低雄激素通过抑制大鼠阴茎海绵体组织Tie2的表达抑制勃起功能

目的:研究雄激素是否通过富含内皮的外膜内皮细胞激酶2(Tie2)/磷酸激酶(AKT)调控大鼠阴茎海绵体组织内皮一氧化氮合酶(eNOS)的表达,并影响阴茎勃起功能。方法:将8周龄雄性SD(Sprague Dawley)大鼠随机分为6组(n=6):假手术组、去势组、睾酮替代组(去势1 d后隔日1次丙酸睾酮3 mg/kg皮下注射)、假手术+Tie2转染组(去势术后4周大鼠阴茎海绵体注射20 ul携带Tie2基因的慢病毒,滴度1×10⁸TU/ml)、去势+Tie2转染组、去势+空载体组。去势术后5周,测定各组大鼠的最大阴茎海绵体内压与平均动脉压比值(ICPmax/MAP)、血清睾酮(T)水平、一氧化氮(NO)含量,以及Tie2、AKT、P-AKT、eNOS、P-eNOS在各组大鼠阴茎海绵体中的表达水平。结果:去势组大鼠的T、阴茎海绵体组织中NO的含量和ICPmax/MAP明显低于假手术组大鼠(P<0.01)。而经过Tie2过表达慢病毒转染后,去势+Tie2组大鼠的NO含量及ICPmax/MAP明显高于去势组大鼠(P<0.01)。去势组大鼠阴茎海绵体组织中Tie...     

雄激素与勃起功能障碍

雄激素对维持性欲是必不可少的,它通过中枢和外周两个层面,以多种机制控制着阴茎勃起的启动、维持和终结.雄激素缺乏不但导致性欲减退,还可造成阴茎的组织结构损害及勃起相关活性物质发生明显改变,如阴茎海绵体平滑肌含量减少、结缔组织增多及白膜下脂肪细胞沉积等.阴茎勃起时,海绵体组织的上述改变可致静脉闭塞不全而发生静脉漏,出现勃起功能障碍(ED).对性腺功能减退的ED患者补充睾酮可收到良好的治疗效果.睾酮替代治疗期间需密切观察,避免不良反应的发生.     

Raf/MEK/ERK1/2信号通路与阴茎勃起功能关系的研究进展

阴茎勃起功能受海绵体平滑肌舒张功能的调控。在细胞外信号调节激酶1/2(ERK1/2)信号通路研究过程中人们认识到Raf/MEK/ERK1/2信号通路与海绵体平滑肌及内皮系统功能关系密切。ERK1/2信号主要通过一氧化氮合酶(NOS)的磷酸化作用参与阴茎勃起功能的调控。目前认为,ERK1/2在勃起过程中磷酸化抑制内皮性NOS(eNOS)活性降低阴茎海绵体平滑肌(CCSMC)舒张及阴茎勃起功能,但其磷酸化抑制的作用位点尚不清楚。在CCSMC及内皮细胞中,ERK1/2信号与其他信号通路之间的相互作用从而调控阴茎勃起功能。本文总结了近年以来Raf/MEK/ERK1/2信号通路的研究进展,阐述其在勃起功能中的作用机制及各信号通路之间的相互作用,为进一步探索阴茎勃起过程的分子机制奠定基础。     

雄激素缺乏与勃起功能障碍

雄激素可通过增强性欲、性唤起等间接促进阴茎勃起,在维持阴茎正常的勃起组织结构方面起重要作用。睾酮缺乏可导致小梁平滑肌减少、细胞外基质增多、白膜下脂肪细胞沉积等。勃起时,阴茎组织的上述改变可引起静脉闭塞不全而发生静脉漏,从而出现勃起功能障碍;睾酮还可影响一氧化氮合酶(NOS)、RhoA/Rho激酶的表达及活性从而直接影响阴茎勃起。对单用磷酸二酯酶5(PDE5)抑制剂无反应的性腺功能减退的ED患者补充睾酮可收到良好的治疗效果。     

