双甲脒对BRL 3A细胞的毒性作用

为了探讨双甲脒对BRL 3A细胞的毒性作用,本试验以0、20、40、60μmol/L的双甲脒分别处理BRL 3A细胞,MTT法检测细胞的相对活力,同时测定细胞培养上清液中细胞乳酸脱氢酶的活性,测定细胞内超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶和细胞内谷胱甘肽还原酶的活性以及还原型谷胱甘肽和活性以及丙二醛的含量。结果表明,与对照组相比,随着双甲脒浓度的升高,细胞活性显著或极显著下降(P0.05或P0.01),细胞培养上清液中LDH活性和细胞内GSH-Px、SOD、CAT的活性显著或极显著升高(P0.05或P0.01),MDA和GSH的含量增加,但差异不显著(P0.05),GR活性先升高后降低,差异显著或极显著(P0.05或P0.01)。结果表明,双甲脒对细胞有明显的毒性,并且氧化应激在其中起到了重要作用。     

莱菔硫烷对镉中毒雄性小鼠生殖机能保护作用的研究

本试验旨在研究莱菔硫烷(SFN)对染镉雄性小鼠生殖机能的保护作用.选择健康的清洁级雄性昆明系小鼠40只,随机分成4组,分别为对照组(H₂O)、镉组(2.3 mg/kg CdCl₂)、莱菔硫烷组(10 mg/kg SFN)、镉+莱菔硫烷组(10 mg/kg SFN+2.3 mg/kg CdCl₂),连续给药10 d,于最后一次给药2 d后脱颈处死,检测小鼠睾丸组织病理学变化、小鼠睾丸及附睾的脏器系数、精液品质及血清睾酮浓度.同时检测睾丸组织内丙二醛(MDA)、谷胱甘肽(GSH)含量及总超氧化物歧化酶(T-SOD)活性的变化.结果表明,与对照组相比,镉组小鼠的睾丸组织发生病理学损伤,睾丸及附睾的脏器系数、精液品质及血清睾酮浓度均极显著降低(P<0.01),睾丸组织T-SOD活性、GSH含量也极显著降低(P<0.01),MDA含量极显著提高(P<0.01);莱菔硫烷组T-SOD活性和GSH含量显著升高(P<0.05),MDA含量与对照组相比差异不显著(P>0.05).与镉组相比,镉+莱菔硫烷组小鼠的精子活率、精子总数极显著增加(P<0.01),睾丸与附睾的脏器系数显著增加(P<0.05),小鼠睾丸组织GSH含量、T-SOD活性极显著提高(P<0.01),MDA含量极显著降低(P<0.01).以上结果指出莱菔硫烷对镉中毒雄性小鼠的生殖机能具有保护作用.     

内毒素诱生自由基致兔血循环系统损伤及CA对其影响

将56只健康白兔随机分组:正常对照组( 组)8只,ET处理组( 组)、CA保护组( 组)各24只,第1、2、3、4和5小时分别采集血液制备血浆样品,检测血浆T-AOC、T-SOD活性和MDA含量,第2和第5小时捕杀并迅速采集心脏制备组织匀浆,检测心脏GSH、GSH-ST、GSH-Px活性,研究内毒素(endotoxin,ET)诱导休克兔产生自由基对血浆和心脏的损伤及阳离子A(cationA,CA)对其影响。结果表明: 组各项指标均保持相对稳定; 组MDA含量比 组升高(P<0.01),T-AOC、T-SOD活性比 组降低(P<0.01),而 组的MDA、T-AOC、T-SOD显著高于 组(P<0.05);除心脏GSH-Px活性第5小时降低不明显外, 组GSH、GSH-Px和GSH-ST活性显著降低(P<0.01), 组比 组GSH、GSH-Px、GSH-ST活性高(P<0.01)。结果提示:CA能减轻ET诱生自由基所致的过氧化损伤,有效缓解内毒素引起的血液循环系统毒性损伤。     

