Effects of grape seed procyanidin on vascular remodeling in renovascular hypertensive rats

AIM: To study the effect of grape seed procyanidin (GSP) on vascular remodeling in renovascular hypertensive (RH) rats. METHODS: The RH rat model was established by two-kidney one-clip method. Two weeks after operation, 28 rats were selected according to the increased tail systolic pressure above 130 mmHg and randomly divided into 4 groups (n=7): RH model group, low GSP treatment group (50 mg·kg^-1·d^-1),high GSP treatment group (200 mg·kg^-1·d^-1) and captopril treatment group (30 mg·kg^-1·d^-1). Meanwhile, 7 rats with sham operation served as controls. Tail systolic pressure, medial thickness (MT), luminal diameter (LD), and the ratio of MT to LD in thoracic aortic wall were determined 6 weeks after treatment. Masson staining and ELISA were used to detect the content of collagen and angiotensin II (Ang II) in aortic tissues. The protein expression of tumor necrosis factor α (TNF-α) in abdominal aortic tissues was determined by Western blotting. RESULTS: Compared with control group, the tail systolic pressure, MT, MT/LD, the content of collagen and the protein expression of TNF-α in aorta were significantly increased in RH model group, but LD decreased. Treatment with GSP and captopril reduced the raised parameters, and increased the LD in RH model rats. These effects were more notable in high GSP treatment group, and equal to captopril treatment group. CONCLUSION: GSP treatment significantly decreases tail systolic pressure in RH rats, and effectively attenuates arterial vascular remodeling by decreasing the content of AngII and reducing the protein expression of TNF-α in aorta.     

Effects of captopril on expression of calcineurin and NF-κB p65 in the heart of hypertensive rats

AIM: To study the effect of captopril on calcineurin and NF-κB p65 in the signal transduction pathway of the cardiovascular remodeling in hypertensive rats. METHODS: Using a animal model of hypertension induced by abdominal aortic banding, the rats were treated with captopril for 10 weeks. The blood pressure was observed with a tail cuff method. The heart weight and heart weight/body weight were measured. The expression of calcineurin and NF-κB p65 were studied by using immunohistochemistry. RESULTS: After treated with captopril, the blood pressure of the model rats was decreased (P<0.01), the heart weight or heart weight/body weight were also decreased (P<0.01). The calcineurin and NF-κB p65 protein overexpression was down-regulated, NF-κB-positive area and area percentage were reduced in the heart of hypertensive rats (P<0.01，P<0.01). CONCLUSION: Captopril reverses the cardiovascular remodeling by affecting the overexpression of calcineurin and NF-κB p65 involved in the cardiovascular remodeling in hypertensive rats.     

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.     

Role of vascular remodeling for urotensin Ⅱ in rat thoracic aorta after balloon injury

AIM: To explore the possible effect of UII in the process of remodeling after vascular injury. METHODS: The rat model of balloon injury in thoracic aorta was established. Male rats were randomized to 4 groups (n=5), including sham injury group, injury group, UII group (UII pumped into the rats after thoracic aorta balloon injury at 1.0 nmol·kg-1·h-1) and urantide group (urantide pumped into the rats after thoracic aorta balloon injury at 10 nmol·kg-1·h-1). At 21 days, the thoracic aortas were taken out to measure the changes of pathology, the expression of UII, the proliferation of VSMC and the expression of collagen. RESULTS: (1) At the 21 days after operations, the systolic blood pressure was higher in UII group than that in injury group ［(140.0±10.0) mmHg vs (132.0±3.4) mmHg, P>0.05］. The systolic blood pressure was also obviously higher than that in urantide group ［(140±10.0) mmHg vs (128.0±2.4) mmHg, P<0.05］. (2) Urotensin Ⅱ was expressed strongly in the injured area after thoracic aorta injury. (3) In contrast to injury group, the intimal thicken in urotensin Ⅱ group enhanced, the decrease in lumen area was marked (0.13±0.05 vs 0.07±0.02, P<0.05), the cell proliferation index was markedly increased (0.74±0.16 vs 0.40±0.11, P<0.01), and the expression of collagen was also markedly increased (counted as IOD, 318±127 vs 78±26, P<0.01). (4) In contrast to injury group, the decrease in lumen area was not abolished (0.09±0.03 vs 0.07±0.02, P>0.05) after chronic infusion of urotensin Ⅱ receptor antagonist urantide, the cell proliferation index was markedly increased (0.73±0.15 vs 0.40±0.11, P<0.01) and the expression of collagen was not statistically increased (counted as IOD, 200±79 vs 78±26, P>0.05). CONCLUSION: Urotensin Ⅱ expresses strongly in the myointimal cells after thoracic aorta injury in rat. The extra UII enhances the proliferation of VSMC and expression of collagen in the myointimal, increases the stenosis of injured vasculature, indicating that UII might take part in the process of repairing after vessel injury.     

