三七皂苷R_1对大鼠脑缺血-再灌注损伤后TNF-α mRNA的影响

The effects of Notoginsenoside R1 on TNF-α m RNA after cerebral ischemia and reperfusion in rats

目的研究脑缺血–再灌注损伤(Ischemia-reperfusion injury,I/R)后脑组织中TNF-α的表达和变化,以及三七皂苷R_1对脑I/R损伤的保护作用。方法将90只雄性SD大鼠随机分为正常组(A组)、实验对照组(B组)、三七皂苷R_1组(C组),每组又分为24、48、72小时3个亚组,每个亚组10只动物。采用TUNEL染色法检测脑细胞凋亡情况。采用逆转录-聚合酶链式反应(Reverse Transcription polymerase Chain Reaction,RT-PCR)技术检测脑组织中TNF-α mRNA的表达变化。结果 TUNEL实验显示:随着时间的延长,A组中凋亡细胞数无明显的改变(P>0.05),B组细胞凋亡数逐渐增加,且各时间点均高于A组(P<0.05)。C组中各时间点脑组织的凋亡数少于B组(P<0.05)。RT-PCR结果显示1A组中各时间点均有TNF-α mRNA的表达,但组内各时间点比较无统计学差异(P>0.05);2随着时间的延长,B组中各时间点脑皮质中TNF-α mRNA含量逐渐增加,与A组比较有统计学意义(P<0.05);3C组在24、48、72小时后脑皮质中TNF-α mRNA的表达低于B组(P<0.05)。结论 1TNF-α因子参与了大鼠脑I/R损伤的发生和发展过程,且随着损伤时间延长,TNFα表达增高,内源性损伤作用变强。2三七皂苷R_1可降低脑组织中TNF-α浓度,减少脑细胞的凋亡,从而在脑I/R后的发挥保护作用。

Objective To investigate the expressional changes of brain tissue TNF-α after cerebral ischemia-reperfusiou injury（I/R） ,and the protective effect of Notoginsenoside R! on brain I/R injury. Methods 90 female SD rats were randomly divided into normal group （group A）, control group （group B）, Notoginsenoside R1 groupg （group C）, each group was further divided into 24h, 48h, 72h group, 10 rats in each group. TUNEL staining was used to detect the apoptosis of brain cells. Reverse transcription-polymerase chain reaction （RT-PCR） was used to detect the expression of TNF-αmRNA in brain tissue. Results The results of TUNEL： with extension of time,the number of apoptotic ceils in A group showed no significant ehange（P〉0.05）. In B, group cells apoptosis increased gradually, and were higher than those in A group at each time point （P〈0.05）. Apoptosis of brain tissue at different time points were higher in the B group than that in the C group （P〈0.05）. The TNF-αmRNA was expressedinA groupat different time points, but had no significant differences between the groups in each time point （P〉0.05）. The content of TNF-αmRNA at each time point inB group in the brain cortex gradually increased, and had statistical significance compared to those in A group（P〈0.05）. Expression of TNF-αmRNA,in C group at 24h, 48h, 72h time point were lower than those in B group （P〈0.05）. Conclusions TNF-αwas participated in the occurrence and development process of cerebral ischemia-reperfusion injury and expressed increased graduallywith the injury developed Notoginsenoside R1 couldreduce the expressionof TNF-α in brain tissue, reduce cerebral cell apoptosis, and protect the brain tissue after I/R.