Changes of expressive proteome of renal tissues in chronic intermittent hypoxic rats

AIM: To obtain the two-dimensional gel electrophoresis maps of renal tissue proteins for identifying the differentially expressed proteins in the chronic intermittent hypoxic rats. METHODS: The rat model of chronic intermittent hypoxia was established. The rats lived in the normoxia environment were used for control. The proteins in the renal tissues underwent two-dimensiona1 gel electrophoresis with immobiline pH gradient isoelectric focusing as the first and vertical SDS-PAGE as the second dimension. Analysis of 2-DE maps was performed to determine the differential expression of proteins between the two groups by the software of ImageMaster 2D Platinum V5.0, in which 4 protein spots expressed differentially were picked up for further identification by MALDI-TOF-MS. RESULTS: Matched and compared with those in control group, 112 protein spots were determined in chronic intermittent hypoxia group. By MALDI-TOF-MS, 4 protein spots with the highest differentially expressive levels were identified as ATP synthase delta subunit mitochondrial precursor, hexokinase, catechol O-methyltransferase and apurinic/apyrimidinic endonudease/redox factor-1. The functions of these identified proteins are involved in cellular energy metabolism, apoptosis, signal transduction, anti-cell injury and hormone metabolism. CONCLUSION: There are obvious differences in expressive proteomes in renal tissue between normoxic and chronic intermittent hypoxic rats. Proteomics can serve as a new approach in the study of obstructive sleep apnea-hypopnea syndrome for discovering new therapeutic targets.     

应用2D-DIGE技术分析柑橘衰退病毒诱导的甜橙差异表达蛋白

【目的】为了研究柑橘衰退病毒(Citrus tristeza virus,CTV)诱导甜橙差异表达蛋白的分离和鉴定,【方法】以接种了CTV的甜橙植株为实验组,健康植株为对照组,利用双向荧光差异凝胶电泳(2D-DIGE)技术分析CTV诱导的甜橙差异表达蛋白。【结果】以t检验P<0.05和相对表达量≥1.5的水平作为判别标准,获得了91个CTV诱导的差异表达蛋白点,在实验组中含量高于对照组的蛋白56个,含量低于对照组的蛋白35个。通过MALDI-TOF质谱分析成功鉴定从中选择的37个蛋白点,其中19个蛋白功能明确,16个为预测或假定具有某种蛋白功能,包括与抗氧化相关的硫氧还蛋白、铁氧还蛋白、超氧化物歧化酶和抗坏血酸过氧化酶,与代谢相关的磷酸甘油酸酯水解酶、木葡聚糖内糖基转移酶、变味蛋白,分子伴侣热激蛋白,折叠酶肽酰脯氨酰顺反式异构酶,以及与光合代谢相关的蛋白等。【结论】CTV的侵染影响到甜橙各种复杂的生物学过程。     

Ischemia reperfusion-induced proteomic changes in rat skeletal muscle

AIM: To investigate ischemia reperfusion (I/R)-induced proteomic changes in rat skeletal muscle. METHODS: Healthy male Wistar rats were randomly divided into two groups as follows (n=6): sham group and I/R group. I/R of right hind limb was induced by 4 h ischemia followed by 24 h reperfusion. The 2-DE was applied to separate the proteins extracted from skeletal muscle tissue at the end of experiment, followed by Coomassie Brillant blue R-250 staining. Computer image analysis was used to determine the differential expression of proteins between the two groups, and 7 protein spots expressed differentially were picked out and subjected to in-gel digest and MALDI-TOP for identification. RESULTS: 354±13 proteins were detected and the match rate was (78.7±1.4)%. 10 proteins displayed significant changes after I/R, of which, 6 proteins increased and 3 proteins decreased in expression. Moreover, 2 spots in I/R group were observed, only 1 spots of which in control. 5 proteins were identified after mass spectrometry. Mitochondrial aldehyde dehydrogenase (ALDH) precursor, heat shock 27 kD protein (HSP27), an unnamed protein product (increased in I/R group), α-actin (decreased in I/R group), and nuclear transport factor 2 (NTF-2) W7a mutant were found in I/R group. CONCLUSION: I/R injury induced differential proteomic changes in rat skeletal muscle. ALDH, α-actin and HSP27 expression, and NTF-2 mutation are involved in I/R injury.     