Comparison of vascular remodeling between small artery and aorta in spontaneous hypertensive rats

AIM: To examine the difference of vascular remodeling between aorta and small artery in sponta-neous hypertensive rats (SHR) and control rats.METHODS: Male SHR (20-week-old) were used as experiment group, and age matched male Wistar-Kyoto (WKY) rats were used as control group. The systolic blood pressure and body weight were measured once a week. At 43 weeks old, the rats were anaesthetized, blood samples were collected, and thoracic aorta and mesenteric small artery tissue were harvested. The morphological changes of the arterial tissue were observed with HE staining. The collagen and elastine fibers were detected by the Sirius red-Victoria blue staining. The protein expression of type I and Ⅲ collagens were analyzed by confocal laser-scanning microscopy and Western blot. The changes of the vascular ultrastructure were imaged by transmission electron microscopy. The expression of proliferating cell nuclear antigen (PCNA) and the cell apoptosis in the arterial wall were examined by immunohistochemical method and TdT-mediated dUTP nick and labeling (TUNEL) detection.RESULTS: The inner diameter (ID) and luminal cross-sectional area (LCSA) of mesenteric small artery were decreased, whereas ratio of wall thickness (WT) to ID (WT/ID) and ratio of wall cross-sectional area (WCSA) to LCSA (WCSA/LCSA) were increased. Meanwhile, adventitia fibroblast migrated to the media, with overload collagens, especially collagen Ⅲ. Proliferation index (PI) and apoptotic index (AI) of the mesenteric small artery wall cells were increased. The ID, LCSA, WT/ID and WCSA/LCSA of the aorta were increased. Moreover, the vascular smooth muscle cells (VSMCs) showed hypertrophy and hyperplasia, with overload collagens. The PI and AI of the aortic wall cells were increased.CONCLUSION: The difference of vascular remodeling between the aorta and small artery is significant. The small artery mainly appears hyperplasia of matrix, especially the adventitial collagen Ⅲ. Meanwhile, the cell apoptosis in the small artery wall is increased. The aorta mainly appears hyperplasia and hypertrophy of media VSMCs.     

Effect of rhynchophylline on TGF-β1/Smad pathway for processing ventricular remodeling in spontaneously hypertensive rats

AIM: To investigate the effect of rhynchophylline (Rhy) on blood pressure, cardiac hypertrophy and myocardial fibrosis in spontaneously hypertensive rats (SHR). METHODS: Spontaneously hypertensive rats were randomly divided into model group, high dose (10 mg·kg^-1·d^-1) and low dose (2.5 mg·kg^-1·d^-1) group of rhynchophylline, captopril group (17.5 mg·kg^-1·d^-1). Wistar-Kyoto rats were used as normal control. Respectively, systolic blood pressure was measured by tail cuff every 2 weeks. After 10 weeks, heart weight index and left ventricular weight index were calculated. The myocardial hydroxyproline and plasma angiotensin Ⅱ were detected. Moreover, basic myocardial histopathological changes and myocardial collagen fibres were observed by HE staining and Masson staining, respectively. The protein expression of TGF-β₁ and Smad3 in the myocardium was measured by the methods of immunohistochemistry and Western blot. RESULTS: Compared with SHR model group, Rhy significantly reduced blood pressure (P<0.05), the levels of HYP in the myocardium (P<0.05) and the levels of AngⅡ in the plasma (P<0.01). The pathological damages of the myocardial tissues and collagen deposition were attenuated. The protein expression of TGF-β₁ and Smad3 was significantly reduced by the treatment with Rhy (P<0.01). CONCLUSION: Rhynchophylline reduces blood pressure and adjusts to improve ventricular remodeling of SHR. The mechanism may be involved in the TGF-β₁/Smad pathway and reducing AngⅡ content.     

