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1.
采用自由饮水的方式于母鼠孕哺期染毒,染毒剂量为0.3、1.0、3.0 g/L,每组10只.用石墨炉原子吸收光谱仪测定仔鼠出生后第21天时血液和海马组织中铅含量.用RT-PCR方法测定21 d不同剂量组仔鼠海马组织中钙结合蛋白-2 (Calsyntenin-2) mRNA的表达、结果显示,各剂量染铅组血液、海马组织中铅含量与对照组相比,明显升高(P<0.05).Y型迷宫试验结果显示,低、中、高剂量铅暴露组21 d的仔鼠,获取记忆的能力(T1)、保持记忆的能力(T2)和记忆保持率(T3)均低于同期对照组仔鼠,差异均有统计学意义(P<0.05).与对照组相比,中、高剂量铅暴露组海马组织中Calsyntenin-2 mRNA的表达明显低于对照组(P<0.05),但是低剂量组与对照组相比,差异无统计学意义(P>0.05).结果表明,孕哺期母体铅暴露使铅在仔鼠体内蓄积,引起仔鼠学习记忆功能损害;铅的神经毒性可能与下调海马组织中Calsyntenin-2 mRNA的表达、影响突触的信息传递有关.  相似文献   

2.
《中国兽医学报》2016,(2):310-313
探讨母体铅暴露对仔鼠海马内钙结合蛋白-1(Calsyntenin-1,Clstn-1)表达的影响。采用自由饮水模式建立铅暴露动物模型,将40只孕小鼠,随机分为对照组和低、中、高剂量染毒组,分别饮用去离子水和1、5、10g/L的乙酸铅水溶液。仔鼠21日龄,分别测其血液和海马组织中铅的含量,然后取其海马组织,分别通过免疫组化方法和Western blot技术检测各组仔鼠海马组织中Clstn-1的蛋白表达情况。结果显示,21d仔鼠血铅、海马铅水平均明显高于对照组(P0.01);免疫组化染色结果显示,Clstn-1免疫组化阳性反应主要定位于胞浆,在仔鼠海马组织的CA1区域中,每个铅暴露组仔鼠Clstn-1免疫组化阳性反应物的面密度与对照组相比增加显著,但平均灰度值明显降低(P0.01);Western blot结果显示,与对照组相比,铅暴露组仔鼠Clstn-1蛋白的表达明显高于对照组,并随染毒剂量增加,表达量增高,差异具有统计学意义(P0.01)。母体铅暴露可能通过上调仔鼠海马内钙结合蛋白的表达进而造成海马损伤,引起神经毒性。  相似文献   

3.
为探讨母体铅染毒对其子一代海马组织中白细胞介素-1β(IL-1β)蛋白表达的影响,将雌性小鼠自妊娠第1天开始经饮水染铅(0.3,1.0,3.0g/L,对照组饮蒸馏水)至仔鼠出生后21d断乳为止。随机抽取各组仔鼠,于出生后7,14,21d分别测其血液和海马组织中铅的含量;于出生后21d,采用Western blot方法测定各组海马组织中IL-1β的表达情况。结果表明,孕哺期不同剂量铅暴露后,出生后7,14,21d的仔鼠血铅、海马铅水平均明显高于对照组(P〈0.05)。Western blot结果显示,IL-1β蛋白在不同剂量铅暴露组海马组织中的表达明显高于对照组(P〈0.05)。相关分析表明,IL-1β蛋白的表达与血铅、海马铅呈显著正相关(P〈0.05)。母体铅暴露使铅在仔鼠体内蓄积,提示仔鼠血铅和海马铅的升高,引起海马组织中IL-1β表达的上调,可能损伤了海马神经元,进而损伤了神经系统。  相似文献   

4.
探讨母体受铅暴露对其后代小鼠的海马组织中突触蛋白表达的影响。铅暴露是从母体妊娠初期至断奶为止。将醋酸铅(PbAc)溶于去离子蒸馏水中,再分别浓缩至0.1%,0.5%和1%3个组作为仔鼠饮用溶液。在出生21d后,用石墨炉原子光谱吸收法测定铅指标,用免疫组织化学和Western blot检测海马组织中突触蛋白的表达。结果表明,所有铅暴露组中海马组织和血液中的铅指标均显著高于对照组(P0.05)。与对照组相比,突触蛋白的表达显著下降(P0.05)。母体铅暴露引起的仔鼠海马组织中突触蛋白过少表达可能是铅神经毒性所致。  相似文献   

