ABSTRACT
ObjectiveTo investigate the effect and mechanism of Yiyi Fuzi Baijiangsan （YYFZBJ） on the apoptosis of colon cancer cell line HCT116. MethodYYFZBJ at different concentrations （0.5， 1， 2， 4， 6， 8， 10， 12， 14， 16 g·L-1） was used to intervene in HCT116 cells for 24， 48， 72 h. The cell counting kit-8 （CCK-8） method was used to determine the effect of YYFZBJ on cell proliferation in vitro. The cells were divided into a blank group， a capecitabine group（1.8 g·L-1）， and low-， medium-， and high-dose YYFZBJ groups （6， 10， and 14 g·L-1） and treated for 48 hours. Flow cytometry was used to detect the apoptosis. Hoechst 33342 staining was used to observe the apoptotic morphology of cells. Mitochondrial membrane potential （MMP） was analyzed by a mitochondrial-targeted deep-red fluorescent probe （Mito-Tracker Red CMXRos）. The expression of proteins related to the mitochondrial apoptosis pathway， such as B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， cytochrome C （Cyt C）， cysteinyl aspartate-specific protease （Caspase）-9， Caspase-3， cleaved Caspase-9， and cleaved Caspase-3 was detected by Western blot. The mRNA levels of Bcl-2， Bax， Cyt C， Caspase-9， and Caspase-3 were determined by real-time polymerase chain reaction （Real-time PCR）. ResultCompared with the blank group， YYFZBJ （8， 10， 12， 14， 16 g·L-1） significantly inhibited the proliferation of HCT116 cells in vitro （P<0.05） in a dose-dependent manner. Compared with the blank group， the medium- and high-dose YYFZBJ groups and the capecitabine group showed increased apoptosis rates of colon cancer cells （P<0.05）. The YYFZBJ groups and the capecitabine group showed reduced number of colon cancer cells with significantly changed cellular morphology and cell apoptosis manifestations， such as strong dark blue fluorescence， nucleus concentration， shrinkage， and fragmentation. With the increase in the mass concentration of YYFZBJ， the blue fluorescence intensity was significantly enhanced. Compared with the blank group， the YYFZBJ groups and the capecitabine group showed reduced MMP in a dose-dependent manner， decreased protein and mRNA levels of Bcl-2， and increased protein expression of Bax， Cyt C， Caspase-9， Caspase-3， cleaved Caspase-9， and cleaved Caspase-3 and mRNA expression of Bax， Cyt C， Caspase-9， and Caspase-3 （P<0.05）. ConclusionYYFZBJ can induce the apoptosis of colon cancer HCT116 cells through the mitochondrial apoptosis pathway.
ABSTRACT
Yiyi Fuzi Baijiangsan is scattered from the Essentials from the Golden Cabinet （《金匮要略》） for the treatment of intestinal carbuncles. The whole prescription is composed of Coicis Semen， Aconiti Lateralis Radix Praeparata， and Patrinia scabiosifolia， with the effect of invigorating spleen， warming Yang， clearing heat， removing dampness， and detoxification， which is a commonly used prescription for the clinical treatment of ulcerative colitis （UC）. UC is a chronic nonspecific inflammatory disease with lesions involving the colorectal mucosa， and the etiology is not yet very clear， which is mostly related to genetics， external and intestinal environment， immunity， infection， and other factors. Animal experiments and clinical studies have shown that Yiyi Fuzi Baijiangsan have the advantages of multi-target and multi-faceted treatment of UC. At present， the research mechanism of the treatment of UC is mainly focused on reducing intestinal inflammatory response， anti-colorectal cancer effect， alleviating oxidative stress， repairing the intestinal epithelial cell barrier， improving intestinal flora disorder， inhibiting apoptosis， maintaining intestinal immune balance， etc. Clinically， the combination of modified Yiyi Fuzi Baijiangsan and western medicine has a satisfactory effect， which can significantly improve the relevant clinical symptoms of patients with UC， delay the condition， and improve the quality of life of patients， with the advantages of high safety and small side effects. Its related research provides theoretical support and data support for the clinical prevention and treatment of UC and the follow-up exploration of the mechanism of Yiyi Fuzi Baijiangsan in the treatment of UC， and is also of great significance to the research on the treatment of UC with Chinese medicine. This paper reviewed the prevention and control mechanism of Yiyi Fuzi Baijiangsan in the treatment of UC.