Effects of sesamin on blood fat,blood glucose and vascular remodeling in rats fed with high-fat and refined-sugar diet

AIM: To explore the effect of sesamin on blood fat, blood glucose and vascular remodeling in rats fed with high-fat, refined-sugar diet. METHODS: A high-fat, refined-sugar diet was given to rats for 24 weeks. Sesamin (120, 60, 30 mg·kg-1·d-1) was given by intragastric administration to the rats at 9th week, which lasted for 16 weeks. After 24 weeks, blood glucose, blood fat, blood pressure, activity of total anti-oxidation capacity (T-AOC) and concentration of hydrogen peroxide in serum and aorta were determined. Changes of histology and collagen fibers were observed in aorta by HE and Masson staining, respectively. Immunohistochemical method was used to examine iNOS protein expression in aorta. In mesenteric arteries, media thickness (M), luminal radius (L) and ratio of media to lumen (M/L) were measured. RESULTS: Compared to model group, sesamin (120, 60 mg·kg^-1·d^-1) obviously decreased the levels of blood glucose, blood fat, blood pressure and concentration of hydrogen peroxide in serum and aorta. Sesamin also markedly enhanced the activity of T-AOC in serum and aorta and reduced collagen deposition and iNOS protein expression in the vascular wall. In addition, proliferation of intima and vascular smooth muscle cells were improved. In mesenteric arteries, sesamin lessened M and M/L and increased L of mesenteric arteries. CONCLUSION: Sesamin ameliorates disorders of glucose and lipid metabolism and inhibits vascular remodeling in rats caused by chronic high-fat, refined-sugar diet.     

Effect of heme oxygenase on vascular remodeling in renal hypertension

AIM: To investigate the effect of heme oxygenase on vascular remodeling in renal hypertension. METHODS: Male Wistar rats were randomly divided into sham-operated, 2K1C （two-kidney one-clip） and hemin-induced groups. Four weeks after the treatments, the thickness of aortic media and HO enzymatic activity of the aorta were determined. Immunohistochemical staining was carried out to detect protein of HO-1 in the aorta. RESULTS: The blood pressure in 2K1C renal hypertension rats started to increase two weeks after the surgery and stabled at a high level at the 4th week. Hemin, an inducer of HO-1, markedly inhibited the increase in blood pressure. Aortic medium thickness of the 2K1C rats at 4th week was 27.5% thicker than that in the sham-operated rats. The thickness of aortic medium of the hemin-induced rats was 16.1% less than that in 2K1C group. At the 4th week after operation, protein level and enzymatic activity of HO-1 in aorta were higher than that in 2K1C group compared to those in the sham-operated group. CONCLUSION: Renal hypertension caused vascular remodeling and the activation of HO-1. HO-1 induction decreased the blood pressure of renal hypertension and reduced vascular remodeling.     

