Earthworm Protein Protects Vascular Endothelial Function in Spontaneously Hypertensive Rats via PI3K/Akt/Nrf2 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo observe the effect of earthworm protein on the expression of phosphatidylinositol 3-kinase/protein kinase B/nuclear factor E2-related factor 2 （PI3K/Akt/Nrf2） pathway in the aorta of spontaneously hypertensive rats （SHR） and explore mechanism of earthworm protein in treating hypertensive vascular endothelial dysfunction （VED）. MethodTen 10-week-old Wistar Kyoto （WKY） rats and fifty SHR rats were selected for a week of adaptive feeding. WKY rats were selected as the normal group， and fifty SHR rats were randomized according to body weight into model， valsartan （8×10-3 g·kg-1·d-1）， and high-， medium-， and low-dose （0.2， 0.1， 0.05 g·kg-1·d-1， respectively） earthworm protein groups. The normal and model groups were administrated with equal volume of double distilled water by gavage. During the drug intervention period， the general situations of rats in each group were observed and their blood pressure was monitored at specific time points every other week before and after administration. After 8 weeks of drug intervention， enzyme-linked immunosorbent assay was employed to measure the levels of angiotensin-Ⅱ （Ang-Ⅱ） and endothelin-1 （ET-1） in the serum of rats in each group. The corresponding kits were used to determine the levels of nitric oxide （NO）， malondialdehyde （MDA）， glutathione peroxidase （GPX）， superoxide dismutase （SOD）， and ferrous ion （Fe2+）. Hematoxylin-eosin （HE） staining was employed to observe the changes in the intima of the aorta. Fluorescence quantitative polymerase chain reaction （Real-time PCR） was employed to measure the mRNA levels of PI3K， Akt， Nrf2， heme oxygenase-1 （HO-1）， and glutathione peroxidase 4 （GPX4） in the aortic tissue. Western blotting was used to determine the protein levels of p-PI3K （Tyr467/199）， PI3K， p-Akt （Ser473）， Akt， Nrf2， HO-1， and GPX4 in the thoracic aorta. ResultCompared with the normal group， the model group had decreased body mass， increased irritability， severe endothelial damage， elevated blood pressure and serum levels of Ang-Ⅱ， ET1， MDA， and Fe2+ （P<0.01）， lowered NO level （P<0.01）， and down-regulated mRNA and protein levels of p-PI3K （Tyr467/199）， PI3K， p-Akt （Ser473）， Akt， Nrf2， HO-1， and GPX4 in the aortic tissue （P<0.01）. Compared with the model group， drug intervention caused no significant change in the body mass， calmed the rats， alleviated the endothelial damage， lowered blood pressure and serum levels of Ang-Ⅱ， ET1， MDA， and Fe2+ （P<0.01）， elevated the NO level （P<0.05）， and up-regulated the mRNA and protein levels of p-PI3K （Tyr467/199）， PI3K， p-Akt （Ser473）， Akt， Nrf2， HO-1， and GPX4 （P<0.05）. ConclusionThe earthworm protein can exert antihypertensive effects by ameliorating VED in SHR. Specifically， it may regulate the PI3K/Akt/Nrf2 signaling pathway to inhibit oxidative stress and ferroptosis.

Involvement of Interleukin-1 β/Insulin-Like Growth Factor 1 in Ameliorating Effects of Electroacupuncture on Myocardial Fibrosis Induced by Essential Hypertension / 中国结合医学杂志

OBJECTIVE@#To investigate the effect of electroacupuncture (EA) at Neiguan (PC 6) on myocardial fibrosis in spontaneously hypertensive rats (SHRs), and to explore the contribution of interleukin-1 β (IL-1 β), insulin-like growth factor 1 (IGF-1), and transforming growth factor β 1 (TGF- β 1) to the effects.@*METHODS@#Nine 12-weeks-old Wistar Kyoto (WKY) male rats were employed as the normal group. Twenty-seven SHRs were equally randomized into SHR, SHR+EA, and SHR + sham groups. EA was applied at bilateral PC 6 once a day 30 min per day in 8 consecutive weeks. After 8-weeks EA treatment at PC 6, histopathologic changes of collagen type I (Col I), collagen type 1 (Col 1) and the levels of IGF-1, 1L-1 β, TGF- β 1, matrix metalloproteinase (MMP)-2 and MMP-9 were examined in myocardial tissure respectively.@*RESULTS@#After 8-weeks EA treatment at PC 6, the enhanced myocardial fibrosis in SHRs were characterized by the increased mean fluorescence intensity of Col I and Col 1 in myocardium tissue (P<0.01). All these abnormal alterations above in SHR + EA group was significantly lower compared with the SHR group (P<0.01). Meanwhile, the increased levels of IL-1 β, IGF-1, TGF-β 1 in serum or myocardial tissue of SHRs, diminished MMP 9 mRNA expression in SHRs were also markedly inhibited after 8 weeks of EA treatment (P<0.05 or P<0.01). Furthermore, the contents of IL-1 β, IGF-1, TGF-β 1 in myocardial tissue were positively correlated with the systolic blood pressure and hydroxyproline respectively (P<0.01).@*CONCLUSION@#EA at bilateral PC 6 could ameliorate cardiac fibrosis in SHRs, which might be mediated by regulation of 1L-1 β/IGF-1-TGF- β 1-MMP9 pathway.

