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N-hexane (n-hexane) as an organic solvent, a wide industrial application. N-hexane in the environment through the respiratory tract, gastrointestinal tract, skin into the body, mainly in the higher fat content of organs such as the brain, kidney, liver, spleen, testes, blood. Acute toxicity grading, n-hexane is a non-toxic class, but because of their volatile, fat-soluble, and accumulation, be considered as highly hazardous poisons; and n-hexane acute and chronic poisoning with different performance: human n-hexane the acute poisoning mainly for acute encephalopathy (central inhibition) and the skin, mucous membrane irritation; chronic poisoning neurotoxicity, mainly caused due to the remote coarse axons of nerve fiber degeneration feeling - around the sport neuropathy. Neurotoxicity of n-hexane poisoning caused yet no specific treatment. Known n-hexane obvious exposed animal liver and nerve tissue lipid peroxide the injury (lipidperoxidation damage LPD) role, causing elevated levels of oxidation products and oxidation - changes in antioxidant enzyme activities, inferred oxidative damage may be one of the n-hexane of the poisoning mechanism. Garlic oil (garlic oil, GO) are extracted from garlic oil immiscible with water, the main ingredient has a strong reduction of the various sulfides, peroxides and oxygen, etc. can be restore, and thus significantly improve the body peroxidase activity and reduce the content of the oxidation products. 2,5 - hexanedione (2,5-hexanedione ,2,5-HD) as active metabolites of the n-hexane, n-hexane neurological toxicity of the main media. Whether garlic oil can antagonize peroxide injury of rat nerve tissue caused by 2,5-HD, and mitigate its due to peripheral nerve toxicity and abroad has not been reported. The experiments on male Wistar rats for six consecutive weeks intraperitoneal injection of 2,5-HD 300mg/kg.bw exposed to manufacture n-hexane chronic poisoning induced neurotoxicity model, and rats exposed to every day for 4-6 hours in advance irrigation the stomach to give 40mg/kg.bw and 80mg/kg.bw garlic oil for preventive intervention, was measured at different time points during administration of the rats the neurological behavior change: hindlimb brace force index, balance index and gait score; determination the experimental endpoint rat brain, spinal cord and sciatic oxidation - antioxidant status: the content of malondialdehyde (malondialdehyde, MDA), glutathione (glutathione, GSH) content, total antioxidant capacity (total antioxidant capability, T-AOC) and inhibit the ability of the hydroxyl radical, lipid 2,5-HD caused to observe the garlic oil over the role of oxidative damage and neurotoxicity. Hexane nerve toxicity is mainly due to its metabolic product (2,5-HD), cytochrome P450 (cytochromeP450, CYP450) a n-hexane/ethyl basic oxidative metabolic enzyme, plays an important role in the metabolic activation process; been numerous reports of garlic oil has a significant inhibitory effect on cytochrome P450, and a variety of animal models of disease, but the garlic oil on the metabolism of n-hexane has not been reported. Studies have shown that the content was positively correlated with the serum level of 2,5-HD in its target organ sciatic nerve, and n-hexane neurotoxicity closely related, thus reducing the serum level of 2,5-HD, will likely reduce the n-hexyl alkyl poisoning occurred, and reflect changes in the levels of n-hexane metabolism. In order to meet this experiment for the determination of serum 2,5-HD claim using ethyl acetate as the extraction agent, and dried over anhydrous potassium carbonate (K 2 CO 3 ) to further improve the efficiency of the extraction, HP-5 capillary GC column separation, hydrogen flame ionization detector (FID) serum 2,5-HD gas chromatography method, and to test the feasibility of the method. On this basis, design once oral administration the Kunming mice n-hexane 3000mg/kg.bw, mice exposed to 2 hours before the intervention group to give 80mg/kg.bw garlic oil, the determination of exposure at different time points after serum 2,5-HD content by different doses of garlic oil (10mg/kg.bw 20mg/kg.bw 40mg/kg.bw 80mg/kg.bw) before and after the intervention and exposure, two hours were given 80mg/kg. bw garlic oil, serum 6 hours after the exposure of 2,5-HD content than that of mice exposed to changes;, and different gender and age mice by the same dose of n-hexane exposure serum 2,5-HD content the determination of the role of n-hexane toxicity, which provide the feasibility of experimental evidence for the prevention of n-hexane poisoning explore the impact of the garlic oil on the metabolism of n-hexane, and gender and age factors. Results 1 body weight changes in normal group stable increase in body weight of rats; model group and garlic oil group weight gain slowed, or even negative growth model rats exposed to 3 weeks after the body weight was significantly lower than the control rats (P <0.01) ; group two weeks after the low dose of garlic oil, garlic oil high dose group after 1 week weight that is significantly lower than the untreated rats (P <0.01 or P <0.05). Rat neural behavior change (1) the gait and scoring: garlic oil is low, the high dose group of rats exposed to 4 weeks to appear motion abnormalities, compared with the model group rats one week in advance; groups gait score model group, garlic oil, the high-dose group than in the control group were significantly increased (P <0.01), and garlic oil high dose group compared with the model group score was significantly higher (P <0.05). (2) hindlimb brace force index: 0 weeks earlier with 2,5-HD exposure model to four weeks, garlic oil, hind legs supporting force and high-dose group index increased by 44%, 50%, 49%, are significantly higher than the normal control group (P <0.01), but the difference was not statistically significant (P> 0.05) between the three groups. (3) Balanced