Altered activity and expression of MMPs/TIMPs are associated with functional and structural left ventricular remodeling in hypertensive rats

AIM: To investigate the relationship between matrix metalloproteinases and tissue inhibitors of matrix metalloproteases imbalance with functional and structural left ventricular (LV) remodeling in the hypertensive rats.METHODS: 6-week-old male stroke-prone spontaneously hypertensive rats (SHR-SPs,n=40) served as the hypertensive heart disease model,and age-matched male Wistar-Kyoto (WKY) rats (n=10) were used as control.After 6 months,the rats in two groups were anesthetized for invasive hemodynamic measurement by Millar pressure-volume (P-V) conductance catheter.Then the rats were sacrificed and hearts were dissected for morphological analysis,gelatin-zymography and Western blotting analysis.RESULTS: Left ventricular (LV) hemodynamic parameters showed the systolic and diastolic dysfunction in SHR-SPs compared with that in control group (P<0.05).Collagen volume fraction,ratio of perivascular collagen area to luminal area,myocardial cross-sectional area and the medial area to luminal area ratio of the SHR-SPs were all increased remarkably (P<0.05).LV matrix metalloproteinase-2 (MMP-2) activities,MMP-2 and tissue inhibitors of matrix metalloprotease-1 (TIMP-1) protein level in SHR-SP were notably higher than those in control group (P<0.05).CONCLUSION: Chronic pressure-overload is capable of inducing imbalances of cardiac ECM and MMPs/TIMPs system,both imbalances induce LV dilation,cardiac systolic and diastolic dysfunction.     

Zacopride attenuates pressure overload-induced ventricular remodeling in rats

AIM:To investigate the protective effect of zacopride (ZAC) on the pressure-overload left ventricular remodeling in the rats induced by coarctation of abdominal aorta. METHODS:Male Sprague-Dawley (SD) rats with pressure overload were induced by the coarctation of abdominal aorta. The model rats were intraperitoneally administered with ZAC, chloroquine (Chlor), and zacopride+chlorquine (ZAC+Chlor). The study duration was 8 weeks. The cardiac structure and function were assessed by echocardiography. The heart weight/body weight (HW/BW) ratio and the left ventricular weight/body weight (LVW/BW) ratio were calculated. The changes of structure and shape in myocardial tissue were observed with HE staining. The ultrastructure of the myocytes was observed under transmission electron microscope. The inward rectifier potassium channel (I_K1) protein expression was determined by Western blot. The mRNA expression of Kir2.1 was detected by RT-PCR. RESULTS:Compared with vehicle group, ZAC improved cardiac function, as indicated by the decreased left ventricular end-diastolic dimension (LVEDD) and left ventricular end systolic dimension (LVESD) (P<0.05), and the increased left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) (P<0.01). The HW/BW and LVW/BW ratios were significantly decreased, and the cross-sectional area of the cardiomyocytes was significantly less in ZAC group than that in vehicle group (P<0.01). The ultrastructure of the myocytes was significantly improved. Chlor blocked the protective effect of zacopride on the pressure-overload left ventricular remodeling. The protein level of I_K1 and mRNA expression of Kir2.1 in the cardiac tissues in ZAC group were significantly increased compared with vehicle group (P<0.01). CONCLUSION:I_K1 agonist ZAC significantly attenuates pressure overload-induced ventricular remodeling in rats.     

Role of TGF-β1/Smads and ERK expression in vascular remodeling induced by high-salt diet in Wistar rats