Effect of high-intensity interval training on the skeletal muscle of spontaneously hypertensive rats

Abstract Aim: This study aimed to evaluate the effect of High-Intensity Interval Training (HIIT) on the skeletal muscle of Spontaneously Hypertensive Rats (SHR). Method: In total, 20 male rats, SHR, 12 months old, were used, distributed into 2 groups: Control Group (C) and Training Group (HIIT). The training lasted approximately 50 minutes/day, 5 days/week, for 8 weeks. Systolic blood pressure (BP) was measured at the beginning and end of the study. Analysis: The medial gastrocnemius muscle was used to measure the smallest fiber diameter, after which the Shapiro-Wilk normality test was performed, followed by the Mann Whitney test to compare the medians and interquartile intervals (IQI) of the muscle fibers and Student t-test for performance. For analysis of BP, Analysis of Variance - ANOVA was used, followed by Tukey's post-test. All procedures adopted a significance value of 5% (p < 0.05). Results: The median values for the variable "smallest diameter" of muscle fibers were 29.48 (IQI: 9.96) µm in the C group and 33.45 (IQI: 9.44) µm in the HIIT group (p < 0.05). Also, the performance was increased in the trained animal group and blood pressure values decreased significantly at the end of the experiment (p < 0.05). Conclusion: The HIIT intensity promoted an increase in the median values of the muscle fibers and performance. Finally, a significant decrease was observed in blood pressure variation values.

Tyrosine hydroxylase and ß2-adrenergic receptor expression in leukocytes of spontaneously hypertensive rats: putative peripheral markers of central sympathetic activity

The sympathetic nervous system (SNS) plays a fundamental role in the pathophysiology of cardiovascular diseases, including primary arterial hypertension. In this study, we aimed to investigate whether the expression of the rate-limiting enzyme in catecholamine synthesis, tyrosine hydroxylase (TH), and the β2-adrenergic receptor (β2-AR) in immune cells from peripheral blood, reflect central SNS activity in spontaneously hypertensive rats (SHR). TH expression in the lower brainstem and adrenal glands and β2-AR expression in the lower brainstem were analyzed by western blot analyses. In the leukocytes, TH and β2-AR expression was evaluated by flow cytometry before and after chronic treatment with the centrally-acting sympathoinhibitory drug clonidine. Western blot analyses showed increased TH and β2-AR expression in the lower brainstem and increased TH in adrenal glands from SHR compared to normotensive Wistar Kyoto rats (WKY). Lower brainstem from SHR treated with clonidine presented reduced TH and β2-AR levels, and adrenal glands had decreased TH expression compared to SHR treated with vehicle. Flow cytometry showed that the percentage of leukocytes that express β2-AR is higher in SHR than in WKY. However, the percentage of leukocytes that expressed TH was higher in WKY than in SHR. Moreover, chronic treatment with clonidine normalized the levels of TH and β2-AR in leukocytes from SHR to similar levels of those of WKY. Our study demonstrated that the percentage of leukocytes expressing TH and β2-AR was altered in arterial hypertension and can be modulated by central sympathetic inhibition with clonidine treatment.

