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 angiotensin II receptor antagonist on remodeling of renal arterioles in hypertension

AIM: To investigate the effects of angiotensin II receptor antagonist on remodeling of renal arterioles in hypertension. METHODS: Eighteen 4 weeks old male rats were divided into three groups: Wistar-Kyoto rats (WKY) for normotensive group, and spontaneously hypertensive rats (SHR) for hypertensive group, and SHR treated with losartan orally (15 mg·kg^-1·d^-1). The rats were raised to 16 weeks old. The morphometric parameters of the renal arterioles, and the widths of vascular smooth muscle cells (VSMC) and intercellular space were studied on kidney slices by light microscope and electromicroscope respectively, combined with computer-assistant image analysis system. The minimal renal vascular resistance (RVR_min) was studied by isolated kidney perfusion system. RESULTS: The systolic blood pressure of the tail artery, wall thickness, wall area, ratio of wall thickness to inner diameter, width of VSMC of renal arterioles and RVR_min were all smaller or lower in losartan group than those of SHR.     

Cellular mechanisms of losartan on reversing left ventricular hypertrophy in spontaneously hypertensive rats

AIM:To investigate the role of proliferation and apoptosis in hypertensive left ventricular hypertrophy (LVH) and the effect of AT₁ blockade with losartan. METHODS:Left ventricles (LV) from 12, 24-week-old SHR (SHR₁₂, SHR₂₄), 24-week-old SHR treated with losartan (15 mg·kg^-1·d^-1, SHR-L₂₄) during 12 weeks, and age-matched WKY rats (WKY₁₂, WKY₂₄) were studied. Expression of PCNA was examined by immunohistochemistry. Apoptotic cells in LV sections were assessed by TUNEL method. Levels of fas mRNA were quantitated by RT-PCR. RESULTS: Compared with age-matched WKY, SHR₁₂ and SHR₂₄ showed increased LV hypertrophied index (HI), increased apoptotic index (AI) of myocytes (P＜0.01), but decreased AI of fibroblasts (P＜0.05). Moreover, SHR₁₂ exhibited increased PCNA labeling of myocytes (P＜0.05) with similar positive rates of fibroblasts.It was also showed that losartan reversed HI, significantly reduced the AI of myocytes and significantly increased the AI of fibroblasts. RT-PCR disclosed that levels of fas mRNA positively correlated with the frequency of apoptosis in LV of either SHR or WKY (r=0.52, P＜0.05). CONCLUSION:The cellular changes of LVH in adult SHR manifest as the imbalance between proliferation and apoptosis of myocytes, and insufficient apoptosis of fibroblasts. The mechanisms of losartan on reversing LVH may be mediated through adjusting the abnormal amount of cells and the expression of fas gene.     

Therapeutic effects of losartan and astragalus membranaceus on diabetic nephropathy

AIM: To observe the protective effects of losartan and astragalus membranace on the kidney of diabetic rats, and to study their possible mechanisms. METHODS: The diabetic rats were induced by a single intraperitoneal injection of streptozotocin. At the end of 12th week,changes in urinary albumin excretion, urinary β₂-MG excretion, Ccr,NO,ET-1 levels in blood, urinary and renal tissue were observed. Serum and urinary TGF-β₁ concentration,average volume of glomeruler,average thickness of glomerular basement membrane were also measured. RESULTS: In the treated diabetic rats, urinary albumin excretion, urinary β₂-MG excretion, Ccr, urinary and renal tissue NO, urinary TGF-β₁, average volume of glomeruler, average thickness of glomerular basement membrane decreased obviously as compared with diabetic untreated rats. These effects were enhanced when losartan was combined with astragalus membranace. CONCLUSION: Losartan or astragalus membranace reversed the injury of renal structure and function in STZ-induced diabetic rats. The protective effects were enhanced when losartan was combined with astragalus membranace. The decrease in NO,ET,TGF-β₁ concentration in renal tissue may be one of mechanisms for this action.     