AIM: To investigate the role of transforming growth factor β₁ (TGF-β₁)/Smads and extracellular signal-regulated kinase(ERK) expression in vascular remodeling induced by high-salt diet in Wistar rats. METHODS: Wistar rats were randomly divided into 3 groups: normal control group (n=13), high salt (8%) model group and high salt+telmisartan group (n=13). Tail-cuff arterial pressure was determined every 2 weeks. After 24 weeks, the rats in high salt model group were divided into model animals with hypertension group (MH, n=12) and model animals without hypertension group (MN, n=12). The remodeling of aorta and mesenteric artery was observed by HE and Masson staining. In addition, the techniques of immunohistochemistry and real-time PCR were applied to detect the expression of proliferating cell nuclear antigen (PCNA), TGF-β₁, p-Smad2/3, p-ERK1/2 and Smad7 at both protein and mRNA levels. RESULTS: Compared with normal control group, blood pressure in MH group was much higher, and media thickness (MT) and collagen volume fraction (CVF) of arteries in MH and MN groups were higher.The mRNA expression of TGF-β₁, Smad2 and Smad7 in the aorta was significantly increased, and the protein levels of PCNA, p-ERK1/2, TGF-β₁ and p-Smad2/3 in the aorta and mesenteric artery media were elevated, but Smad7 decreased. After telmisartan treatment, MT and CVF were much lower,and the protein levels of PCNA, TGF-β₁, p-Smad2/3 and p-ERK1/2 were significantly reduced, whereas Smad7 was increased. CONCLUSION: The abnormal expression of TGF-β₁/Smads and ERK may be involved in the mechanism of remodeling of aorta and mesenteric artery induced by high-salt diet. Telmisartan prevents the vascular remodeling via regulating TGF-β₁/Smads and ERK signal pathways mediated by angiotensinⅡ type 1 (AT₁) receptor, at least in part.     

Intermedin inhibits pulmonary collagen synthesis in rats with pulmonary hypertension induced by high pulmonary blood flow

AIM: To explore the regulatory effect of intermedin (IMD) on pulmonary collagen synthesis and accumulation in rats with pulmonary hypertension induced by high pulmonary blood flow.METHODS: Healthy male SD rats (n=20) were randomly divided into control group (n=7), shunt group (n=7) and shunt with IMD group (n=6). The shunting of abdominal aorta and inferior vena cava was produced in rats of shunt group and shunt with IMD group. After 8 weeks, IMD was administered into the rats of shunt with IMD group subcutaneously by mini-osmotic pump for 2 weeks. Mean pulmonary artery pressure (mPAP), relative medial thickness (RMT) of pulmonary arteries, contents of hydroxyproline, collagen type I and III, bone morphogenetic protein-2 (BMP-2), and the mRNA expression of procollagen I and III in lung tissues were measured and compared. RESULTS: Compared with control group, mPAP and RMT of medium and small pulmonary arteries in the rats of shunt group were significantly increased. Meanwhile, the lung hydroxyproline, collagens I and III and BMP-2 contents, and the mRNA expression of lung procollagen I and III were all significantly increased compared with control group. However, IMD significantly decreased mPAP, alleviated the changes of pulmonary vascular micro-structure, decreased the collagen accumulation and pulmonary tissue homogenate BMP-2 contents, and inhibited the mRNA expression of procollagen I and III in the lung tissue of shunting rats.CONCLUSION: IMD plays a protective role in the development of pulmonary hypertension and pulmonary vascular structural remodeling induced by high blood flow by inhibiting pulmonary collagen synthesis and accumulation, possibly in association with the BMP-2 pathway.     

Role of carvedilol on energy metabolic reversion and remodeling of pressure overload-induced left ventricular hypertrophy in rats

AIM: To study changes of medium chain acyl-CoA dehydrogenase (MCAD), muscle carnitine palmitoyltransferase I (M-CPT-I) and colligin mRNA/protein expression, to elucidate molecular mechanism of the recapitulation of fetal energy metabolism and ventricular remodeling and the effects of carvedilol during the development of pressure overload-induced left ventricular hypertrophy in rats. METHODS: Male Wistar rats of hypertrophy induced by constriction of abdominal aorta （CAA） were randomized into 2 groups (n=12, each group): 4-week group (CAA4 weeks group) and 12-week carvedilol intervention group (CAR group). Additional rats (n=12) underwent abdominal cavity incision without ligation to serve as age-matched sham operated controls (SH). Hemodynamics, ventricular remodeling parameters and free fatty acid （FFA） both in blood serum and myocardium were measured. RT-PCR analysis of the expression of mRNA of M-CPT-I, MCAD and collagen binding protein (colligin) were investigated. The protein expression of colligin was analyzed by Western blotting in the experimental animals and sham operation.RESULTS: LVM/BW and MAP in CAA group were increased more significantly than in sham group. There were progressive increases in FAA both in blood serum and myocardium in CAA group than in sham group, accompanied with downregulation of gene expressions of M-CPT-I and MCAD and colligin mRNA/ protein upregulation in LV in CAA group, while changes of all of these parameters in CAR group were attenuated.CONCLUSIONS: (1) The down-regulated expression of cardiac FAO enzyme genes (M-CPT-I and MCAD) in the hypertrophied heart may be responsible for “the recapitulation of fetal energy metabolism” during the development of pressure overload-induced left ventricular hypertrophy in rats. (2) Carvedilol attenuates the reversion of the metabolic gene expression back towards fetal type. (3) Carvedilol is effective in regressing the left ventricular remodeling by inhibiting colligin protein expression. A molecular mechanism by which carvedilol may confer cardioprotective effects in heart failure may be, in part, via preserving of the adult metabolic gene regulation and regressing left ventricular remodeling.     