补骨生髓方对去势骨质疏松大鼠Smad/ERK信号通路的影响

目的探讨补骨生髓方对去势骨质疏松大鼠Smad/ERK信号通路的影响。方法通过采取去性腺法(摘除睾丸法)建立骨质疏松症大鼠实验模型,造模成功后按照实验方案对大鼠进行灌胃治疗,2个月后处死各组大鼠,HE常规染色,并通过免疫组织化学染色的方法检测正常对照组及各中药干预组去势大鼠ERK、p-ERK、Smad4蛋白的表达。结果与模型组比较,补骨生髓方高剂量组与中剂量组ERK蛋白的表达差异有统计学意义(P0.05),而低剂量组与模型组表达差异无统计学意义(P0.05);与模型组比较,补骨生髓方高剂量组与中剂量组p-ERK蛋白的表达差异有统计学意义(P0.05),而低剂量组与模型组表达差异无统计学意义(P0.05);与模型组比较,补骨生髓方高剂量组与中剂量组Smad4蛋白表达差异有统计学意义(P0.05),而低剂量组与模型组比较差异无统计学意义(P0.05)。结论补骨生髓方在一定程度上能够上调ERK、p-ERK、Smad4蛋白的表达水平,其可能是通过调控ERK蛋白磷酸化介导的Smad/ERK信号通路来发挥作用;ERK、Smad4可能是今后治疗骨质疏松的潜在作用靶点,但仍需要进一步开展更深入的研究。     

雄激素对阿朴吗啡和电刺激诱导的去势大鼠勃起功能的影响

目的 :探讨应用安雄进行雄激素替代对去势大鼠勃起功能的影响。方法 :取40只成年雄性 SD大鼠 ,分为去势、高、低剂量安雄及假手术 4组。治疗 4周后采用阿朴吗啡 ( APO)皮下注射与电刺激海绵体神经诱导大鼠勃起 ,对其勃起功能进行评价。结果 :高、低剂量安雄组与假手术组大鼠 APO诱导的勃起成功率、勃起次数与电刺激诱导的海绵体内压 ( ICP)均较去势组大鼠为高或多 ,统计学处理差异有显著性 ( P<0 .0 1 )。结论 :通过 APO皮下注射和电刺激海绵体神经证实 ,去势导致大鼠勃起功能明显下降 ;采用安雄进行雄激素替代可恢复其勃起功能 ;去势既影响了药物诱发的勃起反应 ,也损伤了外周电刺激诱导的勃起反应     

淫羊藿苷上调NRF2改善SHR大鼠勃起功能

目的:研究淫羊藿苷是否通过调节核因子类红细胞2-相关因子2(NRF2)通路改善自发性高血压大鼠(SHR)的勃起功能.方法:随机将10周龄健康雄性WKY大鼠(WKR)与雄性SHR大鼠分为4组(每组6只,共24只):WKY对照组、WKR+淫羊藿苷组[10 mg/(kg·d)灌胃]、SHR对照组,SHR+淫羊藿苷组[10 m...     

大鼠勃起功能障碍模型的研究进展

勃起功能障碍(erectile dysfunction,ED)是指男性持续不能达到和/或维持足够的勃起以完成满意的性生活.阴茎勃起是一个受多系统调节的复杂过程,正常的阴茎勃起依赖于正常的性心理反应,正常的生理结构,正常的神经、内分泌和血管功能.任何一个环节的异常都可能导致勃起功能障碍.由于影响阴茎勃起的因素较多,研究者们针对不同的病因设计了不同的大鼠ED模型,根据ED的临床分类将大鼠ED模型叙述如下.     

不同月龄大鼠勃起功能状况的实验研究

目的:探讨不同月龄组大鼠勃起功能的变化规律,为建立增龄性大鼠ED模型提供动物实验依据。方法:80只雄性Wistar大鼠,分为3、6、12和18月龄组,分别灌胃枸橼酸西地那非进行阴茎勃起功能实验;20只3月龄雌性Wistar大鼠,随机分为4组,建立发情雌鼠模型,观察不同月龄雄鼠的勃起状况与勃起次数。结果:3、6、12和18月龄组Wistar大鼠勃起率及勃起次数分别为85%、75%、40%、30%及(2.27±0.80)、(2.00±0.61)、(1.40±0.51)、(1.29±0.49)次,不同月龄勃起率与勃起次数比较,差异有统计学意义(P<0.05),且随着月龄的增加,各组大鼠勃起功能有降低的趋势。分别将6、12、18月龄组的勃起率及勃起次数与3月龄组大鼠做两两比较,发现6月龄组与12月龄组较3月龄组的差异无统计学意义(P>0.05);18月龄组较3月龄组有明显降低,且差异有显著性(P<0.05)。结论:衰老是大鼠发生ED的主要危险因素,18月龄大鼠已具备建立增龄性大鼠ED模型的条件。     