Effects of irbesartan on expression of integrin-linked kinase and podocyte injury in rats with early diabetic nephropathy

AIM: To determine the influence of irbesartan (Irb) on the podocyte injury and of integrin-linked kinase (ILK) expression in early experimental diabetic nephropathy (DN).METHODS: The hyperglycemic DN rat model was induced by intraperitoneal injection of streptozotocin (STZ,35 mg/kg) in spontaneously hypertensive rats (SHR).The diabetic SHR were randomly divided into 2 groups: DN SHR (DN,n=8) and DN SHR treated with irbesartan (50 mg/kg per day by gavage for 8 weeks,DN+Irb,n=9).Non-diabetic normotensive Wistar-Kyoto rats (WKY,the animals of normal blood pressure have the same genetic background with SHR) were used as control (control,n=11).The biochemical parameters and pathological changes were analyzed.Moreover,the influence of irbesartan on the expression of ILK and podocyte injury in SHR with DN was examined.RESULTS: Compared with control,DN SHR were associated with hyperglycaemia,hypertension,hyperlipidaemia,insulin resistance and albuminuria,which were similar to those of human T2DN.In addition,these DN SHR showed expansion of mesangial matrix,loss of podocyte and podocyte injury.Importantly,the expression of ILK in DN SHR was upregulated as compared to control.Treatment with irbesartan significantly decreased this overexpression of ILK,along with a decrease in albuminuria,reduction of blood pressure,prevention of podocyte injury,inhibition of mesangial matrix expansion and restoration of podocyte numbers.CONCLUSION: ILK is involved in the podocyte injury in DN.Irbesartan downregulates the expression of ILK and decreases podocyte injury.     

Changes and significance of ATPase and free radical in aged rats with brain ischemia-reperfusion

AIM: To study the mechanism of brain ischemia-reperfusion injury from ATPase activity and free radical metabolism in aged rats. METHODS: The young rats (5 months) and the aged rats (more than 20 months) were divided into young control group(YCG), young model group(YMG), aged control group(ACG) and aged model group(AMG). The ATPase and SOD activities and the contents of MDA, Ca^2＋, Na^＋ and K^＋ were measured in the rats with 30 min brain ischemia followed by 60 min reperfusion. RESULTS: The Ca^2＋content in the AMG was higher than that in the YMG and the ACG. The Na^＋-K^＋-ATPase activity in the ACG was lower than that in the YCG,was lower in the AMG than that in the YMG. The Ca^2＋-ATPase activities in the YCG was higher than that in the ACG, was lower in the AMG than that in the YMG and was higher than the ACG's. The serum and brain tissue SOD activities in the ACG was lower than that in the YCG, was lower in the AMG than YMG 's. The serum and brain tissue MDA/SOD ratio in the AMG was higher than that in the ACG.CONCLUSION:The brain tissue ischemia-reperfusion injury was related with calcium overload and free radical injury.The pathological changes were obvious and had some characteristics in the aged rats compared with the young rats because of the brain t issue aging changes in ATPase,calcium content and free radical metabolism in the aged rats.     

Effects of benazepril on cardiac function,free oxygen radicals,sarcoplasmic reticulum Ca2+-ATPase following cardiac ischemia-reperfusion in spontaneously hypertensive rats

AIM: To investigate the effects of angiotensin converting enzyme inhibitor (ACEI), benazepril (B), on cardiac function , free oxygen radicals, sarcoplasmic reticulum(SR) Ca²⁺-ATPase following ischemia-reperfusion in sportaneously hypertensive rats (SHRs). METHODS: Thirty 10-week-old female SHRs were randomly assigned into two groups: group SHR was control; The animal in group SHR+B was given with 10 mg/kg of benazepril per day. Another 15 Wistar rats with the same age and sex were normal control (group Wistar). After 12 weeks of pretreatment, all rats in each group were subjected to 30 min of left anterior descending coronary artery occlusion and 30 min of reperfusion. Hemodynamic parameters, left heart-to-body weight ratio (LVW/BW), myocardial malondialdehyde (MDA) concentration, superoxide dismutase (SOD) activity, and SR Ca²⁺-ATPase activity were measured. RESULTS: Compared to group Wistar, the rats in group SHR had higher blood pressure, LVW/BW and myocardial MDA concentration, more serious left cardiac function injury and lower myocardial SOD activity and SR Ca²⁺-ATPase activity; group SHR+B had lower myocardial MDA concentration, higher myocardial SOD activity, but no difference in blood pressure, LVW/BW, the degree of left cardiac function injury and myocardial SR Ca²⁺-ATPase activity. CONCLUSION: Benazepril can attenuate ischemia-reperfusion-induced cardiac function injury by regression of left ventricular hypertrophy (LVH), improving SR Ca²⁺-ATPase activity and decreasing oxygen free radicals injury in SHRs.     

