茵陈苓桂剂对非酒精性脂肪肝病大鼠脂代谢及氧化—抗氧化系统的影响

目的探讨茵陈苓桂剂对高脂饮食诱导的非酒精性脂肪肝模型大鼠脂代谢及氧化应激的影响。方法将60只SD大鼠随机分为正常组10只,造模组50只,正常组给予普通饲养,造模组给予高脂饲料。第8周末,造模组随机抽取2只大鼠做肝组织病理切片提示造模成功,将其随机分为多烯磷脂酰胆碱组,以及茵陈苓桂剂高、中、低剂量组,每组10只,剩余为模型组。各给药组给予相应药物灌胃治疗,正常组、模型组给予等体积蒸馏水,干预4周后,HE染色观察肝组织病理并计算非酒精性脂肪肝病活动度积分(NAFLD activity score,NAS),生化法检测血清天门冬氨酸基转移酶(aspartate aminotransferase,AST)、丙氨酸氨基转移酶(alanine aminotransferase,ALT)、甘油三酯(triglyceride,TG)、总胆固醇(total cholesterol,TC),ELISA测定血清及肝组织丙二醛(malondialdehyde,MDA)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-PX)、超氧化物歧化酶(super oxide dismutase,SOD)的水平。结果(1)与正常组比较,模型组大鼠血清ALT、AST、TC、TG水平显著升高(P<0.01),大鼠血清、肝组织MDA水平均显著升高(P<0.05或P<0.01),而SOD、GSH-PX水平降低(P<0.05或P<0.01)。(2)与模型组比较,西药组与中药高、中剂量组大鼠血清ALT、AST、TC、TG水平均降低,中药低剂量组大鼠血清AST、TC显著降低(P<0.05或P<0.01)。(3)与模型组比较,西药组与中药中、低剂量组血清、肝组织GSH-PX均升高(P<0.01),西药组与中药低剂量组血清SOD升高(P<0.05),而西药组肝组织SOD升高(P<0.01),中药高剂量组血清、肝组织MDA降低(P<0.05),GSH-PX及SOD水平升高(P<0.05或P<0.01)。(4)与西药组比较,中药高剂量组血清、肝组织GSH-PX显著升高(P<0.01)。(5)肝组织病理结果发现:与模型组相比,各给药组肝组织脂肪变、炎细胞浸润、肝小叶结构均有所改善,NAS均降低(P<0.05或P<0.01)。结论茵陈苓桂剂可有效调节非酒精性脂肪肝病大鼠的血脂和肝酶,降低大鼠血清、肝组织MDA水平,提高SOD、GSH-PX活性,因此能保护肝功能,调节脂代谢,改善肝脏病理损伤,减少脂质过氧化物的产生,增强抗氧化能力,从而恢复氧化—抗氧化系统平衡,这可能是其防治非酒精性脂肪肝病的作用机制之一。

