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Objective:To explore the effect of Anmeidan （AMD） on the learning and memory levels of sleep deprived rats through mitochondrial mediated hippocampal neuronal apoptosis pathway. Method:Forty-eight SD rats were randomly divided into blank group， model group， low， medium， high-dose AMD groups （4.86， 9.72， 19.44 g·kg-1·d-1） and estazolam group （0.1 mg·kg-1·d-1）. Insomnia model was prepared by self-made sleep deprivation box for 14 days. Morris water maze was used to detect learning and memory levels， enzyme-linked immunosorbent assay （ELISA） was used to detect the expressions of cytochrome C （Cyt-C）， cysteine aspartic acid protease-3 （Caspase-3） in hippocampus. Transmission electron microscopy （TEM） was used to observe the morphological structure of mitochondria in hippocampus. Protein and mRNA expressions of Cyt-C， Caspase-3， Bcl-2， Bax were detected by immunofluorescence （IF） and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） respectively. Result:In the model group， the incubation period of the platform and the total distance of swimming and the time of first arriving platform were prolonged， the number of platform crossing and the time of target quadrant movement were reduced， protein and mRNA expressions of Bcl-2 dropped， protein and mRNA expressions of Bax increased （P<0.01）， and mitochondrial structure was abnormal with crista fracture， swelling and deformation. And protein and mRNA expressions of Cyt-C， Caspase-3 increased significantly （P<0.01）. Low， medium and high-dose AMD groups could improve levels of space exploration and navigation of SD rats （P<0.01）， increase protein and mRNA expressions of Bcl-2， decrease protein and mRNA expressions of Bax， improve the damage of mitochondria， and decrease the protein and mRNA expressions of Cyt-C， Caspase-3 （P<0.01）. Conclusion:AMD can improve the learning and memory levels of SD rats， the effect is related to the mitochondrial mediated hippocampal neuronal apoptosis pathway and decrease of Cyt-C and Caspase-3 expressions.
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Objective: To explore the effect of modified Si Junzitang (MSJZT) drug serum on the expression of apoptosis-related molecules of gastric cancer cell SGC-7901 and further its anti-tumor mechanism.Method: A total of 40 SD rats were randomly divided into four groups:low-dose,middle-dose,high-dose MSJZT (0.213,0.426,0.853 g·kg-1) groups and normal group (n=10).The treatment groups were administrated through gastric perfusion,and the normal group was given the equivalent volume of normal saline for 10 days.1.5 h after the last treatment,chloral hydrate peritoneal anesthesia was performed,blood was collected from heart,and different doses of serum were separated to prepare drug-containing serum of low-dose,middle-dose,high-dose MSJZT groups,in order to incubate SGC-7901 gastric cancer cell.Early and late apoptosis rates were detected with flow cytometry.Afterwards,the tumor suppressor gene p53,c-nucleoprotein gene (c-Myc),cysteine-aspartic acid protease-3(Caspase-3),B-cell lymphoma-2(Bcl-2) mRNA expressions were confirmed by fluorescence quantitative polymerase chain reaction (Real-time PCR).The protein expressions of p53,c-Myc,Caspase-3,Bcl-2 were detected by immunofluorescence.Result: Compared with the normal group,the high-dose MSJZT group could obviously increase the apoptosis rate to 22.58%(PPPPPPConclusion: MSJZT drug serum could exert an anti-tumor effect by inhibiting the expression of the anti-apoptotic protein Bcl-2,and promoting the expressions of pro-apoptotic-related molecules p53,c-Myc,Caspase-3.
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Objective To establish oxidative stress model of hydrogen peroxide treatment by using human umbilical vein endothelial cells (ECS) as cell model to study the protective mechanism of anti oxidative stress and determine the signal transduction pathway of remifentanil.Methods Primary cultured human umbilical vein endothelial cells which were incubated with 0.1 M hydrogen peroxide to establish injury model to study remifentanil protection and related pathways.The experiment was divided into nine groups: control group (group C), Hydrogen peroxide group (group H1), Hydrogen peroxide+SP600125 group (group H2), Hydrogen peroxide+SB203580 group (group H3), Hydrogen peroxide+PD98059 group (group H4), hydrogen peroxide+remifentanil group (group HR1), hydrogen peroxide+remifentanil+SP600125 group (group HR2), hydrogen peroxide+remifentanil+SB203580 group (group HR3), hydrogen peroxide+remifentanil+PD98059 group (group HR4).Groups H1, H2, H3 and H4 only performed MAPK pathway blockade experiments.Groups HR1, HR2, HR3 and HR4 individually added remifentanil 10 ng/ml to protect 1 h.SOD activity, MDA level, Caspase-3 activity were detected and anti oxidative stress of remifentanil observed to confirm preliminary transduction pathway;Using RT-PCR expression levels of c-Jun before was observed before and after treated with remifentail 10 ng/ml.The aim was to determine the transduction pathway of the signaling molecules.Results Compared with group C, SOD activity were decreased significantly, MDA performance level were increased significantly in groups H1, H2, H3 and H4 (P<0.05).Compared with group H1, SOD activity was increased significantly, MDA performance level was decreased significantly in group HR1 (P<0.05).SOD activity difference and MDA performance level of groups HR2 and H2 had no statistical significance.Compared with group H3, SOD activity was increased significantly, MDA performance level was decreased siginificantly in group HR3 (P<0.05).Compared with group H4, SOD activity was increased significantly, MDA performance level was decreased significantly in group HR4 (P<0.05).Caspase-3 activity of groups H1, H2, H3 and H4 were higher significantly than that of group C (P<0.05).The level of C-Jun mRNA in group H1 was significantly higher than that of group C;But it was higher in group HR1 than that of group C, it was significantly lower than that of group H1 (P<0.05).Conclusion By activating the JNK pathway and its downstream signaling molecule c-Jun, remifentanil 10 ng/ml has the effect of increasing SOD activity, reducing the level of MDA expression and playing a role in anti oxidative stress.