5.
《中国兽医学报》2016,(1):148-151
雌性小鼠自妊娠第1天开始经饮水染铅(0.1%、0.5%、1%浓度溶解在去离子水中,对照组饮蒸馏水)至仔鼠出生后21d断乳为止。在出生后的第21天,分别采用石墨炉原子吸收光谱法测定血铅和海马组织中铅的水平,免疫组织化学和免疫印迹法检测海马组织中胰岛素样生长因子1(insulin-like growth factor 1,IGF1)的表达。结果显示:与对照组相比,铅暴露组血铅水平和海马组织中铅的水平显著增高(P0.05);铅暴露组IGF1的表达比对照组明显下降(P0.05)。结果表明:母体铅暴露使铅在仔鼠体内蓄积,使仔鼠海马组织中IGF1蛋白的表达下调,从而造成海马神经元损伤,引起神经毒性。  相似文献   

6.
《中国兽医学报》2017,(2):308-311
探讨母体铅染毒对其仔鼠大脑皮层组织中IL-1β和TNF-α表达量的影响,揭示铅神经毒性的潜在机制。母鼠采用自由饮水的方式自妊娠1d开始经饮水染铅(0.1%,0.5%和1.0%的浓度溶解在去离子水中,对照组饮蒸馏水)至仔鼠出生后21d断乳为止,每组10只。于出生后21d,用石墨炉原子吸收光谱仪测定仔鼠血液和大脑皮层组织中铅含量。用免疫组织化学染色法和Western blot方法测定大脑皮层组织中IL-1β和TNF-α蛋白表达情况。结果表明,各剂量染铅组血铅和脑铅含量与对照组比较明显升高(P<0.05);与对照组相比,随着试验组染铅浓度的升高,IL-1β和TNF-α的表达量显著增加(P<0.05)。铅可能通过诱导大脑皮层中IL-1β和TNF-α的高表达,影响学习记忆从而造成神经系统损害。  相似文献   

7.
为了观察母体怀孕及哺乳期铅摄入致仔鼠海马c-fos的变化,试验采用RT-PCR的方法分别检测母鼠怀孕期铅暴露后仔鼠和仔鼠出生早期经乳汁染铅后脑海马组织c-fos的变化。结果表明:出生后给铅,仔鼠脑海马组织c-fos的表达2天时低剂量组c-fos的表达明显增强,高剂量组的表达也增多,且均较对照组表达多;4天时c-fos的表达较2天时有所下降,其中低剂量组受到抑制而高剂量组表达较对照组明显增多;10天时3个组均增强,高剂量组比对照组表达明显增强;20天时c-fos的表达均下降,但2个剂量组的表达仍较对照组多。孕晚期给铅,4天时高剂量组的c-fos表达受到严重抑制;生后10天时3个组表达迅速增加;20天时c-fos的表达又迅速下降,但低剂量组表达仍比对照组高而高剂量组表达低于对照组。孕早期给铅,仔鼠生后2天时2个剂量组c-fos表达比对照组多,其中高剂量组表达比对照组明显增加;但4天时表达急剧减少;10天时c-fos的表达又急剧增加,但高剂量组的表达比对照组和低剂量组少;20天时c-fos的表达又急剧下降。说明母鼠不同的铅暴露时间和强度对仔鼠脑海马组织c-fos的表达变化不同。  相似文献   

