1.Inhibition of expression of P-selectin by antioxidant in cholesterol-fed rats.
Choong Sik LEE ; Jeung Mok CHOI ; Dae Hyun PARK ; Dae Young KANG ; Thomas C REGISTER ; Michael R ADAMS
Journal of Korean Medical Science 1999;14(1):8-14
Butylated hydroxytoluene (BHT) can inhibit experimental atherosclerosis in animals. Although the agent is an antioxidant, the exact mechanism of the reaction in atherosclerosis is still unknown. To investigate the effects of BHT on expression of P-selectin (PADGEM, GMP-140), intercellular adhesion molecule-1 (ICAM-1) and class II MHC (Ia) antigen, we proposed an experiment on rats. Male rats (n=18 per group) were fed either a normal cholesterol control diet, a normal cholesterol diet containing 0.5% BHT (BD), a high cholesterol diet containing 1.5% cholesterol and 0.1% sodium cholate (CD), or the CD diet containing 0.5% BHT (BCD). Rats were sacrificed after 3 days, and after 1, 2, 4, 10, and 17 weeks of dietary treatment. Although there was no gross or light microscopic atherosclerotic lesions, scanning electron microscopy revealed monocytic adhesion to aortic endothelium and mild endothelial injuries in CD and BCD groups. Immunohistochemically, the addition of BHT to a high cholesterol diet inhibited P-selectin expression but not in ICAM-1 and Ia antigen. These findings suggest that in rats, high cholesterol diets induce expression of ICAM-1, P-selectin and Ia antigen. In addition, the antiatherogenic effect of BHT may play a role in the inhibition of P-selectin.
Animal
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Antioxidants/pharmacology
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Antioxidants/metabolism*
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Aorta, Abdominal/ultrastructure
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Aorta, Abdominal/pathology
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Aorta, Thoracic/ultrastructure
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Aorta, Thoracic/pathology
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Butylated Hydroxytoluene/pharmacology
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Butylated Hydroxytoluene/metabolism*
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Cholesterol/metabolism
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Cholesterol, Dietary/metabolism*
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Male
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Microscopy, Electron, Scanning
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P-Selectin/biosynthesis*
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Rats
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Rats, Sprague-Dawley
2.Aortic artery elastic lamina degradation, collagen remodeling, oxides stress and inflammation in the apolipoprotein E deficient mice with or without aortic banding.
Li CHEN ; Wen-li CHENG ; Yuan-nan KE ; Yong WANG ; Tong-liang ZHOU ; Lin PAN ; Hong LI ; Jian GUO
Chinese Journal of Cardiology 2012;40(6):505-510
OBJECTIVETo explore the elastic lamina degradation and the collagen remodeling of aortic artery as well as oxides stress and inflammation of the apolipoprotein (Apo E) deficient mice with or without experimental hypertension.
METHODSEighty male Apo E deficient mice were fed with high-fat diet beginning at six weeks of age. At 8-week old, they were randomly divided into hypertension group and control group (n=40 each), the mice in hypertension group were subjected the suprarenal aortic constriction operation and then randomly divided into two subgroups: 15 weeks age and 30 weeks age groups. At the end of experiment, the vascular elastic lamina degradation and the content of collagen were determined by morphological method, plasma ICAM-1 level was measured by ELISA, and the rennin activity measured by radioimmunoassay, the superoxide anion detected by fluorescence, the MOMA-2 observed by immunofluorescence in all animals. mRNA expression of NF-κB P65 and MMP9 was detected by real-time PCR.