Losartan improves alveolar bone dynamics in normotensive rats but not in hypertensive rats

Abstract Hypertension is one of the main causes of premature death in the world; also, it is associated with several bone alterations. Preclinical studies have demonstrated delayed alveolar bone healing in hypertensive rats. However, losartan has been favorable for consolidation of bone grafts and reduction in active periodontitis. Therefore, losartan is suggested to be effective in bone formation stages, as well as in the synthesis of matrix proteins and mineralization. Objectives: To evaluate the alveolar bone dynamics in hypertensive rats treated with losartan by laser confocal microscopy and histological analysis. Methodology: Thirty-two rats, 16 spontaneously hypertensive rats (SHR) and 16 Wistar albinus rats, treated or not with losartan (30 mg/kg/day) were used. Calcein fluorochrome at 21 days and alizarin red fluorochrome at 49 days were injected in rats (both 20 mg/kg). The animals were submitted to euthanasia 67 days after treatment, and then the right maxilla was removed for laser confocal microscopy analysis and the left maxilla for histological analysis. Results: This study showed a greater calcium marking in normotensive animals treated with losartan in relation to the other groups. Laser confocal microscopy parameters showed higher values of bone volume formed, mineralized surface, active surface of mineralization and bone formation rate in normotensive animals treated with losartan. However, a smaller mineralized surface was observed in all hypertensive animals. Conclusion: Losartan can improve bone mineralization parameters under normal physiological conditions, but the same anabolic effect does not occur under hypertension.

Icariside II improves left ventricular function by improving endoplasmic reticulum stress and caspase 12 signaling in SHR / 中国药理学通报

Aim: To explore the mechanism of icariside II (ICS II) on improving left ventricular function based on endoplasmic reticulum stress and caspase-12 signaling in spontaneously hypertensive rats (SHRs). Methods: Thirty 14-week-old male SHRs were divided into model group, ICS II low, middle and high dose groups and positive drug group (n = 6). WKY was used as control group (n =6). ICS II groups were respectively given ICS 114, 8, 16 mg · kg-1(ig, qd), and positive drug group was given losartan (20 mg · kg-1). At the end of 26th weeks, anesthetized rats were measured by ultrasound for detection of the left ventricular function, RT-PCR was used to determine the level of GRP78 mR- NA in the left ventricle tissue, and Western blot was used to assess the levels of GRP78 and cleaved-caspase- 12/9/3 protein in the left ventricle tissues. Results: Compared with WKY group, the internal diameter and posterior wall thickness of the left ventricular end diastolic increased, while the ejection fraction and fractional shortening decreased in SHR group. GRP78 mRNA and protein levels were up-regulated, and the levels of cleaved-caspase-12/9/3 protein were raised in left ventricle (P < 0.05). Compared with SHR group, the internal diameter and posterior wall thickness of the left ventricular end diastolic increased in ICS II medium and high dose groups and positive drug group (P <0. 05), while the ejection fraction and fractional shortening decreased (P<0.05). GRP78 mRNA and protein levels were down-regulated, the levels of cleaved-caspase-12/ 9/3 protein declined in left ventricle (P <0.05). Conclusions: ICS II could improve left ventricular function in SHRs, and its mechanism may be related to improving left ventricular endoplasmic reticulum stress and down-regulating the elevated caspase-12 signaling.

Effect of Mangiferin on Morphology of Relevant Vessels and MCP-1/CCR-2 Pathway in Spontaneously Hypertensive Rats / 中国实验方剂学杂志

Objective:To observe the morphological changes of carotid artery, thoracic aorta and superior mesenteric artery in spontaneously hypertensive rats(SHR), in order to further study the effect of Mangiferin on the expressions of inflammatory factors and monocyte chemoattract protein-1 (MCP-1)/c-chemokine receptor type 2 (CCR-2) pathway in SHR. Method:Forty spontaneously hypertensive rats were randomly divided into model group, benazepril group (10 mg·kg-1·d-1) and low, medium and high-dose mangiferin groups (25, 50, 100 mg·kg-1·d-1). Eight male WKY rats of the same age were selected as normal control group. Systolic blood pressure was observed every two weeks after eight weeks of administration. Morphology of carotid artery, thoracic aorta and superior mesenteric artery was observed by hematoxylin-eosin (HE) staining. Immunohistochemical assay (IHC) and Western blot were used to detect MCP-1 and CCR-2 protein expressions in thoracic aorta. MCP-1 and CCR-2 mRNA expression levels in thoracic aorta were detected by Real-time quantitative fluorescence PCR (Real-time PCR). Result:Compared with the normal group, the inflammatory cells in the model group increased significantly, the systolic blood pressure was significantly higher than that in the WKY group (PPPPConclusion:There are inflammation damages in carotid artery, thoracic aorta and superior mesenteric artery of spontaneously hypertensive rats. Mangiferin has an anti-inflammatory effect by possibly inhibiting the expressions of MCP-1/CCR-2 pathway in SHR vessels.

