Effects of osthole on renal oxidative stress injury in aged spontaneously hypertensive rats by regulating PI3K/Akt/mTOR signaling pathway / 中国临床药理学与治疗学

AIM: To explore the mechanism of osthole on elderly spontaneously hypertensive rats. METHODS: 20-month-old spontaneously hypertensive rats (SHRs) and healthy Wistar-Kyoto (WKY) rats were purchased. SHRs were treated with osthole (i.g.) for 8 weeks. The systolic blood pressure and diastolic blood pressure of rats were monitored. Hematoxylin-eosin staining (H&E), periodic acid-schiff staining (PAS) and Masson staining were used to observe the pathological changes of rat kidney tissues. The activity of superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione (GSH) in rat kidney was detected by ELISA kit. PI3K/Akt/mTOR signaling pathway related proteins were detected by western blot. RESULTS: Osthole reduced the systolic and diastolic blood pressure of SHRs, improved the histopathological changes of SHRs kidney, reduced the activity of MDA in SHRs kidney, and increased the activity of SOD and GSH. Osthole reduced the levels of p-PI3K, p-Akt and p-mTOR. CONCLUSION: Osthole reduces the activity of PI3K/Akt/mTOR signaling pathway and exerts a protective effect on renal oxidative stress injury in aged spontaneously hypertensive rats.

Effect of electroacupuncture on myocardial fibrosis in spontaneously hypertensive rats based on cholinergic anti-inflammatory pathway / 中国针灸

OBJECTIVE@#To observe the effect of electroacupuncture (EA) at "Neiguan" (PC 6) on myocardial fibrosis in spontaneously hypertensive rats (SHR), and explore preliminarily the mediating role of cholinergic anti-inflammatory pathway (CAP) and its downstream nuclear factor κB (NF-κB) signaling pathway.@*METHODS@#Six 12-week-old WKY male rats were employed as the normal group. Eighteen 12-week-old SHR were randomly divided into 3 groups, i.e. a model group, an EA group and a blocking group (EA after blocking α7 nicotinic acetylcholine receptor [α7nAchR]), with 6 rats in each one. In the EA group, EA was delivered at "Neiguan"(PC 6) and the site 0.5 cm from its left side, with disperse-dense wave, 2 Hz/15 Hz in frequency and 1 mA in current intensity. One intervention took 30 min and was given once every 2 days, lasting 8 weeks. In the blocking group, prior to each EA, the α7nAchR specific blocker, α-bungartoxin was injected intravenously in the tails of the rats. After EA intervention, the systolic blood pressure (SBP), the diastolic blood pressure (DBP) and the mean arterial pressure (MAP) were measured with non-invasive blood pressure monitor. Using echocardiogram, the left ventricular (LV) anterior wall end-diastolic thickness (LVAWd) , LV posterior wall end-diastolic thickness (LVPWd) and the LV end-diastolic internal diameter (LVIDd) were measured. The level of hydroxyproline (Hyp) in the myocardial tissue was determined by using alkaline hydrolysis, and that of acetylcholine (Ach) was detected by ELISA. With the real-time PCR adopted, the mRNA expression of NF-κB p65, tumor necrosis factor α (TNF-α), interleukin (IL)-1β and IL-6 were determined.@*RESULTS@#Compared with the normal group, SBP, DBP, MAP, LVAWd and LVPWd were increased (P<0.01), and LVIDd was decreased (P<0.01) in the rats of the model group. SBP, DBP, MAP and LVAWd were dropped (P<0.01, P<0.05), and LVIDd rose (P<0.01) in the EA group when compared with those in the model group. The differences in the above indexes were not statistically significant between the blocking group and the model group (P>0.05). Compared with the normal group, Hyp level and the mRNA expression of NF-κB p65, TNF-α, IL-1β and IL-6 in the myocardial tissue increased (P<0.01, P<0.05) and Ach level decreased (P<0.01) in the model group. Hyp level, the mRNA expression of NF-κB p65, TNF-α, IL-1β and IL-6 in the myocardial tissue were reduced (P<0.05, P<0.01) and Ach level rose (P<0.01) in the EA group when compared with those in the model group. These indexes were not different statistically between the blocking group and the model group (P>0.05).@*CONCLUSION@#CAP may be involved in ameliorating the pathological damage of myocardial fibrosis during EA at "Neiguan"(PC 6). The underlying effect mechanism is associated with up-regulating the neurotransmitter, Ach and down-regulating mRNA expression of NF-κB p65 and pro-inflammatory factors such as TNF-α, IL-1β and IL-6 in myocardial tissue.