Role of estrogen on expression of TNF-α and MMP-9 in intracranial arterial vascular wall of spontaneous hypertensive rats

AIM: To study the effects of estrogen on the inflammatory response and vascular remodeling of intracranial artery in rats.METHODS: Thirty-two female spontaneous hypertensive rats (SHR) were randomized into 4 groups: spontaneous hypertensive group(sham-operated), ovariectomized group, ovariectomized+17 beta-estradial group and ovariectomized+vehicle group (8 rats in each group).On day 14, estradiol was detected by radioimmunoassay.The pathological changes were observed under light microscope.The protein expression of tumor necrosis factor alpha (TNF-α) and matrix metalloproteinase-9 (MMP-9) in vascular wall of Willis circle was detected by Western blotting.RESULTS: The estrogen level was lower in ovariectomized group than that in sham-operated group (P<0.01).The estrogen level was higher in ovariectomized rats treated with 17 beta-estradial than that in ovariectomized rats treated with vehicle (P<0.01).Advanced aneurysm was not found in all groups.Early aneurysmal change was not found in sham-operated group.Early aneurysmal changes in some rats were observed in ovariectomized group (2 rats), ovariectomized+vehicle group (3 rats) and ovariectomized+17 beta-estradial group (1 rat).The protein levels of TNF-α and MMP-9 in the vascular wall of Willis circle in sham-operated group were lower than that in ovariectomized group (P<0.01).Additionally, the protein levels of TNF-α and MMP-9 in the vascular wall of Willis circle of ovariectomized rats treated with 17 beta-estradial were lower than those of ovariectomized rats treated with the vehicle (P<0.01).CONCLUSION: Estrogen can influence the vascular remodeling of intracranial artery by inhibiting the inflammatory response and degradating MMP-9 in the vascular wall.     

Effect of angiotensin Ⅱ on the remodeling of aorta in spontaneously hypertensive rats

AIM: To investigate the effects of a 10-weeks treatment with angiotensin Ⅱ (Ang Ⅱ) subtype I receptor antagonist losartan on vascular remodeling of thoracic aorta in male spontaneously hypertensive rats (SHR). METHODS: SHR were treated from 16 to 26 weeks of age with losartan at 15 mg/kg·d^-1 or 0.75 mg/kg·d^-1. RESULTS: Losartan (15 mg/kg·d^-1) treatment significantly decreased systolic blood pressure compared with the control group, while losartan (0.75 mg/kg·d^-1) had no the effect, losartan(15 mg) prevents the development of aortic hypertrophy by preventing hypertrophy of vascular smooth muscle cells (VSMC). In the losartan 0.75 group, these parameters were not changed. But in the losartan 15 and losartan 0.75 groups, the collagen content of the aortic media decreased significantly. CONCLUSION: It is inferred that the effect of Ang Ⅱ on stimulating VSMC growth of the aorta in SHR is dependent on arterial pressure, while the effect on collagen fibers is through pressure independent mechanism.     