Effects of ischemic preconditioning on oxidative stress and mitochondrial function in young and old rat myocardium with ischemia/reperfusion

AIM: To study the protective effect of ischemia preconditioning (IPC) on ischemia/reperfusion (IR)-damaged myocardium in young and old rats. METHODS: Male Wistar rats aged at 3 months (young) and 20 months (old) were used to establish myocardial IPC model and IR model with the method of Langendorff heart perfusion. The rats were divided into young ischemia/reperfusion (YIR) group, young ischemic preconditioning (YPC) group, old ischemia/reperfusion (OIR) group and old ischemic preconditioning (OPC) group. Transmission electron microscopy was used to observe the ultrastructural changes of myocardial tissue and myocardial mitochondria. The myocardial infarction area was determined by TTC staining. The lactate dehydrogenase (LDH) content in coronary effluent fluid and the levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in myocardial tissues were detected by the method of colorimetry. The levels of nitrated and carbonylated proteins in myocardial tissue were measured by ELISA. The myocardial cell apoptosis was analyzed by TUNEL assay. The mitochondrial respiratory function and mitochondrial permeability transition pore opening induced by calcium load were evaluated by oxygen electrode method. RESULTS: Compared with YIR group, the myocardial infarction area in YPC group was obviously smaller, SOD activity in myocardial tissues increased, LDH activity in coronary effluent fluid and the content of MDA decreased, and the levels of nitrated and carbonylated proteins in the cardiac tissues reduced. In YPC group, the mitochondrial membrane structure appeared intact, cristae of the mitochondria showed close arrangement, and the matrix was compressed under the electron microscope. Myocardial mitochondrial respiratory control rate, state Ⅲ oxygen consumption and the P/O ratio in YIR group all significantly increased, proton leak decreased, mitochondrial swelling induced by calcium distinctly reduced, and myocardial apoptosis rate declined. No significant difference of the above indexes between OIR group and OPC group was observed. Compared with YPC group, myocardial ultrastructural damage increased clearly, cardiac oxidative stress increased, mitochondrial respiratory function declined, and cell apoptosis and necrosis increased in OPC group. CONCLUSION: Ischemic preconditioning has protective effect against myocardial IR injury in young rat hearts, while old rat hearts were less sensitive to ischemic preconditioning, leading to bluntness of cardioprotection with IPC in aging hearts. This may be related to mitochondrial injury and severe cellular apoptosis caused by increase of cardiac oxidative stress levels in the aging ischemic preconditioning heart.     

红果肉与白果肉杨梅花青苷和糖代谢途径的差异蛋白研究

 以红果肉杨梅‘大叶梅’和白果肉杨梅‘水晶’为研究材料,运用2-DE 和MALDI-TOF-TOF 质谱技术,比较分析两个品种成熟期果肉花青苷和糖代谢途径相关蛋白的差异表达,共获得41 个2 倍差 异表达蛋白,其中33 个差异蛋白得到质谱鉴定。与白果肉的‘水晶’相比,有3 个蛋白质在红果肉的‘大 叶梅’中特异表达,29 个蛋白质上调表达,1 个下调表达。将这些差异蛋白按照功能进行分类,大部分 涉及花青苷代谢（12%）、糖和能量代谢（30%）、蛋白质代谢（18%）、防御应答（15%）等生理过程。发 现花青素合酶（ANS）、苯基丙氨酸解氨酶（PAL）、查尔酮合酶11（CHS11）、二磷酸尿核苷葡萄糖︰类 黄酮3–O–葡糖基转移酶（UFGT）是花青苷合成密切相关的关键酶,这些酶在红果肉的‘大叶梅’中 明显上调表达;同时,还发现9 个糖代谢相关蛋白在红果肉的‘大叶梅’中也显著上调表达。花青苷和 糖代谢途径关键蛋白对杨梅果肉花青苷合成和颜色的形成起到重要作用。     

Proteomic study of myocardial mitochondria with ischemia/reperfusion injury and pinacidil postconditioning in isolated rat hearts

