| 1.PurposeThis experiment for viral hepatitis, using the pathogenesis of immunological mechanisms BCG plus LPS polysaccharides joint induction reexposed liver damage model, by means of measuring the animal serum ALT, AST value, liver tissue superoxide dismutase (SOD) vigor, lipid peroxide malondialdehyde (MDA), liver tissue pathology form and ultrastructure, peroxidation damage and immune adjusting etc through, and do statistical analysis, evaluation NHS on immunity liver injury protection effect, and discuss the mechanism for NHS capsule resistance to damage the liver immunity provides scientific basis for NHS and participation in clinical application and development of capsule to provide the basis.2.MethodsKunming mice only 60 were randomly divided into normal group, BCG add LPS model group, Biphenyl Dicarboxylate group, NHS capsule of high,medium and low doses groups of 10 only. Methods:according to the weight will be 60 mice dungenzhai yishu 1-60#, with DPS7.55 statistical software for processing:experimental design and the complete randomized-60, grouping experiments sample number of groups 6, OK.Laboratory feeding a week, reference method made modules, except normal control group outside, the rest group first day all tail intravenous BCG2.5mg/0.2ml. one (about 5×107 living bacterium) made moulds. Normal control group, BCG add LPS model group,tap water 0.5ml. ig 10 days; only, Biphenyl Dicarboxylate group 12mg/kg doses, ig 10 days. NHS capsule is tall, medium, low doses groups were given NHS 3400mg/kg,1700mg/kg,850mg/kg gastric lavage 10 days. Besides the normal control group outside, each animal in all the end time after the treatment, tail intravenous LPS 7.5μg/0.2 ml one. Methods:first mouse fixed in mice holder, let the tail to set all dew outside, with 75% ethanol is wiped the tail, make its hyperemia. Choose a root most plentiful vein, left-handed means and index finger knead tail end, right hand holding the syringe needle with 4, with 30 degree Angle were venipuncture push note solution frictionless feeling, and visible along the veins appear a white line, indicates that needle indeed inside blood-vessel, can continue to push finish solution. After completion, uproot needle injection, lightly in injection site bleeding. After injection fasting not water, after 10 hours for model processing.Mice with 0.03g/10g hydration after anesthesia, orbit chlorine aldehyde take blood 1.0 ml, with sterile tubes collection, centrifugal serum, blood biochemistry analyzer, the automatic detection ALT, AST level. Former says mice weight, blood collection after death mice, anatomical dislocated take liver and spleen weighing, calculation, liver and spleen visceral index. Mice executed after take liver tissue, using formaldehyde is fixed, the conventional 20% paraffin embedding slice, did HE histopathological staining light microscopy, consult immunity liver injury experimental study relevant literatures and liver tissue morphological related research literature, according to the liver tissue inflammation and necrosis and haemorrhage degree, will the liver tissue pathology change five levels.Cut take left medial liver tissue 0.5-1.0g, cold saline flush filter paper blot, weighing, ice bath of liver homogenate made by 10%,4000 turn/min, cryogenic centrifugal 15min, fetch the supernatant fluid. According to the instruction using glucosinolate and detect malondialdehyde (mda), determining SOD activity. Its operation step by kit instructions. TNF-cactivity, IL-6 active detection by ELISA, press kit that operation. Experimental data with mean+standard deviations (X±S) said, and in groups; t test comparison:Comparison:analysis of variance between groups. P<0.05 think differences are statistically significant. With SPSS 13.0 statistical software analysis.3.Results(1) The treatment group ALT, AST value significantly lower than that in model group(P<0.05),indicating that NHS capsules can effectively prevent the immunity liver damage caused by the liver function decreases.(2) The treatment group pathological structure obviously better than the model group(P<0.05), proof NHS capsules can effectively prevent the BCG add LPS of mouse liver tissue structure reexposed damage.(3) Various treatment group liver tissue homogenate SOD was obviously higher than model group, MDA content obviously lower than model group,there is a significant difference significance(P<0.05),indicating that NHS capsule of antioxidation, scavenging free radicals role.(4) Each treatment group TNF-α;IL-6 value obviously lower than model group,there is a significant difference significance(P<0.05), indicating that NHS capsule resistant to damage the liver function probably with its inhibition cytotoxic factor TNF-α,IL-6 the release of the relevant.(5) Medium NHS capsule group all indexes of double ester group compared with biphenyl non-significance difference, explain various aspects and Biphenyl Dicarboxylate curative effect quite.4.Conclusion(1) NHS capsules can reduce BCG and LPS immunity liver damage on mice hepatic pathology structural damage and to protect its liver function.(2) NHS capsule dose and Biphenyl Dicarboxylate curative effect quite. |
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