| In recent years, the alcoholism patients grow in number. Among them, aboutone-to two-thirds have impairment of muscle function. The toxic effect of ethanol isthe most important etiological factor for alcoholic myopathy since there's not ethanoldehydrogenase in skeletal muscle cells. In the experiments in chapter 1, we study thedirect effect of acute treatment of ethanol at different concentrations on theproliferation of mouse C3H muscle myoblasts, C2C12 cells, and the cell cycle ofC2C12 myoblasts treated with 100mM ethanol for 24 hours.Many reports have confirmed that both low intensity laser irradiation andmonochromatic light had noticeable effects on the biological systems which are awayfrom homeostasis. It could initiate cellular signaling transduction pathway no matterby visible light or by infrared light. Light emitting diode array (LED) ofpredominance is most possible to take the place of laser irradiation in the biomedicinedomain in the future. In the experiments in chapter 2, red light (640±17nm) fromLED (RLED) was used to irradiate ethanol-treated C2C12 myoblasts to studyphotobiomodulation (PBM) of low intensity RLED on ethanol-inhibited cellproliferation.1. The effect of ethanol on the C2C12 myoblast proliferationEthanol-inhibited myoblast proliferation might be one pathogenesis of alcoholicmyopathy. The C2C12 myoblasts were incubated with different concentration ethanolfor 4, 8, 16 or 24 hours, respectively. The cell proliferation was evaluated by3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Theresults showed that the higher the concentration of the ethanol used to treat C2C12myoblasts, the more significant the inhibition of cell proliferation, when C2C12myoblasts were treated for 4, 8, 16 and 24 hours. The ethanol at 150mM or 200mMsignificantly inhibited cell proliferation when the incubation time was 4 hours(P＜0.05). The ethanol at all concentrations significantly inhibited cell proliferationwhen the cells were treated for longer than 8 hours (P＜0.05). The C2C12 myoblastswere incubated with ethanol at 50, 100, 150 and 200 mM for different time, respectively. The results showed that the most significant inhibition of cellproliferation when cells were incubated for 8 hours (P＜0.01). It demonstrated thatC2C12 myoblast proliferation would be inhibited significantly by acute ethanoltreatment.C2C12 myoblasts treated with ethanol at 100mM for 24 hours were stained withpropidium iodide, and then evaluated with Flow Cytometry. The results indicated thatG0/G1 phase, S phase and G2/M phase had not significant difference compared withcontrol.2. Photobiomodulation of low-intensity red light from light emitting diode array onthe proliferation of ethanol-treated C2C12 myoblastsThe function of C2C12 myoblasts in homeostasis is normal when they are nottreated with ethanol, and the cell proliferation could not affected by low intensityRLED. Ethanol might disrupt cellular homeostasis so that PBM could rehabilitate thecellular dysfunction.When C2C12 myoblasts were treated with ethanol at 150mM for 4 hours or 8hours, the inhibited proliferation could be rehabilitated significantly by RLED at1134mJ/cm2 (1.26mW/cm2, 900s) (P＜0.05). When C2C12 myoblasts were treatedwith ethanol at 100mM for 16 hours, the inhibit proliferation could be rehabilitatedsignificantly by RLED at 252 mJ/cm2 (0.28mW/cm2, 900s) (P＜0.05). When C2C12myoblasts were treated with ethanol at 100mM for 24 hours, the inhibit proliferationcould be rehabilitated significantly by RLED at both 495mJ/cm2 (0.55mW/cm2, 900s)(P＜0.05) and 576mJ/cm2 (0.64mW/cm2, 900s) (P＜0.01), and there was a linearcorrelation between the PBM of the cell proliferation and RLED intensity (from0.28mW/cm2 to 0.64mW/cm2) (P＜0.05). The experiments results supported thebiological information model of PBM.Conclusions: The C2C12 myoblast proliferation could be significantly inhibitedby acute ethanol treatment, and then be significantly rehabilitated by RLED.
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