AIM: To investigate the protective effect of pinacidil postconditioning on rat myocardium suffering ischemia/reperfusion injury by mitochondrial proteomics. METHODS: Langendorff apparatus was used to establish the model of myocardial ischemia/reperfusion injury. Sprague-Dawley rats were randomly divided into 2 groups: pinacidil postconditioning group (Pina group) and ischemia/reperfusion injury group (I/R group). After 20 min of perfusion with K-H solution, the perfusion was suspended for 40-min (global ischemia) follow by 60 min of reperfusion in I/R group. In Pina group at the end of 40 min global ischemia, the isolated hearts were perfused with K-H solution containing pinacidil (50 μmol/L) for 2 min followed 58-min perfusion with regular K-H solution. Total proteins extracted from the mitochondria were applied to the two-dimensional gel electrophoresis (2-DE). The differentially expressed protein spots over 2 times were evaluated by a software. Then they were subjected to in-gel digestion, and analyzed by spectrometry. RESULTS: The expression levels of NDUFA10, NDUFS2 and NDUFV2 were elevated but those of IDHA and ECH1 were decreased in Pina group compared with I/R group. Interestingly, 2 spots in the 2-DE map were identified as ATPase subunit δ. The expression levels of one spot was elevated, while the other was decreased. CONCLUSION: Pinacidil postconditioning may decrease the degree of increased expression levels of NDUFA10, NDUFS2 and NDUFV2, promote the expression of IDHA and ECH1, and induce the phosphorylation of ATPase subunit δ, which may be related to the protective mechanism of pinacidil postconditioning.     

Energy metabolism and apoptotic effect of microwave radiation on rat myocardial cells

AIM: To investigate the effect of microwave radiation at different intensities on the rat myocardium and its possible mechanism.METHODS: The rats were radiated by the intensity of 500, 1 000, 1 500 and 2 000 W/m² with 2 450 MHz microwave for 6 min. The heart tissue was collected 6 h after microwave radiation. ATP and mitochondria complex Ⅳ and Ⅴ were measured. The changes of the tissue structures were observed under transmission electron microscope. The apoptosis of the myocardial cells was detected by a cell analyzer. The protein level of cleaved caspase-3 was determined by Western blotting.RESULTS: The concentration of ATP and activity of mitochondria complex Ⅳand Ⅴ signi-ficantly decreased compared with control group in the cardiac tissues. The decreased number, morphological abnormalities such as dissolved cavitation, matrix and obvious tumefaction of mitochondria were observed under transmission electron microscope. The microwave radiation induced the apoptosis of myocardial cells in the rats. The cell apoptotic rate and the protein level of cleaved caspase-3 increased with increasing intensity of microwave radiation(P<0.05).CONCLUSION: Microwave radiation has obvious injury effect on the rat heart, which can cause cardiac energy metabolism dysregulation and cardiac myocyte apoptosis.     

Gene and protein alterations in the hippocampus of rats with temporal lobe epilepsy

AIM:To examine the expression profiles of both genes and proteins in hippocampus of rats with temporal lobe epilepsy (TLE) for revealing the molecular mechanisms of TLE and looking for the candidate targets and new therapeutic approaches in clinical practice.METHODS:Rat temporal lobe epilepsy was induced by administration of lithium chloride and pilocarpine (LiCl-PILO).The expression spectra of genes and proteins were constructed through the techniques of cDNA microarray,two-dimensional (2D) electrophoresis and Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).Subsequently,the differentially expressed genes and proteins were identified and analyzed.RESULTS:There were 192 genes of differential expression observed in hippocampal tissues of LiCl-PILO-induced temporal lobe epilepsy,and 159 genes have been registered in Genbank database,in which 84 genes were up-regulated while 75 genes were down-regulated.78 protein spots of differential display were screened out,in which 31 proteins were detected to be down-regulated and 47 were up-regulated.Finally,5 proteins were identified.CONCLUSION:These genes and proteins found in our study may play pivotal roles in the pathogenic mechanisms of epilepsy and may promise new therapeutic targets for refractory epilepsy in the future.     

Differential analysis of serum proteomics in Crohn's disease treated with infliximab

AIM: To identify the serum proteins that might serve as biomarkers for predicting mucosal healing (MH) in the patients with Crohn's disease (CD) treated with infliximab (IFX). METHODS: We collected serum samples before treatment (0 week, group A) and 14 weeks after treatment (group B) from 7 CD patients with IFX treatment who had achieved MH, as well as the serum samples from 7 CD patients who had not achieved MH (0 week, group C; 14 weeks, group D). Two-dimensional fluorescence difference gel electrophoresis was applied to analyze and compare the results of serum profiles between groups A and B, C and D, A and C, B and D. Matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry and bioinformatics tools were utilized to preliminarily identify and figure out the differentially expressed proteins. RESULTS: (1) In total, there were 44 differentially expressed spots, 36, 3, 10 and 31 differentially expressed spots were detected while comparing A with B, C with D, A with C and B with D, respectively. (2) Among those spots, 17, 2, 2 and 15 proteins were identified, respectively. In total, there were 19 differentially expressed proteins, including apolipoprotein E, apolipoprotein A-I, complement factor H, and so on. (3) Protein functional association networks were carried out based on STRING database. CONCLUSION: The serum protein profiles obviously change after IFX treatment in MH CD patients, and the serum protein profiles of MH patients are different from that of non-MH patients after IFX treatment. The 19 proteins we identified may serve as potential biomarkers for predicting MH in CD patients with IFX treatment.     

