EFFECT OF ANGIOTENSIN II RECEPTOR ANTAGONIST AND ENDOTHELIN RECEPTOR ANTAGONIST ON NITROGLYCERIN TOLERANCE IN RATS

INTRODUCTION Nitroglycerin (Nit) and other nitrovasodilators have been widely used to treat angina pectoris and congestive heart failure. The vasorelaxing effect of Nit is mediated by elevating intracellular cGMP levels, which results from the activation of soluble guanylate cyclase (sGC) by the active intermediate metabolite of Nit— nitric oxide (NO) in vascular smooth muscle cells(1).However, during the long- term or chronic administration,the hemodynamic and anti- ischemic efficacy…     

TGF-β1-induced LPP expression dependant on Rho kinase during differentiation and migration of bone marrow-derived smooth muscle progenitor cells

Lipoma preferred partner(LPP) has been identified as a protein which is highly selective for smooth muscle progenitor cells(SMPCs) and regulates differentiation and migration of SMPCs,but mechanisms of LPP expression are not elucidated clearly.The aim of the present study was to discuss the mechanisms by which LPP expression is regulated in the differentiation and migration of SMPCs induced by TGF-β1.It was found that TGF-β1 could significantly increase the expression of LPP,smooth muscle α-actin,smooth muscle myosin heavy chain(SM-MHC),and smoothelin in SMPCs.Moreover,inactivation of Rho kinase(ROK) with ROK inhibitors significantly inhibited LPP mRNA expression in TGF-β1-treated SMPCs and mouse aortic smooth muscle cells(MAoSMCs).At the same time,LPP silencing with short interfering RNA significantly decreased SMPCs migration.In conclusion,LPP appears to be a ROK-dependant SMPCs differentiation marker that plays a role in regulating SMPCs migration.     

Effects of Atractylodes Macrocephala on the Cytomembrane Ca^2＋-activated K^＋ Currents in Cells of Human Pregnant Myometrial Smooth Muscles

The study examined the inhibitory effect of Atractylodes macrocephala(AM) on the uterine contraction during premature delivery and explored its electrophysiological mechanism by studying the effects of AM on the Ca2 -activated K currents of pregnant human myometrial smooth muscle cells with or without the treatment with interleukin-6.Single cells were acutely isolated from pregnant human myometrial smooth muscles.Whole-cell Ca2 -activated K currents were recorded by using an Axopatch1-D amplifier.The cells were divided into three groups: group A in which AM was added into perfusate,group B,in which interleukin-6 was added into perfusate) and group C in which AM was added into perfusate after addition of interleukin-6.IL-6 10 ng/mL inhibited BKca by 36.9%±13.7% as compared with control(P<0.01).AM at 2 mg/mL raised BKca by 36.7%±22.6% or 45.2%±13.7% with or without the treatment of IL-6,respectively(P<0.01).It is concluded that AM was able to enhance the BKca of pregnant human myometrial smooth muscle cells treated or un-treated with interleukin-6 and its effect on the BKca IL-treated cells was stronger that its effect on BKca of untreated cells.Our results suggested that AM can help to maintain the membrane potentials and the resting status of pregnant human myometrial smooth muscle cells.     

Lovastatin increases nitric oxide synthesis in IL- 1β- stimulated smooth muscle cells

Objective Nitric oxide (NO) production by inducible NO synthase (iNOS) may play an importa nt role in the pathogenesis of atherosclerosis. Although lovastatin has been s hown to reduce the progression of atherosclerosis, it is not known whether it re gulates NO production. We investigated the effects of lovastatin on NO synthesi s and the mechanisms by which lovastatin exerts its effects in rat vascular smoo th muscle cells. Methods Primary cultures of the vascular smooth muscle cells were obtained from the medi a of the thoracic aorta of Sprague Dawley rats (200-250 g). Nitrite levels in the culture medium of rat vascular smooth muscle cells were determined colorimet rically. Results Lovastatin (10(-5) mol/L) significantly increased interleukin- 1β (IL- 1β , 10 ng/mL)- induced nitrite accumulation in a time (0-24 hours)- dependent man ner. Exogenous mevalonate and geranylgeranyl- pyrophosphate completely reverse d the stimulatory effects of lovastatin on nitrite production. Furthermore, in hibition of Rho by C3 exoenzyme mimicked the increase in IL- 1β- induced nitrit e accumulation induced by lovastatin in the vascular smooth muscle cells. Conclusion These results demonstrate that lovastatin up- regulates NO formation in rat vasc ular smooth muscle cells stimulated by IL- 1β, and the effect may be associated with the inhibition of Rho activity.     