The effects of moderate intensity exercise on left ventricular remodeling in spontaneously hypertensive rats / 中华物理医学与康复杂志

Objective:To observe any effect of moderate-intensity exercise on left ventricular remodeling (such as cardiomyocyte hypertrophy, apoptosis and proliferation) in spontaneously-hypertensive rats (SHRs) and explore the possible mechanisms.Methods:Thirty 4-month-old female SHRs were randomly divided into a sedentary group ( n=15) and an exercise group ( n=15). Fifteen Wistar Kyoto rats served as the control group. The exercise group underwent daily 60-min moderate-intensity treadmill exercise 5 days per week for 12 weeks, while the sedentary and control groups were raised quietly in cages for the same period. After the 12-week intervention, the caudal artery blood pressure was measured using a non-invasive blood pressure monitor. The rats were then sacrificed and their hearts were sampled for morphometric measurement. Cardiomyocytes were isolated and underwent DAPI staining to measure their length, width and area. Apoptosis cardiomyocytes was detected by using terminal-deoxynucleoitidyl transferase mediated nick end labeling and their proliferation was assessed using immunofluorescent staining. The number of cardiac progenitor cells was detected by flow cytometry, while the expression of the cardiac calcineurin Aβ subunit (CNAβ) and phosphorylated Akt (p-Akt) protein were measured using western blotting. Results:Compared with the control group, a significant increase was observed in the heart weight, heart mass index (HMI), systolic blood pressure, diastolic blood pressure, myocardial thickness of the left ventricular wall (anterior wall, posterior wall and septal wall), cardiomyocyte morphology (length, width and area), cardiomyocyte apoptosis rate, proliferation rate, number of cardiac progenitor cells and protein expression of CNAβ in the sedentary group. Compared with the sedentary group, the average heart weight, HMI, myocardial thickness of the left ventricular wall (anterior wall, posterior wall and septal wall), cardiomyocyte morphology (length, width and area), cardiomyocyte proliferation rate, number of cardiac progenitor cells and p-Akt protein expression had increased significantly in the exercise group. The average systolic blood pressure, diastolic blood pressure, apoptosis rate and CNAβ protein expression had decreased significantly.Conclusions:Moderate-intensity exercise can induce physiological cardiac hypertrophy in SHRs, relieve apoptosis, increase the number of cardiac progenitor cells and promote cell proliferation, thereby inhibiting cardiac remodeling.

Effect of electroacupuncture on renal fibrosis in spontaneously hypertension rats and its related mechanisms / 针刺研究

OBJECTIVE: To observe the effect of electroacupuncture (EA) on blood pressure, renal fibrosis and expression of tissue inhibitors of metalloproteinase-1 (TIMP-1), plasminogen activator inhibitor 1 (PAI-1), and alpha smooth muscle actin (α-SMA) in spontaneous hypertension rats (SHR), so as to explore its mechanisms underlying improving hypertensive renal damage. METHODS: Forty male SHR (15 weeks in age) were randomly divided into 5 groups: model, medication (Losartan), Shenshu, Geshu, and Shenshu＋Geshu groups(n=8 rats in each group), and the same age-old male 8 Wistar-Kyoto (WKY) rats were used as the normal control group. Rats of the medication group were treated by gavage of Losartan potassium solution (3 mg/mL, 30 mg·kg-1·d-1, once a day for 12 weeks), and those of the 3 EA groups treated by EA stimulation of bilateral "Shenshu" (BL23), "Geshu"(BL17) or both BL23 and BL17 (2 Hz/100 Hz, 1 mA, 15 min each time, once every other day for 12 weeks). The systolic blood pressure of the tail artery was measured before, and 4, 8 and 12 weeks after the intervention. The expression of TIMP-1, PAI-1 and α-SMA proteins of the right kidney tissue was measured by immunohistochemistry. Histopathological changes of the right renal tissue were observed under light microscope after H.E. stain. RESULTS: The blood pressure was significantly higher in the mo-del group than those in the normal control group (P<0.01), and considerably decreased at the 4th , 8th, and 12th week of the interventions in the medication and 3 EA groups (P<0.01). The expression levels of renal TIMP-1, PAI-1 and α-SMA proteins were notably higher in the model group than those in the normal control group and considerably decreased at the 12th week of the interventions in the medication and 3 EA groups than in the model group (P<0.01). H.E. staining of the renal tissue showed disordered arrangement of the renal cells, congestion and dilation of capillaries with thickened vascular wall, renal tubule atrophy and lumen stenosis with some necrosis of renal tubules, protein tubule and cell tubules, increase of some glomerular mesangial matrix and hyperplasia of fibrous tissue in the model group, which was re-latively milder in the medication and 3 EA groups. CONCLUSION: EA of BL23 and BL17 can reduce the blood pressure in SHR, which may be related to its function in down-regulating expression of TIMP-1, PAI-1 and α-SMA proteins.