RESULTIn 15-week old group, the elastic lamina degradation Grade II and the intima-media thickness in the hypertension group were significantly higher than in the control group [(5.4±3.3)% vs. (8.9±2.5)%, P<0.05; (98.66±18.90) µm vs. (70.08±11.71) µm, P<0.05]. In 30-week old group, the elastic lamina degradation Grade III, the III type of collagen and the intima-media thickness were also significantly higher than in the control group [(15.2±3.7)% vs. (8.1±3.3)%, P<0.01; (23.00±7.73)% vs. (11.00±3.82)%, P<0.05; (147.31±22.60) µm vs. (103.98±17.21) µm, P<0.01]. The level of ICAM-1 in hypertension group was significantly higher than that of control group in both 15-week old and in 30-week old mice [(46.3±3.7) µg/ml vs. (40.6±5.7) µg/ml, P<0.05; (56.0±3.1) µg/ml vs. (45.2±2.8) µg/ml, P<0.05]. The superoxide anion, the MOMA-2, mRNA expression of NF-κB P65 and MMP9 in the hypertension group were significantly higher than in the control group in both 15-week old and in the 30-week old mice. The increase in hypertension group was more pronounced in the 30-week old mice than in the 15-week old mice.
CONCLUSIONThe elastic lamina degradation and the collagen remodeling of aortic artery as well as oxides stress and inflammation are more significant in the Apo E deficient mice with hypertension than in control Apo E deficient mice.
Animals ; Aorta ; physiopathology ; Apolipoproteins E ; genetics ; Collagen ; metabolism ; Hypertension ; metabolism ; pathology ; Inflammation ; pathology ; Male ; Mice ; Mice, Knockout ; Oxidative Stress
3.Expression of connective tissue growth factor and pathological remodeling in ascending thoracic aortic aneurysm.
Yan-hai MENG ; Chuan TIAN ; Lei LIU ; Liang WANG ; Wen-zhi LIU ; Qian CHANG
Chinese Journal of Surgery 2011;49(3):261-265
OBJECTIVETo study the expression of connective tissue growth factor (CTGF) and its significance in sporadic ascending thoracic aortic aneurysm (AAA), and initially to investigate the mechanisms of pathological remodeling in AAA.
METHODSAAA specimens were taken from 18 patients during elective surgical intervention, and 18 control specimens of ascending aorta were obtained from patients undergoing coronary artery bypass surgery. Specimens were stained with HE and Masson to evaluate the arrangement and aggregation of cells and collagen types I and III; immunohistochemistry staining was performed using antibodies directed against markers of CTGF; real-time PCR analysis was performed to quantify the expression level of CTGF and collagen types I and III.
RESULTSPathological results show degradation of elastin and hyperplasia of collagen fibers as well as disordered arrangement of smooth muscle cells in AAA. When compared with controls, protein levels of CTGF were significantly increased [(44 ± 4)% vs. (33 ± 5)%, P < 0.01]. Similar patterns were shown in mRNA levels of CTGF (P < 0.01). Using real-time PCR method, elevated levels (relative expression ratio of mRNA: 10.54/3.8 and 1.79/1.19, respectively; P < 0.01, both) of collagen types I and III were observed. CTGF expression had a correlation with both collagen fibers and aortic aneurysm diameter (r = 0.784, P < 0.01; r = 0.793, P < 0.01).
CONCLUSIONSThese results indicate increased expression of aortic collagen types I and III as well as CTGF in AAA specimens, which is likely to be responsible for the aortic wall pathological remodeling. The expression of CTGF was positively correlated with the aortic diameter. As a cytokines factor can stimulate collagen synthesis, CTGF may be involved in the pathogenesis and progression of AAA.
Aged ; Aorta ; metabolism ; pathology ; Aortic Aneurysm, Thoracic ; metabolism ; pathology ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Connective Tissue Growth Factor ; metabolism ; Female ; Humans ; Male ; Middle Aged
4.Effects of PPAR-gamma agonist and MMP-2 on formation of atherosclerosis plaque in rabbits.
Feng LUO ; Zhao-hui WANG ; Lin-lin DU ; Jue WANG
Chinese Journal of Pathology 2007;36(8):556-557
Animals
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Aorta
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metabolism
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pathology
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Atherosclerosis
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etiology
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metabolism
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pathology
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Cholesterol, Dietary
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Male
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Matrix Metalloproteinase 2
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metabolism
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PPAR gamma
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agonists
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Rabbits
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Random Allocation
5.Molecular mechanism of effect of wenban humai granule on stability of atheromatous plaque.
