摘要
目的：探讨健脾益肺方对hSOD1-G93A转基因小鼠p38丝裂原活化蛋白激酶（p38 mitogen-activated protein kinase,p38 MAPK）蛋白及炎症因子表达的影响。方法：将24只8周龄的SPF级hSOD1-G93A转基因小鼠随机分为模型组、健脾益肺方（2.86,5.72,11.44 g·kg（-1））组,每组6只。8周龄SPF级同窝出生正常野生型小鼠6只作为正常组。模型组和正常组予生理盐水灌胃,药物组予不同剂量的中药灌胃,每日1次,共120 d。用免疫荧光法（immunofluorescence）检测小鼠脊髓小胶质细胞的活化情况,p38 MAPK,p-p38 MAPK蛋白的荧光强度核浆比,蛋白质免疫印迹法（Western blot）检测小鼠脊髓p38MAPK,p-p38 MAPK,肿瘤坏死因子-α（TNF-α）及环氧化酶-2（COX-2）蛋白的表达。结果：与正常组比较,模型组、健脾益肺方（2.86 g·kg（-1））组小胶质细胞激活数明显增多（P〈0.05,P〈0.01）,p38 MAPK,p-p38 MAPK荧光强度核浆比显著升高（P〈0.01）,与模型组比较,健脾益肺方（5.72,11.44 g·kg（-1））组小胶质细胞激活数及所占总数比例显著减少（P〈0.01）,p38 MAPK,p-p38 MAPK荧光强度核浆比显著下降（P〈0.01）;与正常组比较,模型组、健脾益肺方（2.86 g·kg（-1））组p38 MAPK,p-p38 MAPK,COX-2,TNF-α蛋白表达明显升高（P〈0.05,P〈0.01）,与模型组相比,健脾益肺方（5.72,11.44 g·kg（-1））组p38 MAPK,p-p38 MAPK,COX-2,TNF-α蛋白表达显著降低（P〈0.01）。结论：健脾益肺方可能通过介导p38MAPK通路抑制p38 MAPK蛋白磷酸化,以浓度依赖性抑制小胶质细胞的活化,下调p38 MAPK蛋白及炎症因子的表达,从而发挥神经保护作用。
Objective： To investigate the effect of Jianpi Yifei prescription on p38 mitogen-activated protein kinase（p38 MAPK） protein and inflammatory factors expression in hSOD1-G93 A transgenic mice.Method： Twenty-four SPF class 8 weeks old hSOD1-G93 A transgenic mice were randomly divided into model group,low concentration Jianpi Yifei prescription group（2. 86 g·kg（-1））,medium concentration Jianpi Yifei prescription group（5. 72 g·kg（-1））,and high concentration Jianpi Yifei prescription group（11. 44 g·kg（-1））,n = 6 in each group. 6 normal SPF class wild mice of 8 weeks old were selected as normal group. The mice in model group and normal group were given with normal saline,while different concentrations of Jianpi Yifei prescription was given in the treatment groups once a day,for continuously 120 days. The activation of spinal cord microglia,p38 MAPK and p-p38 MAPK protein fluorescence intensity karyoplasmic ratio were detected by immunofluorescence（IF）,and Western blot（WB） was used to detect the protein expression levels of p38 MAPK,p-p38 MAPK,tumor necrosis factor-α（TNF-α） and cyclooxygenase-2（COX-2）. Result： As compared with normal group,the activation number of microglia was increased significantly in model group and the low concentration Jianpi Yifei prescription group（P 〈 0. 05,P 〈 0. 01）; the fluorescence intensity karyoplasmic ratio of p38 MAPK and p-p38 MAPK was increased significantly（P 〈 0. 01）. As compared with model group,the number of microglia activation and the proportion of the total number was significantly reduced in the medium and high concentration Jianpi Yifei prescription groups（P 〈 0. 01）; and the fluorescence intensity karyoplasmic ratio of p38 MAPK and p-p38 MAPK was significantly reduced（P 〈 0. 01）. As compared with normal group, the protein expression levels of p38 MAPK,p-p38 MAPK, COX-2, and TNF-α were significantly increased in model group and the low concentration Jianpi Yifei prescription group（P 〈 0. 05,P 〈 0. 01）. As compared with model group,the protein expression levels of p38 MAPK,p-p38 MAPK,COX-2,and TNF-α were significantly reduced in the high,medium concentration Jianpi Yifei prescription groups（P 〈 0. 01）. Conclusion： Jianpi Yifei prescription may inhibit p38 MAPK protein phosphorylation by mediating p38 MAPK pathway,and plays a neuroprotective role through inhibiting the activation of microglia in a concentration-dependent manner and down-regulating the expression of p38 MAPK protein and inflammatory factors.
作者
潘显梅
杨碧莹
杜宝新
郑瑜
李惠珍
PAN Xian-mei;YANG Bi-ying;DU Bao-xin;ZHENG Yu;LI Hui-zhen(Guangzhou University of Chinese Medicine, Guangzhou 510006, China;Guangdong Province Hospital of Chinese Medicine, Guangzhou 510006, China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2018年第15期155-160,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
广东省中医药局科研项目(20161080)
广东省中医院中医药科学技术研究专项(YN2014PJ06)
