|
Cruciferous vegetables in lead poisoning prevention, the experimental study of the cruciferous vegetable extract (the main component of the glucose glucosinolate referred glucosinolate, GS) antagonism for lead toxicity. Previous studies have shown that 4 - methyl the sulfur oxygen butyl glucosinolate (RAA) metabolite sulforaphane (SUL), all kinds of the glucosinolate metabolites (mainly isothiocyanate antioxidant activity ITC) is the most prominent one of the indole GS on the body may have a harmful effect, so the content of the two indicators to filter the extraction of raw materials as a screening cruciferous vegetables. HPLC-UV detection of three cruciferous vegetables GS component content, select RAA content, the lower levels of the indole GS kale flowers as the extraction of raw materials. Subsequently, kale flower extract (the main component of GS) and its degradation products (the main component of ITC) and selenium yeast as an experimental material to explore the GS and ITC antagonistic by to establish preventive lead poisoning mouse model, lead toxicity effect and both effects difference; GS and ITC antioxidant effect and both the effect of differences; synergy GS and selenium antagonistic lead toxicity and antioxidant synergy. First observed antagonistic role of lead toxicity, cruciferous vegetables and broaden its applications. Specifically: 1 extract raw materials screening 1.1 method using high performance liquid chromatography of broccoli sprouts mature broccoli, kale flower GS identification and quantitative analysis of three kinds of cruciferous vegetable material, select one of the RAA content higher and lower levels of the indole GS a vegetable as the extraction of raw materials. 1.2 Results 1) total GS content: broccoli sprouts maximum (1509.42μmol/100g FW ~ 2315.42μmol/100g FW), followed by mature broccoli strains (636.82μmol/100g FW to 969.75μmol/100gFW), the lowest kale, flowers (245.91μmol/100gFW ~ 433.92μmol/100g FW). 2) the GS content indole: three materials indole GS content were lower than 15% of the total of GS, and the lowest for broccoli mature strains, to its indole GS content is almost close to zero. 3) RAA content: the broccoli sprouts groups RAA content is generally higher than broccoli mature strains group and kale flowers (25% to 41%), but its group differences among the varieties; broccoli mature strains group RAA almost not detected; the kale group content than buds group (245.91μmol/100gFW ~ 433.92μmol/100g FW), but the RAA total GS content, a higher proportion (15% ~ 25%). 1.3 Conclusion broccoli sprouts group GS content (1509.42μmol/100gFW 2315.42μmol/100g FW), RAA content of total the GS high proportion (25% to 41%), the total content of the indole GS GS low proportion ( 2% to 5%), most suitable as a raw material for extraction, but for practical reasons, not ultimately used as the raw material for extraction. Broccoli mature strains, although higher levels of total GS (636.82μmol/100gFW 969.75μmol/100g FW) and of indole GS content to total the the GS lower proportion (0 to 3%), but because of its RAA content is almost zero, Therefore, it can not be used as raw material for extraction. Kale flowers readily available raw materials, RAA content of total GS higher proportion (15% to 25%), and low levels of total GS (245.91μmol/100gFW ~ 433.92μmol/100g FW), and still comply with the requirements of the extraction of raw materials, final selection the kale flowers as the extraction of raw materials. 2 extract was 2.1 Method kale flowers were freeze dried powder, with 70% methanol as the extraction agent, ultrasonic extracted twice, each time 30min, concentrated by rotary evaporation and freeze-dried to obtain a crude extract. HPLC-UV quantitative analysis, HPLC-ESI-MS/MS the GS structural identification. 2.2 The results obtained kale flower extract 279g, total GS content for 77.44μg / g, 18% of the the RAA total GS content. 