Effect of Berberine on the mRNA Expression of Nitric Oxide Synthase （NOS） in Rat Corpus Cavernosum

Penile erection is a complex neurovascularprocess that involves relaxation of the corpus cav ernosum smooth muscle[1]. Relaxation of cavern osal smooth muscle leads to engorgement of bloodin the corpus cavernosum that results in an in crease in intracavernosal pressure and thus in tu mescence. Many studies have shown that nitric ox ide (NO) released from cavernous nerves and endo thelium is a key factor for penile erection. TheNO cGMP signaling pathway is…     

Inhibitory Effect of Sodium Ferulate on the Cardiomyocyte Hypertrophy Induced by AngiotensionⅡ

Objective:To investigate the effects of sodium ferulate(SF) on myocardial hypertrophy of rat and explore the protective mechanism. Methods:The myocardial hypertrophy was induced by 0.1 μmol·L~(-1) Ang II. The cytoactive was detected by MTT. The cultured cardiomyocytes from Sprague Dawley neonate rats were randomly divided into normal,model,L-arginine(L-arg 1 000 μmol·L~(-1)) group and SF(50,100,200 μmol·L~(-1)) group.To observe whether SF had nonspecific injurious effect on the cells,SF 200 μmol·L~(-1) was added into the normal cardiomyocytes and to determine whether the effect of SF on cardiomyocyte hypertrophy was associated with NO release,another two groups were established. NG-nitro-L-arginine-methyl ester(L-NAME) 1 500 μmol·L~(-1) combined with SF 200 μmol·L~(-1) or L-arg 1 000 μmol·L~(-1),respectively. Cardiomyocyte hypertrophy was confirmed by observing the histological changes and the measurements of cell diameter,protein content and ANF and β-MHC m RNA expression of the cells.The levels of NO,NOS and eNOS activity,the contents of cGMP and cAMP. The expression of eNOS were detected by Real time PCR and Western blotting. Results:(1) SF(50,100,200 μmol·L~(-1)) had no obvious side effect on cultured neonatal rat cardiomyocytes in vitro. In the group added 0.1 μmol·L~(-1) Ang Ⅱ alone,the cells displayed swollen,with undistinguishable border;the diameter and protein content of cardiomyocytes was increased remarkably,and the expression of ANF and β-MHC m RNA were up-regulated by Ang Ⅱ. SF and L-arg could ameliorate the cardiomyocyte hypertrophy which can be inhibit by L-NAME.(2) Compared with normal group,0.1 μmol · L~(-1) Ang II could decrease the NO content,NOS and eNOS activity in supernatant of cultured cardiomyocytes,decrease the content of cGMP and increase the content of cAMP incardiomyocytes,upregulation the expression of eNOS.SF-H and L-arg administrated could siginificantly ameliorate these changes. Conclusion:SF can inhibit cardiomyocyte hypertrophy induced Ang Ⅱ in rats. The probable mechanism involved to promote NO-cGMP signaling pathway and up-regulate the expression of eNOS.     

Ultrastructural study of cerebral arteries in Moyamoya disease.