Relationship between microangiogenesis and astrocytes after cerebral infarct in SHR-SP rats / 西安交通大学学报(医学版)

Objective: To investigate the possible role of astrocytes after brain infarction in stroke-prone, spontaneously hypertensive (SHR-SP) rats and the association with angiogenesis and the architecture. Methods: We maintained SHR-SP rats on high sodium water starting to accelerate the stroke onset. The 3D quantification of microvasculatures (diameter, branch number) by cofocal microscope after FITC-dextran was injected into the rats via the left femoral vein. Glial fibrillary acidic protein (GFAP) expression and microvessel density (MVD) using counting the number of factor -positive endothelial cells were evaluated by immunofluorescence and immunohistochemistry, respectively. Results: Cerebral infarction occurred at week 7 after high sodium water intake (13 g/L NaCl) in SHR-SP group. When compared with the non-infarcted contralateral hemisphere and SHR-SP on normal sodium intake and WKY rats, GFAP expression and MVD were significantly increased, respectively, and the diameter and the branch number of vessels were decreased, respectively, in cerebral infarcts with boundary zones of SHR-SP rats (P<0.01). Linear correlation analysis showed that GFAP expression was positively correlated with MVD and the diameter and the branch number of vessels in cerebral infarcts in SHR-SP (P<0.01). Conclusion: Astrocytes hyperplasia may be associated with increased regional angiogenesis and the changes of architecture in SHR-SP rats with high sodium water (13 g/L NaCl) that induces focal cerebral infarcts.

Lentiviral vectors carrying siRNA inhibit S1PR3 gene expression in the corpus cavernosum smooth muscle cells of rats with spontaneous hypertension / 中华男科学杂志

Objective@#To screen lentiviral vectors carrying siRNA which can specifically down-regulate the gene expression of the sphingosine-1-phosphate receptor 3 (S1PR3) in the corpus cavernosum smooth muscle (CCSM) cells of rats with spontaneous hypertension (SHT) and investigate the influence of the vectors on the signaling pathways of ROCK1, ROCK2 and eNOS in the CCSM cells of SHT rats.@*METHODS@#Using the S1PR3 mRNA sequence of the rat as an interfering target, we designed and synthesized three pairs of siRNA sequences (siRNA1, 2 and 3) targeting S1PR3 and one pair of negative control, and then constructed and packaged them into lentiviral vectors. We cultured the CCSM cells of SHT and Wistar-Kyoto (WKY) rats in vitro and randomly divided them into groups A (SHT untransfected control), B (SHT transfected and carrying negative control virus), C (SHT transfected and carrying siRNA1 targeting S1PR3), D (SHT transfected and carrying siRNA2 targeting S1PR3), E (SHT transfected and carrying siRNA3 targeting S1PR3), and F (WKY untransfected control). With the multiplicity of infection (MOI) = 60, we transfected the CCSM cells of the SHT rats with the lentiviral vector and then determined the expression of the green fluorescent protein (GFP) as well as the mRNA and protein expressions of S1PR3, ROCK1, ROCK2 and eNOS in the CCSM cells of the SHT and WKY rats by RT-PCR and Western blot.@*RESULTS@#Gene sequencing proved the successful construction of the lentiviral vector. The transfection efficiency of the CCSM cells of the rats was >80% in groups B, C, D and E. Compared with group A, the mRNA and protein expressions of S1PR3, ROCK1 and ROCK2 exhibited no significant difference in group B but were remarkably decreased in groups C, D, E and F (P0.05) but remarkably lower than those in group F (P0.05) but markedly increased in groups A, B, C and D (P< 0.05), while those of eNOS remarkably decreased in groups A, B, C, D and E (P< 0.05).@*CONCLUSIONS@#The three constructed lentiviral vectors carrying siRNA targeting different loci of the S1PR3 gene could significantly inhibit the expression of S1P3 as well as RhoA/Rho kinase signaling pathways in the CCSM cells of SHT rats, and the vector carrying siRNA3 exhibited the highest inhibitory effect.