尖叶假龙胆不同组分对酒精性肝损伤模型大鼠的保护作用研究

目的研究尖叶假龙胆不同组分拮抗酒精性肝损伤(alcoholic liver disease,ALD)效果并分析其机理,确定尖叶假龙胆治疗ALD的最佳疗效组分。方法采用渗漉法对尖叶假龙胆水、30%、60%和90%乙醇提取液的不同极性部位进行分离富集。健康雄性SD大鼠随机分为空白组、模型组、阳性药组、全草组、水洗脱组、30%乙醇洗脱组、60%乙醇洗脱组和90%乙醇洗脱组8组。实验组大鼠给予梯度酒精灌胃,建立慢性ALD模型,空白组给予等量生理盐水8周。从第5周起,以尖叶假龙胆水煎液及渗漉法水洗脱部位及30%、60%、90%乙醇洗脱部位对大鼠进行灌胃,阳性药对照组给予等量复方蛋氨酸胆碱片灌胃,第8周后取材。肝脏称重计算肝指数;电镜观察肝组织病理变化;酶联免疫吸附测定法检测血清中谷草转氨酶(aspartate aminotransferase,AST)、谷丙转氨酶(alanine aminotransferase,ALT)、甘油三酯(triglyceride,TG)、总胆固醇(total cholesterol,TC)、白介素6(interleukin-6,IL-6)、白介素8(interleukin-8,IL-8)、白介素10(interleukin-10,IL-10)、转化生长因子-β1(transforming growth factor-β,TGF-β1)及肝组织匀浆中丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽(glutathione,GSH)、乙醇脱氢酶(alcohol dehydrogenase,ADH)、乙醛脱氢酶(acetaldehyde dehydrogenase,ALDH)含量;免疫组化法检测细胞间黏附分子-1(intercellular cell adhesion molecule-1,ICAM-1)及肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)蛋白表达。结果(1)相较模型组,全草组ALT、AST、TG、TC、IL-6、IL-8、TGF-β1、MDA含量和ICAM-1、TNF-α蛋白表达降低,IL-10、SOD、GSH、ADH、ALDH含量升高,肝脏指数降低,均具有显著差异(P<0.05)。(2)相较模型组,水洗脱组TG、ALT、IL-6、TGF-β1和MDA含量降低,ADH含量升高,均具有显著差异(P<0.05),其余指标无明显差异(P>0.05)。(3)相较模型组,30%乙醇洗脱组TG、ALT、IL-6、TGF-β1和MDA含量降低,GSH、ADH、ALDH含量升高,均具有显著差异(P<0.05),其余指标无明显差异(P>0.05)。(4)相较模型组,60%乙醇洗脱组ALT、AST、TG、TC、IL-6、IL-8、TGF-β1、MDA含量以及ICAM-1、TNF-α蛋白表达降低,IL-10、SOD、GSH、ADH、ALDH含量升高,肝脏指数降低,均具有显著差异(P<0.05)。(5)相较模型组,90%乙醇洗脱组ALT、AST、TG、TC、IL-6、IL-8、TGF-β1、MDA含量以及ICAM-1、TNF-α蛋白表达降低,IL-10、SOD、GSH、ADH和ALDH含量升高,肝脏指数降低,均具有显著差异(P<0.05)。(6)在降低TG、MDA和升高ADH方面全草组与90%乙醇洗脱组具有显著差异(P<0.05),在降低TG、MDA、IL-6和升高ADH、ALDH方面60%乙醇洗脱组与90%乙醇洗脱组具有显著差异(P<0.05)。结论尖叶假龙胆组分具有拮抗ALD效果,不同组分作用效果有所差异,其中最优组分为90%乙醇洗脱组,其拮抗ALD机制可能是通过调节脂代谢系统,平衡氧化—抗氧化系统,提高酒精代谢率,抑制促炎细胞因子的生成,促进抗炎细胞因子生成以及抑制TNF-α、ICAM-1蛋白表达从而发挥作用。

Effects of Relative Soil Water Content on Antioxidant Enzyme System in Malus sieversii(Lebed.) Roem

By pot experiment under artificially simulated water stress conditions, soluble protein content, MDA content and SOD, POD, CAT and APX activities in Malus sieversfi leaves were determined to reveal the response mechanism of M. sieversii to changes of relative soil water content. According to the results, with the decrease of relative soil water content, MDA content in M. sieversii leaves increased by mem- brane lipid peroxidation. Cells resist water stress-induced membrane lipid peroxidation and clear the increased reactive oxygen species by improving soluble protein content and SOD, POD, CAT and APX activities. However, various enzymes were involved in the response to water stress under different moisture conditions. In addition, the results indicated that M. sieversii had a good adaptability to higher relative soil water contents.

The influence of cadmium on the antioxidant enzyme activities in polychaete Perinereis aibuhitensis Grube(Annelida:Polychaeta)

The infaunal polychaete Perinereis aibuhitensis Grube,distributed widely along Asian coasts and estuaries,is considered a useful animal model in ecotoxicological tests and a promising candidate in biomonitoring programs.This paper deals with the activities of antioxidant enzymes including superoxide dismutase(SOD),catalase(CAT),and glutathione peroxidases(GSH-Px) in infaunal polychaete P.aibuhitensis exposed to a series of sublethal water-bound cadmium(Cd) concentrations(0,0.34,1.72,3.44,6.89,and 17.22 mg L-1) under a short-term exposure(1-8 d).The results indicate that the SOD and GSH-Px activities in P.aibuhitensis are stimulated first and then renewed to the original level.The CAT activity of worms decreases at an earlier exposure time but increases to the control values at a later exposure time.Our study suggests that Cd can interfere with the antioxidant defense system of P.aibuhitensis.However,the changes in antioxidant enzyme activities for this species do not show the best promise as biomarkers in Cd biomonitoring of estuarine and coastal zones because weak or non-dose-effect relationships between the antioxidant enzymes activities and Cd levels are found.

