Photobiomodulation utilizes monochromatic (or quasimonochromatic) light in the electromagnetic region of 600~1000 nm for the treatment of soft tissues in a nondestructive and nonthermal mode. It is conceivable that photobiomodulation is based upon the ability of the light to alter cell metabolism as it is absorbed by general hemoproteins and cytochrome c oxidase (COX) in particular. Recently it has been suggested radiation of visible and infrared (IR) activates retrograde signaling pathway from mitochondria to nucleus. In this review, the role of COX in the photobiomodulation will be discussed. Further a possible role of water as a photoreceptor will be suggested.
Electron Transport Complex IV ; Magnets ; Metabolism ; Mitochondria ; Phototherapy* ; Water

Apoptosis ; Down-Regulation ; Electron Transport Complex IV ; chemistry ; genetics ; metabolism ; Humans ; Mitochondria ; metabolism ; Mutation ; Neoplasms ; genetics ; metabolism ; pathology

OBJECTIVETo investigate the mechanisms of Yinxing Pingchan recipe (YXPC) and its components, i.e. the components for detoxicating (A), for calming liver (B) and for dissolving blood stasis(C), in preventing and treating Parkinson's disease, and the path of its inhibition on nigrostriatal dopaminergic neuron (DAn) apoptosis in model mice of Parkinson's disease.

METHODSMale C57BL/6J mice were divided into the normal group, the model group and four Chinese medicinal groups, that is, the YXPC group, and Group A, B and C, treated with YXPC and its components A, B and C respectively. Mouse model of Parkinson's disease was established by intraperitoneal injection with 1-methl-4-phenyl-1,2,3,6-tetrahydropyridin (MPTP). All mice were sacrificed in 2 batches at the 14th and the 28th day respectively. The activity of mitochondrial enzyme complex I, II and IV (MEC I, II and IV) in the brain of mice were measured, respectively.

RESULTSAs compared with the normal group, the activity of MEC I and IV in brain was significantly lower (P < 0.05 or P < 0.01), and that of MEC II had no obvious change in the model group. As compared with the model group, the activity of MEC I was significantly higher in YXPC group and Group C at the 14th day (P < 0.05), while the activity of MECII in Group A at the 14th day, Group B at the 28th day and Group C at both 14th and 28th day was significantly lower (P<0.05 or P<0.01). Activity of MEC IV in the four Chinese medicinal groups at the 14th day all significantly increased (P<0.05 or P<0.01), and retained at high level in Group B and Group C at the 28th day (P<0.05).

CONCLUSIONYXPC and its components can maintain the mitochondrial function by partial inhibiting the activity of its enzyme complex, preventing DAn apoptosis to slow down the progress of Parkinson's disease.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine ; Animals ; Brain ; enzymology ; Drugs, Chinese Herbal ; pharmacology ; Electron Transport Complex I ; metabolism ; Electron Transport Complex II ; metabolism ; Electron Transport Complex III ; metabolism ; Electron Transport Complex IV ; metabolism ; Enzyme Activation ; drug effects ; Male ; Mice ; Mice, Inbred C57BL ; Mitochondria ; enzymology ; Parkinson Disease ; drug therapy ; enzymology ; etiology ; Random Allocation

OBJECTIVETo investigate the effects of estrogen on the activity and subunits expression of cytochrome c oxidase (COX) in mitochondria from genioglossus of chronic intermittent hypoxia (CIH) rat.

METHODSForty-eight male Sprague-Dawley (SD) rats were randomly divided into three groups: the normal control group (NC), the chronic intermittent hypoxia group (CIH) and estrogen plus hypoxia group (E + CIH). Rats in the latter two groups were used to build CIH models (8 h/d, 5 w). In the mean time, rats in NC group and CIH group were injected with sterile olive oil (0.2 ml each time, twice a week), and rats in E + CIH group were injected with estrogen (0.2 mg/kg, twice a week), respectively. At the end of the treatment, the genioglossus of the rat was removed and the mitochondria were isolated by centrifugation program. COX activity was measured with a spectrophotometer. The protein content of COX subunit I and IV was detected by Western blotting analysis, and mRNA expression levels of COXI and COXIV were determined by real-time polymerase chain action.