哮喘对大鼠勃起功能影响的实验研究

目的:探讨哮喘对大鼠勃起功能的影响及相关蛋白的表达.方法:使用卵清蛋白溶液构建大鼠哮喘模型,然后颈部注射阿扑吗啡100 μg/kg,观察30 min内勃起次数.以未勃起或勃起次数明显减少的大鼠为模型组,随机选取6只正常大鼠作为对照组.使用苏木精和伊红(HE)比较阴茎海绵体的组织结构.采用实时荧光定量逆转录-聚合酶链反应...     

Ganoderma lucidum polysaccharide ameliorated diabetes mellitus-induced erectile dysfunction in rats by regulating fibrosis and the NOS/ERK/JNK pathway

BackgroundDiabetes mellitus-induced erectile dysfunction (DMED) is a frequent complication of diabetes mellitus (DM), with limited therapy at present. This study aimed to explore the role and mechanism of Ganoderma lucidum polysaccharide (GLP) on DMED.MethodsDMED was induced in the experimental rats [male 12-week-old Sprague-Dawley (SD) rats] by treatment with streptozotocin (60 mg/kg) and apomorphine (APO). Next, rats in the GLP low dose (GLP-L)/GLP high dose (GLP-H) groups were treated with GLP (100 or 400 mg/kg/d, respectively) for 8 weeks. Subsequently, erectile function was assessed by APO and electrostimulation of the cavernous nerve (CN). Serum or penile testosterone (T), luteinizing hormone (LH), follicle-stimulating hormone (FSH), and cyclic guanosine monophosphate (cGMP) contents were evaluated by enzyme-linked immunosorbent assay (ELISA). The levels of oxidative stress indicators in the corpus cavernosum (CC) were measured by corresponding kits, and histological changes in the CC were observed by hematoxylin-eosin (HE) and Masson staining. Additionally, the apoptosis index, caspase-3, caspase-9, and eNOS expression, and mitochondrial membrane potential (MMP) were also detected. Furthermore, quantitative polymerase chain reaction (qPCR) and western blot assays were conducted to determine the NOS, TGF-β1 mRNA expression, ERK1/2, eNOS, JNK phosphorylation, and arginase II protein expression.ResultsThe erectile function test revealed that erectile dysfunction (ED) was alleviated in the DMED rats following treatment with GLP. Moreover, GLP upregulated the T and cGMP content, improved the oxidative stress and histological injuries of CC, and also inhibited the apoptosis and MMP loss of penile tissues in DMED rats. Furthermore, GLP treatment enhanced the mRNA expression of NOS and TGF-β1 and suppressed the phosphorylation of ERK1/2, eNOS, and JNK, as well as the protein expression of arginase II in DMED rats.ConclusionsGLP ameliorated DMED by repairing the CC pathological damage and upregulating NOS expression and ERK/JNK phosphorylation, indicating that GLP may be a candidate drug for DMED therapy.     

针刺通过调节DKK1通路调控骨代谢改善去卵巢模型大鼠骨质疏松的机制研究

目的 探讨针刺通过调节DKK1通路调控骨代谢对于去卵巢模型大鼠(ovariectomized rats,OVX)骨质疏松的影响及其机制.方法 采用经典的双侧卵巢切除进行雌性SD大鼠的原发性骨质疏松造模,阿伦磷酸盐阳性药物对照组服用阿伦磷酸盐治疗,各期针灸组采取针刺双侧"肾腧"穴.观察针刺治疗对OVX大鼠全身骨量、生物力...     

芦荟素通过激活ERK1/2-Runx2信号通路介导对去卵巢大鼠骨量流失的保护作用

目的 观察芦荟素对去卵巢大鼠骨量的影响,并探讨可能的机制.方法 将30只雌性Sprague Dawley大鼠随机分为3组:假手术组(Sham)、去卵巢组(OVX)及去卵巢大鼠+芦荟素治疗组(LHS,去卵巢大鼠每天接受50 mg/kg芦荟素治疗,持续12周).12周后取双侧股骨进行微型计算机断层扫描(Micro-CT)检...     