Changes of Bax, Bcl-2, CCR-2, MCP-1, and TGF-β1 genes in the left ventricle of spontaneously hypertensive rat after losartan treatment / 소아과

PURPOSE: Increased apoptosis was recently found in the hypertrophied left ventricle of spontaneously hypertensive rats (SHRs). Although the available evidence suggests that apoptosis can be induced in cardiac cells by various insults including pressure overload, cardiac apoptosis appears to result from an exaggerated local production of angiotensin in adult SHRs. Altered expressions of Bcl associated X (Bax), Bcl-2, chemokine receptor (CCR)-2, monocyte chemoattractant protein (MCP)-1, transforming growth factor (TGF)-β1, phosphorylated extracellular signal-regulated kinases (PERK), and connexin 43 proteins, and kallikrein mRNA were investigated to explore the effects of losartan on the SHR model. METHODS: Twelve-week-old male rats were grouped as follows: control (C), SHR (hypertension: H), and losartan (L; SHRs were treated with losartan [10 mg/kg/day] for 5 weeks). Western blot and reverse transcription polymerase chain reaction assays were performed. RESULTS: Expression of Bax, CCR-2, MCP-1, TGF-β1, PERK, and connexin 43 proteins, and kallikrein mRNA was significantly increased in the H group compared to that in the C group at weeks 3 and 5. Expression of Bax, CCR-2, MCP-1, TGF-β1, and connexin 43 proteins and kallikrein mRNA was significantly decreased after losartan treatment at week 5. PERK protein expression was significantly decreased after losartan treatment at weeks 3 and 5. Bcl-2 protein expression was significantly decreased in the H group compared to that in the C group at weeks 3 and 5. CONCLUSION: Losartan treatment reduced expression of Bax, CCR-2, MCP-1, TGF-β1, PERK, and connexin 43 proteins, and kallikrein mRNA in SHRs, along with decreased inflammation and apoptosis.

Effects and mechanism of total phenols of Magnolia officinalis combined with Maijunan Tablets on blood pressure of spontaneous hypertensive rats / 中草药·英文版

Objective: Maijunan (MJA) Tablets is a protected variety of traditional Chinese medicine (TCM) consisted of Pueraria lobata, hydrochlorothiazide (HTCZ), Uncaria rhynchophylla (366:1:980) and excipient. In the present work, MJA was consisted of the total flavones of P. lobata, HCTZ and total alkaloids of U. rhynchophylla (40:11:75). The combination of MJA and the total phenols of Magnolia officinalis (M-MJA) was consisted of the total flavones of P. lobata, the total phenols of M. officinalis, HCTZ and the total alkaloids of U. rhynchophylla (40:40:11:75). The aim of this work was to examine the effect and mechanism of M-MJA on the blood pressure of spontaneous hypertensive rats (SHRs). Methods: Adult male SHRs were randomly divided into control group, MJA group (180 mg/kg·d), and the M-MJA group (218 mg/kg·d) (n = 5). SHRs were orally administered with M-MJA and MJA respectively once a day for 8 weeks, the blood pressure of SHRs was measured every two weeks, and the biochemical indicators related to blood pressures were detected at the last dosing. Results: After oral administration of M-MJA to SHRs once a day for 8 weeks, the systolic and diastolic blood pressures of SHRs were deceased significantly. M-MJA affected renin-angiotensin-aldosterone system by decreasing the levels of Ren, Ang II and ALD, affected the endothelial function by decreasing the levels of ET-1 and 20-HETE, and increasing the level of eNOS, affected the oxidative stress by increasing the protein expression of Nrf2 and the activities of HO-1 and GSH-Px, and decreasing the protein expression of CYP2E1 and CYP4A, as well as the content of MDA. Conclusion: These results indicated that M-MJA could regulate the renin-angiotensin-aldosterone system, improve endothelial function, and inhibit CYP4A activity to reduce the production of 20-HETE, alleviate the oxidative stress disorder of the visceral organs, and eventually exert antihypertensive effect. Additionally, the anti-oxidant ability, regulating the renin-angiotensin-aldosterone system and improving endothelial function of M-MJA are more powerful than that of MJA, suggesting that M-MJA may have a better anti-hypertensive effect than MJA.