RESULTSCOX activity in CIH group [(0.143 +/- 0.029) microkat/mg] was lower significantly than that in NC group [(0.273 +/- 0.058) microkat/mg, P < 0.01]; In E + CIH group [(0.203 +/- 0.073) microkat/mg], COX activity was higher than that in CIH group (P < 0.05), but still lower than that in NC group (P < 0.05). Compared with NC group (47.325 +/- 7.502), decreased COXI expression was observed in CIH group (10.789 +/- 8.144, P < 0.01) and E + CIH group (25.593 +/- 11.108, P < 0.01). While in the mean time, the protein content of COXI in E + CIH group was higher than that in CIH group (P < 0.05). There was no significant difference in protein content of COXIV among the three groups (P > 0.05). COXI and COXIV mRNA levels had no significant change among the three groups (P > 0.05).

CONCLUSIONSCIH exposure could decrease mitochondrial COX protein expression and activity in the genioglossus of the rat, while estrogen administration might be beneficial to the recovery of those.

Animals ; Electron Transport Complex IV ; metabolism ; Estrogens ; pharmacology ; Hypoxia ; Male ; Mitochondria ; metabolism ; Muscle, Skeletal ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tongue ; metabolism

This study was intended to evaluate the effects of hypoxic exposure on gene expression and coordination of cytochrome oxidase (COX) subunits I (COX I) and IV (COX IV) encoded by mtDNA and nDNA respectively in rat cerebral cortex. Male Wistar rats were exposed to hypoxia in a hypobaric chamber simulating high altitude at 5000 m for 2, 5, 15 and 30 d. Control rats were fed outside the hypobaric chamber (the height was 300 m above sea level). Rats were sacrificed and mitochondria from cerebral cortex were isolated by differential centrifugation at each time point. COX I and COX IV proteins in isolated rat cerebral cortex mitochondria were detected by Western blot analysis and mRNA in the cerebral cortex by RT-PCR. The ratios of protein and mRNA were used to estimate the coordinative expression of two subunits. The results showed that COX I mRNA increased significantly at 2 and 5 d, and decreased to the control level at 15 and 30 d; COX IV mRNA remarkably increased at 2, 5 and 15 d, and dropped below the control level at 30 d. The mRNA ratio of COX IV to COX I reached a peak at 15 d, but showed no differences between other time points. The Western blot analysis of COX I and COX IV in isolated rat cerebral cortex mitochondria showed no obvious changes during hypoxic exposure. Our findings demonstrate that hypoxia can affect mRNA expression of COX I and COX IV and their coordination, while protein expression of both subunits are stable and coordinative. This study suggests that the expression of COX I and COX IV proteins during hypoxic exposure is coordinately regulated by post-transcriptional mechanisms.
Animals ; Cerebral Cortex ; metabolism ; Electron Transport Complex IV ; metabolism ; Gene Expression Regulation, Enzymologic ; Hypoxia ; metabolism ; Male ; Mitochondria ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo identify the binding site position of the hepatitis B virus (HBV) X protein (HBx) functional interaction with the cytochrome C oxidase subunit III (COX III, a key regulator of mitochondrial function) by using a yeast two-hybrid system.

METHODSTwo fragments of HBx mutants (X1 1-72aa and X2 1-117aa) were amplified by PCR and inserted into the bait plasmid pAS2-1.The resultant mutant plasmids were transfected into yeast cells using the lithium acetate-method.PCR and gene sequencing were used to confirm that the mutant fragments were expressed properly in yeast cells.Western blotting was used to verify that the mutant proteins were translated accurately in the yeast cells.Filter assay was used to exclude autoactivated mutants.Hybridization in solid medium and beta-gal activity detection were used to determine the precise position of the binding site for HBx and COX III interaction.

RESULTSThe two mutant plasmids containing HBx 1-72aa and 1-117aa respectively were successfully constructed and the mutants were both properly expressed and translated in yeast cells; no autoactivated mutants were detected throughout the experimental process.The binding site of HBx and COX III was found to be encompass the amino acids 72 through 117 of HBx.

CONCLUSIONAmino acids 72 through 117 of HBx are the key domain of the HBx functional interaction With COX III; this domain may represent a useful target for molecular-based therapies to treat HBV-related diseases.