Analysis of serum biomarkers in human knee osteoarthritis

AIM: To analyze the proteomic components of the sera from knee osteoarthritis patients and normal people, and to search proteins that might serve as serum biomarkers for osteoarthritis diagnosis, treatment or pathogenesis. METHODS: Two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) was applied to analyze the sera obtained from the patients with knee osteoarthritis (n=4) and normal controls (n=4). The differentially expressed proteins were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Western blotting analysis was applied to confirm the results. RESULTS: Comparative proteomic data of serum from the patients with osteoarthritis was successfully obtained. Eight differentially expressed protein spots were observed. Five up-regulated and 3 down-regulated proteins were identified. Western blot analysis confirmed that α₂-macroglobulin was increased. CONCLUSION: There are significant differences between serum proteins obtained from the patients with knee osteoarthritis and normal controls. α₂-Macroglobulin might be utilized as potential biomarkers for the diagnosis and treatment of osteoarthritis.     

Effects of ginsenoside Rb1 on proteome from hippocampus of rats with epilepsy induced by Coriaria lactone

AIM: To explore the mechanism of development of Coriaria lactone (CL)-induced epilepsy and the neuroprotective effects of ginsenoside Rb1 (GRb1) on the process of epilepsy by identifying the proteins related to CL and GRb1 in rat hippocampus with two-dimensional electrophoresis (2-DE) technology.METHODS: The rat model of epilepsy was induced by CL. The rats in control group, CL epileptic group and GRb1 +CL epileptic group were decapitated and the hippocampus were collected. Two-dimensional electrophoresis was applied to separate the proteins from each group. Analysis of 2-DE maps was used to determine differential expression of proteins by ImageMaster 2D Platinum 5.0, and some differentially expressed protein spots were picked up for further identification by matrix-assisted laser sorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS).RESULTS: Six proteins were successfully identified. Among these, the expression of brain creatine kinase, endophilin-A1 and UPF0628 protein C10orf96 homolog was lower in GRb1+CL epileptic group than those in CL epileptic group. The expression of cytochrome P-450, phosducin-like protein and bridging integrator 3 protein was lower in GRb1+CL epileptic group than those in control group.CONCLUSION: The protein expression profiles are significantly different among control group, CL epileptic group and GRb1+CL epileptic group. The identified proteins may be related to the neuroprotective effects of GRb1. Among these, brain creatine kinase, endophilin-A1 and UPF0628 protein C10orf96 homolog may be related to CL-induced seizures.     

Analysis of differential protein expression during the early stage of in vitro tuberization in taro

Tuberization is a complex and multilevel developmental process. Many important metabolic changes in the early stage of tuberization are crucial to the tuber differentiation and development. In this study, we attempted to identify proteins differentially expressed in the early stage of in vitro tuberization in taro (Colocasia esculenta var. antiquorum) by two-dimensional polyacrylamide gel electrophoresis (2-DE) and mass spectrometry (MS). Protein samples from shoot tips cultured in 8% sucrose media at 0, 2, 4, 6 and 8 d were separated with 2-DE. A total of 13 differentially expressed proteins were analyzed with MS. Four proteins via, glyceraldehyde-3-phosphate dehydrogenase, alcohol dehydrogenase, chloroplast protein synthesis elongation factor (EF-Tu), and ankyrin repeat protein HBP1 were successfully identified during in vitro tuberization of taro. This implies that important metabolic changes, including sucrose metabolism, signal transduction and cell defense, occurred in the early stage of in vitro tuberization in taro.     