8.
为了研究孕鼠在孕期暴露双酚A(bisphenol A,BPA)对仔鼠生殖激素及相关基因的影响,试验将40只昆明孕鼠随机分为A、B、C、D共4组,每组10只。其中A组为对照组,饲喂普通鼠粮;B、C、D组孕鼠整个妊娠期(妊娠1 d至分娩)分别按每只鼠每天50、500、2 500 mg/kg体重给予BPA,待孕鼠自然分娩,观察记录仔鼠死亡情况。至仔鼠性成熟(56日龄)剖杀仔鼠,摘取睾丸或卵巢称重并计算脏器指数,HE染色观察卵巢或睾丸组织结构的变化,ELISA试剂盒分析仔鼠血清睾酮(T)、促卵泡素(FSH)及雌二醇(E2)水平,免疫组织化学方法检测仔鼠睾丸或卵巢Bax、Bcl-2蛋白的表达,实时荧光定量PCR检测仔鼠睾丸StAR、CYP11a或卵巢AMH、Kitlg mRNA表达。结果显示,孕鼠暴露BPA极显著增加了仔鼠死亡率(P<0.01),显著降低了仔鼠睾丸重(P<0.05)。ELISA检测结果表明,孕鼠暴露BPA极显著降低了仔鼠T(♂)及FSH(♀)含量(P<0.01),极显著升高了仔鼠(♀) E2水平(P<0.01)。HE染色结果显示,随BPA剂量增加,仔鼠睾丸组织损伤严重,间质细胞减少;卵巢组织结构随BPA剂量增大,空泡逐渐增多,黄体颗粒数量逐渐减少。免疫组化结果显示,孕鼠暴露BPA增加了仔鼠睾丸或卵巢组织中Bax阳性蛋白表达,减少了Bcl-2阳性蛋白表达,显著降低了雄性仔鼠StAR mRNA表达量(P<0.05);B、D组雄性仔鼠CYP11a mRNA表达量极显著降低(P<0.01),而C组CYP11a mRNA表达量极显著升高(P<0.01);C、D组雌鼠Kitlg mRNA表达极显著降低(P<0.01),AMH mRNA表达量显著升高(P<0.05)。本试验结果表明,孕鼠妊娠期暴露不同剂量BPA增加了仔鼠死亡率,扰乱了生殖激素平衡和睾丸/卵巢中相关凋亡蛋白及生殖基因表达。  相似文献   

9.
为观察母鼠围产期铅暴露对仔鼠各组织微量元素含量的影响及N-乙酰半胱氨酸(NAC)的保护效应,采用电感耦合等离子体质谱法(ICP-MS)测定低水平铅暴露后的仔鼠血液、大脑、骨骼、肝脏和肾脏等组织中的铅、锌、铜、铁、锰和硒等微量元素的含量。结果表明,与对照组相比,染铅组所测组织铅含量均极显著升高(P<0.01),锌含量均极显著降低(P<0.01);肝脏铁含量、肾脏硒含量及骨骼铁、铜、硒含量均显著降低(P<0.01或P<0.05)。NAC拮抗组与染毒组比较,各组织铅含量显著降低(P<0.01或P<0.05),血液和肾脏锌含量显著升高(P<0.05),其他元素含量无明显差异(P>0.05)。  相似文献   

10.
通过检测基质金属蛋白酶(MMPs)在正常及患病奶牛子宫内膜组织中的表达变化,探讨奶牛子宫内膜炎的发病机理。采集正常及患子宫内膜炎奶牛的子宫内膜组织,半定量PCR法检测其中MMP-2、MMP-9 mRNA的表达,明胶酶谱法检测MMP-2、MMP-9的酶活性。结果显示,患病组织中明胶酶的mRNA表达量显著上升,酶活性显著提高,说明在炎症发生时,明胶酶增多以降解被破坏的细胞外基质,从而修复子宫内膜。  相似文献   

11.
为探讨基质金属蛋白酶(MMPs)-2、MMP-9在奶牛子宫内膜炎中的作用机制,选用产后6~10d健康及患急性化脓性子宫内膜炎的中国荷斯坦奶牛各10头,分别为对照组和试验组,通过ELISA检测PGF2a、孕酮浓度;免疫组化检测子宫内膜Ⅰ型胶原、Ⅳ型胶原的表达;SYBR Green I荧光定量PCR检测子宫内膜MMP-2和MMP-9mRNA表达。结果表明:试验组PGF2a浓度极显著低于对照组(P<0.01),孕酮含量显著高于对照组(P<0.05),PGF2a与孕酮浓度呈显著负相关(r=0.893);试验组子宫内膜中Ⅰ型胶原、Ⅳ型胶原的表达量均高于对照组,差异极显著(P<0.01);MMP-2和MMP-9mRNA水平则显著低于对照组(P<0.01)。结果提示:产后子宫内膜炎病牛子宫内膜MMP-2、MMP-9低表达,导致Ⅰ型和Ⅳ型胶原降解受阻,使细胞外基质为子宫内膜细胞提供的刺激信号不足,子宫分泌PGF2a量少,黄体消退障碍,血清孕酮含量高;说明子宫内膜MMP-2、MMP-9mRNA表达对产后子宫内膜炎的发生和发展起促进作用。  相似文献   

