荞麦种子萌发期多种抗氧化酶活性的研究

通过研究荞麦种子萌发期(0～7 d)超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)和抗坏血酸过氧化物酶(ASP)四种抗氧化酶的酶活性变化,以及对自由基等的清除效果,分析荞麦种子内在抗氧化酶系统在萌发期对细胞膜结构的修复及保护作用。实验结果表明:荞麦种子在萌发期产生的活性氧、自由基等对四种抗氧化酶均有激活效应,其中超氧化物歧化酶(SOD)活性最先上升,其他三种酶随其后被激活,四种酶的氧化反应存在一定关联性和协同作用。苦荞的抗氧化酶活性高于甜荞。     

外源蔗糖和Ca2+对荞麦幼苗耐盐性的影响

以盐敏感荞麦品种‘TQ-0808’为实验材料,在100mmol·L-1NaCl胁迫下研究添加不同浓度蔗糖和Ca2+处理对荞麦幼苗耐盐性的影响。不同浓度的外源蔗糖和Ca2+处理均能明显降低盐胁迫下荞麦叶片的质膜透性和丙二醛(MDA)含量,明显增加荞麦叶片的超氧化物歧化酶(SOD)和抗坏血酸过氧化物酶(APX)活性及净光合速率,且外源Ca2+处理的增加效果明显好于外源蔗糖处理。说明外源蔗糖和Ca2+处理能明显改善盐胁迫下荞麦幼苗的耐盐性,对盐胁迫具有较好的缓解作用,外源蔗糖和Ca2+处理的最适浓度分别为6mmol·L-1和20mmol·L-1。     

Ca~(2+)通过膜脂影响肌质网Ca~(2+)-ATP酶的活性与Ca~(2+)转运功能

重建在大豆磷脂脂质体上的兔骨骼肌肌质网Ca~(2+)—ATP酶在ATP驱动下可将溶液中的Ca~(2+)转运到脂酶体内部;外加EGTA则可除去脂酶体外部的Ca~(2+),由此可得到四种含Ca~(2+)状态不同的脂酶体:(1)内、外都无Ca~(2+);(2)仅外部有Ca~(2+);(3)内、外都有Ca~(2+);(4),仅内部有Ca~(2+).用DPH和AS系列萤光探针对这四种含Ca~+状态不同的脂酶体的膜脂流动性进行了测定,结果表明:脂酶体外部加入Ca~(2+),脂双层外表面的流动性降低.当Ca~(2+)进入脂酶体内部后,内表面膜脂的流动性也降低,而且外层膜脂流动性进一步降低.脂酶体内、外的Ca~(2+)含量不同时,Ca~(2+)—ATP酶功能状态也不同.转运到脂酶体内部的ca~(2+)积累到一定浓度后,通过Ca~(2+)泵向内转运的Ca~(2+)及Ca~(2+)—ATP酶活力都受到了抑制.转运进行到第四分钟时的酶活只有第一分钟的9%.但在相同的实验条件下,失去了完整的膜结构的纯化的Ca~(2+)—ATP酶蛋白没有被抑制.这提示完整的膜结构是这种抑制作用所必需的,而且膜两侧Ca~(2+)浓度的梯差可通过影响膜脂来调节Ca~(2+)—ATP酶的功能.     

邻苯二甲酸二(2-乙基己基)酯长期暴露对鲤鱼毒理学指标的影响

以吐温80和水为对照组,用5 mg·L-1、20 mg·L-1、80 mg·L-1、160 mg·L-1的邻苯二甲酸二(2-乙基己基)酯(DEHP)对鲤鱼暴露染毒20 d,用试剂盒等方法研究了DEHP对鲤鱼肝脏抗氧化防御系统、肝解毒酶、能量代谢酶和肝损伤标志酶的影响。结果表明,与水空白对照组和吐温80组相比,各浓度DEHP组,肝脏超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活力和抗超氧阴离子自由基能力的差异均无统计学意义,谷胱甘肽过氧化物酶(GSHPX)活力表现为低促高抑,160 mg·L-1组抗羟自由基能力显著降低,丙二醛(MDA)含量显著升高。研究结果还表明,在DEHP实验浓度范围内,谷胱甘肽硫转移酶、葡萄糖-6-磷酸脱氢酶活力表现为低促高抑,而160 mg·L-1组7-乙氧基-3-异吩噁唑酮-脱乙基酶、Na+K+-ATPase、单胺氧化酶(MAO)活力和各DEHP组Ca2+Mg2+-ATPase活力均受到了显著抑制,20 mg·L-1以上组肝γ-谷氨酰转肽酶活力则显著升高。结果显示,一定浓度的DEHP能损伤肝脏组织,并对肝组织的毒理学指标产生影响。     

