ABSTRACT
OBJECTIVES: To investigate the neuroprotective effect of electroacupuncture (EA) at "Quchi"(LI11) and "Zusanli"(ST36) in the rats with cerebral ischemia reperfusion injury and its influence on programmed necrosis of cerebral cortical neurons. METHODS: Sixty male SD rats were randomly divided into sham-operation group, model group, EA group and inhibitor group, with 15 rats in each group. Left middle cerebral artery occlusion model was established using the modified thread embolism method. In the sham-operation group, the carotid artery was exposed and dissociated in each rat. EA was applied to "Quchi"(LI11) and "Zusanli"(ST36) on the right side for 30 min each time, once daily for 7 days in the rats of the EA group. The rats in the inhibitor group were intraperitoneally injected with norstatin-1 (0.6 mg/kg) for consecutive 7 days. The neurological deficit score of rats in each group was observed. HE staining was adopted to detect the degree of pathological damage of the cerebral cortex in the infarction area. Using TUNEL staining, the apoptosis of cortical neurons in the infarction area was determinedï¼the contents of tumor necrosis factor α (TNF-α), interleukin (IL)-1ß and IL-6 were detected by ELISAï¼the mRNA and protein expression of the receptor interacting protein-1 (RIP1), the receptor interacting protein-3 (RIP3) and the substrate mixed lineage kinase like protein (MLKL) were detected by fluorescence quantitative PCR and Western blot, respectively. RESULTS: In comparison with the sham-operation group, the neurological deficit score in the model group was higher(P<0.01)ï¼HE staining showed that there was the pathological damage in the infarction areaï¼the neuron apoptosis rate, the contents of TNF-α, IL-1ß and IL-6, and the mRNA and protein expressions of RIP1, RIP3 and MLKL increased(P<0.01) in the model group. In the EA group, the neurological deficit score was reduced(P<0.01)ï¼HE staining showed that the pathological damage was ameliorated in the infarction areaï¼the neuron apoptosis rate, the contents of TNF-α, IL-1ß and IL-6, and the mRNA and protein expressions of RIP1, RIP3, MLKL decreased(P<0.05, P<0.01) when compared with those in the model group. CONCLUSIONS: EA can attenuate cerebral ischemia reperfusion injury and display its neuroprotective effect probably through inhibiting programmed necrosis of cerebral cortical neurons in the rats.
Subject(s)
Brain Ischemia , Electroacupuncture , Neuroprotective Agents , Reperfusion Injury , Rats , Male , Animals , Rats, Sprague-Dawley , Tumor Necrosis Factor-alpha/genetics , Brain Ischemia/genetics , Brain Ischemia/therapy , Interleukin-6 , Reperfusion Injury/genetics , Reperfusion Injury/therapy , Neurons/metabolism , Interleukin-1beta/genetics , Interleukin-1beta/metabolism , Necrosis , Apoptosis , Infarction , RNA, Messenger , Protein KinasesABSTRACT
Ajuga lobata D. Don is a medicinal plant rich in 20-hydroxyecdysone (20E), alkaloids, and other active substances. In this study, the cell suspension was incubated for 7 days, followed by the analysis on the effects of abscisic acid (ABA) on the regulation of 20E synthesis. Then A. lobata suspension cells treated with 0.15 mg/l ABA were used as material, with the Illumina technology applied for transcriptome sequencing. Digital analysis on the gene expression profile was carried out on ABA treated and control samples, respectively. Finally, transcriptomics was applied to assess the molecular response of A. lobata induced by ABA through applying transcriptomics by evaluating differentially expressed genes. The results suggested that ABA promoted 20E accumulation, while longer processing time caused cell browning. A total of 154 genes were significantly regulated after ABA treatment, with 99 up-regulated and 55 down-regulated, respectively. In addition to 20E-related pathways, the genes belonged to the ko00900 (terpenoid backbone biosynthesis) pathway (six differentially expressed genes [DEGs]), ko00100 (steroid biosynthesis) pathway (four DEGs), and ko00140 (steroid hormone biosynthesis) pathway (six DEGs). Providing a better understanding of the 20E biosynthetic pathway and its regulation, in particular in plants, this study is necessary.