The cerebral arteries in seven cases of Moyamoya disease, demonstrated by angiography were observed by light, transmission and scanning electron microscopy. In two cases, the lumen of the terminal portion of the internal carotid artery was completely occluded by proliferated connective tissue. The characteristic pathologic change was extensive destruction of the smooth muscle cells and the elastica interna. Degeneration and necrosis of smooth muscle cells were obvious in the media. Many condensed organelles and other elements of cellular destruction were dispersed throughout the interstitium. The media was thinned. The proliferated smooth muscle cells in the intima showed the same destructive changes as in the media. The elastica interna was excessively infolded and was thickened or thinned, disrupted or disintegrated in some segments. Identical, but less severe and more localized changes were seen in three middle cerebral arteries (M2 and M5 segments) and two collateral arteries at the base of brain. These changes suggest that Moyamoya disease is a progressive disorder characterized by repeated destruction of smooth muscle cells occurRing over a long time period. The pathogenesis of these changes is discussed.

     

Studies on Relationship between Serum Nitric Oxide and Plasma Cyclic Guanosine Monophosphate and Prolonged Bleeding after Medical Abortion as well as Prophylaxis and Treatment of Bleeding with Traditional Chinese Medicine

Objectives To study the relationship between serum nitric oxide(NO and plasma cyclic guanosine monophosphate(cGMP)and prolonged bleeding after medical abortion.Methods A total of 120women having received medical abortions at random were recruited and divided into two groups:the one(Group A,n=60) taking “Gong-Fu Mixture(Uterus-Recovering Mixture)”and the other(Group B,n=60)not taking it after abortion.On d 10,20 and 30 after medical abortion,serum NO and plasma cGMP were tested before and after mifepristone administration and 10 d later by Gresis reac-tion method and radioimmunoassay respectively.Results NO concentration in serum and cGMP concentration in plasma decreased signifi-cantly after taking mifeprlstone given(P＜0. 05).Ten days later,the number of thos ewith bleeding discontinuation in the group A was significantly greater than that in the group B(P＜0.05).Serum NO level and plasma cGMP level in the group A de-creased more significantly than those in the group B(P＜0. 05).Conclusion The slow decrease of serum NO and plasma cGMP is closely related to prolonged bleeding after medical abortion.“Gong-Fu Mixture(uterus-recovering mixture)”is effective in prevention and treatment of prolonged bleeding.     

Effects of cholesterol liposomes on cytoskeleton and proliferation of rabbit sphincter of Oddi cells in culture

Objective: To discuss the relationship between hypercholesterolemic disease and the functional and structural changes of Sphincter of Oddi (SO) by the study of effect of Cholesterol Liposome (CL) on structural and quantitative changes of SO cells. Methods: Rabbit SO was isolated for primary cell culture and subculture. After subcultured with different concentration of CL culture medium for 20 h, the structural and quantitative changes of SO cells were analyzed and detected by MTT-test, flow cytometer (FCM), electronic microscope and electrophoresis technique respectively. Results: CL contributed a prominent stimulus to SO cells proliferation at middle concentration (<0. 5 - 0. 8 mg/ml), which could be confirmed by FCM analysis which indicated the number of SO cells in S-phase increasing remarkably; however, high concentration of CL inhibited SO cells' proliferation (>1. 0 mg/ml) and induced apoptosis of SO cells. Swelled mitochondria and dilated endoplasmic reticulum as well as disjoined and diminished     

Adenovirus mediated gene transfer of vascular smooth muscle cells and endothelial cells in vitro.

Introducing foreign gene(s) into vascular smooth muscle cells (VSMCs) and endothelial cells (ECs) is the pre-require-ment of gene therapy for cardiovascular diseases. We have explored the use of adenoviral vectors (Adv-CMV / LacZ) to transfer LacZ gene into cultured VSMCs and ECs. Our results demonstrated that adenoviral vectors transfered foreign gene into VSMCs and ECs high-efficiently with dose-dependent response pattern. The frequencies of transfection reached 100% at the viral titer of 109 pfu / ml. Comparing the sensitivities of VSMCs and ECs to adenoviral vectors, we found that ECs were more sensitive than VSMCs, of which the frequencies of transfection in ECs reached 80% while in VSMCs only 40% for 8 hrs after transfection. In addition, the transfection of ECs and VSMCs with adenoviral vectors was partly blocked by monoclonal antibodies to Fiber and Core protein of the adenoviral capsid, but not by monoclonal antibody to Hcxon protein. It is suggested transfection of ECs and VSMCs with adenovirus     

