目的 探讨高血压小鼠铅染毒后小脑铁死亡变化。方法 选取25只健康C57雄性小鼠腹腔注射血管紧张素Ⅱ(Ang 11)0。05 mg/kg连续7 d的方法构建高血压模型，当收缩压≥140 mm Hg后，取20只高血压小鼠随机分为高血压对照组、高血压铅染毒组。另取20只正常血压的C57小鼠随机分为血压正常对照组与血压正常铅染毒组。血压正常对照组与高血压对照组小鼠自由饮水，血压正常铅染毒组和高血压铅染毒组小鼠饮用含250 mg/L乙酸铅饮水，高血压对照组与高血压铅染毒组小鼠每两天一次腹腔注射Ang Ⅱ，共12周。采用旷场实验和平衡木实验检测小鼠运动功能障碍情况;应用试剂盒检测不同组别小鼠小脑中二价铁(Fe2+)、丙二醛(malondialdehyde，MDA)和谷胱甘肽(glutathione，GSH)含量;采用 Western blot 法测定小鼠小脑组织中溶质载体家族7成员11(solute carrier family 7 member 11，SLC7A11)、谷胱甘肽过氧化物酶4(glutathione peroxidase 4，GPX4)、核受体共激活因子 4(nuclear receptor coactivator 4，NCOA4)、微管相关蛋白 1 轻链 3B(microtubule associated protein light chain 3B，LC3B)、铁蛋白重链 1(ferritin heavy chain 1，FTH1)蛋白表达。结果 旷场实验结果显示，高血压铅染毒组小鼠活动距离(1013。04 cm)显著低于高血压对照组(1351。18 cm)和血压正常铅染毒组(1287。35 cm);且高血压铅染毒组通过平衡木时间也延长，为29。40 s(P＜0。05)。此外，高血压铅染毒组小鼠小脑中Fe2+含量为3。33 μmol/g prot，是高血压对照组的1。54倍、血压正常铅染毒组的1。14倍;MDA含量为4。71 nmol/mg prot，高于高血压对照组和血压正常铅染毒组;GSH含量为5。36 μmol/g prot，低于高血压对照组与血压正常铅染毒组(P＜0。05)。Western blot结果显示，与高血压对照组及血压正常铅染毒组比较，高血压铅染毒组SLC7A11和GPX4蛋白表达均显著下降(P＜0。05)。此外，与血压正常对照组相比，高血压对照组和血压正常铅染毒组小鼠小脑中NCOA4、LC3B蛋白表达均上升，FTH1蛋白表达均下降(P＜0。05)。高血压铅染毒组NCOA4、LC3B蛋白表达高于高血压对照组和血压正常铅染毒组，FTH1蛋白表达降低(P＜0。05)。结论 铅染毒可加剧高血压小鼠小脑组织中铁死亡，铁自噬参与了其发生发展。
Cerebellar ferroptosis of hypertension mice following lead exposure
OBJECTIVE Exploring the changes in cerebellar ferroptosis in hypertensive mice after lead exposure.METHODS Twenty-five healthy C57 male mice were selected to construct a hypertensive model by intraperitoneal injection of angiotensinⅡ(Ang Ⅱ)at a concentration of 0.05 mg/kg for 7 consecutive days.After a systolic blood pressure of 140 mmHg,20 hypertensive mice were randomly divided into a hypertensive control group and a hypertensive lead exposure group.Twenty C57 mice with normal blood pressure were randomly divided into a blood pressure normal control group and a blood pressure normal lead exposure group.The mice in the normal blood pressure control group and the hypertensive control group drank water freely.Mice in the lead exposure group with normal blood pressure and the lead exposure group with hypertension drank lead acetate water containing 250 mg/L.Ang Ⅱ was injected intraperitoneally every two days in the hypertensive control group and hypertensive lead exposed group mice.Each group of mice was poisoned for 12 weeks.Using open field experiments and balance beam experiments to detect motor dysfunction in mice.Using a reagent kit to detect the levels of divalent iron(Fe2+),malondialdehyde(MDA),and glutathione(GSH)in the cerebellum of different groups of mice.Western blot was used to determine the protein expression of member 11 of the solute carrier family 7(SLC7A11),glutathione peroxidase 4(GPX4),nuclear receptor coactivator 4(NCOA4),microtubule associated protein 1 light chain 3B(LC3B),and ferritin heavy chain 1(FTH1)in mouse cerebellar tissue.RESULTS The result of the open field experiment showed that the activity distance(1013.04 cm)of mice in the hypertensive lead exposure group was significantly lower than that of the hypertensive control group(1351.18 cm)and the lead exposure group with normal blood pressure(1287.35 cm).And the lead exposure group with hypertension also extended the time through the balance beam,which was 29.40 seconds(P＜0.05).In addition,the Fe2+content in the cerebellum of mice in the hypertensive lead exposure group was 3.33 μmol/g prot,which was 1.54 times that of the hypertensive control group and 1.14 times that of the lead exposure group with normal blood pressure.The MDA content was 4.71 nmol/mg prot,higher than that of the hypertensive control group and the lead exposure group with normal blood pressure.The GSH content was 5.36μmol/g prot,lower than that of the hypertensive control group and the lead exposure group with normal blood pressure(P＜0.05).Western blot result showed that compared with the hypertensive control group and the lead exposure group with normal blood pressure,the protein expression of SLC7A11 and GPX4 in the hypertensive lead exposure group was significantly reduced(P＜0.05).In addition,compared with the control group with normal blood pressure,the expression of NCOA4 and LC3B proteins in the cerebellum of mice in the hypertension control group and lead exposure group with normal blood pressure increased,while the expression of FTH1 protein decreased(P＜0.05).The expression of NCOA4 and LC3B proteins in the hypertensive lead exposure group was higher than that in the hypertensive control group and the lead exposure group with normal blood pressure,while the expression of FTH1 protein decreased(P＜0.05).CONCLUSION Lead exposure can exacerbate iron death in the cerebellar tissue of hypertensive mice,and iron autophagy may be involved in its occurrence and development.
NETL
NSTL
万方数据