Mn~(2+)和Cu~(2+)与脂质体和脂酶体相互作用的ESR研究

用ESR实验研究了Mn~(2 )、Cu~(2 )与DOPC,DPPC,SPL,DOPA,DPPA脂质体及其与H~ -ATP酶复合体重组的脂酶体的相互作用.通过Mn~(2 )—ESR谱线强度以及Cu~(2 )—ESR谱g因子的测量得出,磷脂分子头部不同的化学组成及其脂酰链的不同状态决定了Mn~(2 )、Cu~(2 )与膜脂结合的强弱程度,通过脂质体和脂酶体中自旋标记物5NS—ESR谱的测量进一步得出Mn~(2 )的结合增大了膜脂排列的序参数,而酶复合体的嵌入都导致与膜脂结合的Mn~(2 )比例减小.因而,当Mn~(2 )与脂酶体相互作用时,膜脂的排列最终达到一个平衡状态.在中性磷脂脂酶体的膜与Mn~(2 )之间,这种相互作用不明显.     

重金属Zn~(2+)胁迫下米氏凯伦藻(Karenia mikimotoi)的生长生理响应研究

采用室内培养试验方法,以米氏凯伦藻(Karenia mikimotoi)为生物材料,设置不同浓度梯度的重金属Zn~(2+)(0、1、5、10、15和20mg·L~(-1)),主要测定藻细胞密度、光合色素、光合效率、抗氧化酶类、丙二醛(malondialdehyde,MDA)等相关指标,探讨重金属Zn~(2+)胁迫下米氏凯伦藻(Karenia mikimotoi)的生长和生理特征。结果发现,在1和5 mg·L~(-1)的Zn~(2+)浓度下,米氏凯伦藻细胞依然保持较好生长繁殖,表明其具有一定的耐受性,而随着重金属Zn~(2+)浓度的提高,细胞生长受到毒害抑制。光合色素含量呈现动态变化,试验结束时(96 h)叶绿素a、叶绿素b和胡萝卜素含量有各自不同的变化趋势,最大光能转化效率(Fv/Fm)表现出先升后降的趋势。Zn~(2+)浓度为10mg·L~(-1)时,超氧化物歧化酶(superoxidedismutase,SOD)活性显著高于对照;过氧化氢酶(catalase,CAT)活性呈现出先上升后下降的趋势,且5、10、15和20mg·L~(-1)Zn~(2+)浓度下米氏凯伦藻的过氧化氢酶(CAT)活性均显著高于对照;10、15mg·L~(-1)Zn~(2+)浓度下米氏凯伦藻的总抗氧化能力(total antioxidantcapacity,T-AOC)显著高于对照,而20mg·L~(-1) Zn~(2+)浓度下的显著低于对照。丙二醛(MDA)呈现出随着Zn~(2+)浓度提高而增加的趋势,5、10、15和20mg·L~(-1)Zn~(2+)浓度下米氏凯伦藻的丙二醛(MDA)均显著高于对照。结果可为了解重金属对海洋微藻的毒性作用提供数据参考。     

Pb~(2+)、Cd~(2+)和Ce~(3+)对猪胰α-淀粉酶活性的影响

分别研究了Pb2 + 、Cd2 + 和Ce3 + 对Ca (Ⅱ )α 淀粉酶活性影响及对其Ca2 + 的竞争作用 .结果表明三种金属离子低浓度情况下 (0 5～ 5mmol/L)对α 淀粉酶具有激活现象 ,而较高浓度则抑制酶活力 .Pb2 + 、Cd2 + 和Ce3 + 竞争置换α 淀粉酶中Ca2 + 能力的大小是 :Pb2 + >Cd2 + >Ce3 + ,其抑制酶活作用大小 :Pb2 + >Cd2 + >Ce3 + .     