Vasoactive intestinal polypeptide, an erectile neurotransmitter, improves erectile function more significantly in castrated rats than in normal rats

What’s known on the subject? and What does the study add? Vasoactive intestinal polypeptide (VIP) is an important erectile neurotransmitter, and our previous study found that the mRNA expression of VIP was independent of androgens. The present study further investigated the vivo effect of VIP on erection. We found that not only the expression of VIP was independent of androgens, but also the effect of VIP on erection was independent of androgens. In fact, we found that VIP played a more significant role on erection in castrated rats than in normal rats.

OBJECTIVE

• ? To investigate the regulatory role of androgen in VIP‐mediated erectile effect. Androgen is essential for physiological erection. Vasoactive intestinal polypeptide (VIP) is an important erectile neurotransmitter. While previous studies demonstrated that VIP expression in the penis was androgen‐independent, it remains controversial whether androgen has any effect on VIP‐mediated erection.

MATERIALS AND METHODS

• ? Male SD rats were divided into a control group, a castration group, and a castration‐with‐testosterone‐replacement group. Four weeks later, each group was subdivided into low and high‐dose VIP subgroups and subjected to intracavernous injection of 0.5 and 2 µg VIP, respectively.
• ? Erectile function was tested by recording intracavernosal pressure (ICP) and mean arterial blood pressure (MAP) before and after VIP injection.
• ? The expressions of the VIP‐receptor (VPAC2), G‐protein stimulatory and inhibitory alpha subunits (Gs‐α, Gi‐α), and PDE3A in rat corpus cavernosum (CC) was qualified by real‐time PCR and Western blot analysis.

RESULTS

• ? Castration reduced erectile function while testosterone restored it. VIP improved erectile function in a dose‐dependent manner.
• ? High‐dose VIP significantly enhanced erectile function in castrated rats and there was no difference of ICP/MAP among three groups after injection of high‐dose VIP.
• ? Low‐dose VIP also resulted in a higher improvement of erectile function in castrated rats, although the ICP/MAP was lower in these rats than in the other two groups. VPAC2 and Gs‐α were up‐regulated while Gi‐α and PDE3A were down‐regulated in CC of castrated rats.

CONCLUSION

• ? VIP improves erectile function much more significantly in hypogonadal condition, mainly due to the higher expression of VPAC2, Gs‐α, and lower expression of Gi‐α and PDE3A in CC of castrated rats. Androgen may negatively regulate the erectile effect of VIP.

     

Androgen deficiency impairs erectile function in rats through promotion of corporal fibrosis

The aim of this study was to investigate the underlying mechanism of androgen deficiency inducing corporal fibrosis, thereby causing erectile dysfunction (ED). Forty 12‐week‐old healthy male rats were divided randomly into four groups: normal control group (Control); castration group (Castration); the other 20 rats were castrated followed by testosterone (T) (orally) each day: castration + 10mg/kg T group (Castration + 10T) and castration + 20 mg/kg T group (Castration + 20T). After 8 weeks' treatment, the main outcome measures were the following: serum levels of T; the ratios of intracavernous pressure (ICP) to mean arterial pressure (MAP); histologic changes in penile smooth muscle cells; the Smad and non‐Smad pathways; and extracellular matrix (ECM) protein deposition. Castration group showed lower level of T and ratio of ICP/MAP, reduced ratio of penile smooth muscle cells/collagen, increased extracellular matrix protein deposition, and a higher expression of the Smad and non‐Smad pathways. Castration + 10T partially preserved erectile function and histology stabilisation. However, the Castration + 20T group showed significantly better erectile function and molecular changes. Better efficacy could be expected with ART of adequate dose. Androgen deficiency induces corporal fibrosis through activation of the Smad and non‐Smad pathways, and accumulation of ECM proteins.     

