Effect of Modified Shaoyao Gancao Decoction on Ultrastructure and Apoptosis of Human Adenomyosis Cells in Vitro
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Modified Shaoyao Gancao Decoction altered adenomyosis cell ultrastructure and increased apoptosis in vitro, suggesting a potential therapeutic mechanism.
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Abstract
Objective To observe the effect of modified Shaoyao Gancao Decoction(MSGD) on the ultrastructure and apoptosis of human adenomyosis cells in vitro. Methods Focal human adenomyosis tissues were digested with collagenase I for primary culture, and then the isolated adenomyosis cells were identified by immunocytochemistry.Adenomyosis cells were seperately treated with MSGD at the concentration of 40, 20, 10 mg/mL(MSGD groups),DMEM(blank control group), or mifepristone at the concentration of 1×10-5mol/L( positive control group) for24 and 48 hours. The ultrastructure of adenomyosis cells after intervention by MSGD and mifepristone was observed by transmission electron microscope, and the apoptotic rate was evaluated by flow cytometry(FCM).Results Under transmission electron microscope, adenomyosis cells had condensed chromatin, shrinked nucleus,increased lysosome count in cytoplasm, dilated mitochondria and endoplasmic reticulum in MSGD groups and mifepristone group. After treatment with mifepristone and MSGD for 24 and 48 hours, the apoptotic rate was significantly increased(P0.05 compared with blank control group). The apoptotic rate in mifepristone group and high-concentration MSGD group were significantly higher than that in medium- and low-concentration MSGD groups after culture for 24 and 48 hours(P0.05). MSGD groups after culture for 48 hours had higher apoptotic rate than that after 24 hours(P0.05), showing certain dose- and time-dependent effect. Conclusion The therapeutic mechanism of MSGD is probably related with the alteration of ultrastructure of human adenomyosis cells and with the increase of their apoptotic rate.
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