1.Inhibitory effect of lentivirus targeting interference Cx26 on proliferation and migration of human highly metastatic hepatocellular carcinoma HCCLM3 cells
Chinese Pharmacological Bulletin 2014;(7):937-941
Aim To investigate effects of Cx26 on pro-liferation,apoptosis,migration of human highly meta-static hepatocellular carcinoma HCCLM3 cells.Meth-ods The HCCLM3 cells were infected with lentiviral vector targeting interference of Cx26 and the stable transfectants were selected by puromycin.The interfer-ence efficiency of Cx26 was detected by Real-time PCR and Western blot.Gap junction function was assessed by “parachute”dye-coupling assay. The effects of Cx26 interference on proliferation,apoptosis,migra-tion of HCCLM3 cells were determined by MTT assay, flow cytometry,transwell migration assay,respective-ly.Results The mRNA and protein expression of Cx26 in LV-Cx26 group was significantly lower than the LV-NC group and wide group (P<0.01),and GJ function in LV-Cx26 group also significantly reduced compared with LV-NC group and wild group (P <0.01 ).The Cx26 interference significantly inhibited the proliferation (P<0.01)),promoted the apoptosis (P <0.01 ),and decreased migration of HCCLM3 cells in vitro (P <0.01 ).Conclusion Lentiviral vector targeting interference Cx26 expression of HC-CLM3 cells can significantly reduce its GJ function,in-hibit the proliferation and migration,promote apopto-sis,and reduce its malignant properties.
2.The coordinated regulation between nuclear DNA and mitochondria DNA
Chinese Journal of Pathophysiology 2000;0(08):-
The overall cellular gene expression is well realized by the coordinated regulation between nuclear DNA and mitochondria DNA. Coordinated regulation between two genomes plays a key role not only in mitochondria biogenesis and protein synthesis but also more importantly in the regulation of gene expression of mitochondria respiratory chain subunits in order to mediate the mitochondrial respiratory function. In the present review, we focus on the mechanism of coordinated regulation between two genomes.
3.Changes of uncoupling proteins activity and the efficiency of oxidative phosphorylation in hypoxic exposed rat brain mitochondria in vitro
Chinese Journal of Pathophysiology 2000;0(11):-
AIM:To observe the effect of GDP on uncoupling proteins(UCPs) activity and the efficiency of oxidative phosphorylation in hypoxic exposed rat brain mitochondria.METHODS: Adult SD rats were randomly divided into three groups (control, acute hypoxia and chronic hypoxia groups). The animals were placed into a hypobaric chamber simulated 5 000 m high altitude for 0, 3 and 30 d, respectively. The mitochondria from rat brain were isolated by centrifugation. The activity of UCPs was detected by the method of [H3]-GTP binding with UCPs specifically. The maximal binding content (Bmax) and the dissociation constant (Kd) were determined by Scatchard plot. The mitochondrial potential was measured by rhodamine 123 method. Oxidative respiratory consumption was measured by Clark electrode. The experiments were conducted under the conditions with or without GDP (1 mmol/L), respectively. RESULTS: For exposed to hypoxia, Bmax and the oxidative consumption of uncoupling respiration were increased. Kd, MMP and RCR were decreased. UCPs activity was inhibited by GDP in three groups. Kd was increased 61.01%, 83.13% and 71.52% and Bmax was decreased 23.18%, 35.20% and 33.38%, respectively. The values in the acute hypoxic group were changed markedly. The sensitivity of UCPs to GDP was elevated significantly by hypoxia. With the reducing of UCPs activity, oxidative consumption of uncoupling respiration was decreased whereas RCR and MMP were increased. The results elucidated increase in the efficiency of oxidative phosphorylation.CONCLUSION: GDP increases the mitochondrial membrane potential and decreases the oxygen consumption of uncoupling respiration in hypoxic exposed rat brain mitochondria by inhibiting UCPs activity. The results suggest that the change in UCPs activity is one of the factors of mitochondrial dysfunction in oxidative phosphorylation induced by hypoxia.