磷酸二酯酶同工酶PDE5的纯化及性质研究

目的 :为了研究磷酸二酯酶 (phosphodiesterase ,PDE ,EC 3 ,1,4,17)同工酶的性质及其临床意义。方法 :我们采用快速蛋白液相层析系统 (fastproteinliquidchromatography ,FPLC)分离纯化PDE5。结果 :从牛阴茎海绵体组织和海绵体血管中均得到 3个蛋白峰 ,它们分别是PDE5、PDE2和PDE3。PDE5是哺乳类海绵体组织中发现的主要PDE同工酶 ,在海绵体血管含量尤为丰富。结论 :PDE5作为药物治疗阳痿症的靶点日益引人注目。为进一步研究PDE5及其抑制剂作用机制打下了基础。     

Effects of buried penis on the structure and function of corpus cavernosum in a rat model

Background While the abnormal appearance of the concealed penis has been well recognized, the effect of buried penis on the structure and function of corpus cavernosum has not been well studied. To explore this issue, we established a rat model and evaluated the effect of buried penis on cavernosum weight, contents and ultrastructure of tissue, and nitric oxide synthase (NOS) activity.Methods Two hundred and ten rats were randomly divided into 3 equal cohorts for 2, 4 and 6 months study (groups A, B and C). Each group was randomly divided into buried group (n=40), control group (n=15), and normal group (n=15), respectively. Intra-purse-string suture of the root of the penis was used to establish the model. Macroscopic development was judged by measuring the weight of the corpus cavernosum. Masson's trichrome staining was performed for observing microstructure while a transmission electron microscope was used for observing ultrastructure. The NOS activity was detected by a NOS activity assay kit.Results Buried penis had no significant influence on the appearance and weight of the corpus cavemosum. Buried penis resulted in decreased smooth muscle content (P>0.05 in group A, and P<0.05 in groups B and C) and increased fibrous connective tissue content (P >0.05 in groups A and B, and P <0.05 in group C) compared with the normal and control groups. Ultrastructural abnormalities of corpus cavernosum were observed in the 6-month buried group. Moreover, there was decrease of NOS activity in groups B and C (P<0.05 in group B and P<0.01 in group C) when compared with the normal and control groups.Conclusion Buried penis affects the structure and function of corpus cavemosum in rats and the effect is positively correlated with the buried time, but there is no significant effect on the macroscopic development.     

Effects of buried penis on the structure and function of corpus cavernosum in a rat model

Background While the abnormal appearance of the concealed penis has been well recognized, the effect of buried penis on the structure and function of corpus cavernosum has not been well studied. To explore this issue, we established a rat model and evaluated the effect of buried penis on cavernosum weight, contents and ultrastructure of tissue, and nitric oxide synthase (NOS) activity.Methods Two hundred and ten rats were randomly divided into 3 equal cohorts for 2, 4 and 6 months study (groups A, B and C). Each group was randomly divided into buried group (n=40), control group (n=15), and normal group (n=15), respectively. Intra-purse-string suture of the root of the penis was used to establish the model. Macroscopic development was judged by measuring the weight of the corpus cavernosum. Masson's trichrome staining was performed for observing microstructure while a transmission electron microscope was used for observing ultrastructure. The NOS activity was detected by a NOS activity assay kit.Results Buried penis had no significant influence on the appearance and weight of the corpus cavemosum. Buried penis resulted in decreased smooth muscle content (P>0.05 in group A, and P<0.05 in groups B and C) and increased fibrous connective tissue content (P >0.05 in groups A and B, and P <0.05 in group C) compared with the normal and control groups. Ultrastructural abnormalities of corpus cavernosum were observed in the 6-month buried group. Moreover, there was decrease of NOS activity in groups B and C (P<0.05 in group B and P<0.01 in group C) when compared with the normal and control groups.Conclusion Buried penis affects the structure and function of corpus cavemosum in rats and the effect is positively correlated with the buried time, but there is no significant effect on the macroscopic development.     

