Yanliang Liu’s research while affiliated with Shandong University of Traditional Chinese Medicine and other places

Ethnopharmacological relevance Mahuang Xixin Fuzi Decoction (MXF), as a classical prescription of traditional Chinese medicine (TCM), has been used to treat the symptoms of fever, nasal congestion and headache in elderly people for almost a thousand years. Aim of the study The purpose of this study was to evaluate the effects and possible mechanisms of MXF on thermal stimulation-induced mouse cardiac myocytes (MCM) cell apoptosis. Materials and methods The apoptosis of the MCM cell model was induced by a PCR-calculated temperature control system with a gradual heating pattern at 43 °C for one hour. The cytotoxic effects were determined using real-time cell analyzer (RTCA) technology. Annexin V-FITC/7-AAD staining, and JC-1 fluorescence were used to assess apoptosis. Specific substrates, enzyme-linked immunosorbent assays (ELISAs), and Western blotting were used to identify proteins in the mitochondrial-mediated pathway. The identification of chemical components in the mouse heart was performed by ultra-performance liquid chromatography–electrospray ionization tandem mass spectrometry analysis. Results MXF inhibited apoptosis through the mitochondrial-mediated signaling pathway, including ameliorating MMP reduction, blocking mitochondrial Cyt C release, reducing Bax levels and increasing Bcl-2 levels, suppressing caspase-9 and caspase-3 activation in cytoplasmic fractions. Moreover, the components of MXF that act on the heart are mainly ephedra alkaloids and aconitine alkaloids. Conclusions The findings demonstrated that MXF treatment markedly reduced MCM cell apoptosis induced by thermal stimulation, which may be ascribed to the mitochondrial-mediated signaling pathway.

Background: Thermal stimulation is a potent inducer of apoptosis in many cell lines. At present, water bath heating is often used to thermal stimulation-induced apoptosis, but this method has poor reproducibility and low efficiency. The aim of this study was to build a simple and reproducible apoptosis model caused by thermal stimulation. Methods and Results: The apoptosis and necrosis rate were detected under different thermal stimulation conditions by flow cytometer. We also analyzed the difference of growth curves between the thermal stimulation model group and the normal group. According to the cell growth curve, the cell re-culture time after thermal stimulation was preliminarily detected, and the reasonability of the time was verified by flow cytometer. The conditions for establishing the model are: PCR calculated temperature control system with a gradual heating pattern, 37°C, 2 min; 38°C, 2 min; 39°C, 2 min; 40°C, 2 min; 41°C, 2 min; 42°C, 2 min; 43°C, 60 min; 42°C, 2 min; 41°C, 2 min; 40°C, 2 min; 39°C, 2 min; 38°C, 2 min; 37°C, 2 min. In this way, the proportion of cell apoptosis is higher than that of cell necrosis with satisfied reproducibility. Conclusions: The present work provide a new method to induce cell apoptosis by thermal stimulation.