镉对体外培养鸡脾淋巴细胞膜抗氧化功能的影响

以终浓度为10μmol/L CdCl2对体外培养淋巴细胞进行染毒,分别培养12、24、36、48和72h时收集细胞,提取体外培养脾淋巴细胞膜,采用试剂盒比色的方法,分别检测了细胞膜GSH-Px、SOD的活性、膜MDA和SA含量及培养液LDH的活性,来观察镉性细胞损害时细胞膜的抗氧化功能和通透性。结果显示,随着镉作用时间的延长,淋巴细胞膜GSH-Px、SOD的活性降低,MDA生成量增加,试验组与对照组差异显著（P〈0.05）或极显著（P〈0.01）;培养液中LDH活性升高,膜SA含量降低。表明镉可使淋巴细胞膜的抗氧化功能降低,膜完整性被破坏。     

镉对PC12细胞的氧化损伤及α-硫辛酸的保护效应

为研究α-硫辛酸（α-lipoid acid,α-LA）对镉致PC12细胞氧化损伤的保护效应,通过不同浓度的醋酸镉染毒PC12细胞,并用100μmol/Lα-LA联合作用,测定PC12细胞内超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GSH-Px）活性和丙二醛（MDA）含量。结果表明：与对照组相比,10μmol/L醋酸镉处理24h组细胞MDA含量和CAT活性极显著升高（P〈0.01）,SOD和GSH-Px活性极显著降低（P〈0.01）;不同浓度镉处理组细胞内MDA含量和CAT活性显著或极显著升高（P〈0.05或P〈0.01）,10和20μmol/L组SOD和GSH-Px活性极显著降低（P〈0.01）。α-LA保护组与相应染毒组相比,MDA含量和CAT活性显著降低（P〈0.05）,SOD和GSHPx活性有升高趋势,但组间差异不显著（P〉0.05）。说明镉可致PC12细胞发生氧化应激,α-LA可以提高PC12细胞的抗氧化能力,对镉引起的氧化损伤有一定的保护作用。     

银杏叶提取物对热应激致鸡心肌细胞氧化损伤的保护作用研究

本试验旨在研究银杏叶提取物（Ginko biloba extract,GBE）对热应激致鸡心肌细胞氧化损伤的保护作用。取12日龄AA肉鸡心脏制备鸡原代心肌细胞,向原代心肌细胞培养板中分别加入0、5、10、50、100、150 mg/mL GBE,CCK-8试剂盒检测心肌细胞存活率;将原代心肌细胞以1×10⁴个/孔接种于96孔板,培养48 h,分别加入0、5、10、50、100、200、500、1 000 μg/mL GBE,Hoechst 33258检测心肌细胞凋亡率,确定最适GBE浓度。42℃热应激处理（0、0.5、1、2和4 h）建立心肌细胞损伤模型,ELISA试剂盒检测乳酸脱氢酶（LDH）、细胞丙二醛（MDA）和谷胱甘肽过氧化物酶（GSH-Px）含量,以及超氧化物歧化酶（SOD）活性;Western blotting检测Hsp72蛋白表达。结果显示,高浓度GBE具有细胞毒性,50、100、150 mg/mL GBE能极显著抑制细胞生长（P<0.01）,50 μg/mL GBE能够极显著提高细胞增殖率（P<0.01）。热应激后细胞发生凋亡,细胞内MDA、LDH含量增加,SOD活性、GSH-Px含量减少,产生氧化损伤;GBE可以降低热应激时细胞LDH和MDA的含量,增加SOD活性和GSH-Px的含量,热应激2、4 h,GBE处理效果明显。此外,GBE可增加心肌细胞中Hsp72蛋白的表达。综上所述,GBE能降低热应激状态下心肌细胞的氧化损伤,其机制可能与提高抗氧化酶活性及提高Hsp72的表达有关。     