Effects of phytosterol ester on aortic aging and expression of related genes in rats

AIM: To explore the protective effect of phytosterol ester (PSE) on aortic aging in rats. METHODS: The female SD rats (12 months old, n=42) were randomly divided into control group, model group and PSE group. During the experiment, the rats in control group, model group and PSE group were treated with basic feed, high-fat diet (HFD) and HFD with 2% PSE (W/W) for 6 months, respectively. The morphological changes of the aorta were observed by HE staining and Masson staining, and the absolute area of smooth muscle cells and collagen fiber in the vascular wall were measured by image analysis. The levels of advanced glycosylation end products (AGEs), malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) in the plasma were detected. The expression of silent information regulator 1 (SIRT1) and peroxisome proliferator-activated receptor γ (PPARγ) at mRNA and protein levels in the vascular tissue was determined by real time PCR and Western blot, respectively. RESULTS: PSE significantly lowered plasma TC and LDL-C, and increased plasma HDL-C level (P<0.05), but had no effect on plasma TG level. PSE significantly attenuated the thickening of intima and media of aging aortic, and decreased the migration of vascular smooth muscle cells (VSMC) and the amount of VSMC and collagen fiber in the aorta (P<0.05). PSE significantly reduced the contents of AGEs and MDA (P<0.05), but had no effect on the activity of SOD and CAT in the plasma. PSE also down-regulated the expression of PPARγ and up-regulated the expression of SIRT1 (P<0.05). CONCLUSION: PSE is able to attenuate the senescence process in the aorta by reducing the production of reactive oxygen species in plasma, and activating SIRT1, or inhibiting the expression of PPARγ in vascular tissues.     

Effects of Xinkang recipe on myocardial miR-25-3p expression and SERCA2a activity in heart failure rats

AIM: To investigate the effects of Xinkang recipe on myocardial miR-25-3p expression and sarcoplasmic reticulum calcium ATPase 2a (SERCA2a) activity in heart failure rats. METHODS: Male SD rats were randomly divided into normal group, sham group, model group, Xinkang recipe group (Xinkang group), and captopril group. The heart failure rat model was induced by intraperitoneal injection of doxorubicin. Distilled water, Xinkang recipe and captopril were administrated by gastric gavage for 35 d, respectively. The indexes of cardiac function and plasma level of brain natriuretic peptide (BNP) were measured. The SERCA2a activity was determined by the inorganic phosphorus method. The myocardial protein expression of SERCA2a and phospholamban (PLB) was detected by Western blot. The myocardial expression of miR-25-3p was detected by stem-loop RT-qPCR. RESULTS: Cardiac output (CO), left ventricular fractional shortening (LVFS) and left ventricular ejection fraction (LVEF) in Xinkang group and captopril group were significantly higher while the plasma levels of BNP were significantly lower than those in model group (P<0.01). The myocardial expression levels of miR-25-3p in Xinkang group and captopril group were significantly lower while the myocardial protein le-vels of SERCA2a and PLB were significantly higher than those in model group (P<0.01). The SERCA2a/PLB ratio and SERCA2a activity in Xinkang group were significantly higher than those in model group (P<0.05), and no significant change was observed between captopril group and model group. CONCLUSION: Xinkang recipe therapy may improve cardiac function in heart failure rats, which may be related to inhibiting the expression of miR-25-3p, increasing the SERCA2a/PLB ratio and enhancing SERCA2a activity in the myocardium.     