Hg~(2+)对苜蓿叶片的毒害效应

研究了Hg2 + 对苜蓿叶片的毒害效应。结果表明 :随着Hg2 + 浓度的增加和处理时间的延长 ,叶绿素含量下降 ,电导度上升 ;低Hg2 + 浓度及短时间处理 ,超氧阴离子自由基 (O·2 )和丙二醛 (MDA)含量增加 ,SOD、POD和CAT等保护酶活性升高 ,表明膜系统受到了伤害 ;高Hg2 +浓度及长时间处理 ,O·2 和MDA含量下降 ,SOD、POD和CAT等保护酶活性降低 ,表明细胞结构和功能受到了不可逆的伤害。     

Cu~(2+)胁迫条件对涡虫体内过氧化氢酶活性的影响

实验以0 .2 5、0 .5、1.0、2 .0mg·kg- 1 4种质量浓度的CuSO4 溶液培养东亚三角头涡虫(Dugesiajaponia) 4 8h后,提取其体内蛋白质并测定其过氧化氢酶(CAT)的活性。测定结果:涡虫体内蛋白的CAT酶活性依次为12 .5 9、2 3.74、18.37、18.72ū·mg- 1 。以自来水(实验室常规培养方法)为对照,其相应的酶活性为18.78ū·ｍｇ- 1 。实验结果表明:低浓度下Cu2 + 刺激CAT酶活性增加,而在高浓度下,由于涡虫耐受力的存在而使涡虫的CAT酶活性维持在正常水平(与对照组差异不大)。当Cu2 + 浓度达到4mg·kg- 1 时所培养的涡虫不到2 4h全部死亡。     

木麻黄幼苗对模拟酸雨胁迫的响应和Ca2+的调节作用

随着酸雨pH值的下降,木麻黄幼苗中超氧化物歧化酶(SOD)活性、叶绿素含量和叶绿素a/b值逐渐下降,过氧化物酶(POD)活性、可溶性蛋白质含量、丙二醛(MDA)含量和超氧阴离子自由基(O2-·)含量逐渐增加,过氧化氢酶(CAT)活性和脯氨酸含量先升后降.在同一强度酸雨胁迫下,经0.1和1 mmol·L-1Ca(NO3)2处理的SOD活性、CAT活性、叶绿素含量和叶绿素a/b值相对较高,POD活性、蛋白质含量、MDA含量、O2-·含量和脯氨酸含量相对较低,Ca2 可缓解酸雨对木麻黄幼苗的伤害.     

Hg2+、Cd2+和Cu2+对菹草光合系统及保护酶系统的毒害作用

研究了Hg2 、Cd2 、Cu2 对高等水生沉水植物菹草光合系统及保护酶系统的毒害作用。结果表明 :3种离子均使菹草叶片叶绿体自发荧光强度、叶绿素含量、光合速率降低。Hg2 、Cd2 、Cu2 对菹草保护酶系统存在不同的影响 ,短时间低浓度条件下 ,诱导超氧化物歧化酶 (SOD )、过氧化氢酶 (CAT)、过氧化物酶(POD)活性上升 ,随着污染时间的延长、污染浓度的增加 ,酶活性下降 ,其中SOD活性上升持续时间最长 ,下降最慢。 3种离子均使菹草可溶性蛋白含量减少。Hg2 、Cu2 毒性较强 ,Cd2 毒性较弱。Hg2 、Cu2 对菹草的致死浓度为 0 .5～ 1mg/L ,Cd2 为 1～ 2 .5mg/L     

