Apoptotic and Anti-Inflammatory Effects of Eupatorium japonicum Thunb. in Rheumatoid Arthritis Fibroblast-Like Synoviocytes

Figure 3

CHOP was involved in EJT extract-mediated apoptosis in MH7A cells. (a) MH7A cells were pretreated with 37.5 μg/ml EJT extract for the indicated time. The cell lysates were subjected to SDS-PAGE and analyzed by immunoblotting for antibodies specific to ATF4, CHOP, and GAPDH. (b) MH7A cells were pretreated with 5 mM NAC for 2 h and then treated with 37.5 μg/ml EJT extract for the indicated time. Immunoblot analyses were performed using specific antibodies. The fold increase in p-eIF2α expression is the ratio of the p-eIF2α to eIF2α. (c, d) The relative mRNA levels of ATF4 and CHOP were measured by using real-time quantitative PCR. Significant differences were indicated by or . (e) MH7A cells were transfected with EGFP- or CHOP-specific shRNA. The cells were treated with indicated concentrations of EJT extract for 24 h and an MTT assay was performed to determine cell viability. Significant differences were indicated by . (f) The cell lysates were subjected to SDS-PAGE and analyzed by immunoblotting with specific antibodies.

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