4.Changes of the myocardium energy metabolism and adenine nucleotide translocase activity in rats after hypoxic exposure
Bing LI ; Junze LIU ; Lifen CHEN
Chinese Journal of Pathophysiology 1986;0(03):-
AIM: To explore the changes of myocardial energy metabolism and adenine nucleotide translocase (ANT) activity in mitochondria in rats exposed to hypoxia. METHODS: Adult male Wistar rats were exposed to simulated high altitude at 5 000 m for control (0 d), 1 d, 5 d, 15 d, 30 d in hypobaric chamber. Myocardial mitochondria were isolated by centrifugation. Mitochondria respiratory function was measured by Clark oxygen electrode. The size of adenine nucleotides pool (ATP, ADP, AMP) in mitochondria were separated and measured by HPLC. ANT activity was measured by [3H]-ADP incorporation. RESULTS: Compared to control, mitochondria state Ⅲ respiratory (ST_3) and RCR decreased and ST_4 increased sharply at 1 d, 5 d and 15 d, ST_3 still lower than that in control at 30 d, while RCR level restored. ATP contents and ANT activity decreased at 1 d and 5 d, then restored to control level at 15 d, then decreased again at 30 d. CONCLUSION: The inhibition of mitochondria respiratory function is the main reason that makes ATP contents decrease during hypoxic exposure. ANT activity and ATP content change cooperatively.
5.Effect of CAP-administration on subunits expression of cytochrome oxidase in mitochondria from brain cortex of rats and its mechanism
Rong SONG ; Junze LIU ; Lifeng CHEN
Chinese Pharmacological Bulletin 2003;0(12):-
Aim To understand the changing aspects of CAP-administration on expression of cytochrome C oxidase(COX) subunits I and IV and their mechanisms regulated by gene expression encoded by mtDNA and nDNA. Methods Wistar rats were randomly divided into control and CAP groups, Rats were administrated CAP(100 mg?kg -1, intraperitoneal injection) every 12 hours for 7 days before sacrificed by decapitation. Rat brain was removed and the cerebral cortex mitochondria was isolated by centrifugation programme. The protein content of COX subunitⅠand Ⅳ in mitochondria and NRF-1 in cerebral tissues was detected by Western blot analysis. And mRNA state levels of COXⅠ, COXⅣ, mtTFA and NRF-1 in tissues were determined by RT-PCR.Results Compared with C group, a decreased protein content of COX subunitⅠand an elevated ratio of subunit Ⅳ/Ⅰwas observed in CAP group, The protein content of COX subunit Ⅳ and NRF-1 as well as COXⅠ,Ⅳ,NRF-1 and mtTFA mRNA state level was not unchanged between the two groups. Conclusion The change of content of COX subunitⅠprotein in mitochondria from cerebral cortex showed there is no regulation of feedback to mitochondrial and nuclear transcription. The nuclear genomes expression does not correspond to mitochondrial expression in CAP-administrated rats.
6.Effect of CAP-administration on respiratory function and cytochrome c oxidase avtivity in mitochondria from brain cortex of rats exposed to hypoxia
Rong SONG ; Junze LIU ; Lifeng CHEN
Journal of Third Military Medical University 2003;0(14):-
Objective To understand the changing aspects of CAP-administration on mitochondrial oxidative phosphorylation function and cytochrome c oxidase (COX) activity during hypoxia exposure and their mechanisms. Methods Except the control group (C group), adult male Wistar rats were exposed to a hypobaric chamber simulated 5 000-meter high altitude for 23 h every day for 0, 1, 5, 15, 30 d (H_ 0, 1, 5, 15, 30 ) respectively and administrated CAP (100 mg/kg, intraperitoneal injection) every 12 h for 7 d before sacrificed by decapitation. Mitochondrial respiratory function and COX activity were measured by Clark oxygen electrode and polarography, respectively. Results As compared with C group, mitochondrial state 3 respiration (ST_3) and respiratory control rate (RCR) and oxidative phosphorylation rate (ORP) and COX activity in H_0+CAP group all decreased significantly, but by prolonging hypoxia exposure increased and restored to the control level. Conclusion Mitochondrial respiratory function, oxidative phosphorylation efficiency and COX activity in rat brain could improve by administrating CAP during hypoxia exposure and almost reach to the control level by prolonging hypoxia exposure.
