorothiazide(HCZ) concentrations in human plasma and to be used for a bioequivalence study of IBST/HCZ tablets.METHODS Plasma samples were pre-treated by protein precipitationIBST-d4 and chlorothiazide(MCTZ) were used as the internal standardThe analytes were retained and eluted by using a Waters ACQUITY UPLC∼?XTERRA RP_(18)(4.6 mmx100 mm, 3.5 μm) column, with a binary mobile phase system consisting of acetonitrile(A) and 5 mmol·L∼(-1) ammonium acetate solution(B)at a flow rate of 0.85 mL·min∼(-1).The analytes were detected in an electrospray ionization source under neg. ion mode, using the ion transitions of m/z 427.1→193.1(IBST), m/z 431.2→193.1(IBST-d4), m/z 295.9→268.9(HCZ) and m/z 357.8→322.0(MCTZ).The quantitation method was applied to a bioequivalence study of IBST/HCZ tablets(150 mg/12.5 mg) in healthy subjects.RESULTS Endogenous substances in plasma did not interfere with the quantification of the substance to be measured and no interference and transformation on and from IBST, HCZ, IBST-d4 and MCTZ.The quant. ranges of IBST and HCZ in human plasma were 10-4 000 ng·mL∼(-1) and 1-400 ng·mL∼(-1), resp., with good linearity.The extraction recoveries of IBST and IBST-d4 were between 95.7% and 105.4%, HCZ and MCTZ were between 95.0% and 106.4%;the internal standard normalization matrix effect factors of IBST and HCZ were recorded as 101.2%-102.6%, and 79.8%-93.6% all with coefficient of variation(CV)<5%, resp.Intra-and inter-batch accuracy and precision were all within 15%, CV<10%.The concentrations of IBST and HCZ determined by the method and the pharmacokinetic parameters calculated thereafter were in accordance with those found in previous reports, and the difference of concentrations generated in the incurred sample reanal. assay and the original measurement was <10%.CONCLUSION The method is rapid, specific, sensitive, accurate, and reproducible requiring a small sample volumn, and fully meets the requirements of high-throughput quant. anal. of IBST/HCZ tablets bioequivalence studies.