黄藤素对山羊子宫内膜上皮细胞的体外毒性研究

为了探究黄藤素的体外细胞毒性,采用不同浓度的黄藤素作用山羊子宫内膜上皮细胞（EECs）后,采用MTT法绘制细胞生长曲线;倒置显微镜下观察细胞生长状态;瑞氏-姬姆萨染色法观察细胞形态学变化并通过测定乳酸脱氢酶（LDH）含量检测细胞膜的完整性。结果显示,当黄藤素的浓度为10~227 μg/mL时,黄藤素对EECs有促增殖的作用;当浓度大于227 μg/mL时出现细胞增殖抑制现象,药物对细胞的半数增殖抑制率（IC₅₀）为360 μg/mL;瑞氏-姬姆萨染色结果显示,100、200 μg/mL的黄藤素作用于EECs时均未对其细胞核、细胞浆的形态产生影响;LDH结果显示,药物浓度≥250 μg/mL时细胞LDH的释放量极显著高于正常细胞组（P<0.01）。综上所述,黄藤素作用于山羊EECs后呈现一定的毒性作用,且呈现显著的剂量依赖效应,作用于EECs的安全剂量范围为≤250 μg/mL。     

镉对原代培养大鼠肝细胞的损伤作用

用活体肝脏灌注、胶原酶消化法分离大鼠肝细胞并作原代培养,以CdCl2(0～250或500umol/L）处理,测定培养上清液中LDH、ALT、ARG和细胞内GSH-Px、GSH、MDA等的含量,探讨镉对肝细胞的损伤作用和脂质过氧化状况。结果当Cd≥10和5umol/L时,细胞内LDH、ARG泄漏分别明显增加,GSH-Px活力显著受到抑制,GSH含量和MDA产生随着镉的剂量增大而增加（Cd≥50 umol/L）。认为镉对肝细胞的损伤和对细胞膜完整性的破坏,与脂质过氧化有关;镉处理后细胞中GSH增加可能与镉抑制了还原型谷胱甘肽向氧化型谷胱甘肽的转化有关。镉可直接抑制ALT本身的活性,因而认为以细胞内ALT泄漏作为镉诱导细胞损伤的评价指标时要予以注意。     

原花青素B2对青霉酸致TM3细胞氧化损伤的修复作用

以TM3细胞为研究对象,探讨原花青素B2对青霉酸致TM3细胞氧化损伤的修复作用。结果显示,在给予青霉酸基础上的同时添加原花青素B2,与单独添加青霉酸相比,TM3细胞内的SOD、CAT和GSH-Px的活性显著升高,细胞内MDA的含量显著降低,TM3细胞的活力则显著增加。表明原花青素B2可以通过修复细胞内的氧化损伤来拮抗青霉酸对TM3的细胞毒性。     

连翘酯苷对LPS诱导的肉鸡法氏囊中NO及抗氧化酶活性的影响

本试验研究了连翘酯苷对内毒素(LPS)诱导的肉鸡法氏囊中一氧化氮(NO)、总抗氧化能力(T-AOC)、总超氧化物歧化酶(T-SOD)、还原型谷胱甘肽(GSH)和丙二醛(MDA)含量的影响.结果表明:连翘酯苷组中T-AOC、T-SOD和GSH活性明显升高,与正常组比较差异显著(P＜0.05)；LPS组中NO和MDA含量明显升高,T-AOC、T-SOD和GSH活性明显降低,与正常组比较差异显著(P＜0.05)；连翘酯苷治疗组中NO和MDA含量明显降低,T-AOC、T-SOD和GSH活性明显升高,与LPS组比较差异显著(P＜0.05).研究结果表明:连翘酯苷可提高肉仔鸡抗氧化功能,并且可以抑制由LPS引起的氧化反应.     

