L/N型钙通道阻滞剂对于小鼠肾脏足细胞损伤的保护作用

Protection of L/N-type calcium channel blocker on injured mouse podocyte.

目的探讨给予L/N型钙通道阻滞剂处理小鼠后,小鼠受损足细胞的损伤变化情况。方法采用随机数字表法将雄性野生型BALB/C小鼠60只分为空白对照组、阳性对照组、硝苯地平组、西尼地平组,每组各15只。空白对照组无特殊处理,阳性对照组、硝苯地平组、西尼地平组采用经眼周静脉丛注射阿霉素15 mg/kg引起小鼠足细胞损伤,硝苯地平组小鼠在足细胞损伤建模后给予L型钙通道阻滞剂(硝苯地平15 mg/kg,口服给药,每日1次)处理;西尼地平组小鼠在足细胞损伤模型建立后给予L/N型钙通道阻滞剂(西尼地平10 mg/kg,口服给药,每日1次)处理。药物处理小鼠4周后,比较4组小鼠收缩压、24 h蛋白尿、足细胞损伤标记podocin的mRNA相对表达量及肾脏组织的病理变化。结果与空白对照组[(121.87±3.80)mmHg]、阳性对照组[(123.07±3.67)mmHg]比较,硝苯地平组[(104.47±3.94)mmHg]与西尼地平组[(105.4±4.57)mmHg]小鼠的收缩压均有所降低,但硝苯地平组与西尼地平组小鼠的收缩压比较差异无统计学意义(P>0.05)。与空白对照组[(17.88±0.69)mg/dL、(27.73±3.18)μg]比较,阳性对照组小鼠的血尿素氮[(73.36±2.17)mg/dL]、24 h蛋白尿含量[(626.73±31.50)μg]明显升高,肾脏的组织podocin的相对表达量(0.23±0.05)明显降低,差异均有统计学意义(P<0.05);与阳性对照组比较,硝苯地平组小鼠的血尿素氮[(73.39±2.50)mg/dL]、24 h蛋白尿含量[(631.80±25.26)μg]、肾脏的组织中podocin的相对表达量(0.29±0.06)无显著改善,差异无统计学意义(P>0.05),而西尼地平组小鼠血尿素氮[(53.63±2.39)mg/dL]、24 h蛋白尿[(389.86±45.19)μg]含量明显降低,肾脏的组织中podocin的相对表达量(0.65±0.11)显著升高,差异均有统计学意义(P<0.05)。与空白对照组比较,阿霉素引起小鼠肾脏较为严重的肾小球硬化,硝苯地平组小鼠较阳性对照组肾脏的病理改变不明显,而西尼地平组小鼠的肾脏肾小球硬化明显缓解。免疫组化检测显示,与空白对照组比较,阿霉素引起肌间蛋白的表达量明显增加;硝苯地平组小鼠较阳性对照组肌间蛋白表达变化不显著,而西尼地平组小鼠肌间蛋白的表达明显降低。结论L/N型钙通道阻滞剂西尼地平能有效保护小鼠肾脏足细胞损伤,降低小鼠蛋白尿的产生,值得临床推广应用。

Objective To explore the effect of L/N-type calcium channel blocker on mouse podocyte injury induced by doxorubicin.Methods Sixty wild-type BALB/C mice were divided into a blank control group,a positive control group,a nifedipine group,and a cilnidipine group according to the random number table method,with 15 mice in each group.The blank control group had no special treatment.The positive control group,nifedipine group and cilnidipine group were injected with adriamycin 15 mg/kg via the periocular venous plexus to cause podocyte damage in mice.The nifedipine group mice were treated with an L-type calcium channel blocker(nifedipine 15 mg/kg,orally administered once a day)after podocyte injury model established;the cilnidipine group were treated with L/N calcium channel blockers(cilnidipine 10 mg/kg orally administered once a day)after the podocyte injury model established.After 4 weeks of drug treatment,the systolic blood pressure,24 h proteinuria,the relative expression of podocin mRNA of the podocyte damage marker and the pathological changes of kidney tissue in the 4 groups were compared.Results Compared with the blank control group[(121.87±3.80)mmHg]and the positive control group[(123.07±3.67)mmHg],the blood pressure of mice in the nifedipine group[(104.47±3.94)mmHg]and the cilidipine group[(105.4±4.57)mmHg]was reduced,but there was no significant difference in blood pressure between the nifedipine group and the cilnidipine group(P>0.05).Compared with the blank control group[(17.88±0.69)mg/dL,(27.73±3.18)μg],the blood urea nitrogen of mice in the positive control group[(73.36±2.17)mg/dL],24 h proteinuria[(626.73±31.50μg]were significantly increased,the relative expression of podocin in the kidney tissue of mice in the positive control group(0.23±0.05)was significantly reduced(P<0.05),the differences were statistically significant(P<0.05);compared with the positive control group,the blood urea nitrogen of mice in the nifedipine group[(73.39±2.50)mg/dL],24 h proteinuria[(631.80±25.26)μg],the relative expression of podocin in the kidney tissue(0.29±0.06)did not significantly improve,the difference was not statistically significant(P>0.05),while the blood urea nitrogen[(53.63±2.39)mg/dL],24 h proteinuria[(389.86±45.19)μg]in the cilidipine group were significantly reduced,while the relative expression level of podocin in the podocytes of mice in the cilnidipine group(0.65±0.11)was significantly increased,and the differences were statistically significant(P<0.05).Compared with the blank control group,doxorubicin caused more severe glomerular sclerosis in the kidneys of mice.Compared with the blank control group,adriamycin caused more severe glomerulosclerosis in the kidneys of mice.The pathological changes of the kidneys in the nifedipine group were less obvious than those in the positive control group,while the glomerulosclerosis in the cilnidipine group was obviously alleviated.Immunohistochemical tests showed that,compared with the blank control group,doxorubicin caused a significant increase in the expression of intermuscular protein;mice in the nifedipine group did not change significantly compared with the positive control group,while the expression of muscle protein in the cilnidipine group was significantly reduced.Conclusion These data suggest that L/N type calcium channel blocker suppressed the albuminuria and protected against podocyte injury induced by doxorubicin in mice.