Captopril alleviates hypertension-induced renal damage, inflammation, and NF-κB activation

Hypertensive renal damage generally occurs during the middle and late stages of hypertension, which is typically characterized by proteinuria and renal inflammation. Captopril, an angiotensin-converting enzyme (ACE) inhibitor, has been widely used for therapy of arterial hypertension and cardiovascular diseases. However, the protective effects of captopril on hypertension-induced organ damage remain elusive. The present study was designed to explore the renoprotective action of captopril in spontaneously hypertensive rats (SHR). The 6-week-old male SHR and age-matched Wistar-Kyoto rats were randomized into long-term captopril-treated (34 mg/kg) and vehicle-treated groups. The results showed that in SHR there was obvious renal injury characterized by the increased levels of urine albumin, total protein, serum creatinine, blood urea nitrogen, renal inflammation manifested by the increased mRNA and protein expression of inflammatory factors including tumor necrosis factor-α, interleukin (IL)-1β, IL-6, and inducible nitric oxide synthase, and enhanced nuclear factor-κB (NF-κB) activation. Captopril treatment could lower blood pressure, improve renal injury, and suppress renal inflammation and NF-κB activation in SHR rats. In conclusion, captopril ameliorates renal injury and inflammation in SHR possibly via inactivation of NF-κB signaling.

Chiral Separation of Manidipine Enantiomers and the Pharmacodynamics of the Hydrochlorides / 中国药学杂志

OBJECTIVE: To investigate the chiral separation of the manidipine enantiomers by the proposed synthetic route, and the blood pressure effect of the hydrochlorides. METHODS: Based on the solubility differences of the salts which are the reaction products of the manidipine enantiomers and the different chiral resolution agents[(+)-Di-p-toluoyl-D-tartaric acid/(-)-Di-p-toluoyl-L-tartaric acid] in the specific solvents, the S and R manidipine enantiomers were obtained respectively, which is called the chemical salting out crystal method. The changes of systolic pressure, diastolic pressure, and mean arterial pressure of spontaneously hypertensive rats(SHR) before and after manidipine hydrochloride medications administration were monitored through implantable physiological signal remote sensing pressure monitoring system (data science international, DSI). RESULTS: The chiral separation process is simple, maneuverability is strong and the ee values and purity of the S and R enantiomorphism isomers are both higher in the synthetic route; in the SHR model, the antihypertensive effect was equivalent between manidipine hydrochloride group and S-manidipine hydrochloride group; R-manidipine hydrochloride did not have the antihypertensive effect. CONCLUSION: The chiral resolution of manidipine enantiomers has certain scientific value, which can improve the efficacy and safety of drug, and which is of great significance to develop manidipine single enantiomeric new drugs.

Effects of Acupuncture on the Level of Inflammatory Cytokines and Toll-like Receptor 4 in Hypothalamic Paraventricular Nucleus of Spontaneously Hypertensive Rats / 中国中医药信息杂志

Objective To observe the effects of acupuncture on the level of inflammatory cytokines and toll-like receptor 4 in hypothalamic paraventricular nucleus (PVN) of SHR; To investigate the mechanisms of acupuncture in reduction of mean arterial blood pressure (MAP) in SHR. Methods Thirty 10-week old SHR were randomly divided into SHR group, acupuncture group and non-acupoint group, with 10 rats in each group. 10 WKY rats were set as control group. Acupuncture group received bilateral acupuncture in "Taichong" acupoint, and twisting and diarrhea method was used to stimulate; non-acupoint group received acupuncture at the back of feet, and soothing and diarrhea with twisting method was used to stimulate. Materials were taken two weeks later. The mean arterial pressure of rats was detected every day; the expression of TLR4 mRNA in PVN was detected by RT-PCR; The expression of TLR4 protein in PVN was detected by Western blot; The levels of TNF-αa nd IL-6 were detected. Results Compared with control group, the mean arterial pressure of SHR group increased; TLR4 mRNA and protein expression in PVN increased significantly; the level of TNF-α and IL-6 increased significantly (P<0.05). Compared with SHR group, the mean arterial pressure of acupuncture group decreased significantly;TLR4 mRNA and protein expression in PVN decreased significantly; the level of TNF-α and IL-6 decreased significantly (P<0.05). There was no significant change in non-acupoint group. Conclusion Acupuncture in"Taichong" acupoint can attenuate blood pressure of SHR by inhibiting expression of TLR4 in PVN and reducing the levels of TNF-α and IL-6.