重金属在拟水狼蛛体内的分布及对其体内抗氧化酶活性的影响

为了探明重金属在拟水狼蛛Pirata subpiraticus体内的分布及对其体内抗氧化酶活性的影响,本研究于2009年6月在河南南阳地区5种不同生境下,共采集50份土壤样本和300头拟水狼蛛样本,采用原子吸收光谱法测定了5种不同生境下雄雌拟水狼蛛体内重金属的分布、GSH含量以及GST,CAT和SOD的活性。结果表明：5个采集样点（S1, S2, S3, S4和S5） 拟水狼蛛体内重金属（Cd,Pb,Cu和Zn）的含量差异显著（P<0.05）。同一地点拟水狼蛛体内重金属（Cd,Pb,Cu和Zn）的含量不同身体部位差异显著（P<0.05）,头胸部>足部>腹部,同一地点雄性拟水狼蛛重金属含量显著高于雌性（P<0.05）。在重金属胁迫下,重金属含量高（S1,S2,S3和S4）的拟水狼蛛体内GSH 含量显著高于参照组（S5）,雄性GSH 含量高于雌性（P<0.05）。对于不同身体部位,GSH 含量差异显著,头胸部GSH 含量最高,其次为足部,腹部含量最低;GSH 含量与重金属含量显著正相关（r2=0.9854,P<0.05）。对于GST,CAT和SOD,重金属含量高则酶活性低,雄性酶活性显著低于雌性,不同身体部位酶活差异不显著;GST,CAT和SOD酶活性与重金属（Cd和Pb或者Pb）含量显著负相关。因此检测拟水狼蛛不同身体部位的酶活性变化就可知环境中重金属的污染程度,拟水狼蛛可以作为重金属污染的重要监测指示生物。     

Molecular defense systems are expressed in the king bolete (Boletus edulis) growing near metal smelters

The induction of defense systems against metal exposure was investigated in 48 wild-growing fruiting bodies of the king bolete (Boletus edulis) from two areas polluted with several transition metals from smelters, as well as five reference areas. To determine the degree of metal exposure, cadmium (Cd), zinc (Zn), and copper (Cu) were determined in caps of fruiting bodies by atomic absorption spectrophotometry (AAS), whereas mercury (Hg) was determined by cold vapor atomic fluorescence spectrometry (CVAFS). Caps were analyzed further with respect to relative activities of the antioxidant enzymes, superoxide dismutase (SOD) and catalase (CAT), as well as concentrations of total glutathione (GSHTOT = GSH + GSSG) and relative concentrations of heat shock protein 70 kDa (HSP70). The results showed that concentrations of the four metals, as well as SOD, CAT and HSP70, were significantly elevated in the exposed group (Mann-Whitney, P < or = 0.001). In contrast, GSHTOT was significantly lowered in the exposed group (P < or = 0.05). Significant positive correlations were established between concentrations of Cd, Zn, Hg, or Cu and activities of SOD (Spearman's P < or = 0.01 for the association between SOD and Cd, P < or = 0.001 for all other metal exposure parameters), CAT (P < or = 0.001 for all exposure parameters), or expression of HSP70 (P < or = 0.001 for all exposure parameters). Significant negative correlations were found between total GSH and Cd (P < or = 0.001), Zn (P < or = 0.001), or Hg (P < or = 0.05). We conclude that antioxidant enzymes are induced in wild-growing B. edulis exposed to environmentally relevant concentrations of potentially toxic transition metals; whereas the net consumption of GSH that occurs with increasing metal exposure may reflect GSH consumption by mechanisms of metal detoxification. Finally, the induction of HSP70 suggests that the antioxidant response and the mechanisms in which GSH is consumed are insufficient for protection against the harmful effects of severe metal stress.     

维生素C多聚磷酸酯对小鼠肝脏脂质过氧化物和抗氧化物酶的影响

为研究维生素C多聚磷酸酯对小鼠肝脏脂质过氧化物和抗氧化物酶的影响 ,我们设置了 4个实验组 ,采用 2 4只小鼠 ,饵料中 35 %维生素C多聚磷酸酯的添加量依次为 0、 5 0 0、 2 5 0 0和 5 0 0 0mg/kg ,喂食 4周后取其肝脏 ,用硫代巴比妥酸分光光度测脂质过氧化物的含量 ,用亚硝酸盐形成法测定超氧化物歧化酶的活性 ,用分光光度法测过氧化氢酶和谷胱甘肽过氧化物酶的活性。结果表明 ,维生素C多聚磷酸酯对小鼠肝脏脂质过氧化物没有明显影响 ,但随着维生素C多聚磷酸酯添加量的增加 ,脂质过氧化物有减少的趋势。维生素C多聚磷酸酯添加量为 2 5 0 0和 5 0 0 0mg/kg的两组 ,其超氧化物歧化酶的活性明显高于对照组和维生素C多聚磷酸酯添加量为 5 0 0mg/kg组 ;过氧化氢酶的活性明显高于对照组。维生素C多聚磷酸酯添加量为5 0 0 0mg/kg组 ,其谷胱甘肽过氧化物酶的活性明显高于其它三组。表明高剂量的维生素C多聚磷酸酯能促进小鼠抗氧化物酶的活性 ,但促进不同抗氧化物酶活性所需的维生素C多聚磷酸酯的量不同     