Effects of UV-B irradiation on isoforms of antioxidant enzymes and their activities in red alga Grateloupia filicina(Rhodophyta)

Macroalgae in a littoral zone are inevitably exposed to UV-B irradiance. We analyzed the effects of UV-B on isoenzyme patterns and activities of superoxide dismutase （SOD）, peroxidase （POX）, catalase （CAT）, and ascorbate peroxidase （APX） of red algae Grateloupia filicina （Lamour.） C. Agardh. The activities of SOD, CAT, and APX changed in response to UV-B in a time- and dose-dependent manner. POX activity increased significantly under all three UV-B treatments. The enzymatic assay showed three distinct bands of SODI （Mn-SOD）, SODII （Fe-SOD）, and SODIII （CuZn-SOD） under a low （Luv） and medium （Muv） dose of UV-B irradiation, while SODI and SODIII activities decreased significantly when exposed to a high dose of UV-B irradiation （Huv）. The activity of POX isoenzymes increased significantly after exposure to UV-B, which is consistent with the total activity. In addition, a clear decrease in activity of CATIV was detected in response to all the three doses of UV treatments. Some bands ofAPX isoenzyme were also clearly influenced by UV-B irradiation. Correspondingly, the daily growth rate declined under all the three exposure doses, and was especially significant under Muv and Huv treatments. These data suggest that, although the protection mechanisms of antioxidant defense system are partly inducible by UV-B to prevent the damage, G.filicina has incomplete tolerance to higher UV-B irradiation stress.

Response of enzymatic and non-enzymatic antioxidant defense systems of Polygonum hydropiper to Mn stress

The response of enzyme and non-enzymatic antioxidants of Mn hyperaccumuator, Polygonum hydropiper （P. hydropiper）, to Mn stress was studied using hydroponics culture experiments to explore the mechanism of Mn tolerance in this species. Results showed that both chlorophyll and carotenoid contents significantly （p〈0.05） decreased with increasing Mn treatment levels （0, 0.5, 1, 2, 4, and 8 mg/L） in hydroponics. The concentrations of malondialdehyde （MDA） and hydrogen peroxide （H202） in the root and shoot of P hydropiper were accumulated under Mn stress. Meanwhile, the anti-oxidative functions of several important enzymes, including superoxide dismutase （SOD）, catalase （CAT）, ascorbate peroxidase （APX） and peroxidase （POD） in plants were stimulated by Mn spike in leaves and roots, especially at low Mn stress; while sulfhydryl group （--SH） and glutathion （GSH） were likely involved in Mn detoxification ofP. hydropiper under high Mn stress.

Acute Toxicity of Cadmium on the Antioxidant Defense Systems and Lipid Peroxidation in the Juveniles of Genetically Improved Farmed （GIFT） Tilapia Oreochromis Niloticus

Heavy metals pose a potential threat to aquatic organisms. In this study, a static-renewal acute toxicity test was conducted to investigate the effects of cadmium on the antioxidant defense systems （both enzymatic and non-enzymatic） and lipid peroxidaton in liver and gill tissues of juvenile GIFT tilapia Oreochromis niloticus. After 8 days of exposure to Cd （0, 0.016, 0.08, 0.4 and 2 mg/L）, livers accumulated significantly more Cd than gills. Catalase （CAT）, superoxide dismutase （SOD） and glutathione S-transferase （GST） activities were stimulated only at the highest concentration tested （2 mg/L）. Glutathione peroxidase （GPx） activity was stimulated in the gill while inhibited in the liver, these alternations in gill and liver showed a strong relationship with Cd levels in these tissues. This may indicate either a tissue-specific response of GPx to Cd or, most probably, a hormetic effect of Cd on GPx. Cd increased GSH levels and decreased the ratio GSSG/GSH in fish livers at 2 mg/L. Cd exposure resulted in an elevated level of MDA in the livers of fish at 2 mg/L, indicating that Cd caused lipid peroxidation. Taken together, the results demonstrated that Cd altered the enzymatic and non-enzymatic defensive systems and caused lipid peroxidation in O. niloticus at relatively high concentrations （compared to environmentally relevant concentrations）. In addition, the results implied that O. niloticus could tolerate high level of Cd in sites polluted by Cd.

Effect of Lead Toxicity on Germination and Enzymatic Activity of Durum Wheat Triticum durum Desf.

The aim of this work is to identify the effect of lead on germinal parameters and the antioxidant enzyme activities （lipase, peroxidase and catalase） in durum wheat Triticum durum Desf. cv （waha, vitron and gta） exposed to the concentrations of 0, 0.15, 0.25 and 0.3 g/L of Pb （NO3）2 during germination process. The obtained results showed that lead reduced the germination, root and aerial biomass. The concentration of 0.3 g/L inhibited completely the germination of the three varieties. It also slowed lipase activity, the degradation of lipids of the seed＇s reserves and disrupted the metabolism of peroxidase and catalase. Concerning the behavior of the three varieties studied, it appears that the Vitron is the best predisposed variety to stand against lead stress by its strong antioxidant defense system.

