v There exists a terrific require to set up powerful herbal formula for colorectal cancer treatment. According for the TCM theories and clinical observa tions, the pathogenesis of colorectal cancer is linked to damp heat, toxicity accumulation, definitely and spleen deficiency. Based mostly over the therapeutic system of clearing heat toxicity, eliminating dampness and tonifying Pi, as well as the contemporary principle of anticancer and anti angiogenesis, and TCM clinical practices, we have established an eight herbs composed formula for colorec tal cancer remedy, that's Teng Long Bu Zhong Tang. We now have demonstrated TLBZT may well inhibit proliferation, activate Caspases to induce apoptosis, upregulate p16 and p21 and downregulate RB phos phorylation to induce cell senescence in colon carcin oma cells in vitro.
In present review, we evaluated the anticancer results of TLBZT, made use of alone and in mixture with reduced dose of five Fluorouracil, in CT26 colon carcinoma in vivo. Methods Supplies DMEM medium and fetal bovine serum was obtained from Hyclone. five Fu injection was bought kind Xudong Haipu Pharmaceutical Co, Ltd. FragEL DNA Fragmentation Detection Kit was purchased from EMD Millipore. Senescence B Galactosidase Staining Kit and PARP anti physique were from Cell Signaling Engineering. Caspase three, Caspase eight and Caspase 9 Activity Assay Kit have been obtained from Beyotime Institute of Biotech nology. Antibody against p21 was pur chases from Boster Bio engineering Constrained Company. XIAP, Survivin, GAPDH and pRB antibodies were purchased from Bioworld Technologies. Antibody towards p16 was obtained from Proteintech.
Antibodies against CD31 and VEGF were the item of from Santa Cruz Biotech nology. Planning of TLBZT The herbs utilised in TLBZT formula are the roots of Actinidia chinensis 30 g, Solanum nigrum 15 g, Duchesnea indica 15 g, Atractylodes macro cephala Koidz 9 g, Poria cocos 15 g, Coix seed thirty g, Mistletoe 15 g, and Scutellaria barbata 30 g. All these herbs had been in the herb retail outlet in Longhua Hospital in accordance to your unique proportion, and decocted twice with eight fold volume of distilled water for one hour. The de coction have been collected, filtered, merged and concen trated to one. 5 g mL, and stored at four C. For Fuel chromatography mass spectrometry analysis, TLBZT have been even further extracted with dichloromethane and diethyl ether, and passed by 0. 22 um filter.
GC MS analysis of TLBZT extract was carried out by GCMS6800 equipped having a DB 5ms column. Helium was made use of as carrier fuel at a constant movement fee of 1 mL min. An injection volume of 1 uL was employed in splitless mode. Injector and ion supply have been maintained at 280 C and 230 C, respectively. The mass scan selection was 50 500. The GC MS profile of TLBZT is presented in More file 1 Figure S1. Cell culture and animal model Murine colon carcinoma CT26 cells had been obtained from obtained from Cell Financial institution of Type Culture Assortment of Chinese Academy of Sciences.