加味脑泰方对去势脑缺血大鼠海马ATF4/CHOP/Puma通路的影响

摘要

AIM:To investigate the effects of Jiawei-Naotai formula (JWNTF) on ATF4/CHOP/Puma pathway in hippocampal neurons of ovariectomized female rats with cerebral ischemia.METHODS:The female rats were randomly divided into sham group, model group, JWNTF group and positive control group.The rats, expect in the sham group, were ovariectomized.The rats in each group were intragastric administration 11 days after ovariectomy.The rats in sham group and model group were given a gavage of 0.9％Na Cl, while the rats in other groups were administrated by corresponding therapy intragastrically for 3 d.The regional cerebral ischemia model was established by middle cerebral artery occlusion (MCAO) suture method 14 days after ovariectomy.The behaviors of the rats were evaluated 24 h after cerebral ischemia.The mRNA levels of Bax, Bcl-2 and caspase-3 were detected by RT-qPCR, and the protein expression of Bax, Bcl-2, caspase-3, ATF4, CHOP and Puma was determined by Western blot.RESULTS:Compared with sham group, the neurobehavioral scores significantly increased in other groups (P<0.05).Compared with model group, the neurobehavioral scores were significantly decreased in positive control group and JWNTF group (P<0.05).The protein expression of Bax, caspase-3, ATF4, CHOP and Puma, and the mRNA expression of Bax and caspase-3 in the hippocampus were much higher, and Bcl-2 was lower in model group than those in sham group (P<0.05).JWNTF significantly reduced the protein expression of Bax, caspase-3, ATF4 and CHOP, and the mRNA expression of Puma, Bax and caspase-3, and markedly increased the expression of Bcl-2 at mRNA and protein levels compared with model group.CONCLUSION:The JWNTF protects against brain damage induced by cerebral ischemia, which may be related to inhibitiing the expression of ATF4/CHOP/Puma pathway-related molecules at mRNA and protein levels.%目的:研究加味脑泰方对去卵巢脑缺血大鼠海马活化转录因子4 (ATF4) /C/EBP同源蛋白 (CHOP) /p53上调凋亡调控因子 (Puma) 通路的影响.方法:将雌性大鼠随机分为假手术组、模型组、加味脑泰方组和阳性对照组, 除假手术组外, 其它大鼠均行去卵巢术;术后11 d, 阳性对照组和加味脑泰方组给予相应药物灌胃, 连续灌胃3 d后行脑缺血术.术后24 h进行行为学评价, 取海马组织行RT-qPCR检测Bax、Bcl-2和caspase-3的mRNA表达, Western blot检测各组大鼠海马组织中Bax、Bcl-2、caspase-3、ATF4、CHOP和Puma蛋白表达.结果:与假手术组比较, 各组的海马神经功能评分明显增高;与模型组比较, 加味脑泰方组与阳性对照组的神经功能评分明显降低 (P <0.05) .与假手术组比较, 模型组大鼠海马Bax、caspase-3、ATF4、CHOP和Puma的蛋白表达及Bax和caspase-3的mRNA表达明显升高, 而Bcl-2的mRNA和蛋白表达明显降低 (P <0.05) ;与模型组比较, 加味脑泰方明显降低Bax、caspase-3、ATF4、CHOP和Puma表达, 提高Bcl-2表达 (P <0.05) .结论:加味脑泰方可能通过抑制ATF4/CHOP/Puma通路相关mRNA及蛋白的表达而减轻脑缺血损伤.