Index: In addition to the control group, the rats in each group balance index showed varying degrees of reduction to four weeks exposed to the model group, the garlic oil is low, the high-dose group were reduced by 30%, 45%, 68% are significantly lower than the control group (P <0.05 or 0.01), and the the garlic oil high dose group compared with the model group with significant difference (P <0.05). Rat nerve tissue oxidation - indicators of change (1) MDA content: model group, the brain, spinal cord and sciatic nerve MDA content, respectively, compared to the normal control rats increased by 26%, 25% and 17%, respectively ( P <0.05 or 0.01), a decrease of 32% to 43% of the garlic oil group compared with the model group (P <0.01) and 32% ~~ 52% (P <0.01). T-AOC, nerve tissue change (2) T-AOC: model rats has been enhanced compared to the control group, but the groups were no statistical differences (P> 0.05). (3) GSH content change: in the nerve tissue, the model rats GSH content is lower than the normal control group, in addition to the low-dose group rat sciatic nerve in garlic oil content is lower than the model group, other nerve tissue garlic oil group higher than the model group, but these changes were not statistically significant (P> 0.05). (4) ability to inhibit the hydroxyl radical: Compared with the normal control group, model group the ability to inhibit the hydroxyl radical weakened by 34% in the brain, spinal cord and sciatic nerve, respectively, 16% and 28% (P <0.05 or 0.01); garlic oil varying degrees of antagonizing this trend, garlic oil in the low-dose group rats were enhanced by 22% compared with the model group, 28% and 28% (P <0.01), the garlic oil high dose group compared with 50%, 46% and 40 % (P <0.01). Serum 2,5-HD content change (1) once exposed metabolism: the disposable oral administration of n-hexane after 4000mg/kg.bw, mouse serum 2,5-HD content over time while increased to 10 hours and reached its peak after gradually reduced, and almost can not be detected after 20 hours. Serum 2,5-HD content change (2) after different doses of n-hexane exposure: mouse serum 2,5-HD content with the increasing doses of n-hexane exposure increased blank mouse serum were not detected; of 2000,4000 and 6000mg/kg.bw, groups of mice exposed to 8 hours after serum 2,5-HD content were 8.04,16.68 and 22.38μg/ml, showed a dose - response relationship. (3) Garlic oil of metabolism in mice exposed to n-hexane: n-hexane time after 3000mg/kg.bw gavage exposure group intervention serum levels of 2,5-HD content with garlic oil trend results are consistent with 4 (1), are to extend the first and then decreased with time; garlic oil intervention mice at each time point serum levels are lower than those exposed mice after exposure 4,6,8,10 hours than exposure group 2,5-HD was significantly lower (P <0.05 or 0.01). (4) different doses of garlic oil on serum 2,5-HD content: two hours before the exposure given different doses of garlic oil, six hours after exposure of mouse serum 2,5-HD content more exposure group decreased by 16.2%, 20.8% and 22.8% (P <0.05) and 32.1% (P <0.01), show a significant dose - response relationship. (5) different dosages of garlic oil on serum 2,5-HD content: n-hexane exposure before and after? Mice 80mg/kg were given garlic oil, their serum 2,5-HD content The simple exposure groups were significantly lower (P <0.05); exposure dose group than exposure lower dose group, but the difference between the two groups was not statistically significant (P> 0.05). (6) 2,5-HD content in the different age serum: different week-old mice by the same dose of n-hexane is a one-time eight hours after exposure, serum 2,5-HD showed different levels (P <0.05): 5 week-old group (22.83μg/ml) higher than the 4-week-old (19.59μg/ml) and 6-week-old the mice (16.42μg/ml). (7) different gender serum 2,5-HD content: different gender mice by a single n-hexane exposure, female serum 2,5-HD content (13.22μg/ml) was significantly higher than males (10.34μg / ml), increased to 27.9% (P <0.05). 5. The mouse serum 2,5-HD gas chromatographic (1) sample processing method selection and method specificity: three extractant recovery rate comparison: dichloromethane> ethyl acetate> ethyl ether, combined with volatile and toxicity considerations, ethyl acetate was chosen as the method of extraction agent; compare adding anhydrous Na the 2 SO 4 , anhydrous K 2 CO 3 and ethyl acetate without salt extraction efficiency, found that adding the right amount of salt can significantly improve the efficiency of the ethyl acetate extraction of 2,5-HD, which anhydrous K 2 < / sub> CO 3 better; separation method extractant ethyl acetate (Rt = 2.26min) and 2,5-HD (Rt = 3.33min) completely, and n-hexane (Rt = 2.266min) there is no interference of the determination of the target 2,5-HD. (2) detection methodology: Methods minimum detectable concentration 0.03μg/ml, linear range 0.03μg/ml 80μg/ml; recoveries of 83.8 to 99.9%; relative standard deviation (RSD) days from 3.3% to 5.2% the daytime 3.3% to 8.0%; 4 ° C sample to 24 hours can be saved and can be stored for one week, -20 ℃. Conclusion 1. Garlic oil could be antagonized by 2,5-HD-induced rat nerve tissue lipid peroxidation injury, but can not improve rat sensory motor nerve dysfunction, which indicated that oxidative damage may not be in the 2,5-HD The primary mechanism of action of toxic neuropathy. 2 garlic oils inhibit the n-hexane exposure to 2,5-HD in mice produce serum 2,5-HD levels were significantly lower, and the inhibitory dose of garlic oil and time; factors age and gender n-hexane metabolism also has a certain impact, more than for us to prevent and reduce the n-hexane poisoning effective experimental basis. 2,5-HD 3 The present experiments established gas chromatographic method, accurate and reliable, and has a good reproducibility, can be used for the determination of serum 2,5-HD.
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