Bao-ting ZHANG ; Qian-lin YAN ; De-xin YAN ; Zhi LI ; Yong-chun YU ; Guo-ping HUANG ; De-sheng TANG ; Xin YE
Chinese Journal of Integrated Traditional and Western Medicine 2005;25(2):154-159
OBJECTIVETo explore the molecular mechanism of wenban humai granule (WHG) in stabilizing atheromatous plaque, by observing its effect on the collagen degradation and synthesis imbalance manner in the fibrous cap of the plaque.
METHODSAtherosclerosis (AS) rabbit model established by feeding high fat diet. The changes of protein and mRNA expression of macrophage CD68, metalloproteinase-1 (MMP-1), alpha-smooth muscle actin (alpha-SMA) and collagen I (C-I) in model rabbits' neo-genesic intima were determined by immunohistochemical stain and in situ hybridization methods before and after treatment as well as before and after modeling.
RESULTSAfter being fed with high fat diet for 7 weeks, the protein and mRNA expression of macrophage CD68, MMP-1 in neo-genesic intima of aorta in the model rabbits significantly increased, these changes could be significantly restored after 8 weeks treatment with WHG or simvastatin. At the same time, the expressions of alpha-SMA protein and C-I protein and mRNA slightly increased due to the immigration of SMC in aortic media to neo-genesic intima, these expressions could be further increased after WHG treatment but showed a reducing trend after simvastatin treatment (P < 0.05 and P < 0.01). In the whole course, positive correlation was shown between protein expressions of CD68 and MMP-1 (r = 0.952, P < 0.01) and also between these of alpha-SMA and C-I (r = 0.793, P < 0.01).
CONCLUSIONWHG affects the collagen degradation and synthesis imbalance in the fibrous cap of the plaque to stabilize plaque through bi-directional regulation, up-regulating synthesis thesis factors and down-regulating degradation factors, while simvastatin perform its action on plaque stability by down-regulating degradation factors alone.
Actins ; metabolism ; Animals ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Aorta ; pathology ; Arteriosclerosis ; drug therapy ; metabolism ; pathology ; Collagen ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Macrophages ; metabolism ; Matrix Metalloproteinase 1 ; metabolism ; RNA, Messenger ; metabolism ; Rabbits ; Random Allocation
6.Suppressive effect of hydrogen sulfide donor on endothelin-1 production in aorta of atherosclerotic rats.
Wei LI ; Junbao DU ; Hongfang JIN ; Email: JINHONGFANG51@126.COM.
Chinese Journal of Pediatrics 2015;53(6):448-452
OBJECTIVETo examine the effect of H2S donor, sodium hydrosulfide (NaHS), on ET-1 level in plasma and aorta in rats with atherosclerosis (AS).
METHODThirty male rats, weighting 200-220 g, were randomly divided into AS, AS+NaHS and control groups, n = 10 in each group.Rats were given a single dose of vitamin D3 (700 000 U/kg) in the first three days and fed with a high-cholesterol diet for 8 weeks to induce AS. Rats in AS+NaHS group were intraperitoneally injected with an H2S donor NaHS, at a dose of 56 µmol/(kg·d) for 8 weeks. At the end of the experiment for 8 weeks, all the rats were sacrificed. The plasma was collected and the aorta and coronary tissues were isolated. The atherosclerotic lesions in both aorta and coronary arteries were detected using oil red O method. H2S concentration in plasma was determined with sulfide-sensitive electrode method. ET-1 levels in plasma and aorta were calculated by radioimmunoassay kit and the localization of ET-1 in the aorta was detected by immunohistochemistry. Plasma nitric oxide synthase (NOS), endothelial NOS (eNOS), inducible NOS (iNOS) were detected with colorimetry.