Identification of serum differential proteins in colorectal adenoma and early malignantly transformed adenoma by proteomics

AIM: To investigate the associated proteins and sensitive biomarkers for early diagnosis of colorectal adenocarcinoma by comparing the results of differential proteomic analysis between colorectal adenoma and early malignantly transformed adenoma. METHODS: Two-dimensional gel electrophoresis was used to define patterns of protein expressions of colorectal adenoma and early malignantly transformed adenoma. Proteins expressed differentially among groups were detected, cut out and analyzed by MALDI-TOF/TOF mass spectrometry. RESULTS: Two-dimensional protein maps of colorectal adenoma and early malignantly transformed adenoma were analyzed with gel-analysis software, an average of 1 672 spots in adenoma, 1 732 in early malignantly transformed adenoma were observed. 28 spots of a 1.5-fold change were found, including 15 proteins down-regulated and 13 up-regulated in early malignantly transformed adenoma, in which 23 proteins were identified by mass spectrometry, the rate of identification was 82.14%. 13 differential proteins were attained, 8 were up-regulated and 5 were down-regulated, which was classified to 6 categories, including protease inhibitor, complement, immunoglobulin, keratoproteins, signal transduction protein and function-unknown proteins. CONCLUSION: The changes of serum proteins in early malignantly transformed adenoma from adenoma can be identified by proteomic technology. Proteins detected in the study may provide new biomarkers correlated with biological behavior of colorectal adenocarcinoma.     

Proteomics analysis of sera from Crohns disease patients and healthy individuals

AIM: To explore differentially expressed proteins of sera from Crohn's disease patients with comparative proteomics technology.METHODS: Two-dimensional differential in-gel electrophoresis was used to define patterns of protein expression in serum cross-labeled with variant CyDye from 4 patients and matched health adults.Proteins that showed differential expressions were analyzed by matrix-assisted laser desorption/ionization time of flight mass spectrometry.RESULTS: Two-dimensional cross-labeled with variant CyDye protein maps of Crohn's disease patients and the health adults were gained successfully.Gel-analysis software identified differently expressed 29 spots Crohn's disease compared with that of health adults,22 of which were identified by using mass spectrometry,including serine/threonine-protein kinase ATR,leukocyte common antigen precursor CD45,60 kD SS-A/Ro ribonucleo-protein,etc.CONCLUSION: Proteomic analysis can identify the serum proteins with variance of Crohn's disease versus health,the different expressed proteins may providing probable new biomarkers correlated with biological behavior of Crohn's disease.     

Exercise training improves heart function through inducing autophagy and inhibitingapoptosis in aged rats

AIM：To study the mechanism of exercise training in improving old rat cardiac functions, and the effect of gradient exercise training on autophagy and apoptosis in aged rats. METHODS：The rats were randomly divided into 3 groups: young, old and old+exercise (old+Ex). Ultrasonic cardiogram was employed to determine the cardiac functions in the rats. Transmission electron microscope was applied to observe the changes of cardiomyocyte ultrastructure, autophagosome formation and mitochondrial morphology. Western blotting was used to observe the protein expression of Atg5, Beclin 1, microtubule-associated protein 1 light chain 3 (LC3) in cardiac tissues and cytochrome C (Cyt C) in the myocardial mitochondria. TUNEL was adopted to test the apoptosis and spectrophotometry was used to detect the opening of calcium-induced mitochondrial permeability transition pore (mPTP). RESULTS：(1) Compared with young group, the observation in old hearts under transmission electronic microscope found irregular arrangement in myofibrils, loose mitochondria matrix, rupture in mitochondrial membrane and mass deposition of lipofuscin granular in myofilament. In old group, the protein expression of Atg5 and Beclin 1 in the cardiac tissues decreased, the ratio of LC3Ⅱ to LC3Ⅰdropped, mitochondrial Cyt C expression declined, apoptotic index rose, and mitochondrial mPTP opening increased. Noticeable increases were found in left ventricular end-systolic diameter and left ventricular end-diastolic diameter, but left ventricular ejection fraction and left ventricular fractional shortening were decreased. (2) The ultra-structure of the hearts in old +Ex group showed clear sacromere structure, dense matrix and increased number of mitochondria, more autophagosomes and distinct decrease in lipofuscin granular deposition. In addition, the protein expression of Beclin 1 and Atg5 rose, conversion from LC3 I to LC3 II increased, apoptotic index decreased, mPTP opened less, the expression of mitochondrial Cyt C up-regulated, and a significant improvement was observed in left ventricle functions in old+Ex group as compared with old group. CONCLUSION：Exercise training may improve the heart functions in aged rat by upgrading cardiomyocyte autophagy and inhibiting cell apoptosis.     

