Berberine and S allyl cysteine mediated amelioration of DEN+CCl4 induced hepatocarcinoma

• Published in: Biochimica et biophysica acta Vol. 1840; no. 1; pp. 219 - 244
• Main Authors: Sengupta, Dipanwita, Chowdhury, Kaustav Dutta, Sarkar, Avik, Paul, Soumosish, Sadhukhan, Gobinda Chandra
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.01.2014
• Subjects: Alkylating Agents - toxicity; animal models; Animals; Antineoplastic Agents - pharmacology; apoptosis; Apoptosis - drug effects; Berberine; Berberine - pharmacology; Blotting, Western; Carbon tetrachloride; Carbon Tetrachloride - toxicity; Carcinoma, Hepatocellular - chemically induced; Carcinoma, Hepatocellular - metabolism; Carcinoma, Hepatocellular - prevention & control; Caspase 3 - metabolism; cell proliferation; Cells, Cultured; cysteine; Cysteine - analogs & derivatives; Cysteine - pharmacology; Cytochrome P-450 CYP2E1 - metabolism; Cytochromes c - metabolism; Di ethyl nitrosamine; diethylnitrosamine; Diethylnitrosamine - toxicity; Flow Cytometry; fluorescence; Hepatocytes - cytology; Hepatocytes - drug effects; Hepatocytes - metabolism; herbal medicines; Histone Deacetylase 1 - metabolism; Immunoenzyme Techniques; Immunoprecipitation; liver neoplasms; Liver Neoplasms - chemically induced; Liver Neoplasms - metabolism; Liver Neoplasms - prevention & control; Male; Membrane Potential, Mitochondrial - drug effects; Mice; Mitochondrial Membrane Transport Proteins; mitogen-activated protein kinase; Oxidative stress; Oxidative Stress - drug effects; p53 acetylation; physiology; precipitin tests; Reactive Oxygen Species - metabolism; S allyl cysteine; therapeutics; NADPH; Oxidative stress; pMdm2; B; PCNA; PT; HDAC1; JNK; p53 acetylation; Carbon tetrachloride; FACS; HO1; S+B; BSA; FITC; S; Nrf2; S allyl cysteine; NaCN; Berberine; DTNB; SOD; PP2A; GR; MMP; NBT/BCIP; POT; ROS; DCFDA; Di ethyl nitrosamine; CYP2E1; fluorescence activated cell sorter; berberine treated group; hemoxygenase1; phosphorylated Mdm2; nuclear factor erythroid 2 related factor 2; glutathione reductase; protein phosphatease 2A; SAC treated group; fluorochrome iso-thiocyanate; mitochondrial membrane potential; post treated group; 5,5-dithiobis (2 nitrobenzoic acid); proliferating cell nuclear antigen; superoxide dismutase; cytochromeP4502E1; nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl phosphate; bovine serum albumin; histone deacetylase type1; sodium cyanide; 2′,7′-Dichlorofluorescein diacetate, TCA, tri choloro-acetic acid; c-Jun N terminal kinase; reactive oxygen species; co-treated group; nicotinamide adenine dinucleotide 2′-phosphate reduced; treatment with combination of SAC and berberine;NS, Non significant
• DOI: 10.1016/j.bbagen.2013.08.020
• ISSN: 0304-4165; 0006-3002

Diethylnitrosamine (DEN) and carbon tetrachloride (CCl4) have been used as initiator and promoter respectively to establish an animal model for investigating molecular events appear to be involved in development of liver cancer. Use of herbal medicine in therapeutics to avoid the recurrence of hepatocarcinoma has already generated considerable interest among oncologists. In this context studies involving S-allyl-cysteine (SAC) and berberine have come up with promising results. Here we have determined the individual effect of SAC and berberine on the biomolecules associated with DEN+CCl4 induced hepatocarcinoma. Effective therapeutic value of combined treatment has also been estimated. ROS accumulation was analyzed by FACS following DCFDA incubation. Bcl2-Bax and HDAC1‐pMdm2 interaction were demonstrated by co-immunoprecipitation. Immunosorbent assay was performed to analyze PP2A and caspase3 activities. MMP was determined cytofluorimetrically by investigating JC-1 fluorescence. AnnexinV binding was demonstrated by labeling the cells with AnV-FITC followed by flow cytometry. CytochromeP4502E1 mediated bioactivation of DEN+CCl4 induced Akt dependent pMdm2‐HDAC1 interaction that led to p53 deacetylation, probable cause of its degradation. In parallel, oxidative stress dependent Nrf2‐HO1 activation increased Bcl2 expression which in turn stimulated cell proliferation. SAC in combination with berberine inhibited Akt mediated cell proliferation. Activation of PP2A as well as inhibition of JNK resulted in induction of apoptosis after 30days of treatment. Extension of combined treatment reverted tissue physiology towards control. Co-treated group displayed normal tissue structure. SAC and berberine mediated HDAC1/Akt inhibition implicates the efficacy of combined treatment in the amelioration of DEN+CCl4 induced hepatocarcinoma. •SAC+berberine reduced Bcl2/Bax heterodimerization after 30days post-treatment.•Treatment restrained Akt dependent pMdm2–HDAC1 interaction.•Retention of p53 acetylation provoked Bax mediated cell death.•60days post-treatment reduced PCNA expression and restored hepatic structure.•SAC+berberine co-treatment maintained liver tissue alike control animal.