Study on relationship between expression of PKCα in glomeruli and development of nephropathy in diabetic rats

AIM:To observe the dynamic changes of expression of PKCα, TGF-β₁ and α-SMA in glomeruli of diabetic rats induced by the alloxon and to invesitigate their roles in the diabetic nephropathy(DN).METHODS:Rats were randomly divided into four groups: normal control group (group A), diabetic group of one week (group B), diabetic group of one month (group C), diabetic group of two months (group D). Immunohistochemistry and Western blotting were used to detect the expression of PKCα, TGF-β₁ and α-SMA in renal tissue of all groups. Blood glucose, triglycerides, cholesterol, creatinine and urine protein were analysed by chemical methods. The morphological changes of renal tissue were checked through microscopy.RESULTS:The expression of PKCα and TGF-β₁ in renal tissue of diabetic groups were increased comparing with those of nomal control group(P<0.05). The mesangial cells expressed α-SMA in two months group. Chronologically the expression of PKCα, TGF-β₁ and α-SMA were positively correlative with each other and the impairment of kidney was also observed.CONCLUSIONS: During the DN process the expression of PKCα increased. PKCα raised GFR and the permeability of glomerular filtration membrane which enhanced urinary albumin excretion. PKCα also increased expression of TGF-β and therefore to induce the expression of α-SMA. The appearance of α-SMA was a marker of the phenotypic transform of renal cells.     

The effects of calcium and reactive oxygen species in rat kidney during ischemia and reperfusion period

AIM and METHODS: Electron cytochemical methods were used to study the changes of calcium and reactive oxygen species in rat kidney during ischemia and reperfusion period.RESULTS:By the end of 1h ischemia, intra-cellular calcium increased. There were no H₂O₂ generation at this time. In the early reperfusion period, large amount of H₂O₂ generated. At this time, there were no evident changes of intra-cellular calcium compare with 1h ischemia group. In the later reperfusion period, less H₂O₂ generated. Intra-cellular calcium increased continuously.CONCLUSION: Calcium and reactive oxygen species all participated in ischemia-reperfusion injury, but the time they participated and their effects were different.     

Role of endogenous angiotensin II in the pathogenesis of aortic calcification in rats

AIM: To explore the effects of angiotensin II on aortic calcification in the rat. METHODS: Arterial calcification of Sprague-Dawley rats was induced by vitamin D₃ plus nicotine. Calcification was confirmed by Von Kossa staining, measurement of calcium content, [⁴⁵Ca²⁺]accumulation and alkaline phosphatase (ALP) activity of vascular tissue. RESULTS: The results showed that calcium content, [⁴⁵Ca²⁺]accumulation and ALP activity in calcified arteries increased significantly compared with those of control. Ang Ⅱ levels in plasma and aortic tissues and the amount of angiotensinogen mRNA in calcified aorta were also increased as compared with control. Captopril (inhibitor of ACE) and losartan (Ang Ⅱ receptor inhibitor) decreased significantly the content of calcium, [⁴⁵Ca²⁺] uptake and ALP activity in calcified aorta. Ang Ⅱ levels in plasma and aortic tissues and the amount of angiotensinogen mRNA in aortic tissue were down-regulated by captopril. The amount of angiotensinogen mRNA and the content of Ang Ⅱ in the calcified aorta were also decreased by losartan. CONCLUSION: The captopril and losartan significantly alleviate the vascular calcification.     