Influence of Irrigation with Reclaimed Water on Antioxidant System of Maize and Soybean

Irrigation with reclaimed water is an important way to deal with water shortage of agricultural production and solve the problem of sewage pollution. The response of crop antioxidant enzyme system is an early-warning indicator of environmental factors changes. Compared with raw wastewater and used fresh water as controls, this research studied peroxides （POD）, superoxide dismutase （SOD） activities and malondialdehyde （MDA） content in leaves from maize （Zea mays L.） and soybean （Phaseolus vulgaris L.） with pot culture under irrigating with reclaimed water which with secondary and third treatment. The results showed that secondary reclaimed water had some adverse effects on antioxidant system of maize in seedling stage, and the influence decreased in later stage of maize; effect of the third reclaimed water on antioxidant system of maize was not significant. Irrigation with reclaimed water has little influence on antioxidant system of soybean in seedling and reaping stage, but it could bring some oxidative stress in blossom stage. As a result, irrigating maize with second or third reclaimed water is safe to the growth of maize and soybean, but it is suggested that second reclaimed water should not be used in seedling stage of maize and reclaimed water should not be used in the second stage of soybean.

Antioxidant Defense System in Wheat （Triticum aestivum L.） Seedlings under Heat Stress and Revival Conditions

The present investigation was carried out to investigate the effect of heat stress and revival on some antioxidative enzymes and metabolites in leaves of the wheat （Triticum aestivum L.） seedlings of heat susceptible （cv. WH 147 and HS 277） and heat tolerant （cv. WH 1021 and HW 2045） cultivars. Seven days old seedlings grown at 25 ℃ were exposed to 40 ℃ for 6 h and these seedlings were again brought to 25 ℃. The observations were recorded in the leaves of control, stressed and revived seedlings on 2nd and 4th day of revival. For the selection ofthermo-tolerant cultivars, screening of the thirty-six cultivars was done based on wilting of primary leaf and values of chlorophyll fluorescence. The MDA （malondialdehyde） and H2O2 concentration in leaves of wheat seedlings increased at the high temperature. There was enhancement in the activities of antioxidative enzymes, viz. CAT （catalase）, POX （peroxidase）, GR （glutathione reductase） and APX （ascorbate peroxidase） in leaves of the tolerant and susceptible cultivars under heat stress, however, higher percent increase was observed in tolerant cultivars. Heat stress increased the SOD （superoxide dismutase） activity in tolerant cultivars but activity declined in susceptible cultivars. On revival, the activities of the CAT, POX and GR declined in comparison to stressed seedlings but remained higher as compared to control. Ascorbate peroxidase activity remained higher on 2nd day and 4th day of revival in all the cultivars.

Investigation of the redox status in H22 hepatocellular carcinoma xenografts treated by a novel anticancer drug-ethaselen

We investigated the redox status of H22 hepatocellular carcinoma xenografts treated with various doses of ethaselen, a novel anticancer drug targeting thioredoxin reductase （TrxR）. The concentrations of low molecular weight antioxidant g!utathione （GSH） and malondialdehyde （MDA）, a product of lipid peroxidation, as well as the activities of important antioxidant enzymes were measured for elucidating the redox status of H22 tumor tissues. We found that the decreased GSH level, decreased thioredoxin reductase and superoxide dismutase （SOD） activities as well as increased MDA content were closely related to the tumor growth inhibition and ethaselen doses. Glutathione peroxidase （GPx） and glutathinne reductase （GR） activities are also affected by ethaselen treatment. However, the catalase （CAT） activity remains unchanged. Finally, we studied the relationship of tumor growth inhibition caused by ethaselen with these redox factors. This study showed that ethaselen could elevate the oxidative stress to suppress the H22 tumor growth in mice model.

Antioxidant effect of indirect moxibustion on healthy subjects:a pilot study

OBJECTIVE:Moxibustion has long been thought to promote vital energy or immunity through clinical experience.This study aimed to investigate the clinical effects of indirect moxibustion on antioxidant and immunomodulation in a normal population.METHODS:Twenty-five healthy volunteers with no objective or subjective disorders were recruited.The participants were treated with indirect moxibustion on acupoints Guanyuan(CV 4) and Shenque(CV 8) three times per week for 4 weeks.The serum levels of reactive oxygen species(ROS) and malondialdehyde(MDA),the total antioxidant capacity(TAC),the activities of catalase and superoxide dismutase(SOD),and the total glutathione content were determined before and after the 12th moxibustion.Lymphocyte subpopulations and 42 cytokines in the peripheral blood were analyzed using flow cytometry and antibody array,respectively.RESULTS:Compared with the initial time point,the serum levels of ROS and MDA were significantly lowered by moxibustion,while TAC was increased(P<0.01 for all).A significant increase was observed in catalase activity(P<0.05),but not in SOD or total glutathione.There were no significant changes in lymphocyte subpopulations or cytokines in the peripheral blood.Fifteen of 25 participants reported at least one symptom in which they felt subjective improvement after moxibustion.CONCLUSION:Indirect moxibustion on acupoints CV 4 and CV 8 improved the antioxidant defense system,which may be a mechanism explaining the clinical effects of moxibustion.