Electron Transport Complex IV ; metabolism ; Hepatitis B virus ; metabolism ; Plasmids ; Polymerase Chain Reaction ; Protein Binding ; Trans-Activators ; metabolism ; Transfection ; Two-Hybrid System Techniques

OBJECTIVE: To investigate the relation between injury time and the expression of cytochrome c oxidase subunit VIc (COX6C) mRNA in skeletal muscle of rat after contusion. METHODS: A total of fifty-four SD rats were divided into the control group and the contusion groups (0.5, 1, 6, 12, 18, 24, 30, and 36 h after contusion), randomly. The contusion model was established by free fall drop of gravity hammer. At corresponding time point after contusion, the regular histology was examined and expression level of COX6C mRNA was tested by real-time PCR after extraction of total RNA from the tissues. RESULTS: The main pathological features of 6 h after injury included edema and hemorrhage in myocytes with no inflammatory cells found. After 6 hours, the findings included myocyte degeneration and necrosis, inflammatory cells infiltration, and fibrous connective tissue proliferation in the contused zone. The expression level of COX6C mRNA was higher than that of the control group within 6 h after contusion. The expression level was lower than that of the control group from 6-36 h after contusion. CONCLUSION The level of COX6C mRNA expresses in a regular way after contusion. It may be useful for estimating wound age in combination with the results of pathological features.
Animals ; Contusions/metabolism* ; Electron Transport Complex IV/metabolism* ; Muscle, Skeletal/metabolism* ; RNA ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Time Factors

OBJECTIVETo clone and identify MF-1 gene which responded to extremely low frequency magnetic fields(ELF MF) in Daudi cells, and explore the response universality of MF-1 gene in several MF-sensitive cell lines, so as to provide experimental basis for revealing the mechanism of biological effects induced by magnetic field.

METHODSThe DNA fragment of MF-1 was cloned and sequenced; the mRNA level of MF-1 gene were analysed in MF-sensitive cell lines(HL-60, L1210 and CHL) by Northern blot after these cells being treated with 0.1 mT and 0.8 mT MF for 20 minutes and 24 hours, respectively.

RESULTSThe MF-1 cDNA sequence had 100% homology with cytochrome oxidase subunit 1 gene(CO1) by searching Gene Bank database; the transcription of CO1 in HL-60, L1210 and CHL cell lines which exposed to 0.1 mT and 0.8 mT MF for 20 minutes were significantly lower(0.38 +/- 0.12 and 0.37 +/- 0.04) than that of control(0.58 +/- 0.12) and so did for 24 hours exposure(0.46 +/- 0.09 and 0.45 +/- 0.09 vs 0.65 +/- 0.06) (P < 0.05).

CONCLUSIONCO1 is a MF-responsive gene. Cytochrome oxidase activity may be affected through low level of CO1 transcription by magnetic fields, thus induce bioeffects in organisms.

Animals ; Cricetinae ; Electron Transport Complex IV ; genetics ; metabolism ; radiation effects ; HL-60 Cells ; Humans ; Leukemia L1210 ; Magnetics ; Mice ; Protein Subunits ; RNA, Messenger ; analysis ; Transcription, Genetic ; radiation effects

OBJECTIVETo investigate the effects of taurine on myocardial mitochondria and their enzyme activities in rats with severe burn.

METHODSOne hundred and twenty healthy adult Wistar rats were subjected to 30% TBSA full-thickness burn. They were randomly divided into burn group (B, with intraperitoneal injection of isotonic saline), treatment group (T, with intraperitoneal injection of taurine, 200 mg/kg),with 60 rats in each group . Ten rats with sham scald were used as control (S group). The myocardial tissue samples in B and T groups were harvested at 1, 3, 6, 12, 24 and 48 postburn hours (PBH) for determination of activity respectively of succinate dehydrogenase (SDH), cytochrome oxidase (CCO), the superoxide dismutase (SOD), Ca2+ -ATPase in mitochondria, and contents of cytochrome c (Cyt c), cytochrome aa3 (Cyt aa3), malondialdehyde (MDA), and Ca2+ in mitochondria and cytoplasm . The myocardial tissue samples of controls were harvested at 1 PBH for determination of above indices.