Losartan inhibits LPS-induced GFAP expression via AMPK activation in mouse hippocampus

AIM: To investigate the effects of losartan on lipopolysaccharide (LPS)-induced glial fibrillary acidic protein (GFAP) expression, and to determine whether adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) activation is involved in the mechanism.METHODS: Adult male KM mice were divided into control group, LPS model group, losartan treatment group, and losartan and Compound C co-treatment group. To establish a model of central nervous system inflammation, the mice received daily intracerebroventricular injection of LPS (24 μg/d) for 2 d. Daily losartan administration (0.5, 1 or 5 mg·kg^-1·d^-1, ip) initiated at 14 d prior to LPS injection. Compound C (10 mg/kg, ip), a selective AMPK inhibitor, started to be injected daily at 2 d prior to LPS injection. The hippocampal tissues in each group were isolated at 3 d after the last LPS injection, and then the protein levels of GFAP, AMPK, p-AMPK, mammalian target of rapamycin (mTOR) and p-mTOR were determined by Western blot.RESULTS: Twice LPS injections significantly increased the expression of GFAP in the hippocampus (P<0.01). Losartan inhibited LPS-induced GFAP expression in a concentration-dependent way, and losartan at 5 mg·kg^-1·d^-1 significantly inhibited GFAP expression and AMPK activation (P<0.05), but it had no obvious effect on mTOR activation. Furthermore, Compound C significantly reversed the effect of losartan treatment on LPS-induced GFAP expression and AMPK phosphorylation (P<0.05).CONCLUSION: Losartan inhibits LPS-induced GFAP expression in the mouse hippocampus, and AMPK activation but not mTOR, is involved in the mechanism.     

Screening of lentiviral vectors carrying effective siRNA targeting S1P2 gene

AIM:To screen the lentiviral vector carrying siRNA with higher efficiency of suppressing the sphingosine-1-phosphate receptor 2(S1P2) gene expression in the primarily cultured corpus cavernosum smooth muscle cells of spontaneously hypertensive rats (SHR).METHODS:SHR and SD rats (n=5 each) were used for primarily culturing corpus cavernosum smooth muscle cells.The cells were randomly divided into 6 groups:SHR siRNA-1,SHR siRNA-2,SHR siRNA-3,SHR GFP,SHR control (SHR non-transfection group),and SD control (SD rat control group).Each group had 5 samples with 1.0×10⁵ cells of each sample.At 72 h after transfection (MOI=60) with lentiviral vectors carrying S1P2 siRNA into the SHR corpus cavernosum smooth muscle cells,the expression of GFP was observed under fluorescence microscope.The protein expression of S1P2,ROCK1,ROCK2 and eNOS in the corpus cavernosum smooth muscle cells,and the mRNA expression of S1P2,ROCK1 and ROCK2 were determined by by Western blot and RT-PCR.RESULTS:The transfection efficiency of the corpus cavernosum smooth muscle cells in SHR siRNA-1,SHR siRNA-2,SHR siRNA-3 and SHR GFP groups were>80%.Compared with SHR control group,the mRNA levels and the protein expression of S1P2,ROCK1 and ROCK2 in SHR GFP group showed no remarkable changes,while those in SHR siRNA-1,SHR siRNA-2,SHR siRNA-3 and SD control groups were significantly lower than those in SHR control group (P<0.05).The protein expression of eNOS in SHR siRNA-1,SHR siRNA-2,SHR siRNA-3 and SHR GFP groups were not significantly changed as compared with SHR control group,but that in SD control group was significantly higher than that in SHR control group.CONCLUSION:Three groups of siRNA lentiviral vectors targeting S1P2 inhibit the expression of S1P2 in the corpus cavernosum smooth muscle cells of SHR,and by silencing the S1P2 expression,the expression of ROCK1 and ROCK2 is inhibited.Among them,siRNA-1 has the highest inhibitory efficiency.     