Effects of renal denervation on blood-pressure response to hemorrhagic shock in spontaneously hypertensive rats / 中华创伤杂志(英文版)

<p><b>PURPOSE</b>Renal denervation (RD) has been demonstrated to be an effective approach to reduce blood pressure for those with resistant hypertension. Yet, we aimed to explore the effect and possible mechanism of RD on blood-pressure response to hemorrhagic shock in spontaneously hypertensive rats.</p><p><b>METHODS</b>A total of 48 male spontaneously hypertensive rats were randomized to three groups: study group, sham-operation group and control group. RD was achieved by cutting off renal nerves and swabbing phenol on it. Ten weeks after RD, 8 rats in each group were sacrificed to collect the kidney and heart tissues. The remaining rats were subjected to an operation to induce hemorrhagic shock which would lead to 40% loss of total blood volume, and observed for 120 min. The serum concentration of norepinephrine was measured before and three weeks after RD.</p><p><b>RESULTS</b>The blood-pressure and norepinephrine levels were reduced significantly after RD (p < 0.05). Systolic blood pressure and diastolic blood pressure of the surgery group were higher than those in the sham and control groups at 15, 30 and 45 min after hemorrhagic shock (p < 0.05), while no significant difference was observed at 60, 90 and 120 min (p > 0.05). Additionally, the beta-1 adrenergic receptor (β1-AR) in the study group was significantly higher than those in the other two groups (p < 0.05) after hemorrhagic shock.</p><p><b>CONCLUSION</b>This study demonstrated that RD could to some extent improve blood-pressure response to hemorrhagic shock in an established model of severe hemorrhagic shock in spontaneously hypertensive rats. The mechanism might be associated with up-regulation of β1-AR.</p>

NMDAR in paraventricular nucleus mediates enhanced sympathetic activities caused by pro-inflammatory cytokines in spontaneously hypertensive rats / 中国比较医学杂志

Objective To investigate whether pro-inflammatory cytokines ( PICs) in the paraventricular nucleus ( PVN) regulate the enhanced sympathetic activities in spontaneously hypertensive rats ( SHR) , and whether N-methyl-Daspartate receptor ( NMDAR ) in PVN mediate the effects of PICs on sympathetic activities. Methods SHR and normotensive wistar-Kyoto( WKY) rats were used in this experiment. TNF receptor and IL-1β receptor ( IL-1RI) protein levels were measured by Western blot. PICs, including TNF-α and IL-1β levels were measured by ELISA. Rats were placed in a stereotaxic instrument to complete the microinjection of drugs. The coordinates for the PVN were determined according to the Paxinos and Watson rat atlas. The raw RSNA and integrated RSNA were simultaneously recorded on a PowerLab data acquisition system. The right carotid artery was cannulated for recording of mean arterial pressure ( MAP) . Results TNF-α receptor p55TNFR, p75TNFR and IL-1βreceptor IL-1RI protein expression and TNF-αand IL-1βlevels in PVN were all increased in SHR compared with WKY rats (P< 0. 05). Bilateral microinjection of etanercept or IL-1ra into PVN to block the effects of TNF-αor IL-1βdecreased the sympathetic activities in SHR rats significantly (P< 0. 05). Bilateral microinjection of NMDAR blockers, both DL-2amino-5-phosphonovaleri acid ( APV) and MK-801 ( Dizocilpine) into PVN decreased the RSNA and MAP in both SHR and WKY rats. APV or MK 801 caused greater decreases in RSNA and MAP in SHR than WKY rats. In addition, pretreatment with APV or MK 801 attenuated the increased RSNA and MAP caused by microinjection of TNF-αor IL-1βinto PVN to a lower level in SHR than in WKY rats (P< 0. 05). Conclusions TNF and IL-1βreceptor protein as well as TNF-αand IL-1βcytokines levels in PVN are all increased in SHR rats. NMDAR in PVN mediates enhanced sympathetic activities and elevated blood pressure caused by TNF-αand IL-1βin SHR.