RESULTAS plaque area in root of aorta of rats in AS group, AS+NaHS group and control group were (11.6±3.3)%, (1.6±1.1)%, (0.0±0.1)% respectively. The difference in AS plaque area in root of aorta among the three groups was statistically significant (F=97.675, P < 0.05). AS plaque area in coronary artery of rats in AS group, AS+NaHS group and control group were (21.4±5.7)%, (4.8±2.5)%, (0.0±0.0)% respectively. The difference in AS plaque area in coronary artery among the three groups was statistically significant (F=97.519, P < 0.05). Plasma H2S level in rats of AS group ((22.0±3.1) µmol/L) was significantly lower than that of control group ((27.9±1.0) µmol/L) and AS+NaHS group ((33.3±6.2) µmol/L, all P < 0.05). Compared with control group ((70.0±10.7) ng/L), plasma ET-1 in rats of AS group ((89.6±14.2) ng/L) and AS+NaHS group ((93.1±15.5) ng/L, P both < 0.05) were increased. However, there was no significant difference in plasma ET-1 content in rats between AS+NaHS group and AS group (P > 0.05). Compared with control group ((3.8±1.2) ng/g), ET-1 content in aorta in rats of AS group ((11.9±4.9) ng/g) and AS+NaHS group ((8.2±2.5) ng/g, both P < 0.05) were increased, and ET-1 content in aorta in rats of AS+NaHS group was decreased compared with AS group (P < 0.05). Immunochemistry results showed that ET expression in cytoplasm in aortic endothelial cells in rats of AS group was strengthened, while ET expression in rats of control group and AS+NaHS group was weak. NOS activity of rats in control group, AS group and AS+NaHS group was (25.4±5.6), (51.8±10.0) and (27.6±6.5) U/ml, eNOS activity (15.3±6.2), (4.5±2.7) and (8.7±3.9) U/ml, and iNOS activity (9.9±4.0), (47.3±10.7) and (19.0±5.2) U/ml, respectively.Differences among the three groups were statistically significant (NOS activity: F=37.231, P < 0.05, eNOS activity: F=14.600, P < 0.05, and iNOS activity: F=72.131, P < 0.05).
CONCLUSIONH2S donor NaHS reduced the AS plaque in AS rats. The mechanisms might involve the protective effect of H2S on the vascular endothelial cell, decreasing ET-1 production in aortal endothelium of atherosclerotic rats.
Animals ; Aorta ; metabolism ; pathology ; Atherosclerosis ; metabolism ; pathology ; Coronary Vessels ; pathology ; Disease Models, Animal ; Endothelin-1 ; blood ; metabolism ; Hydrogen Sulfide ; pharmacology ; Male ; Nitric Oxide Synthase Type II ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Random Allocation ; Rats ; Sulfides ; pharmacology
7.Effects of phlorizin on vascular complications in diabetes db/db mice.
Lin SHEN ; Bei-An YOU ; Hai-Qing GAO ; Bao-Ying LI ; Fei YU ; Fei PEI
Chinese Medical Journal 2012;125(20):3692-3696
BACKGROUNDDiabetic macrovascular complications are important causes of cardiovascular and cerebrovascular diseases and also one of the major causes of morbidity and mortality in patients with type 2 diabetes mellitus (T2DM). Phlorizin has been reported to be effective in reducing the blood glucose level in diabetic mellitus, while little is known about its effects on vascular complications. This study aimed to observe the effects of phlorizin on the aorta of diabetes db/db mice and explore its mechanism.
METHODSDiabetic db/db mice (n = 16) and age-matched db/m mice (n = 8) were divided into three groups: normal control group (CC group, db/m mice, n = 8), untreated diabetic group (DM group, db/db mice, n = 8) and diabetic group treated by phlorizin (DMT group, db/db mice, n = 8). Phlorizin (20 mg/kg body weight) was given in normal saline solution intragastrically for 10 weeks. Animals were weighed weekly. At the 10th weekend, all mice were fasted overnight and then sacrificed. Fasting blood was collected, and the aortas were dissected. The blood samples were analyzed for fasting blood glucose (FBG), serum advanced glycation end products (AGEs), malondialdehyde (MDA) and superoxide dismutase (SOD) activity, the aortic ultrastructure was studied.
RESULTSThe weight and serum concentration of FBG, AGEs, and MDA in the DM group were higher than that in the CC group (P < 0.01), and they were significantly lower in the DMT group (P < 0.05). Serum SOD activity was lower than that in the CC group (P < 0.01), and it is significantly higher in the DMT group (P < 0.05). The severity of aorta damage in the DMT group was less than that in the DM group.