G蛋白耦联受体激酶结合蛋白1通过调节细胞外调节激酶1/2的活性促进成骨细胞迁移

目的探索G蛋白耦联受体激酶结合蛋白1(GITI)在成骨细胞迁移中的作用,并分析其机理。方法通过Western blot方法检测GIT1蛋白在鼠的成骨细胞内的表达；用免疫荧光染色方法确定：在血小板衍生生长因子(PDGF)不刺激和刺激的条件下,GIT1和细胞外调节激酶1/2(ERK1/2)在成骨细胞内的位置；用共同免疫沉淀的方法测定GIT1和ERK1/2相互结合,并且用免疫荧光双染的方法确定这两种蛋白相互结合的位置；用包含GIT1-RNA发夹结构的腺病毒感染成骨细胞后,用免疫荧光染色方法确定磷酸化ERK1/2(pERK1/2)在成骨细胞内的位置,用划痕愈合法检测在PDGF刺激下的迁移能力。结果在成骨细胞内,PDGF刺激导致了GIT1和ERK1/2的相互结合,并且这种结合发生在成骨细胞的局部粘附内。包含GIT1-RNA发夹结构的腺病毒明显抑制了pERK1/2招募至成骨细胞局部粘附内以及PDGF所刺激的成骨细胞的迁移。结论在PDGF刺激下,GIT1招募pERK1/2至成骨细胞的局部粘附内,从而促进成骨细胞的迁移。     

Probucol improves erectile function by regulating endoplasmic reticulum stress in rats with streptozotocin-induced diabetes

This study was to explore the effect and mechanism of Probucol on STZ-induced erectile dysfunction in diabetic rats. Thirty SD male rats aged 12 weeks were given intraperitoneal injection of STZ after fasting for 12 hr. Diabetic rats were haphazardly partitioned under two assemblies and administered 0 or 500 mg/kg probucol by oral gavage to 12 weeks. Control group was intraperitoneally injected with physiological saline, and saline was administered by oral gavage daily. Intracorporeal pressure was used to evaluate erectile function. Levels of proteins were detected using immunohistochemistry and Western blotting. α-SMA and vWF were detected using immunofluorescence staining. After treatment, erectile function in probucol group was significantly improved. Endoplasmic reticulum stress-related proteins were expressed higher in DM group than in sham group, while expression of these proteins decreased significantly in probucol group. However, α-SMA and vWF were expressed at lower levels in DM group than in sham group, and probucol treatment reversed this phenomenon. Finally, Bax and Caspase3 were expressed at higher levels and Bcl-2 was expressed at lower levels in DM group, while the opposite result was obtained in probucol group. In conclusions, probucol improves erectile function by reducing endothelial dysfunction and inhibiting PERK/ATF4/CHOP pathway in STZ-induced diabetic rats.     

Effects of icariin on erectile function and expression of nitric oxide synthase isoforms in castrated rats

Aim： To investigate the effect of icariin on erectile function and the expression of nitric oxide synthase （NOS） isoforms in castrated rats. Methods： Thirty-two adult male Wistar rats were randomly divided into one shamoperated group （A） and three castrated groups （B, C and D）. One week after surgery, rats were treated with normal saline （groups A and B） or oral icariin （1 mg/[kg·day] for group C and 5 mg/[kg·day] for group D） for 4 weeks. One week after treatment, the erectile function of the rats was assessed by measuring intracavernosal pressure （ICP） during electrostimulation of the cavernosal nerve. The serum testosterone （ST） levels, the percent of smooth muscle （PSM） in trabecular tissue, and the expression of mRNA and proteins of neuronal nitric oxide synthase （nNOS）, inducible nitric oxide synthase （iNOS）, endothelial nitric oxide synthase （eNOS） and phosphodiesterase V （PDES） in corpus cavernosum （CC） were also evaluated. Results： ICP, PSM, ST and the expression of nNOS, iNOS, eNOS and PDE5 were significantly decreased in group B compared with those in group A （P 〈 0.01）. However, ICE PSM and the expression of nNOS and iNOS were increased in groups C and D compared with those in group B （P 〈 0.05）. Changes in ST and the expression of eNOS and PDE5 were not significant （P 〉 0.05） in groups C and D compared with those in group B. Conclusion： Oral treatment with icariin （〉 98.6 % purity） for 4 weeks potentially improves erectile function. This effect is correlated with an increase in PSM and the expression of certain NOS in the CC of castrated rats. These results suggest that icariin may have a therapeutic effect on erectile dysfunction.