Involvement of glutathione and enzymatic defense system against cadmium toxicity in Bradyrhizobium sp. strains (peanut symbionts)

In this study, the effects of cadmium (Cd) on cell morphology and antioxidant enzyme activities as well as the distribution of the metal in different cell compartments in Bradyrhizobium sp. strains were investigated. These strains were previously classified as sensitive (Bradyrhizobium sp. SEMIA 6144) and tolerant (Bradyrhizobium sp. NLH25) to Cd. Transmission electron micrographs showed large electron-translucent inclusions in the sensitive strain and electron-dense bodies in the tolerant strain, when exposed to Cd. Analysis of Cd distribution revealed that it was mainly bounded to cell wall in both strains. Antioxidant enzyme activities were significantly different in each strain. Only the tolerant strain was able to maintain a glutathione/oxidized glutathione (GSH/GSSG) ratio by an increase of GSH reductase (GR) and GSH peroxidase (GPX) enzyme activities. GSH S-transferase (GST) and catalase (CAT) activities were drastically inhibited in both strains while superoxide dismutase (SOD) showed a significant decrease only in the sensitive strain. In conclusion, our findings suggest that GSH content and its related enzymes are involved in the Bradyrhizobium sp. tolerance to Cd contributing to the cellular redox balance.     

Effects of cadmium stress upon activities of antioxidative enzymes, photosynthetic rate, and production of phytochelatins in leaves and chloroplasts of wheat cultivars differing in yield potential

We tested the mode of action of Cd on photosynthesis and activities of ATP-sulfurylase (ATP-S), catalase (CAT), superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR), and on contents of phytochelatins (PCs) and glutathione (GSH) in two cultivars of wheat (Triticum aestivum L.) PBW-343 and WH-542 differing in yield potential. Cd treatment increased Cd content and photosynthetic activity in PBW-343 more than in WH-542. The activities of APX, GR, ATP-S, and synthesis of PCs and GSH were also increased by Cd, but the CAT and SOD activities were inhibited in both the cultivars. The efficient functioning of antioxidative enzymes, production of PCs and GSH, helped in counteracting the effects of Cd namely in PBW-343, protected photosynthetic ability, and increased the tolerance to Cd.     

The balance between Cu,Zn-superoxide dismutase and catalase affects the sensitivity of mouse epidermal cells to oxidative stress.

Oxidants are toxic, but at low doses they can stimulate rather than inhibit the growth of mammalian cells and play a role in the etiology of cancer and fibrosis. The effect of oxidants on cells is modulated by multiple interacting antioxidant defense systems. We have studied the individual roles and the interaction of Cu,Zn-superoxide dismutase (SOD) and catalase (CAT) in transfectants with human cDNAs of mouse epidermal cells JB6 clone 41. Since only moderate increases in these enzymes are physiologically meaningful, we chose the following five clones for in-depth characterization: CAT 4 and CAT 12 with 2.6-fold and 4.2-fold increased catalase activities, respectively, SOD 15 and SOD 3 with 2.3-fold and 3.6-fold increased Cu,Zn-SOD activities, respectively, and SOCAT 3 with a 3-fold higher catalase activity and 1.7-fold higher Cu,Zn-SOD activity than the parent JB6 clone 41. While the increases in enzyme activities were moderate, the human cDNAs were highly expressed in the transfectants. As demonstrated for the clone SOD 15, this discordance between message concentrations and enzyme activities may be due to the low stability of the human Cu,Zn-SOD mRNA in the mouse recipient cells. According to immunoblots the content of Mn-SOD was unaltered in the transfectants. While the activities of glutathione peroxidase were comparable in all strains, the concentrations of reduced glutathione (GSH) were significantly lower in SOD 3 and SOD 15. This decrease in GSH may reflect a chronic prooxidant state in these Cu,Zn-SOD overproducers.(ABSTRACT TRUNCATED AT 250 WORDS)     

