Intervention effect of Cigu Xiaozhi prescription on ceramide lipoapoptosis in non-alcoholic fatty liver disease.

OBJECTIVE: To explore the mechanism of the Chinese medicine Cigu Xiaozhi prescription (, CGXZ) in the treatment of the non-alcoholic fatty liver disease (NAFLD) by detoxification and phlegm-reducing, the effect of CGXZ prescription on ceramide-mediated lipid apoptosis in Hep G2 cells with NAFLD. METHODS: The experiment was randomly divided into 6 groups: normal control group, model group, CGXZ prescription medicated serum high, medium, and low dose groups, and pioglitazone positive control group. Using 500 µmol/L free fatty acid (FFA) mixture to induce Hep G2 cells to establish NAFLD cell model, respectively, with 2%, 4%, and 6% concentration of CGXZ prescription medicated serum intervention for 24 h. The changes in organelles and lipid droplet accumulation were observed under the electron microscope. Furthermore, TdT-mediated dUTP Nick-End Labeling method was used to assay hepatocyte apoptosis; Biochemical determination of glutamic-pyruvic transaminase, glutamic oxalacetic transaminase, triglycerides, and FFA levels in Hep G2 cells; the content of ceramide was determined by high-performance thin-layer chromatography. Finally, Western Blot and quantitative real-time polymerase chain reaction (qRT-PCR) were used to determine the protein and gene expression levels, such as inducible nitric oxide synthase (iNOS), nuclear factor κB (NF-κB), B cell lymphoma 2 (Bcl-2) and Bcl-2-associated X (Bax). RESULTS: Under the electron microscope, the cells in the model group showed moderate-to-severe steatosis, and apoptotic bodies could be seen. The model group had greater improvements in the apoptosis rate (P < 0.01), and the levels of ceramide C2 and FFA in the cytoplasm (P < 0.01) than the normal control group. The protein expressions of NF-κB, iNOS, and Bax were significantly up-regulated (P < 0.05), while the Bcl-2 had no significant change (P > 0.05). Compared with the model group, the levels of ceramide C2 and FFA (P < 0.01), the protein expressions of NF-κB, iNOS, and Bax (P < 0.05) in the CGXZ prescription treatment group and pioglitazone positive control group were significantly decreased; Only the Bcl-2 protein was significantly up-regulated in the high-dose Chinese medicine group (P < 0.05). The down-regulation of Bax mRNA expression in each Chinese medicine treatment group was significantly better than in the pioglitazone positive control group (P < 0.01). CONCLUSIONS: The CGXZ prescription, formulated with the method of detoxification and phlegm, can inhibit lipoapoptosis in the NAFLD cell model by down-regulating the levels of ceramide C2 and FFA, which may be achieved by regulating ceramide/iNOS/NF-κB signaling pathway.

[[Technology Surgical Intervention in Ulcerative Colitis Modeling Descending Part of the Colon in Rats Line "Wistar"].]

BACKGROUND: Ulcerative colitis (UC) is a chronic immune inflammatory condition of the colon with an unknown aetiology, leading to disability and reduced quality of life of patients. UC primarily affects young adults. In most cases, inflammatory bowel disease (iBD) debuts at reproductive age. The incidence of UC and severe clinical course has increased overall across the world. The study of the mechanisms of pathogenesis and aetiology of this disease contributes to the development of new effective methods of treatment. OBJECTIVE: The aim of our study was to develop technology of the surgery directed to induction of reversible ischemic damage, the erosive-ulceration of gut mucosae (descending colon) at rats of the WISTAR line. MATERIALS AND METHODS: Experimental research was done using male rats < (n=40), their age was more than 6 months, weight of body is 280 - 350 grams. The animals were separated into 3 groups: 1 - (n=14) Sholimov's operative method, 2 group (n=16) - model using special technology, 3 group (n=10) - Coopr's H. S. induction ulcerative colitis. Observation was been done during 10 days. We offered special surgical technique: parietal ligation on every vasa recta of colon descendens of rats the length 3 cm along colon, on the apex vesicae and then vasa recta are cut. Then during 7 days 1% solution of dextran sulfate sodium (DSN) was given for experimental animals after postoperative period. By the histolog- ical biopsy of colon mucosa the results were estimated. RESULTS/CONCLUSION: we have created erosive-ulceration of gut mucosae (descending colon) of rat with productive inflam- mation, vasculitis and plasmatic impregnation of the vessel walls and intimal thickening. Our model of ulcerative colitis can be used for the development and testing of new methods for the study and treatment of this pathology.