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In addition to its own specific functions, an organelle can also interact with other organelles to complete important physiological functions. The disorders of organelle interactions are closely associated the development and progression of various diseases. In recent years, the role of organelle interactions has attracted more attention in the progression of nonalcoholic fatty liver disease, especially the interactions between mitochondria, lipid droplets, and other organelles.
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Liver fibrosis/cirrhosis is the common pathological process of most chronic liver diseases. Many studies have confirmed that a certain degree of the reversal of liver fibrosis or cirrhosis can be achieved after effective treatment. How to use simple serological markers to evaluate the reversal of liver fibrosis and cirrhosis is a research hotspot at present. This article summarizes the current status of the research on serological markers for the reversal of liver fibrosis and cirrhosis.
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Objective To construct recombinant adeno-associated virus and lentivirus carrying siRNA of TIMP-1 and to investigate the efficiency of infection and short-term inhibitory effect of TIMP-1 gene expres-sion on rat hepatic stellate cells. Methods One pair of siRNA which could effectively inhibit expression of the TIMP-1 gene in HSC-T6 was screened and cloned into AAV vector and lentiviral vector to construct the recombinant AAV/siRNA-TIMP-1/GFP and Lenti/siRNA-TIMP-1/GFP. AAV/GFP and Lenti/GFP as neg-ative control were also obtained. Experiments were assigned to five groups: AAV/siRNA-TIMP-1/GFP, AAV/GFP, Lenti/siRNA-TIMP-1/GFP, Lenti/GFP group and mock group. Rat HSC-T6 cells were infected by these recombinant viruses at a concentration of MOI by 10. To monitor the efficiency of infection, fluores-cence microscope and flow cytometer were used. After 7 d post-infection, Western blot was used to detect the TIMP-1 protein expression. Results HSC-T6 had no significant changes after infection. The efficiency of infection in AAV/GFP and Lenti/GFP group were 72.7% and 70.0%, AAV/siRNA-TIMP-1/GFP and Lenti/siRNA-TIMP-1/GFP group were 64.58% and 61.86%. The protein expression levels of TIMP-1 in HSC-T6 cells at 7 d post-infection by the recombinant AAV and Lentivirus were decreased 40.0% compared with those in mock control and normal HSC-T6 (P<0.05). Conclusion Recombinant AAV/siRNA-TIMP-1/GFP and Lenti/siRNA-TIMP-1/GFP could effectively infect HSC-T6 with similar efficiency and suppress the expression of TIMP-1 in rat HSC-T6 remarkably.
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Rep78/68 is a kind of nonstructural protein from adeno-associated virus. It inhibits a variety of viruses and tumor transformation. In addition, it has significant effect on the proliferation and metabolism of host cells. In this paper, we reviewed the latest research advances of this protein.
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BACKGROUND:After cerebral ischemia/reperfusion injury,as a executioner caspase, procaspase-3 and caspase-3-like activity increased significantly. We observe both the dephosphorylated and phosphorylated procaspase-3, and try to find out their variations during the processes of cerebral ischemia/reperfusion injury.OBJECTIVE:Observe the expression of procaspase-3 in hippocampus following transient forebrain ischemia.DESIGN: A randomized and controlled experiment.DEPARTMENT:Department of Biochemistry and Molecular Biology,Capital University of Medical Sciences.SETTING:Department of Hyperbaric Oxygen of Beijing Chaoyang Hospital Affiliated to Capital University of Medical Sciences.METHODS:Transient forebrain ischemia was induced by bilateral common carotid occlusion (BCCAO) for 20 minutes. Hippocampus was obtained at reperfusion time points of 6 hours, 12 hours,24 hours and 48 hours respec-tively after 20 minutes of BCCAO. Sham-operated group did not occlude bi-lateral common carotid, and hippocampus was obtained at reperfusion time point of 24 hours. Western Blotting was used to detect the level of procaspase-3.MAIN OUTCOME MEASURES:The comparison of total procaspase-3,dephosphorylated procaspase-3 and phosphorylated procaspase-3 level in hippocampus between each group.3 level: Total procaspase-3 level increased in hippocampus at reperfusion time points of 12 hours and 24 hours post-BCCAO (9 133.1 ±2 216.3,dephosphorylated procaspase-3 level increased in hippocampus at reperfusion time point of 24 hours post-BCCAO (7812.0±1625.1, 3825.8±155.6, P was not significant (P ＞ 0.05) as compared with the expression levels in sham-operated mice.CONCLUSION: Procaspase-3 is upregulated after ischemia/reperfusion.The increment of dephosphorylated form of procaspase-3 was higher than that of phosphorylated form of procaspase-3 upon cerebral ischemia/reperfusion injury, which indicates that cerebral ischemia/reperfusion injury possibly induced the dephosphorylation of procaspase-3 and promoted its transforming into activated form.
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BACKGROUND: The level of blood neuron-specific enolase may help predict the severity of brain damage.OBJECTIVE: To define the optimal time window of hyperbaric oxygen(HBO) treatment for brain ischemia based on the dynamical changes in plasma neuron-specific enolase bioactivity.DESIGN: Factorial design.SETTING: Department of Biochemistry and Molecular Biology of Capital University of Medical Sciences.PARTICIPANTS: The experiment was conducted in the Laboratory of the Department of Hyperbaric Oxygen, Beijing Chaoyang Hospital Affiliated to Capital University of Medical Sciences in June 2002. Totally 54 adult female SD rats were randomized into 3 groups, namely sham operation group(n=6), ischemia-reperfusion (IR) group (n=24), and HBO group (n=24), the latter 2 groups further divided into 4 groups according to the reperfusion time of 6, 24, 48 and 96 hours, with 6 rats in each subgroup.METHODS: [1] Rat models of IR was prepared by occlusion of the 4 arteries for 20 minutes followed by reperfusion for different time. [2] The rats in the sham operation received the same operation without blocking the arteries. The rats in HBO group were subjected to HBO treatment (0.2 MPa,pure oxygen for 45 minutes), which was given after a 3-hour reperfusion inthe 6-hour subgroup and scheduled once daily at the same time point in the other 3 subgroups until blood sampling. The rats in IR group and sham operation group were kept under normal pressure without additional oxygen.MAIN OUTCOME MEASURES: Blood samples were collected at the specified time points in IR and HBO groups and at 24 hours of reperfusion in the sham operation group. Enzyme-linked immunosorbent assay (ELISA)was used to determine the activity of plasma neuron-specific enolase.RESULTS: Totally 54 rats enter result analysis after supplementary.Plasma neuron-specific enolase level was significantly lower in the sham operation group (1.97±0.09) μg/L than in 6 and 96-hour subgroups in the IR group [(2.80±0.26), (2.40±0.19) μg/L, respectively, P ＜ 0.05],and was obviously lower in 6-hour HBO subgroup than the 6-hour IR group [(2.04±0.27) μg/L, P ＜ 0.05], which was slightly increased at 24hours after HBO treatment but the difference was of no statistical significance (P ＞ 0.05).CONCLUSION: IR injury may lead to increment of plasma neuron-specific enolase level, which occurred at 6 and 96 hours respectively in IR group, possibly due to acute neuronal necrosis during brain ischemia and subsequent delayed neuronal apoptosis. HBO treatment promotes the recovery of neuron-specific enolase level, with 6 hours of reperfusion as the optimal therapeutic time window.