CONCLUSIONSPhlorizin protected the db/db mice from diabetic macrovascular complications, attributed to the decreasing of blood glucose and AGEs level, and its antioxidant potential. This study may provide a new natural medicine for treating diabetic macrovascular complications.
Animals ; Aorta, Thoracic ; pathology ; Blood Glucose ; analysis ; Diabetic Angiopathies ; drug therapy ; pathology ; Glycation End Products, Advanced ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Phlorhizin ; therapeutic use ; Superoxide Dismutase ; metabolism
8.Analysis on mechanical properties of aortic artery in rats with atherosclerosis.
Ming CHEN ; Shenglin LIU ; Zheng DAI ; Yi WANG ; Yi LIU ; Yuan YU
Journal of Biomedical Engineering 2009;26(1):89-92
To understand the mechanical properties of aortic artery of atheroselerosis (AS), the aortic artery of rat with AS was studied by mechanical test. Wistar rats were used for establishing the model. The mechanical measurements of opening angle in zero-stress state and the vessel loading test were conducted on the isolated aortic arteries of AS rats. Data on the stress-strain of aortic artery were obtained. Determination of percentage of collagen content was made with the use of electron microscope. The relationship between mechanical measurements and collagen concentration was evaluated. The opening angle in the group of AS was significantly smaller than that in control (87.74 degrees +/-9.67 degrees vs. 196.03 degrees +/- 27.76 degrees, P < 0.001). Significant decrease of material constants (alpha0, alpha1, alpha2, b0, b1, b2) in both long axis and radial axis was observed in AS group(campared with control, P < 0.05-0.001). Close relationship between the mechanical constants and the percentage of elastin and collagen content was observed (r = -0.7523 to -0.8423, P < 0.001). In conclusion, mechanical remodeling in aortic artery of AS might be related with histological remodeling.
Animals
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Aorta, Abdominal
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metabolism
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pathology
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Atherosclerosis
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metabolism
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pathology
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physiopathology
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Biomechanical Phenomena
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Collagen
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analysis
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Elastin
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analysis
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Female
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Male
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Random Allocation
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Rats
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Rats, Wistar
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Stress, Mechanical
9.Identification of differentially expressed genes in rats and preliminary analysis in regression of vascular calcification.
Xiu-Juan WU ; Yi LIU ; Wen-Xue SUN ; Xiao-Hong DU ; Tian-Xin CHEN ; Rui-Yang MAO ; Li-Yuan XU
Chinese Journal of Applied Physiology 2011;27(4):465-470
OBJECTIVETo investigate the differentially expressed genes in rat in the process of regression of vascular calcification by using the suppression subtractive hybridization (SSH).
METHODS24 SD male rats which aged 6 weeks and specific pathogen free grade were selected and randomly divided into 3 groups (n = 8): control group, calcification group and regression group respectively. Vascular calcification model (vitamin D3 plus nicotine, VDN) were made from rats in calcification group and regression group, and rats in control group were intragastric administered with normal saline and lavaged with peanut oil. Rats were bred for 8 weeks in calcification group and control group, while rats in regression group were fed for 16 weeks. All rats were killed to measure concentration of calcium in the arterial tissue and examine the pathological lesion changes. Subtractive hybridization among vascular cDNA sequences from calcification group and regression group were established. The cDNA fragments which expressed higher or lower in regression group than those in calcification group were isolated. Differentially expressed genes with cDNA fragment were inserted into PMD18-T plasmid vector and transformed competent DH-5alpha, cDNA libraries of differentially expressed gene between calcification group and regression group were then constructed. Recombinant vectors were analyzed by colony PCR, positive genes were randomly selected for sequencing and analyzed by BLAST. 4 genes were randomly selected for RT-PCR certification combined with semi-quantitative analysis of DNA bands.
