研究目的
To characterize the response of human synoviocyte MH7A cells to visible LED red light in an attempt to elucidate the associated action mechanism.
研究成果
LED red light (630 nm) inhibits proliferation and migration of MH7A cells. The growth-inhibiting effects of LED red light on human synoviocyte MH7A cells appear to be associated with regulation of the TRPV4/PI3K/AKT/mTOR signaling pathway.
研究不足
The specific role of TRPV4 channel in the mechanism of 630-nm LED light inhibiting the proliferation and cytokine secretion of synoviocyte MH7A cells still needs to be further studied.
1：Experimental Design and Method Selection:
Human synoviocyte MH7A cells were treated with 630-nm LED light after stimulation of tumor necrosis factor-α (TNF-α). The effects of light radiation on cell proliferation and migration were detected by MTT assay and scratch test. The expressions of inflammatory cytokines were measured using RT-qPCR. This was followed by detection of the levels of extracellular proteins IL-6 and IL-8 after differential radiation. Furthermore, the expression levels and activation of proteins on PI3K/AKT/mTOR signaling pathway were examined with Western blot.
2：Sample Selection and Data Sources:
Human fibroblast–like synoviocyte line MH7A was purchased from GuanDao Biological Research Center of Shanghai.
3：List of Experimental Equipment and Materials:
Light-emitting diodes (CP-YL3213-630, Truwin Optoelectronic Medical, Beijing, China), absorbance reader (SpectraMax 340PC, Molecular Devices, Sunnyvale, CA), NIKON DS-Fi2-L3 (magnification, × 40), LightCycler? 96 thermal cycler Instrument (Roche Applied Science, Penzberg, Germany), NuPAGE? Novex Gel Electrophoresis system (10% gels, Invitrogen).
4：Experimental Procedures and Operational Workflow:
Cells were exposed through the bottom of plastic culture plates to the 630-nm radiation. The time it takes to radiation once was 5, 10, and 15 min to achieve radiation energies of 13 J/cm2, 26 J/cm2, and 39 J/cm2, respectively.
5：Data Analysis Methods:
Statistical analyses of the data were performed using the GraphPad Prism (version 5.0) statistical program.
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获取完整内容-
Light-emitting diodes
CP-YL3213-630
Truwin Optoelectronic Medical
To deliver 630-nm radiation to human synoviocyte MH7A cells
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absorbance reader
SpectraMax 340PC
Molecular Devices
To measure the absorbance at 490 nm to determine the viability of cells
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NIKON DS-Fi2-L3
NIKON
To take images of scratch healing under magnification, × 40
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LightCycler? 96 thermal cycler Instrument
Roche Applied Science
To carry out PCR on a thermal cycler instrument
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NuPAGE? Novex Gel Electrophoresis system
10% gels
Invitrogen
To perform gel electrophoresis
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