西地那非的作用机制

西地那非(俗名伟哥)是一种治疗阴茎勃起功能障碍的口服药物,其疗效约为70%～80%。阴茎勃起功能障碍的主要原因是阴茎海绵体供血不足,可能与缺少一氧化氮(NO)有关。NO可使鸟苷酸环化酶(GC)活化,在活化的GC作用下,鸟苷酸(GTP)转变成环鸟苷酸(cGMP);cGMP可被磷酸二脂酶(PDE)降解为5′-cGMP,后者无生物学效应,而cGMP则可引起阴茎海绵体平滑肌血管舒张,使阴茎迅速充血而勃起。研究表明,西地那非可抑制PDE的作用,使cGMP聚集,从而发挥其治疗作用。然而,目前也有关于用药不当致死的病例,美国已有69例(老年人占90%以上)因服用西地那非而致死的报告。本文仅就西地那非药理作用机制作一简要介绍。     

致敏后重复抗原攻击的大鼠肺组织环核苷酸磷酸二酯酶活性的变化

目的 :探讨致敏后重复抗原攻击大鼠肺组织 c GMP- PDE、c AMP- PDE活性变化。方法 :HPL C法测定致敏后重复抗原攻击的大鼠 (抗原攻击组 )、致敏不攻击的大鼠 (致敏组 )和正常大鼠 (正常组 )肺组织 c GMP- PDE、c AMP- PDE活性。结果 :抗原攻击组肺组织 c GMP- PDE、c AMP- PDE活性分别为 798.76± 5 2 9.2 3、5 97.5 0± 2 2 0 .75nmol·g-1protein·min-1,其中抗原攻击组大鼠肺组织 c GMP- PDE活性较正常组明显升高 (P<0 .0 5 ) ,抗原攻击组大鼠肺组织 c AMP- PDE活性较致敏组和正常组均明显升高 (P<0 .0 5 )。结论 :大鼠哮喘发病过程中肺组织c GMP- PDE、c AMP- PDE活性明显升高     

Effects of buried penis on the structure and function of corpus cavernosum in a rat model

Background While the abnormal appearance of the concealed penis has been well recognized, the effect of buried penis on the structure and function of corpus cavernosum has not been well studied. To explore this issue, we established a rat model and evaluated the effect of buried penis on cavernosum weight, contents and ultrastructure of tissue, and nitric oxide synthase （NOS） activity. Methods Two hundred and ten rats were randomly divided into 3 equal cohorts for 2, 4 and 6 months study （groups A, B and C）. Each group was randomly divided into buried group （n=40）, control group （n=15）, and normal group （n=15）, respectively. Intra-purse-string suture of the root of the penis was used to establish the model. Macroscopic development was judged by measuring the weight of the corpus cavernosum. Masson＇s trichrome staining was performed for observing microstructure while a transmission electron microscope was used for observing ultrastructure. The NOS activity was detected by a NOS activity assay kit. Results Buried penis had no significant influence on the appearance and weight of the corpus cavernosum. Buried penis resulted in decreased smooth muscle content （P 〉0.05 in group A, and P 〈0.05 in groups B and C） and increased fibrous connective tissue content （P 〉0.05 in groups A and B, and P 〈0.05 in group C） compared with the normal and control groups. Ultrastructural abnormalities of corpus cavernosum were observed in the 6-month buried group. Moreover, there was decrease of NOS activity in groups B and C （P 〈0.05 in group B and P 〈0.01 in group C） when compared with the normal and control groups. Conclusion Buried penis affects the structure and function of corpus cavernosum in rats and the effect is positively correlated with the buried time, but there is no significant effect on the macroscopic development.     

