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发布于:2018-6-13 10:39:08  访问:8 次 回复:0 篇
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Labeled S. S tubes were centrifuged at 20,000 ?g for 10 min at
160 l of 5 TCA was added to each pellet and heated for 15 min at 90 then 320 l of freshly prepared DPA solution was added, vortexed and incubated for 4 hr 37 . Optical density was read at 600 nm with a spectrophotometer (Smart specTM Plus, catalog # 170-2525).DNA Isolations and ladder assayCCl4 is a powerful pulmonary toxin that induces acute and chronic lung toxicity. This study induced pulmonary toxicity by CCl4 and determined the therapeutic effect of C. opaca fruit in CCl4 administered rats. Parameters studied included antioxidant enzymatic levels, genotoxicity and characteristic histological findings of lungs.Effects of C. opaca fruit on enzymatic antioxidant levelsDNA was isolated by using the MedChemExpress GSK343 methods of Wu et al. [21]. 100 mg of tissue in a petri dish was washed with DNA Buffer and PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/21231855 homogenized in 1 ml lysis buffer. 100 l of proteinase K (10 mg/ml) and 240 l 10 SDS, shaked gently, and incubate overnight at 45 in a water bath then 0.4 ml of phenol, was added shaked for 5-10 min, and centrifuge at 3000 rpm for 5 min at 10 . Supernatant was mixed with 1.2 ml phenol, 1.2 ml GSK2256098 price chloroform/isoamyl alcohol (24:1); shaked for 5-10 min, and centrifuged at 3000 rpm for 5 min at 10 . 25 l of 3 M sodium acetate (pH 5.2) and 5 ml ethanol was added with supernatant,Oxidative stress can affect almost every organ including the lungs. ROS production is caused by oxidative stress that directly damages lipids, proteins and DNA. In reaction to this, the body has its own defense system that consists of antioxidant enzymes that limit the level of damage. The protective effects of different fractions of C. opaca fruit against CCl4-induced pulmonary changes of tissue proteins and antioxidant levels is shown in Table 1. CCl4 treatment considerably (p < 0.05) decreased tissue protein, catalase, peroxidase, and superoxide dismutase levels compared with the control group. Co-administration of EFC, HFC and MFC significantly (p < 0.05) increased antioxidant enzyme activity.Effects of C. opaca fruit on TBARS and H2OThe effects of C. opaca fruit on TBARS and H2O2 levels are shown in Table 2. Increased TBARS and H2O2 levels were due to CCl4 toxicity. Post-administration of variousSahreen et al. BMC Complementary and Alternative Medicine 2014, 14:40 http://www.biomedcentral.com/1472-6882/14/Page 5 ofTable 1 Effects of various fractions of C. opaca fruit on tissue proteins and antioxidant enzyme levelsGroup Control Oil + DMSO CCl4 Sily + CCl4 HFC + CCl4 EFC + CCl4 MFC + CCl4 Protein (g/mg tissue) 2.01 ?0.03f 1.96 ?0.03f 0.74 ?0.01a 1.49 ?0.02e 0.89 ?0.03b 1.05 ?0.06c 1.35 ?0.078d CAT (U/min) 4.11 ?0.21d 4.09 ?0.30d 2.09 ?0.24a 3.60 ?0.11c 2.60 ?0.16b 3.02 ?0.07c 3.30 ?0.10c POD (U/min) 10.42 ?0.53d 1I.00 ?0.20d 5.74 ?0.42a 9.64 ?0.23c 6.53 ?0.32b 6.89 ?0.17b 7.80 ?0.17b SOD (U/mg protein) 3.40 ?0.23e 3.25 ?0.21e 1.12 ?0.02a 2.59 ?0.10d 1.46 ?0.10b 1.51 PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/24247322 ?0.08b 2.20 ?0.13cValues are Mean ?SD (06 number).Labeled S.
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