Intervention effect of Captopril on serum of spontaneous hypertension rats based on metabonomic research / 中国药理学通报

Aims To analyze the intervention effect of Captopril on serum endogenous metabolites alternations in spontaneous hypertension rats ( SHR) and to investi-gate possible therapeutic mechanism .Methods The rapid resolution liquid chromatography/quadrupole-time of flight tandem mass spectrometry ( RRLC-Q-TOF-MS) and technology coupled with partial least squares discriminant analysis ( PLS-DA ) processed by SIMCA-P software were used to distinguish significantly different variables and identify potential biomarkers . Results Compared to the normal group , metabolic profiling changed significantly in model group and Cap-toril group .Totally 4 metabolins and their metabolic pathways were detected , which were closely related to the endothelial function .Conclusion Metabolomics reveals possible therapeutic mechanism of Captopril for protecting endothelial function from overall metabolism of the body .It also shows unique potential in terms of interpretation of the complex mechanisms of drugs .

Antihypertensive Effect of the Extract from Compound Prunella Vulgaris L. in Spontaneous Hypertension Rats / 中国药师

Objective:To study the antihypertensive effect of the extract from compound Prunella vulgaris L. in spontaneous hy-pertension(SH)rats. Methods:Forty SH rats were randomly divided into the model group,high,middle and low dose groups of ex-tract from vompound Prunella vulgaris L. ,the compound kendir leaves tablets Ⅰ group with 8 ones in each. Non-invasive blood pres-sure measurement was used to detect the SBP and DBP of the SHR rats. Then the serum NO,AngⅡ ,ET-1 and ANP content were measured after the eight-week treatment. The pathological changes were observed after kidney HE staining in the SH rats. Results:Compared with that in the model group,the blood pressure in high-dose treatment group,middle-dose treatment group and the positive model group was significantly decreased(P < 0. 05). The AngⅡ and ET-1 levels in high-dose treatment group,middle-dose treatment group and the positive model group were decreased(P < 0. 01),and NO and ANP contents in serum were significantly increased when compared with those in the model group(P < 0. 01). The pathological examination showed that the pathological changes in the model group were faster than those in all the drug-treatment groups,the pathological changes included glomerular and renal tubular atrophy, glomerular vascular wall thickening and renal tubular epithelial cell degeneration or necrosis. Conclusion:The extract from compound Prunella vulgaris L. can reduce blood pressure of SH rats. The mechanism may be associated with the level reduction of AngII and ET-1 and content elevation of NO and ANP.

The antihypertensive effect of ethyl acetate extract of radish leaves in spontaneously hypertensive rats

Radish (Raphanus sativus L.) is a cruciferous vegetable, and its leaves have antioxidant and anticancer properties. This study was conducted to evaluate the effects of ethyl acetate extracts from radish leaves on hypertension in 11-week-old spontaneously hypertensive rats (SHRs). The SHRs were randomly divided into 3 groups of 6 rats each on the basis of initial systolic blood pressure (SBP) and were treated with oral administration of radish leaf extract (0, 30, or 90 mg/kg body weight [bw], respectively) for 5 weeks. Six Wistar rats were used as normotensive controls. The amount of the radish leaf extract had no effect on body weight. The SBP of the SHRs showed a decreasing trend with the consumption of the radish leaf extract. In the third week, the SBP of the group fed 90 mg extract/kg bw reduced from 214 mmHg to 166 mmHg and was significantly lower than that of the normotensive and hypertensive controls. The extract did not show a significant effect on the angiotensin-converting enzyme (ACE) activity in the serum, kidney, and lung. The extract increased the concentration of NO in serum and the activities of antioxidant enzymes such as glutathione peroxidase and catalase in red blood cells (RBCs). The serum concentrations of Na+ and K+ were not significantly different between all groups. However, the fecal concentrations of Na+ and K+ increased; the fecal concentrations of Na+ and K+ for the normotensive and hypertensive controls were not different. Urinary excretion of Na+ was higher in the normotensive Wistar rats than in the SHRs, while that of K+ was not significantly different. These findings indicate that consumption of radish leaves might have had antihypertensive effects in SHRs by increasing the serum concentration of NO and fecal concentration of Na+ and enhancing antioxidant activities.