3 Lead animal experiments 3.1 to about 110 weight 20g male Kunming mice were randomly divided into 11 groups. In addition to the the blank group free drink deionized water, the other groups are free to drink 300mg / L lead acetate in water. Intervention factor is applied at the same time: the blank model group daily with deionized water gavage; positive group dimercaptosuccinic acid (DMSA) gavage administered irrigation three days stopped four days; high, medium and low dose group daily irrigation of containing total the GS the amount GS0.133μmol / g BW (H-GS) 0.044μmol / g BW (M-GS), 0.022μmol / g BW (L-GS) kale flower extract solution the stomach, myrosinase on degradation products 0.133μmol / g BW (H-ITC) 0.044μmol/gBW (M-ITC) 0.022μmol / g BW (M-ITC) solution gavage; selenium yeast group to the selenium content 33.2μg solution of / kgBW (Se) gavage; the selenium yeast ehioglucoside mixture group (Se M-GS) the selenium yeast 0.022μmol 33.2μg/kg BW / g the BW of glucosinolate mixed solution gavage. 30 days after the abdominal vein blood, mouse liver, brain, femur, Determination of lead content in the four organs and tissues as well as the content of selenium and detection of whole blood glutathione peroxidase (GPx), superoxide dismutase (SOD), glutathione S-transferase (GST) activity and liver tissue GPx, GST activity and malondialdehyde (MDA) content. The determination of lead by graphite furnace atomic absorption spectrometry; selenium determination by hydride generation - atomic fluorescence spectrometry; antioxidant enzyme activity assay kit. 3.2 Results: 1) antagonized the toxicity of lead role: GS, medium, and low dose group Excretion effect, L-GS measured various organs and tissues were significantly lower than those in the untreated control group (P <0.05), H-GS group The mouse blood lead, liver, brain lead levels, M-GS group blood and femur lead levels were significantly lower than those in the untreated control group (P <0.05). ITC high, medium and low dose group Excretion effect: inter L-ITC mice brain tissue lead levels, lead levels of M-ITC mice, H-ITC mice femur lead levels and model the control group, no significant difference (P <0.05), the ITC three dose groups of mice measured lead in various organs and tissues were significantly lower than those in the untreated control group (P <0.05). 2) antioxidant effect: whole blood GST activity, GST water in addition to the H-ITC group blood was significantly higher than that in the untreated control group (P <0.05), the rest of the group (including the blank group and the positive drug group) with the model control group there was no significant difference (P <0.05). Whole blood GPx activity, in addition to the H-GS, M-ITC model control group no significant differences (P <0.05), the rest of the group were significantly higher than that in the untreated control group (P <0.05). SOD activity of whole blood, except H-GS, H-ITC, the rest of the group were significantly higher than that in the untreated control group (P <0.05). Liver tissue GST activity, in addition to M-ITC, the rest of the group and the model control group showed no significant difference (P <0.05). Liver tissue GPx level, except Se group, the rest of the group and the model control group showed no significant difference (P <0.05). Liver tissue MDA levels in each group and the model control group showed no significant difference (P <0.05). 3) lead levels and the corresponding organ and tissue antioxidant indicators correlation: the animals blood GPx activity, SOD activity, GST activity was negatively correlated with blood lead levels. Which the correlation between SOD activity and blood lead levels was significant (P <0.05). Negative correlation between GPx activity, GST activity in liver tissue lead content in animal liver tissue showed a positive correlation between MDA content in the liver tissue lead content in the liver tissue. Correlation does not have a significant (P <0.05). 3.3 Conclusion 1) kale flower extracts can effectively reduce the animal measured the level of lead content in the organs and tissues, the three dose groups showed a dose-response relationship, in which the low-dose group has both antagonistic role of lead toxicity has antioxidant properties. 2) kale flower extract can effectively improve the in vivo antioxidant system GPx, SOD enzyme activity, and the level of antioxidant enzyme activity in vivo corresponding organ tissue lead level was significantly negatively correlated. 3) kale flower extract and selenium in the the antagonistic lead role and improve activity of GPx, SOD found no synergy. 4) kale flower extract and its vitro degradation product ITC has the same effect in the antagonistic lead toxicity and antioxidant effects, and preliminary findings GS and its degradation products in vivo biological activity consistent.
|