7.Effects of brain mitochondrial uncoupling protein content and activity in rat exposed to hypoxia simulating high altitude on mitochondrial energy synthesis
Pan GAO ; Junze LIU ; Chen XIA
Journal of Third Military Medical University 2003;0(21):-
Objective To observe the changes of rat brain mitochondrial uncoupling protein (UCP) content and activity, and explore the effect of UCP on mitochondrial energy metabolism during hypoxia exposure. Methods Adult SD rats were set randomly into control and hypoxia group (n=8 in each group). The rats of hypoxia group were put into a hypobaric chamber simulating 5000-meter high altitude for 3 days (23 h/d). The brain mitochondria was isolated by centrifugation. Mitochondrial oxidative respiratory function was measured by Clark oxygen electrode. Mitochondrial membrane potential was detected by Rhodamine123 method. The content of adenine nucleotide pool (ATP, ADP, AMP) in mitochondria was measured by high performance liquid chromatography. UCP content and activity were detected by [~ 3 H]-GTP binding method. Results High altitude hypoxia resulted in significant increase of UCP activity and a 2.9-fold rise of UCP content of rats (P
8.Inhibition of GDP on mitochondrial respiratory oxygen consumption and its effect on the mitochondrial membrane potential of rat brain
Journal of Third Military Medical University 1988;0(05):-
Objective To investigate effect of guanosine diphosphate (GDP) on the mitochondrial respiratory oxygen consumption and the mitochondrial membrane potential (MMP) of rat brain and explore the relationship of the change of uncoupling proteins (UCPS) activity with the oxygen consumption and MMP. Methods The mitochondria of rat brain were isolated by centrifugation. Mitochondria oxidative respiratory consumption was measured by Clark electrode after the treatment of GDP at different concentrations so as to calculate mitochondrial state 3 respiration (ST3), mitochondrial state 4 respiration (ST4), respiratory control rate (RCR), and the rate of oxidative phosphorylation (OPR). MMP was detected by Rhodamine 123 method at the different concentrations of GDP. Results With the increase of GDP concentration form 0 to 1.0 mmol/L, the values of ST3, ST4 and OPR were reduced while RCR was elevated. But when the concentration increased to 1.4 mmol/L, the former 3 indexes begun to increase while the later declined. When the GDP concentration reached to 1 mmol/L, the inhibitory rate was only 35.1%, 51.3%, 14.2% to ST3, ST4 and OPR respectively, while RCR was increased to 133.2%. No matter the concentration was over 1 mmol/L or under 1 mmol/L, the ability of inhibition was attenuated. MMP reached to the highest point when GDP exerted the highest inhibitory rate on mitochondrial respiratory oxygen consumption. Conclusion GDP, an inhibitor of UCPS, can regulate the respiratory oxygen consumption and MMP of the isolated rat brain mitochondrial directly in a dose-effect fashion. The change of UCPS activity can affect the respiratory oxygen consumption and MMP.
9.Roles of CAP treatment for respiratory function of mitochondria from cerebral cortex of acute hypoxic exposure rats
Lifeng CHEN ; Junze LIU ; Rong SONG ; Yongming DANG ;
Journal of Third Military Medical University 2003;0(18):-
Objective To investigate the roles of mitochondrial DNA (mtDNA) in the respiratory function of cerebral mitochondria in rats exposed to acute hypoxia by observing the changes of mitochondrial respiratory function and cytochrome C oxidase (COX) activity. Methods The rat cerebral cortex mitochondria were isolated by centrifugation. Mitochondrial respiratory function and COX activity were measured by Clark oxygen electrode. Results ① Compared with the control group (C), hypoxia group (H) showed significantly elevated state 4 respiration (ST4), decreased state 3 respiration (ST3), and respiratory control rate (RCR). ② ST3 in group of treatment with chloramphenicol (CAP) plus hypoxia (MH) was significantly lower than that in Group C, while ST4 in Group MH was significantly lower than that in groups C and H. RCR in Group MH was lower than that in Group C, but higher than that in Group H. ③ COX activity in Group H was significantly lower than that in Group C. In Group MH, COX activity was higher than that in Group H, but was still lower than that in Group C. Conclusion The complete expression of mtDNA may play an important role in mitochondrial respiratory function. CAP treatment might be beneficial to the recovery of rat respiratory function.
10.Kinect Somatosensory Interaction Technology and Its Application in Medical Rehabilitation (review)
Chen DING ; Junze WANG ; Chang QU ; Zhan GAO
Chinese Journal of Rehabilitation Theory and Practice 2013;19(2):136-138
The system structure, working principle and main function of Kinect somatosensory interaction technology are presented in this paper. The feasibility of Kinect sensor using software libraries provided by Kinect forWindows SDK to interact with the application program is discussed. The present situation and development trend of Kinect somatosensory interaction technology in medical rehabilitation are introduced.