Jiuqiang Naoliqing enhancing the expression of CGRP and Synapsin Ⅰ in brain of spontaneous hypertension rats / 中国康复理论与实践

@#ObjectiveTo study the influence of Jiuqiang Naoliqing (JNQ) on the expression of calcitonin gene related peptide(CGRP)and Synapsin Ⅰ in brain of the spontaneous hypertension rats (SHR). MethodsThe rats were randomly divided into 4 groups: Wistar group, SHR group, lower dose of JNQ treated SHR group and higher dose of JNQ treated SHR group. The expression of CGRP and Synapsin Ⅰ in the dentate gyrus, CA1 subfield of hippocampus and cortex were determined by immunohistochemistry after treatment for 3 weeks. ResultsCompared with the Wistar group, the expression of CGRP and Synapsin Ⅰ in the dentate gyrus, CA1 subfield of hippocampus and cortex of SHR group significantly decreased. The treatment with lower dose of JNQ significantly enhanced the expression of CGRP in cortex(P<0.05 vs SHR).The treatment with higher dose of JNQ significantly enhanced not only the expression of CGRP in the dentate gyrus, CA1 subfield of hippocampus and cortex, but also that of Synapsin Ⅰ in the CA1 subfield of hippocampus selectively in comparison with SHR group. ConclusionJNQ may improve the micro circulation in brain by up regulating the expression of CGRP and enhance the modulating function of central nervous system by up regulating the expression of Synapsin Ⅰ in spontaneous hypertension rats.

Effect of Jiuqiang Naoliqing on the TXA_2 and PGI_2 level in spontaneous hypertension rat plasma / 中国康复理论与实践

@#ObjectiveTo investigate the effect of Jiuqiang Naoliqing(JNQ) on the TXA2 and PGI2 level in spontaneous hypertension rat (SHR) plasma.MethodsThe plasma was separated after the SHR and Wistar rats were treated with JNQ at the dose of 0.133g/kg,0.265g/kg,0.530g/kg and 1% carboxymethyl cellulose respectively for 5 weeks. The level of TXB2 and 6 keto PGF1α ,stable metabolin of TXA2 and PGI 2,in SHR plasma was tested by radioimmunoassay.ResultsThe level of TXB2 and the ratio of TXB2/6 keto PGF1α (T/P) in SHR plasma increased significantly(P<0.01),and there was no significant difference in the concentration of 6 keto PGF1α between Wistar rats and SHR plasma(P>0.05). JNQ could increase the generation of 6 keto PGF1α and decrease the level of TXB2 and T/P in SHR plasma after treated with different dosages for 5 weeks.ConclusionJNQ may improve the balance between TXA2 and PGI2 in SHR plasma.

Jiuqiang Naoliqing affords protection to vital organs of spontaneous hypertension rats / 中国康复理论与实践

@#ObjectiveTo investigate the Jiuqiang Naoliqing's (JNQ) histological influence on hearts, brains and kidneys of spontaneous hypertension rats (SHR). MethodsThe rats were randomly divided into four groups: Wistar control group, SHR group, higher dose JNQ treated SHR group(0.530 g/kg) and lower dose JNQ treated SHR group(0.265 g/kg). The treatment lasted five weeks, and the rats' blood pressure were monitored through tail pulse. After the perfusion procedure, rats' hearts, brains and kidneys were rapidly removed in low temperature condition and stored in 10% formalin solution of 4 ℃.Then routine sections were obtained and the slides were stained with HE.ResultsBefore treatment, the blood pressure of SHR groups were distinctly higher than that of the control group(P<0.01), nevertheless, no obvious blood pressure downgrade were observed after three week and five week treatment. Histopathologic study showed: in SHR group,heart with hypertrophic cardiac muscle, proliferative arterial wall and strictured lumina; Cortex with angiostenosis, proliferative vascular wall and large perivascular space; Glomerulus atrophy with hyaline degeneration. These pathologic changes got respective alleviation after five week treatment of JNQ, particularly in higher dose group.ConclusionSHR who got five week treatment of JNQ didn't gain obvious blood pressure downgrade. But the treatment did good to their vital organs and can obviously alleviate hearts, brains and kidneys' pathologic changes.

Effect of atorvastatin on ventricular remodeling in spontaneous hypertension rats / 中国病理生理杂志

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-?B 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-?B 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-?B 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-?B and inhibiting myocardial chronic inflammation.

Study on the Property of TCM Syndrome of Spontaneous Hypertension Rats / 中医杂志

Objective To establish the disease-syndrome integrated animal models suitable for the studies of TCM through differen- tiating the property of TCM syndromes of spontaneous diseased animal models.Methods With the observation on general behaviors, irritable degree,turning endurance time,pain threshold,urine and stools,luster of hair,growing speed of hair,body weight,tongue condition,degree of eyeball protruding,conjunctiva chroma,blood pressure,heart rate,etc.of spontaneous hypertension rats(SHR) and the comparison with normal rats,the study was carried out on the macroscopic description of property of TCM syndrome of SHR (14～18 weeks of age)and their ethology.Results The blood pressure of SHR at the early stage tended to raise with age growing. Compared with the normal group,the heart rate of SHR rats was obviously quicker(P