RESULTSThe activity of CCO in B group was decreased at 1 PBH , especially at 6 ,12 PBH. The activity of SDH in B group was decreased to lowest level at 6 PBH, and its value was lower than that of S group at each time point. The activity of CCO or SDH in T group was not obviously decreased, and the activity of CCO at 3, 6, 12 PBH showed significant difference compared with B group (P < 0.05). The contents of Cyt aa3 and Cyt c in B group at 3, 6, 12, 24 PBH were obviously decreased, which were significantly lower than those in T group (P < 0.05). The activity of SOD in B group at 3, 6, 12 PBH was obviously decreased, the activity of Ca2+ -ATPase at 3, 6, 12 and 24 PBH was decreased to different extent, which was significantly lower than those in T group (P < 0.05). The MDA contents in B and T groups were higher than that in S group at 3-48 PBH ,and it was highest in B group (P < 0.05). The Ca2+ content of mitochondria in B group at 1 PBH was increased (13.7 +/- 1.5), and it was (24.8 +/- 2.6), (29.7 +/- 3.1), (16.3 +/- 1.9) and (13.5 +/- 1.7) at 3, 6, 12, 24 PBH respectively,and they were all higher than that of S group (10.7 +/- 1.6, P < 0.05). The Ca2 contents of cytoplasm in group B at 3 - 24 PBH were also higher than that in S group (P < 0.05). The Ca2+ content of mitochondria in T group at 3, 6, 12, 24 PBH was (16.8 +/- 2.8), (18.7 +/- 1.9), (10.5 +/- 1.8) and (13.3 +/- 1.7)respectively, which were lower than that in B group at every time point.

CONCLUSIONTaurine have protective effect on mitochondria and their enzyme activities in myocardium in rats with severe burn, and it may be attributable to improving the ability of eradicating oxygen free radicals and alleviating Ca2+ overload in the mitochondria.

Animals ; Burns ; drug therapy ; enzymology ; metabolism ; Calcium ; metabolism ; Calcium-Transporting ATPases ; metabolism ; Cytochromes c ; metabolism ; Electron Transport Complex IV ; metabolism ; Female ; Male ; Mitochondria, Heart ; enzymology ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Succinate Dehydrogenase ; metabolism ; Superoxide Dismutase ; metabolism ; Taurine ; therapeutic use

OBJECTIVETo observe the influence of heat shock preconditioning on the expressions of heat shock protein (HSP) 60 and HSP 70 and on the activities of cytochrome oxidase (CCO) and superoxide dismutase (SOD) in mitochondria in gastric mucosa of severely scalded rats, and to investigate its protective mechanism on acute gastric mucosal lesion in rats with severe scald.

METHODSOne hundred and forty-four Wistar rats were randomized into three groups, i. e. scald group ( n = 40, acute gastric mucosal lesion was made after scald, other 8 normal rats without scald were employed as blank control); HS group ( n =40, with heat shock preconditioning 20 h before scald), and other 8 rats preconditioned with heat shock but without scald were employed as experimental control I; actinomycin D group ( n = 40, with intraperitoneal injection of 0. 1 mg/kg actinomycin D 30 min before heat shock preconditioning and other treatment as HS group), and other 8 rats with merely actinomycin D injection were employed as experimental control II. Eight rats in each group were sacrificed and laparotomized at 3, 6, 12, 24 and 48 post-scald hours (PSH) , respectively to determine the index of gastric mucosal lesions (UI ) , the mRNA expressions of HSP70 and protein expression of HSP60 and HSP70, and the changes in the activities of SOD and CCO.

RESULTSUI of the scalded rats increased as the time elapses, reaching the peak (12. 8 +/- 1.9) at 24 PSH. In addition, UI in HS group was significantly lower than that in scald group at each time-point except that at 3 PSH ( P < 0. 05 or 0. 01). The extent of gastric mucosal lesion in rats in actinomycin D group was obviously aggravated compared with that in scald and HS groups ( P <0. 05). The HSP70 mRNA expression in both scald and HS groups was increased at each time-points except for 48PBH, while that in actinomycin D group was increased at 24 PBH and 48PBH. The expressions of HSP70 and HSP60 were greatly increased in HS group compared with those in scald group ( P < 0. 05 or 0. 01) , while those in actinomycin D group were significantly inhibited ( P < 0. 05). The activities of CCO and SOD were gradually decreased in gastric mucosa in scald group, but it was greatly improved by HS preconditioning at 6, 12, 24 PSH ( P < 0. 05 or 0. 01).

CONCLUSIONHeat shock preconditioning is beneficial for the protection of acute gastric mucosal lesion of rats after severe scald, due to increase of HPS60 and HSP70 expression, and increase of CCO and SOD activities in mitochondria.

Animals ; Burns ; metabolism ; pathology ; Chaperonin 60 ; metabolism ; Electron Transport Complex IV ; metabolism ; Female ; Gastric Mucosa ; metabolism ; pathology ; HSP70 Heat-Shock Proteins ; metabolism ; Heat-Shock Response ; Male ; Mitochondrial Proteins ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Superoxide Dismutase ; metabolism