Study on the Effect and Mechanism of Tandospirone Citrate on Blood Pressure in Spontaneously Hyperten-sive Rats / 中国药房

OBJECTIVE:To study the effect and mechanism of tandospirone citrate (TC) on blood pressure in spontaneously hypertensive rats (SHR). METHODS:SHR were divided into model group,positive control group (Levamlodipine besylate tab-lets,2.5 mg/kg),TC high-dose,medium-dose,low-dose groups(TC capsules,40,20,10 mg/kg),8 in each group. Other 8 nor-mal rats were chosen as normal control group. The rats were intragastrically administrated medicines,once a day,for 28 d. Systolic blood pressure(SBP)before first administration and after 0.5,1,1.5,2,3,4 h of administration in each group were measured, and SBP after 1 h of administration were measured once every 7 d. After last administration,nitric oxide(NO)content in serum, endothelin,renin,angiotensin Ⅱ(Ang Ⅱ) and norepinephrine (NE) contents in plasma were detected. RESULTS:In single ad-ministration,compared with model group,SBP in 0.5,1 h in positive control group and TC groups after first administration were obviously decreased (P<0.05 or P<0.01),then SBP were obviously decreased only in positive control group and TC high-dose group (P<0.05 or P<0.01). In multiple administration,compared with model group,SBP in 7th,14th,28th day of administra-tion in positive control group and TC high-dose,medium-dose groups were obviously decreased(P<0.05 or P<0.01). Compared with model group,NO content in serum in positive control group,TC high-dose group were obviously increased;endothelin,re-nin,Ang Ⅱ and NE contents in plasma were obviously decreased (P<0.05 or P<0.01). Compared with positive control group, NO content in serum in TC groups was obviously decreased;endothelin,renin,Ang Ⅱ and NE contents in plasma were obviously increased in TC medium-dose group (P<0.05). CONCLUSIONS:TC can obviously decrease the blood pressure of SHR. The mechanism may be associated with adjusting the balance of NO and endothelin,and decreasing renin,AngⅡand NE contents.

Screening of lentiviral vectors carrying effective siRNA targeting S1P2 gene / 中国病理生理杂志

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.

Changes in amino acid profiling in spontaneously hypertensive rats and regulatory effect of extract from Coreopsis tinctoria / 中国中药杂志

To compare the amino acid metabolic profiling in urine of spontaneously hypertensive rats (SHR) and normal Wistar rats, and investigate the regulatory effect of extract from Coreopsis tinctoria on blood pressure and amino acid metabolic profiling in SHR. Right aged SHR and Wistar rats were housed to fit the new environment for 2 weeks. After that, their systolic pressure(SBP), diastolic pressure(DBP) were measured and urine was collected. Amino acids profiles for SHR and Wistar rats were acquired by using AQC precolumn derivatization HPLC-fluorescence method, and then partial least squares discriminant analysis(PLS-DA) was applied to facilitate differentiation and determine metabolic differences between collected samples from two groups of rats. Consequently, 40 SHR were randomly divided into 5 groups: model group, high, middle, low dosage groups of C. tinctoria extract (3.2, 1.6,0.8 g•kg⁻¹), and captopril group (4 mg•kg⁻¹). They were treated for 4 weeks by ig administration, and then their urine samples were collected to determine the amino acid metabolic profiling in various groups. After treatment for 4 weeks, as compared with Wistar group, serine, alanine, tyrosine, and cystine in the amino acid metabolic profiling were significantly increased in SHR group. As compared with SHR model group, threonine and methionine were decreased significantly in captopril group (P<0.01); amino acid metabolism was changed to different degrees in high, middle, and low dosage groups of C. tinctoria extract, and the threonine in low dose group was significantly decreased (P<0.01); serine and threonine were decreased (P<0.05), and valine, methionine and lysine were significantly decreased (P<0.01) in middle dose group; threonine, valine, methionine and lysine were significantly decreased in large dose group (P<0.01). The results showed that middle and high doses of extract from C. tinctoria could significantly improve disturbance of amino acid metabolism, help to further clarify the drug property research of C. tinctoria, and provide data support for amino acid metabolic pathway abnormalities in hypertension patients.