Effects of valsartan alone or in combination with benazepril on blood pressure and left ventricular hypertrophy in spontaneously hypertensive rats

AIM:To evaluate the effects of different doses of valsartan alone or with concomitant be-nazepril on blood pressure,left ventricular hypertrophy,RAASfunction and endoxi nlevel in spontaneously hy-pertensive rats(SHR).METHODS:Thirty SHR(fourteen-week-old,male)were divi ded into five groups(six rats in each group):SHR control group:fed with normal saline;benazepril group:fed with 1 mg·kg^-1·d^-1benazepril);low dose valsartan group:fed with 8 mg·kg^-1·d^-1valsartan;high dose valsartan group:fed with 24 mg·kg^-1·d^-1valsartan;combination drug therapy group:fed with valsartan(8 mg·kg^-1·d^-1)and benazepril(1 mg·kg^-1·d^-1),all for 8 weeks.WKY control group(n=6):fed with normal saline for 8 weeks.RESULTS:SBP,LVM/BW,TDMof SHR were remarkably lower than those of control after drug i n-tervene,and effect on SBP was most remarkable in high dose valsartan group and i nthe combi nation drug ther-apy group;effect on LVM/BW,TDM were most remarkable in combination drug therapy group.Renin activi-ties in plasma and myocardiumwere remarkably i ncreased in drug i ntervene groups.The levels of AngⅡi nplasma and myocardiumwere remarkably increased in two different dose of valsartan treati ng group,and thelarger dose of valsartan were,the higher levels of AngⅡin plasma and myocardium were;decreased in be-nazepril treati ng group and combination drug therapy group.Na⁺-K⁺-ATPase activities in myocardi umwere remarkably i ncreased and the level of endoxi n i n myocardium were remarkably decreased as SBP de-creased after drug intervene.CONCLUSION:Different dose of valsartan alone or combi ned with benazeprilcan decrease SBP of SHR,have the effect of inhibiti ng progression of ventricular hypertrophy.The effect ofcombination drug therapy group was most remarkable among five groups and can avoi d the si de effect of highAngⅡin plasma and myocardiumduri ng long-termuse of valsartan alone.     

The protection of the hepatic ischemic preconditioning is concerned with the NO/ET-1 system

AIM: To study the relationship between the disturbance of nitric oxide/endothelin-1(NO/ET-1) and hepatic ischemia/reperfusion(I/R) injury as well as the regulation of NO/ET-1 system by hepatic ischemic preconditioning(IPC). METHODS: The changes of NO/ET-1 system and their relationship with hepatic I/R injury were compared between I/R group and IPC+I/R group in a rat hepatic I/R model. Two hours after reperfusion, the liver tissues were detected by RT-PCR to see whether there was inducible nitric oxide synthase (iNOS) mRNA expression. RESULTS:In the acute phase of hepatic reperfusion, the ratio of NO/ET-1 was reduced, which was due to a significant reduction of NO₂^-/NO₃^- (the metabolic product of NO) and significant elevation of ET-1 in the blood plasma. The content of ALT, AST, LDH and TNF-α in blood plasma, and of MDA in liver tissue were increased but ATP in liver tissue was reduced, the hepatic damage was deteriorated. The protection of the hepatic IPC was concerned with the elevation of the ratio of NO/ET-1 caused by the elevation of NO₂^-/NO₃^-, and reduction of ET-1 as well. There was no iNOS mRNA detected in the liver tissues.CONCLUSION: Hepatic I/R injury is related to the disturbance of NO/ET-1. The protection of the hepatic IPC in the acute phase might be conducted by its regulation of NO/ET-1 system. The cNOS rather than the iNOS generated the NO in this situation.     