Research on the Antioxident Function of Sulforaphane on the Leydig Cells of Cadmium Exposured Mice

The study was aimed to explore the antioxident function of sulforaphane (SFN) on the leydig cells of cadmium (Cd) exposured mice.Cd and SFN were added to the cell culture medium of TM3 cell, half inhibitory concentration (IC₅₀) of Cd and SFN safe dose range were determined.The tested model was set up,and the relative survival rate of TM3 cells, lactate dehydrogenase (LDH) activity and cell antioxidant levels were determined to study the antioxident function of SFN to cadmium exposured mice.The results showed that:①With the increase of Cd concentration,the relative survival rate of TM3 cells was decreased,and the IC₅₀ of Cd was 51.4 μmol/L;Within a certain concentration range,SFN could increased the cells survive rate,but there was toxicity when the SFN concentration was more than the scope,and the greater the concentration,the greater the toxicity. At experimental condition,SFN safety concentration were 2.5,5 and 10 μmol/L.②Compared with the control group,the GSH content,the T-SOD and GSH-Px activities in Cd group were significantly or extremely significantly decreased (P < 0.05;P < 0.01),and the LDH activity,the MDA content were increased.Additionally,the LDH activity and MDA content of SFN groups were decreased,while others three indexes were increased. Compared with Cd group,the GSH content,the T-SOD and GSH-Px activities of Cd+SFN groups were significantly or extremely significantly increased (P < 0.05;P < 0.01),however,the LDH activity,MDA content were decreased. The result indicated that SFN had the antagonism effect on toxicity of Cd in TM3 cell.     

Protective Effect of Sulforaphane on Reproductive Function of Male Mice with Cadmium Poisoning

The study was aimed to explore the protective effect of sulforaphane (SFN) on the reproductive function of male mice with cadmium poisoning.40 healthy clean grade male Kunming mice were randomly divided into four groups:control group (H₂O),cadmium chloride group (2.3 mg/kg CdCl₂),sulforaphane group (10 mg/kg SFN),sulforaphane + cadmium chloride group (10 mg/kg SFN+2.3 mg/kg CdCl₂),and continuous administration for 10 d,all mice were executed by dislocated cervical vertebra at 2 d after the last administration,and then the pathologic changes of testicular tissues,organ coefficient of testicle and epididymis,sperm quality and concentration of testosterone were tested.Additionally,the contents of GSH and MDA,and the activities of T-SOD in testis were also detected at the same time. Compared with the control group,pathology damages were observed in cadmium chloride group,organ coefficient of testis and epididymis,sperm quality and levels of testosterone extremely significantly decreased (P<0.01),the activities of T-SOD and GSH content were extremely significantly decreased (P<0.01),and the concentration of MDA was extremely significantly enhanced (P<0.01).Compared with the control group,the activity of T-SOD and concentration of GSH in sulforaphane group were significantly increased (P<0.05),and the concentration of MDA was not significant different between the control group and sulforaphane group (P>0.05).While compared with the cadmium chloride group,the sperm motility rate and sperm total count in sulforaphane and cadmium chloride group were extremely significantly increased (P<0.01),the organ coefficient of testicle and epididymis was increased significantly (P<0.05),the concentration of GSH and activity of T-SOD in testicular tissue were extremely significantly increased (P<0.01),and the concentration of MDA was extremely significantly decreased (P<0.01).The results indicated that sulforaphane had the protection effect on reproduction function of male mice with cadmium poisoning.     

镉胁迫对白三叶的富集能力、叶片显微结构及其生理特性的影响

以白三叶为试验材料,对其进行不同浓度Cd²⁺(0, 100, 200, 300, 400, 500 μmol/L)胁迫7 d,结果表明,随Cd²⁺浓度增加,白三叶各营养器官Cd积累量随之升高,各器官对Cd的富集能力表现为:根>叶>茎;同时,富集在叶片部分的Cd对白三叶叶片产生了毒害作用,使叶肉细胞的细胞壁着色加深,栅栏组织细胞以及海绵组织细胞的形状、大小及细胞排列的疏密程度发生改变。Cd²⁺胁迫后,白三叶叶片叶绿素含量显著降低(P<0.05),叶片的MDA含量呈上升趋势,且与Cd²⁺浓度之间存在剂量效应关系;整个胁迫过程中,SOD,POD和CAT活性则呈先升高后降低的趋势。相关性分析结果表明,Chl a和Chl b含量与Cd²⁺浓度间呈极显著负相关;MDA含量与Cd²⁺浓度间呈极显著正相关;SOD,POD,CAT之间呈极显著正相关,三者发挥协同作用以抵抗逆境毒害,体现了白三叶对重金属Cd的适应性。     

