硫酸铍致小鼠肺线粒体通透性转换孔高通透性开放及其后续效应研究

发布时间：2018-06-22 06:50

本文选题：硫酸铍 + 肺　；参考：《南华大学》2013年硕士论文

【摘要】：目的：通过实验动物体内染毒硫酸铍，观察硫酸铍对小鼠肺细胞线粒体通透性转换孔（PTP）开放的影响及其后续效应，丰富硫酸铍致肺毒性的机制。方法：选择36只6周龄的健康昆明雄性小鼠，随机分成3组，每组12只，设1个空白对照组和2个染毒组：空白对照组按0.1ml/10g（体重）腹腔注射灭菌生理盐水，染毒组分别按0.1ml/10g（体重）腹腔注射1mg/kg和2mg/kg BeSO4灭菌生理盐水溶液。隔天一次，持续一个月，观察实验期间小鼠的饮食、活动、精神状态及生长发育等一般情况。染毒结束后采取颈椎脱臼法处死小鼠，，迅速取出肺脏：用于常规病理组织学检查；采用差速离心法制备肺线粒体悬液，荧光分光光度法检测线粒体活性氧（ROS）含量及线粒体膜电位（Δψm），分光光度法检测线粒体通透性转换孔（PTP）开放程度；运用免疫印迹法（Western Blot）测定凋亡相关蛋白（细胞色素C、Bax、Bcl-2）的表达。结果： 1、染毒期间，对照组小鼠饮食及活动情况正常，BeSO4染毒组小鼠活动较少；一周后，高剂量组小鼠体重低于对照组，染毒第二、三、四周后，低、高剂量组小鼠体重均低于对照组，且差异均有统计学意义（P＜0.05）。 2、病理组织学检查发现，对照组肺组织结构正常；1mg/kg BeSO4染毒组肺组织可见淤血、出血，肺泡腔内少量炎症细胞、支气管周围炎、间质性肺炎、小叶性肺炎；2mg/kg BeSO4染毒组可见支气管扩张出血，支气管周围肺泡扩张，肺泡腔内有大量炎性渗出物，间质性肺炎、小叶性肺炎、融合性肺炎。 3、小鼠肺细胞线粒体中ROS含量、线粒体PTP开放程度、促凋亡蛋白Bax及胞质中Cyt-C的表达均随BeSO4剂量的增加而增加（P0.01），各组间差异均有统计学意义（P0.05）；与对照组比较，线粒体Δψm、抗凋亡蛋白Bcl-2表达及Bcl-2/Bax均降低（P0.01），且随剂量的增加而降低（P0.05）。结论： 1、腹腔注射硫酸铍可引起小鼠肺组织出现以炎症为主的病理组织学改变，具有明显的肺毒性。 2、腹腔注射硫酸铍可致小鼠肺线粒体PTP高通透性开放、膜电位下降、细胞色素C溢出、促凋亡蛋白Bax表达增加、抗凋亡蛋白Bcl-2表达减少，提示肺线粒体PTP的高通透性开放可能是硫酸铍致肺毒性的机制之一。
[Abstract]:Aim: to observe the effect of beryllium sulfate on the opening of mitochondrial permeability transition pore (PTP) in mouse lung cells and its follow-up effects, and to enrich the mechanism of pulmonary toxicity induced by beryllium sulfate. Methods: Thirty-six healthy Kunming male mice aged 6 weeks were randomly divided into 3 groups, 12 mice in each group, one blank control group and two exposed groups: the blank control group was injected with sterilized saline according to 0.1ml/10g (body weight). In the exposed group, 1mg/kg and 2mg/kg BeSO4 were injected intraperitoneally according to 0.1ml/10g (body weight). The diet, activity, mental state, growth and development of the mice were observed every other day for a month. At the end of the poisoning, the mice were killed by cervical dislocations, and the lungs were removed quickly: for routine histopathological examination, and for the preparation of lung mitochondria suspension by differential centrifugation. The contents of reactive oxygen species (Ros) and mitochondrial membrane potential (螖蠄 m),) were detected by fluorescence spectrophotometry, and the expression of apoptosis-related protein (cytochrome CnBaxBcl 2) was detected by Western blot. Results: 1. During the exposure period, the normal diet and activities of mice in the control group were less than those in the control group, and after one week, the body weight of the mice in the high dose group was lower than that in the control group, and the second, third, and fourth weeks after exposure, the body weight of the mice in the high dose group was lower than that in the control group. The body weight of the mice in the high dose group was lower than that in the control group (P < 0.05). 2. Histopathological examination showed that the lung tissue structure of the control group was normal at 1 mg / kg BeSO4. Small amount of inflammatory cells in alveolar cavity, peribronchitis, interstitial pneumonia, lobular pneumonia and 2 mg / kg BeSO4 exposed group showed bronchiectasis, bronchoalveolar dilatation, massive inflammatory exudate in alveolar cavity, interstitial pneumonia. Lobular pneumonia, fusion pneumonia. 3. Ros content in mitochondria of mouse lung cells, mitochondrial PTP opening degree, The expression of apoptotic protein Bax and Cyt-C in cytoplasm increased with the increase of the dose of BeSO4 (P0.01), the difference was statistically significant (P0.05), compared with the control group, the expression of mitochondrial 螖蠄 m, anti-apoptotic protein Bcl-2 and Bcl-2 / Bax decreased (P0.01), and decreased with the increase of the dose (P0.05). Conclusion: 1. Intraperitoneal injection of beryllium sulfate can induce inflammatory pathological changes in the lung tissue of mice, which has obvious pulmonary toxicity. 2. Intraperitoneal injection of beryllium sulfate can induce high permeability of PTP in mouse lung mitochondria. Decreased membrane potential, cytochrome C overflow, increased expression of pro-apoptotic protein Bax and decreased expression of anti-apoptotic protein Bcl-2 suggest that the high permeability of PTP in lung mitochondria may be one of the mechanisms of pulmonary toxicity induced by beryllium sulfate.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R114

【共引文献】