Roles of gap junction in TGF-β1-induced proliferation of vascular smooth muscle cells from spontaneous hypertensive rats

AIM: To investigate whether gap junction participates in transforming growth factor β₁(TGF-β₁)-induced proliferation of spontaneous hypertensive rat (SHR) vascular smooth muscle cells (VSMCs). METHODS: The thoracic aorta of the rats were sampled. The primary SHR VSMCs were isolated and cultured in vitro. The cells were divided into 4 groups: control group, TGF-β₁ group,18α-glycyrrhetinic acid(18α-GA) group and TGF-β₁+18α-GA group. The proliferation of SHR VSMCs was observed by the methods of MTT and flow cytometry. The protein expression and co-localization of connexin(Cx)43 and Cx40 in SHR VSMCs were detected by immunofluorescence staining. The protein levels of Cx43 and Cx40 in the cells were also measured by Western blotting. The method of molecular dye transfer (scrape dye transfer method) was applied to detect the function of gap junction in SHR VSMCs. RESULTS: The protein expression of Cx43 and Cx40 in SHR VSMCs was positive and co-localized in the cytoplasm. Compared with control group, the percentage of S-phase detected by cell cycle and A value detected by MTT in TGF-β₁ group were obviously increased (P<0.05), indicating that the proliferation of the cells was enhanced. However, the proliferation of the cells decreased in 18α-GA group (P<0.05). Compared with TGF-β₁ group, the percentage of S-phase and A value in TGF-β₁+18α-GA group were both significantly decreased (P<0.05), indicating that the proliferation of the cells decreased. Compared with control group, the protein expression of Cx43 in TGF-β₁ group was increased (P<0.05), whereas the protein expression of Cx40 was not changed (P>0.05), and the protein expression of Cx43 and Cx40 in 18α-GA group were decreased (P<0.05). Compared with TGF-β₁ group, the expression of Cx43 in TGF-β₁+18α-GA group was significantly decreased (P<0.05),but no difference of the Cx40 protein levels between the two groups was observed. Compared with control group, the function of gap junction detected by scrape dye transfer method in TGF-β₁ group was enhanced (P<0.05), and weakened in 18α-GA group (P<0.05). Compared with the TGF-β₁ group, the function of gap junction in TGF-β₁+18α-GA group was significantly attenuated (P<0.05). CONCLUSION: TGF-β₁ enhances the function of gap junction to stimulate the proliferation of SHR VSMCs through the expression of Cx43 protein. The expression of Cx40 protein may not play a major role in this process.     

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.     

Effect of atorvastatin on ventricular remodeling in spontaneous hypertension rats

AIM：To explore the effect of atorvastatin on cardiac remodeling in spontaneous hypertension rats (SHR).METHODS：Twelve spontaneous hypertension rats were divided randomly into two groups：group of atorvastatin (atorvastatin 50 mg·kg^-1·d^-1) and group of SHR (0.5% mucilage of arabic gum,10 mL·kg^-1·d^-1).Additionally,six male Wistar-Kyoto rats (0.5% mucilage of arabic gum,10 mL·kg^-1·d^-1) were selected as control group.Systolic blood pressure was assessed with the tail-cuff method.After six weeks,entire heart,and left ventricle were weighed.The left ventricular weight index was calculated and myocardial hydroxyproline and collagen protein concentration were measured.The serum high sensitivity CRP (hs-CRP) was measured by nephelometry.The localization of vascular cell adhesion molecule (VCAM) in myocardium was investigated by immunohistochemistry assays.The level of NF-κB mRNA expression was detected with in situ hybridization.Ultrastructure in cardiac muscle was also observed under transmission electron microscope.RESULTS：The expression of myocardial VCAM and NF-κB in SHR group was stronger than that in WHY group.Compared with SHR group,entire heart weight,left ventricular weight,left ventricular weight index,serum hs-CRP,myocardial hydroxyproline and collagen protein concentration was decreased,the expression of myocardial VCAM and NF-κB in SHR group was weaker than that in atorvastatin treatment group.The myocardial pathological change such as incomplete karyotheca in cardiac muscle cells,no clear of transverse striation and the mess in myofibril alignment,and hyperplasy in interstitial collagen fibre were observed in SHR group and these changes were improved in atorvastatin treatment group.CONCLUSION：The cardiac remodeling in SHR is improved by atorvastatin.The molecular mechanism may be related to its down-regulating the expression of VCAM protein and NF-κB and inhibiting myocardial chronic inflammation.     