TMT-tagged quantitative proteomics analysis of differentially expressed proteins and enrichment of signaling pathways in nasopharyngeal carcinoma cells with or without SATB1 over-expression

AIM: To analyze the effects of special AT-rich sequence binding protein 1 (SATB1) expression on the protein expression profiles in human nasopharyngeal carcinoma (NPC) cells, and to enrich the differential signaling pathways through bioinformatics analysis. METHODS: SATB1 over-expressing lentivirus and negative control lentivirus were used to infect the CNE1 cells, and then the cell lines were obtained by puromycin stressed method. The total proteins of the 2 cells were extracted, and the differentially expressed proteins were screened by TMT-labeled protein quantification technique and tandem mass spectrometry. The mRNA levels of the differential protein-coding genes were verified by RT-qPCR. GO analysis was used to annotate and enrich the differentially expressed proteins, and the KEGG database was used to enrich and analyze the signaling pathways of differential proteins. RESULTS: SATB1 over-expressing CNE1 cells were established through infected with associated lentivirus. Compared with the control group, 278 differentially expressed proteins were identified in SATB1 over-expressing CNE1 cells, in which 115 were up-regulated and 163 were down-regulated. 10 representative differential protein-coding genes were verified by RT-qPCR, which showed the consistence with the proteomic results. GO analysis indicated differentially expressed proteins were mainly involved in cellular processes, single-organism processes, biological regulation, metabolic processes, protein binding and catalysis. Cell components of differentially expressed proteins mainly existed in cell part, cells and organelles. KEGG analysis showed that differentially expressed proteins were involved in signaling pathways closely related to tumors, includeing MAPK, PI3K-Akt, AMPK, JAK-STAT, p53, PPAR, Hippo and HIF-1 signaling pathways. CONCLUSION: Over-expression of SATB1 significantly alters the protein expression profiles in the NPC cells and affects multiple signaling pathways closely related to tumors. Proteomics also provides a possible macro approach to the screening of molecular mechanisms, therapeutic and prognostic targets for NPC.     

Effect of acupuncture on mitophagy of skeletal muscle in rats with exercise-induced skeletal muscle damage

AIM: The effect of acupuncture on mitophagy-related protein expression in skeletal muscle of rats after heavy-load exercise was investigated to explore the role of acupuncture in the repairment of exercise-induced skeletal muscle damage. METHODS: Male adult Sprague-Dawley rats (n=128) were randomly divided into 4 groups:control (C, n=8) group, exercise (E, n=40) group, acupuncture (A, n=40) group, and exercise and acupuncture (EA, n=40) group. The rats in E group and EA group performed an eccentric exercise, and the rats in A group and EA group immediately after exercise received acupuncture treatment. The rats in the latter 3 groups were further divided into 0 h, 12 h, 24 h, 48 h and 72 h sub-groups (n=8), and soleus muscle was collected at each time point. The transmission electron microscopy was used to observe the ultrastructural changes of the mitochondria in skeletal muscle. The content of citrate synthase (CS) was measured by ELISA. The protein expression of skeletal muscle PTEN-induced putative kinase 1 (PINK1), parkin and microtubule-associated protein 1 light chain 3 (LC3) was determined by Western blot. RESULTS: After the heavy-load exercise, the mitochondria swelled and accumulated under cell membrane. The number of mitophagosomes was increased, and the content of CS was significantly decreased (P<0.05). The expression of PINK1, parkin and LC3 was significantly elevated (P<0.05). However, the acupuncture intervention after exercise promoted the recovery of mitochondrial ultrastructure, attenuated mitophagolysosome formation, maintained CS content and down-regulated the expression of PINK1, parkin and LC3 (P<0.05). CONCLUSION: Heavy-load exercise causes the damages of mitochondrial structure and function in the skeletal muscle and activates PINK1/parkin pathway to induce excessive occurrence of mitophagy. Acupuncture intervention after exercise is able to alleviate the damage of mitochondria in the skeletal muscle through decreasing the expression of mitochondrial outer membrane protein PINK1, reducing the recruitment of downstream cytoplasmic protein parkin, thereby affecting the combination of LC3 and mitochondria to inhibit the overactivation of mitophagy.     