Expression of Class I and Class II a/b Histone Deacetylase is Dysregulated in Hypertensive Animal Models

BACKGROUND AND OBJECTIVES: Dysregulation of histone deacetylase expression and enzymatic activity is associated with a number of diseases. It has been reported that protein levels of histone deacetylase (HDAC)1 and HDAC5 increase during human pulmonary hypertension, and that the enzymatic activity of HDAC6 is induced in a chronic hypertensive animal model. This study investigated the protein expression profiles of class I and II a/b HDACs in three systemic hypertension models. SUBJECTS AND METHODS: We used three different hypertensive animal models: (i) Wistar-Kyoto rats (n=8) and spontaneously hypertensive rats (SHR; n=8), (ii) mice infused with saline or angiotensin II to induce hypertension, via osmotic mini-pump for 2 weeks, and (iii) mice that were allowed to drink L-N(G)-nitro-L-arginine methyl ester (L-NAME) to induce hypertension. RESULTS: SHR showed high systolic, diastolic, and mean blood pressures. Similar increases in systolic blood pressure were observed in angiotensin II or L-NAME-induced hypertensive mice. In SHR, class IIa HDAC (HDAC4, 5, and 7) and class IIb HDAC (HDAC6 and 10) protein expression were significantly increased. In addition, a HDAC3 protein expression was induced in SHR. However, in L-NAME mice, class IIa HDAC protein levels (HDAC4, 5, 7, and 9) were significantly reduced. HDAC8 protein levels were significantly reduced both in angiotensin II mice and in SHR. CONCLUSION: These results indicate that dysregulation of class I and class II HDAC protein is closely associated with chronic hypertension.

Effect of Compound Yibazhen Granules on Early Renal Injury in Spontaneous Hypertensive Rats / 医药导报

Objective To observe the antihypertensive effect of compound Yibazhen granules on spontaneous hypertensive rats ( SHR) . Methods Wistar rats were served as normal control group. Sixty SHR were randomly divided into model control group,captopril group,Jane chrysanthemum antihypertension tablet group and compound high dose group,middle dose group and low dose group ( n = 10 each group ) by digital table method. Captopril group was given captopril 30 mg.kg-1 .d-1 ,and Jane chrysanthemum antihypertension tablet group was treated with Jane chrysanthemum antihypertension tablet ( 0. 6 tablet per kg ) , compound Yibazhen granules high dose group, middle dose group and low dose group received compound of 13.18,6.59 and 3.3 mg.kg-1 .d-1 ,respectively. Normal control group and model control group were intragastrically administered with 0.9% sodium chloride solution for 8 weeks. Changes of systolic and diastolic blood pressure of rats and blood urea,creatinine,nitric oxide (NO),nitric oxide synthase (NOS) and angiotensin Ⅱ (Ang Ⅱ) were observed. Results Diastolic pressure of rats in compound Yibazhen granules high dose group, middle dose group and low dose group decreased significantly in 2 weeks. Systolic blood pressure and diastolic blood pressure of compound Yibazhen granules high dose group decreased significantly in 4 weeks,compared with the model control group (P<0.05). Compared with the model control group, concentration of urea and crea in compound Yibazhen granules high dose group, middle dose group and low dose group were significantly lower( P<0.05) . The content of NOS and AngⅡ in rats of compound Yibazhen granules high dose group decreased significantly and the contents of NO increased, which were compared with the model control group ( P<0. 05, P<0. 01 ) . Conclusion The protective effect of compound Yibazhen granules in treating early renal damage in SHR is related to decreasing diastolic blood pressure,concentration of urea,crea and AngⅡ and regulating the levels of NOS and NO.

Effect of Shunaoxin Dropping Pill on blood pressure and cardiac protection in spontaneously hypertensive rats / 中草药

Objective: To evaluate the effect of Shunaoxin Dropping Pill (SDP) on blood pressure and cardiac protection in spontaneously hypertensive rats (SHRs). Methods: The SHRs were randomly divided into three groups, model (M) group, SDP group, and valsartan (VL) group. Non-invasive blood pressure instrument was used to measure blood pressure. Serum content of nitric oxide synthase (NOS) and angiotensin II (Ang II) were measured using kits for 6 weeks after ig administration. Doppler echocardiography was used to evaluate left ventricular weight, and HE staining was used to assess aortic and cardiac morphology. Results: The blood pressure of SHRs decreased to minimum 1 h after ig administration of SDP (800 mg/kg). Long-term use of SDP could stabilize blood pressure of SHRs. Serum content of NOS in SDP group was significantly more than that in M group (P < 0.05). The serum content of Ang II in SDP group was significantly less than that in M group (P < 0.05). The left ventricular weight of SDP group was significantly lower than that in M group (P < 0.05). SDP administered group can improve myocardial and aortic morphology abnormalities caused by hypertension. Conclusion: SDP has a certain therapeutic effect on hypertensive with myocardial hypertrophy rats, it can be used as a potential drug on hypertensive disorders associated with cardiac hypertrophy.