12.
关节滑膜炎是骨关节疾病发病的基本病理过程,试验通过研究马急性滑膜炎关节液中基质金属蛋白酶-3(MMP-3)和基质金属蛋白酶-13(MMP-13)含量的变化,进而研究马滑膜炎机理。试验选用8匹马,在马右侧中间腕关节内注入0.5 ng大肠杆菌脂多糖(LPS)诱导滑膜炎,同时在对侧关节内注入等量的PBS作为对照,于注射药物前和注射药物后的8、24 h和168 h,采马关节液.用ELISA检测马关节液中MMP-3和MMP-13的含量。结果注入LPS的关节液中MMP-3和MMP-13的量在注射后的24 h内持续升高,在24 h达到最大量10.01 ng/mL和6.08 ng/mL,与对照组比较差异性显著,24 h后开始下降,168 h时含量恢复到对照水平。试验结果表明,12'S诱导的滑膜炎模型为一过性的.在急性滑膜炎过程中,MMP-3和MMP-13的表达和释放增加,并且这两种酶在滑膜炎过程中起到重要作用。  相似文献   

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The objective here was to evaluate the acute effects of induced arthritis on synovial fluid (SF) levels of matrix metalloproteinases (MMP) -2, -8 and -9 in horses. To evaluate MMP-2 and -9 activities and the effect of non-steroidal anti-inflammatory drug (NSAID) bufexamac during remission from acute arthritis. Aseptic arthritis was induced in 24 Standardbred horses using 20 mg of amphotericin B as a single intra-articular (IA) injection in the right intercarpal joint. After 1 week and 2 weeks, horses were treated intra-articularly with 10, 20, or 40 mg of bufexamac suspension or with sterile saline solution as control. SF was sampled prior to induction and at weekly intervals for 5 weeks. Fluids were evaluated for MMP-2 and MMP-9 activity by gelatin zymography or for MMP-8 immunoreactivity by Western Blotting. IA injection of amphotericin B consistently resulted in significant increase in the immunoreactivity of MMP-8 and activity of both the latent and the active forms of MMP-2 and -9, among which the active form of MMP-2 increased the most. MMP-9 levels declined to pre-induction levels within 2 weeks, whereas levels of MMP-2 remained still high after 5 weeks. Treatment with bufexamac did not significantly affect levels of gelatinolytic MMP. Results suggest that after acute arthritis of horses, elevated MMP activity is present in the joint, for several weeks, to a degree that could promote cartilage degradation, and treatment with the NSAID bufexamac is not likely to affect that. Furthermore, analysing levels of MMP-9 activity and especially levels of active forms of MMP-2 activity may be valuable to predict the time of occurrence of arthritis in horses.  相似文献   

15.
Although mast cells contribute to host protective immunity against bacterial infections, the exact mechanism of their recruitment at the affected site has been unclear. Recently, we have reported that both mouse and human mast cells are capable of producing matrix metalloproteinase (MMP)-9, a matrix-degrading enzyme necessary for leukocyte transmigration. Here, we demonstrated that bacterial lipopolysaccharide (LPS) enhanced MMP-9 production of mouse bone marrow derived-cultured mast cells. This action of LPS was partially suppressed by the pretreatment of cultured mast cells with a protein kinase C (PKC) inhibitor, indicating the possible involvement of PKC signaling pathways in the production of MMP-9 by LPS. Thus, these suggest the upregulation of mast cell MMP-9 by bacterial components, thereby resulting in their migration at the affected site.  相似文献   