Expression of ERK and MKP-1 in vascular smooth muscle of spontaneously hypertensive rats with different ages

AIM: To investigate the expression of mitogen- activated protein kinase and mitogen-activated protein kinase phosphatase-1 in thoracic aorta smooth muscles of spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY) with different ages and the relationship between those and hypertension. METHODS: The caudal arterial pressure was measured by tail-cuff. Protein expression of p-ERK was detected by Western blotting, and MKP-1 mRNA in thoracic aorta smooth muscle was examined by RT-PCR. RESULTS: (1) The blood pressure of SHR was obviously higher than that of age-matched WKY (P<0.01), elevated with age (P<0.05) and became stable from 14-week-old. (2) The expression of p-ERK and MKP-1 in SHR was higher than that in WKY in 5-week-old rats, and the expression of p-ERK increased with age, while the expression of MKP-1 decreased with age (P<0.05). CONCLUSION: MKP-1 may play an important role in the development of hypertension in SHR. The decrease in the expression of MKP-1 that resulted in the activation of MAPK may induce vascular smooth muscle proliferation and hypertrophy.     

Anti-aging effect of stromal vascular fractions in photoaging skin

AIM: To investigate the anti-aging effect of the stromal vascular fractions (SVF) injection in photo-aging skin of nude rats. METHODS: Six-week-old nude rats were divided into control group and photoaging group. The nude rats in photoaging group were continuously irradiated with ultraviolet B (UVB) for 8 weeks to build the photoaging skin model, and were randomly divided into placebo group, and low-dose (LD), middle-dose (MD) and high-dose (HD) SVF treatment groups. The human SVF was obtained by trypsin digestion from the adipose tissue in healthy females, and was subcutaneously injected into the back skin of photoaging nude rats in LD, MD and HD SVF treatment groups (10⁴,10⁵ and 10⁶ cells in 100 μL suspension, respectively). The animals in photoaging skin model group were treated with placebo. Control group did not have any intervention. The changes in the thickness of epidermis and histology after 7 d and the thickness of the dermis and histology after 28 d were examined. RESULTS: The thickness of the epidermis was thinner in MD and HD SVF treatment groups than that in photoaging skin model group after 7 d (P<0.05). The percentage of stratum corneum and the abnormal proliferation of the epidermal basal cell layer were reduced (P<0.05). After 28 d, MD and HD SVF treatment groups showed that the content of collagen I was higher than that in photoaging skin model group (P<0.05), and the contents of collagen III and MMP3 were decreased (P<0.05). The dermis in HD group was thicker than that in photoaging skin model group (P<0.05).CONCLUSION: SVF may have anti-aging potential in photoaging skin and also has clinical application value.     

Effects of puerarin on apelin-12, angiotensin II,NO and blood pressure in renovascular hypertensive rat

AIM: To investigate the effects of puerarin on blood pressure in renovascular hypertensive rats, and to measure puerarin-induced changes of apelin-12, angiotensin II (Ang II) and nitric oxide(NO), the factors related to development of hypertension. METHODS: Sixty-five male Sprague-Dawley rats were used, of which 8 rats were randomly selected as sham operation group, and the remaining were used to make two-kidney, one-clip model. The rats that met the criterion for Goldblatt hypertensive rat model were randomly allocated into 5 groups: high-, middle- and low-dose puerarin groups, captopril group, and model group. The drugs were administered for 6 weeks. Blood pressure was measured every 2 weeks. Six weeks after treatment, all rats were sacrificed under deep anesthesia. Blood and kidney samples were collected. The level of apelin-12 in serum and kidneys was detected by ELISA. The level of Ang II in plasma and kidneys was measured by radioimmunoassay. NO level in serum was examined by nitrate reductase assay. RESULTS: Puerarin had an antihypertensive effect in a dose-dependent manner. Marked decreases in the level of serum apelin-12 in high- and middle-dose puerarin groups were observed(P<0.01). Puerarin at low dose did not cause obvious change in the content of apelin but still reached significant level (P<0.05). As the dose of puerarin went up, the level of apelin-12 in the kidneys was gradually decreased. Puerarin at high and middle doses obviously reduced the level of AngII in plasma, while purarin at low dose did not produce any significant effects. Puerarin at high and middle doses markedly increased the level of NO in serum, but puerarin at low dose did not induce any significant changes. CONCLUSION: Puerarin has an antihypertensive effect, and its mechanism may be related to inducing the changes of apelin, Ang II and NO, and regulating the balance among those factors.