Nuclear calcium oscillation in cultured neonatal rat myocytes

AIM: To determine whether nuclear Ca²⁺ is independently regulated from the cytosolic Ca²⁺ and nuclear Ca²⁺ oscillation induced by many modulating factors in cultured rat neonatal myocytes and its mechanism. METHODS: Rat neonatal cardiac myocytes were cultured, and fluo-4/AM was loaded as calcium probe. The changes of cytosolic and nuclear Ca²⁺ were observed by confocal laser microscopy. RESULTS: Calcium fluorescent intensity oscillated slightly in myocardiocytes and the average intensity was much higher in the nucleus than that in the cytosole. Ca²⁺ oscillation in nucleus and cytosole induced by norepinephrine, isoproperenol, ATP were completely blocked by Ca²⁺-ATPase antagonist thapsigargin (10^-6 mol/L),L-type Ca²⁺ channel blocker verapermil (500 μmol/L) and KCl (20 mmol/L). Ca²⁺-ATPase antagonist thapsigargin completely blocked the propagation of Ca²⁺ waves and simutaneouly induced a temporary Ca²⁺ increase followed by a magnificient drop and loss of response to norepinephrine. CONCLUSIONS: The results suggested that generation and maintenance of calcium oscillation both in cytosole and nucleus depended on extracellular Ca²⁺ influx, membrane potential, Ca²⁺ release and uptake of cytosolic and nuclear calcium stores. The difference between cytosolic calcium and nuclear calcium indicated that calcium regulating system relatively independent of cytosole may exist in nucleus.     

Relationship between the intracellular calcium concentration changes and left ventricular hypertrophy and function in the spontaneously hypertensive rats

AIM:To elucidate the relationship between the intracellular calcium concentration changes and left ventricular hypertrophy and function in the spontaneously hypertensive rats (SHR).METHODS:Intracellular free calcium concentrations were measured by Fura 2 methodology and left ventricular function quantitated by cardiac catheterization in 20 SHR aged 10, 22, and 34 weeks and 20 age-matched Wistar-kyoto (WKY) rats.RESULTS:(1) The systolic blood pressure(SBP), intracellular calcium concentrations and left ventricular mass / body weight index (LVM/BW) were significantly higher in all three age groups of SHR than the corresponding groups of WKY; (2) Compared with age-matched WKY groups, the peak left ventricular pressure descending rate(-dp／dt_max) decreased while left ventricular relaxation time constant (τ)increased significantly in SHR aged 22 and 34 weeks. The peak left ventricular pressure ascending rate(dp／dt_max) and the left ventricular contractility index were significantly increased only in the 34 weeks SHR; (3) Intracellular calcium concentrations showed a positive correlation with LVM/BW,SBP,-dp／dt_max and τ(r=0.47-0.83,P<0.01)and a negative correlation with dp／dt_max and the left ventricular contractility index (r=-0.46,P<0.05 and r=-0.81, P<0.01).CONCLUSION:Intracellular calcium overload is one of the potential mechanisms in the induction of left ventricular hypertrophy as well as of systolic and diastolic dysfunction.     

Changes of myocardial nuclear membrane Ca2+-ATPase function in ischemia/reperfusion injury

AIM: The changes of myocardial nuclear membrane Ca²⁺ -ATPase function was investigated in ischemia/reperfusion injury. METHODS: The model of myocardial ischemia/reperfusion injury was established in rats. Myocardial nuclei were purified with sucrose density centrifugation, the activity of Ca²⁺ -ATPase was measured and calcium uptake was assayed with [⁴⁵ Ca²⁺ ] . RESULTS: Plasma levels of malondialdehyde (MDA) and free fatty acid (FFA) in myocardial ischemia/reperfusion injury increased significantly( P<0.01 vs control). Ca²⁺ -ATPase activity and [⁴⁵ Ca²⁺ ] uptake was lower than normal at below 10 μmol/L, while higher at 50 μmol/L. CONCLUSION:These data indicate dysfunction of nuclear menbrane calcium pump and [⁴⁵ Ca²⁺ ] uptake function in myocardial ischemia/reperfusion injury.