16.
REASONS FOR PERFORMING STUDY: Airway matrix metalloproteinases (MMPs) increase following inhalation of organic dust. The relative contribution of dust components to this elevation is unknown. OBJECTIVE: To identify components of organic dust responsible for elevated MMP levels in equine airways. METHODS: Bronchoalveolar lavage (BALF) from 7 heaves-susceptible horses, collected 6 h following inhalation challenges with saline, 2 different hay dust suspensions (HDS-1 and -2) and soluble and particulate fractions of HDS-1, were analysed for MMP-2 and -9 using SDS-page gelatin zymography. RESULTS: HDS-1 challenge increased BALF proMMP-9 and total MMP-9. HDS-1 fractions, or the particulate fraction with added lipopolysaccharide, increased BALF proMMP-9 and total MMP-9 in combination, but not when inhaled separately. HDS-2 inhalation elevated BALF complex forms, proMMP-9, active MMP-9, total MMP-9 and total MMP-2. CONCLUSIONS: The results suggest synergistic action of soluble and particulate organic dust components. The fact that HDS-1 and HDS-2 had different glucan concentrations supports a role for moulds in the activation of MMP-9. POTENTIAL RELEVANCE: Activation and release of MMPs in response to inhaled moulds are involved in the aetiopathogenesis of heaves. Endotoxin contributes to the synergistic action of the dust components, but the overall MMP response to organic dust inhalation in heaves-susceptible horses largely reflects the mould content of the dust. In the future, inhibition of MMP production and release may offer therapeutic means for treatment and prevention of heaves and recommendations for acceptable dust levels can be given.  相似文献   

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本研究旨在揭示肌抑素(MSTN)与基质金属蛋白酶(MMPs)的调控关系,探明MSTN对PK15细胞MMP-2/7/9表达的影响。对MSTN单等位基因敲除猪背膘组织进行转录组测序分析,复苏前期制备的MSTN基因敲除PK15细胞系:单等位基因敲除的PK3108细胞系和双等位基因敲除的L18细胞系,通过实时荧光定量PCR和Wesrern blotting分别检测PK15、PK3108和L18细胞系中MSTN、MMP-2/7/9基因的mRNA和蛋白表达水平。结果发现,与野生型猪相比,MSTN单等位基因敲除猪背膘组织转录因子C/EBPδ、MMP-2/7基因mRNA表达量均极显著下调(P < 0.01);细胞外基质中纤连蛋白(FN)和层连接蛋白(LN)含量均极显著增加(P < 0.01)。复苏的PK3108和L18细胞呈现绿色荧光。实时荧光定量PCR结果显示,PK3108和L18细胞中MSTN、MMP-2/7/9的mRNA表达量均极显著低于PK15细胞(P < 0.01);Western blotting结果显示,PK3108和L18细胞中MSTN、MMP-2/7/9的蛋白表达量均明显低于PK15细胞。本研究结果表明,在MSTN基因敲除的PK15细胞中,MSTN功能缺失能显著降低MMP-2/7/9的表达,且MMP-2/7/9蛋白表达降低的趋势与MSTN蛋白表达的趋势相一致。  相似文献   

19.
Activity of matrix metalloproteinase-2 (MMP-2) in canine oronasal tumors   总被引:1,自引:0,他引:1  
Activity of matrix metalloprotease-2 (MMP-2) and the expression of its related molecules were examined in spontaneous canine oronasal tumors. Tissue samples from melanoma and squamous cell carcinoma possessed higher MMP-2 activity, as shown in gelatin zymography, in comparison with acanthomatous epulis and nasal adenocarcinoma. Regional lymph node invasion and distant metastases were more frequently observed in the MMP-2 positive cases. There were no significant differences by RT-PCR examination in the expression of the genes encoding MMP-2, MT1-MMP and TIMP-2 among the tumor histological types. However, the MMP-2/TIMP-2 ratio showed a significantly higher level of the genes in the malignant oral melanoma and squamous cell carcinoma. The MMP-2/TIMP-2 ratio was also positively correlated with MMP-2 activity in gelatin zymography. These results indicate that the MMP-2/TIMP-2 ratio may be of value in evaluating the prognosis in canine oronasal cavity tumors.  相似文献   

20.
探讨原花青素对家兔动脉粥样硬化模型基质金属蛋白酶-9(MMP-9)表达的影响。将24只新西兰白兔随机分为对照组(n=8)、模型组(n=8)和原花青素组(n=8)。模型组给予高脂日粮,原花青素组在模型组的基础上加用原花青素。采集试验前及喂食12周后试验兔的耳缘静脉血,检测静脉血中血脂水平的变化,观察主动脉病理形态学变化情况,用免疫组织化学方法检测MMP-9的表达。结果显示,在高脂日粮喂养的模型组中MMP-9表达明显升高,主动脉的病理形态学变化严重;而原花青素组主动脉病变明显减轻,MMP-9蛋白表达明显下降(P0.05)。表明原花青素对于动脉粥样硬化的发展具有抑制作用,这一作用可能是通过抑制MMP-9的表达来实现。  相似文献   

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