RESULTSVDN model of rats were successfully constructed. Concentration of tissue calcium in calcification group (15.34 mg/g +/- 2.51 mg/g) was significantly increased compared to that in control group (5.20 mg/g +/- 0.75 mg/g, P < 0.001), while in comparison with calcification group (15.34 mg/g +/- 2.51 mg/g), calcium in regression group was relatively lower (12.73 mg/g +/- 1.89 mg/g, P < 0.05). 28 up-regulated genes and 22 down-regulated genes were gained through sequencing and BLAST analysis among positive clones. RT-PCR validation indicated that 4 genes such as prdx3 and Ank2 had increasedly expressed in regression group than those in calcification group, the average fold change was 1.7.
CONCLUSIONRat vascular calcification tissue had characteristic of active regression. Genes in relation to pyrophosphoric acid synthesis, glutamate signal peptides, anti-oxidant and ant-apoptosis were up-regulated, at the same time many genes related to ossification and oxidation activity were down-regulated in the process of calcification regression. Increased expression of calcification suppressor genes accompanying decreased expression of calcification promoting genes might be the intrinsic mechanisms which initiated the active regression of calcified tissues.
Animals ; Aorta ; metabolism ; pathology ; Gene Expression Profiling ; Gene Expression Regulation ; Male ; Rats ; Rats, Sprague-Dawley ; Vascular Calcification ; genetics ; physiopathology
10.Comfortable lifestyle-induced imbalance of neuro-endocrine-immunity network: a possible mechanism of vascular endothelial dysfunction.
Guo-qiang YUAN ; Zhen-hua JIA ; Hai-tao YANG ; Shi-zhen WU ; Huai-lin GAO ; Cong WEI ; Hui-ming ZHU ; Yi-ling WU
Chinese journal of integrative medicine 2010;16(1):54-60
OBJECTIVETo observe the changes of vascular endothelial functions and general neuro-endocrine-immunity (NEI) network under the state of qi-deficiency syndrome induced by excessive idleness and to approach their internal relevance and illuminate initially the pathophysiological mechanism of vascular lesion induced by excessive idleness.
METHODSA total of 100 male Wistar rats were randomly divided into the control group and the qi-deficiency syndrome model group, 50 rats in each group. The qi-deficiency syndrome model was established by feeding the animals with hyper-alimentation diet in combination with restricting movement for 10 weeks. Changes of common chemical signal molecules related to NEI and vascular endothelial functions were measured by the end of the experiment. Furthermore, their internal relevance was analyzed by the method of canonical correlation analysis.
RESULTSThe vascular endothelial structure and function were obviously injured in the model group. Compared with the control group, the chemical signal molecules, such as 5-hydroxytryptamine (5-HT), corticosterone (CORT), triiodothyronine (T3), tetraiodothyronine (T4), angiotensin II (Ang II), interleukin-1 (IL-1), and tumor necrosis factor-alpha (TNF-alpha) in peripheral blood of the model group (n=43) were changed significantly (P<0.05 or P<0.01). Canonical correlation analysis showed that vascular endothelial dysfunction was correlated to the changes of these signal molecules in the NEI network.
CONCLUSIONSComfort-based lifestyle induced not only vascular endothelial dysfunction but also an imbalance of the NEI network. Vascular endothelial dysfunction and the imbalanced NEI network interacted with each other, and an imbalance of the NEI network may be the pathophysiologic basis for the genesis and development of vascular endothelial dysfunction, even diseases of the blood vessel.
Animals ; Aorta ; metabolism ; pathology ; physiopathology ; ultrastructure ; Biomarkers ; analysis ; metabolism ; Cardiovascular Diseases ; etiology ; metabolism ; pathology ; physiopathology ; Disease Models, Animal ; Endothelins ; metabolism ; Endothelium, Vascular ; metabolism ; pathology ; physiopathology ; Immune System ; metabolism ; pathology ; physiology ; Male ; Neuroimmunomodulation ; physiology ; Neurosecretory Systems ; metabolism ; pathology ; physiology ; Nitric Oxide ; metabolism ; Qi ; Rats ; Rats, Wistar ; Sedentary Lifestyle ; Syndrome ; Yin Deficiency ; etiology ; metabolism ; pathology ; physiopathology