亚临床酮病奶牛血清中微量元素代谢水平的研究

本试验旨在研究荷斯坦奶牛亚临床酮病对血清中微量元素的影响,选择40头产后1～15 d的荷斯坦亚临床酮病奶牛,通过应用原子吸收光谱法测定血清中Cu、Fe、Mn、Zn、Se、Co含量,应用分光光度计和比色法测定血清中GSH-Px、T-AOC、T-SOD活力及MDA含量。结果显示,阳性组血清中Cu、Zn、Se、Co的含量在整个检测过程中均极显著低于对照组(P＜0.01);阳性组血清中Mn的含量在第1、5天的检测中显著低于对照组(P＜0.05),第10、15天极显著低于对照组(P＜0.01);阳性组血清中Fe的含量与对照组相比差异不显著（P＞0.05）;阳性组血清中GSH-Px、T-SOD、T-AOC的活性均极显著低于对照组(P＜0.01);阳性组血清中MDA的含量极显著高于对照组(P＜0.01)。结果表明,亚临床酮病直接影响奶牛抗氧化系统GSH-Px、T-AOC、T-SOD的活性,间接地影响血清中微量元素Cu、Mn、Se、Zn的含量。Co是维生素B₁₂的重要组成部分,维生素B₁₂在肝脏内的浓度下降,从而导致亚临床酮病血清中Co的含量降低。亚临床酮病奶牛肝脏和氧化酶受损,破坏Fe的结合,机体内贮存的Fe则可以释放出来,维持机体的需要,血清中Fe的含量没有显著的变化。     

铅、镉胁迫对饲用高粱部分生理指标的影响

采用盆栽试验研究了土壤外源重金属铅、镉及其复合污染对饲用高粱（Sorghum vulgare Pers.）部分生理指标的影响。旨在为植物生长代谢过程中适应和抵御重金属污染的损伤机制提供一定的理论依据。结果表明：随着Pb和Cd单一及复合胁迫浓度的增加,饲用高粱的叶绿素含量降低,丙二醛含量、叶片细胞膜透性、脯氨酸含量均呈上升趋势。Pb和Cd单一处理组中Cd胁迫的毒害作用强于Pb。Cd浓度为5和10 mg·kg^-1的复合处理组与单一处理组间无显著差异,Cd浓度为50 mg·kg^-1的复合处理组以及Cd₁₀Pb₅₀₀复合处理组对饲用高粱幼苗的毒害作用高于单一胁迫,表明Pb加强了高浓度Cd对植物的毒害作用。     

The Study on the Cytotoxicity of Palmatine on Goat Endometrial Epithelial Cells in vitro

To explore the cytotoxicity of palmatine in vitro,goat endometrial epithelial cells (EECs) were stimulated by different concentrations of palmatine,then the growth curve of cell was drew by MTT assay which could detect the cell proliferation. The cell growth state was observed by inverted microscope,and Wright-Giemsa staining was used to observe morphological changes of the cells while LDH test was used to check the membrane permeability of EECs. The results showed that when the concentration was 10 to 227 μg/mL,palmatine had a role in promoting proliferation of the EECs,while when the concentration was more than 227 μg/mL,cell proliferation was inhibited,and 50% inhibitory rate of proliferation (IC₅₀) of this drug on EECs was 360 μg/mL.Wright-Giemsa staining results showed that themorphology of the nucleus and cytolymph were not significantly changed when 100 and 200 μg/mL palmatine effected EECs. LDH results showed that when the palmatine concentration was more than 250 μg/mL,LDH content was extremely significantly higher than that of normal cell group(P<0.01).In conclusion,palmatine presented certain toxic effect on EECs with a significant dose-response effect,and the safe dose range of palmatine was less than 250 μg/mL.