淫羊藿苷对动脉性ED模型大鼠阴茎海绵体压力和一氧化氮合酶亚型表达的影响

目的　建立动脉性勃起功能障碍大鼠模型 (arteriogenicerectiledysfunction ,A -ED) ,研究淫羊藿苷 (Icariin)对A -ED大鼠勃起功能和一氧化氮合酶 (NOS)三种亚型表达的影响。方法　 4 0只Wistar大鼠随机分为假手术组 (A组 )、A -ED组 (B组 )、A -ED低剂量淫羊藿苷治疗组 (C组 )和AED高剂量淫羊藿苷治疗组 (D组 )。结扎大鼠双侧髂内动脉建立A -ED动物模型后 ,C组和D组每天口饲不同剂量的淫羊藿苷 ( 5mg/kg ,10mg/k) ,A组和B组口饲等体积的生理盐水。 30d后 ,通过检测各组大鼠阴茎海绵体压力 (ICP)变化评估淫羊藿苷对大鼠勃起功能的效果 ,切取阴茎海绵体组织 ,通过逆转录 -PCR和Western印迹方法检测N0S亚型的表达变化。结果　建立A -ED模型 30d后 ,B组ICP较手术前显著降低 ,N0S三种亚型的表达水平也显著降低。经淫羊藿苷治疗后 ,C组和D组的ICP比B组明显升高 (P <0 .0 1) ,且D组较C组增高效果明显 ;C组和D组的eNOS表达水平较之B组有所增高 ,与ICP的变化有相同的趋势。C组的iNOS表达水平有一定程度的升高。治疗后nNOS表达水平没有明显增高。结论　经淫羊藿苷治疗后大鼠勃起功能显著改善 ,这种功能改善与eN0S的表达增加有相同的趋势 ,提示淫羊藿苷可能通过恢复eN0s的表达来改善A -ED模型的勃起功能。     

阿霉素诱发肾性水肿病症大鼠模型的建立

目的考察复制大鼠的肾性水肿(肾阴虚、肾阳虚水肿)病症结合模型的可行性。方法采用灌胃甲状腺素片及尾静脉注射阿霉素的方法制造大鼠肾阴虚水肿模型,肌注氢化可的松和尾静脉注射阿霉素方法制造大鼠肾阳虚水肿模型。通过观察模型大鼠体征变化、24 h尿蛋白含量、三碘甲状腺原氨酸(T3)、甲状腺(T4)、环磷酸腺苷(c AMP)、环磷酸鸟苷(c GMP)、雌二醇(E2)及睾酮(T)等指标含量,判断模型是否成功。结果阴虚水肿组动物出现多动、脱毛、大便干结、体重降低和体温升高等临床表现,与空白组比较T3、T4、c AMP、E2、24h尿蛋白含量明显升高,c GMP、T含量降低,c AMP/c GMP比值显著升高,且各项指标均差异具有显著性(P0.05)。阳虚水肿组动物则出现活动减少、大便溏泻、体重和体温下降等体征变化,与空白组相比,T3、T4、c AMP、E2含量显著降低,c GMP、T、24h尿蛋白含量明显升高,c AMP/c GMP比值降低,且各项指标均差异具有显著性(P0.05)。结论分别由甲状腺素片联合阿霉素、氢化可的松注射液联合阿霉素诱导的动物病症结合模型,符合中医证型诊断下肾阴虚、阳虚水肿病症,造模方法可行。     

PDE5抑制剂与黑色素瘤风险的相关性研究进展

5型磷酸二酯酶抑制剂（PDE5抑制剂）是治疗勃起功能障碍的首选药物,其通过选择性抑制PDE5的活性,减少cGMP的降解,扩张阴茎海绵体血管,从而维持男性勃起反应。目前,部分基础研究表明PDE5抑制剂的作用机制可能会影响皮肤细胞的信号通路,导致患者黑色素瘤的风险增加。此外,较多临床研究也对PDE5抑制剂和黑色素瘤发病风险的相关性进行了评估,但目前尚未得出明确一致的结论。本文就上述研究进展进行综述。