Antioxidative stress proteins and their gene expression in brown trout (Salmo trutta) from three rivers with different heavy metal levels

Three populations of brown trout (Salmo trutta) exposed to different metal levels in their natural environments, were studied with respect to antioxidants metallothionein (MT), superoxide dismutase (SOD) and catalase (CAT) as well as for corresponding mRNA levels. In addition, mRNA levels were studied for glutathione peroxidase (GPx) and glutathione reductase (GR). The Cd/Zn-exposed trout (Naustebekken River) had higher accumulated levels of Cd, Cu and Zn in gills, and higher levels of MT (both protein and mRNA) in liver and kidney as well as in gills compared to the Cu-exposed trout (Rugla River) and trout from an uncontaminated reference river (Stribekken River). Less MT found in the Cu-exposed trout may increase susceptibility to oxidative stress, but no higher levels of antioxidant mRNAs were found in gills of these trouts. The data indicated that chronic exposures of brown trout to Cd, Zn and/or Cu did not involve maintenance of high activities of SOD and CAT enzymes in gills, although SOD mRNA levels were higher in the Cd/Zn-exposed trout. In livers, mRNA levels of SOD, CAT and GPx were higher in the metal-exposed trout, but in the case of GR this was only seen in kidneys of Cd/Zn-exposed trout. However, both metal-exposed groups had higher activities of SOD enzyme in liver compared to the unexposed reference trout, and CAT activity was found to be higher in kidneys of Cu-exposed trout. The Cu-exposed trout did not seem to rely on MT production to avoid Cu toxicity in gills, but rather by keeping the Cu uptake at a low level. A coordinated expression of different stress genes may also be important in chronic metal exposure. It may be concluded that the observed metal effects relies on acclimation rather than on genetic adaptation in the metal exposed populations.     

Accumulation and oxidative stress biomarkers in Japanese flounder larvae and juveniles under chronic cadmium exposure

This study investigated how Cd exposure affected oxidative biomarkers in Japanese flounder, Paralichthys olivaceus, at early life stages (ELS). Fish were exposed to waterborne Cd (0–48 µg L^− 1) from embryonic to juvenile stages for 80 days. Growth, Cd accumulation, activities of superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6), glutathione S-transferase (GST, EC 2.5.1.18), and levels of glutathione (GSH) and lipid peroxidation (LPO) were investigated at three developmental stages. Flounder growth decreased and Cd accumulation increased with increasing Cd concentration. In metamorphosing larvae, CAT and SOD activities were inhibited and GSH level was elevated, while LPO was enhanced by increasing Cd concentrations. CAT and GST activities of settling larvae were inhibited but GSH level was elevated at high Cd concentrations. In juveniles, SOD activity and LPO level were increased but GST activity was inhibited as Cd concentration increased. Antioxidants in flounder at ELS were able to develop ductile responses to defend against oxidative stress, but LPO fatally occurred due to Cd exposure. These biochemical parameters could be used as effective oxidative biomarkers for evaluating Cd contamination and toxicity in marine environments: CAT, SOD, GSH, and LPO for metamorphosing stage; CAT, GSH, and GST for settling stage; and SOD, GST, and LPO for juvenile stage.     

外源抗坏血酸对镉胁迫下黑藻抗氧化系统的保护作用

研究了不同浓度抗坏血酸(ascorbic acid,AsA)对5 mg·L^-1Cd²⁺胁迫下黑藻（Hydrilla verticillata）体内超氧阴离子(O₂^-)产生速率、过氧化氢(H₂O₂)含量、抗坏血酸和谷胱甘肽(GSH)含量,以及超氧化物歧化酶(SOD)、超氧化物酶(POD)、过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)活性的影响.结果表明,与单一Cd²⁺胁迫相比,随着外源AsA浓度的升高,黑藻体内活性氧生成速率逐步降低;抗氧化物质AsA先升后降,GSH缓慢升高;抗氧化酶APX和CAT活性先升后降,POD活性逐步下降,并接近正常状态,对SOD活性影响不大.可见AsA能够有效缓解Cd²⁺对黑藻的毒害,且60 mg·L^-1浓度下的缓解效果最好.