Effects of short-chain acyl-CoA dehydrogenase on hypertensive vascular remodeling

AIM: To investigate the changes of short-chain acyl-CoA dehydrogenase (SCAD) in hypertensive vascular remodeling and to explore the relationship between SCAD and vascular remodeling in hypertension.METHODS: The spontaneously hypertensive rats (SHR; 24 weeks old) and Wistar rats (24 weeks old) were used as experimental control groups. The SHR and Wistar rats of 16 weeks old were trained by swimming as experimental groups. The systolic pressure was measured periodically. The thickness of vascular wall and the diameter of the vascular lumen were measured. The contents of ROS and ATP, the enzyme activity of SCAD, and the expression of SCAD at mRNA and protein levels in the aorta were determined. The free fatty acid in the serum and aorta was also measured.RESULTS: Compared with Wistar group, the diameter of vascular lumen decreased in SHR group. The thickness of vascular wall, the ratio of vascular wall and the diameter of vascular lumen, and the blood pressure in SHR group were increased significantly (P<0.05). Compared with SHR group, the diameter of vascular lumen increased in SHR+swim group. The thickness of vascular wall, the ratio of vascular wall and the diameter of vascular lumen, and the blood pressure in SHR+swim group were decreased significantly. Compared with control group, the expression of SCAD at mRNA and protein levels, the enzyme activity of SCAD, and the content of ATP were decreased in SHR group. However, the free fatty acid in the serum and aorta, and the content of ROS in the aorta were increased in SHR group. The expression of SCAD at mRNA and protein levels, the enzyme activity of SCAD, the content of ATP were increased in Wistar+swim group and SHR+swim group. However, the free fatty acid in serum and aorta, and the content of ROS in the aorta were decreased in Wistar+swim group and SHR+swim group.CONCLUSION: Decrease in SCAD expression may be associated with hypertensive vascular remodeling. Swimming training can reverse hypertensive vascular remodeling by increasing the expression of SCAD in the aorta.     

自交不亲和甘蓝亲和花粉授粉早期差异蛋白质分析

 为从蛋白质的角度揭示自交不亲和甘蓝亲和授粉早期花粉与柱头相互作用,将蛋白质组学方法应用于自交不亲和性甘蓝柱头与亲和花粉互作早期差异蛋白质的研究。结果表明,亲和授粉后3 ~ 5 min与1 h对比,有116个差异蛋白质点,而授粉后1 h与2 h差异不显著。按照特异表达与变化量大的蛋白质点优先原则,初步选取差异点中71个点做质谱分析,鉴定出31个可信差异蛋白质。与亲和授粉后3 ~ 5 min组相比,1 h组中特异表达蛋白质有19个,上调表达9个,下调表达3个。31种蛋白质的生物信息学分析表明,有两种蛋白与前人发现的与花粉管在柱头中生长发育相关蛋白相同,其他29种参与包括胁迫与防御应答在内多种生理过程。对早期差异蛋白中这种“既有促进花粉管发育的蛋白,也有与防御相关蛋白”现象分析表明,花粉管生长性侵入柱头的过程有可能伴随着柱头抗性的提高。     

Identification of proteins involved in insulin stimulation in v ascular smooth muscle cells of SHR rats

AIM:To identify proteins involved in insul in stimulation and the molecular mechanism of proliferation and migration in vas cular smooth muscle cells (VSMCs).METHODS:A series of methods,including 2-D electrophoresis,PDQ uest software analysis of 2-DE gels,peptide mass fingerprinting based on matrix -assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TO F-MS) and SWISS-PROT database searching,were used to separate and identify the differentially expressed proteins.The difference of some proteins was proved by Western blotting.Proliferation and migration of VSMCs treated with insulin wer e also observed.RESULTS:DNA synthesis were increased in VSMCs.［3H］-thymidi ne incorporation in VSMCs from SHR (14.40±0.85) was higher than that in VSMCs from WKY (9.21±0.93,P<0.05).Migration rate of VSMCs from SHR was abou t 1.8 times higher than that in VSMCs from WKY (P<0.05).Image analysis re vealed that averages of protein spots detected were 502±32 and 612±39 in contr ol VSMCs and in cells treated with insulin,respectively.Result of western blo tting confirmed that α-SM protein was down-regulated and matrix Gla,OPN protei ns were up-regulated by insulin stimulation.CONCLUSION:The results suggest that the differential proteomic analysis may be useful to study related proteins involved in insulin-regulated p roliferation in VSMCs.