S1P1-3在自发性高血压大鼠阴茎海绵体中的表达

发布时间：2018-07-02 11:34

本文选题：1磷酸鞘氨醇 + 1磷酸鞘氨醇受体1-3　；参考：《泸州医学院》2014年硕士论文

【摘要】：目的：高血压与勃起功能障碍（ED）之间有密切联系，高血压并发ED可能与高血压状态下阴茎海绵体组织内皮功能障碍、eNOS活性及表达下调以及RhoA/Rho激酶信号通路上调有关，而eNOS、RhoA/Rho激酶的上游调控与1磷酸鞘氨醇(S1P)有关，S1P是一类具有生物活性的鞘脂介质，作为一个细胞外配体，与其相应的细胞膜受体—S1P受体(SIP receptor)相结合触发相应的细胞信号转导，与不同的S1P受体结合发挥不同的生物学效应。而高血压状态下S1P与eNOS、RhoA/Rho激酶的关系目前不清楚。本研究在于探讨自发性高血压大鼠（SHR）与正常血压大鼠（WKY）阴茎海绵体内1磷酸鞘氨醇受体1-3（S1P1-3）表达的变化，阐明S1P1-3在阴茎勃起中的作用及其与eNOS/NO、RhoA/Rho激酶信号通路之间的关系。方法：成年健康雄性SPF级SHR与WKY大鼠各10只，14周龄大小，体重约为250-300g，随机分为SHR组、WKY对照组。麻醉前首先称量大鼠体重，以1%戊巴比妥钠30mg/kg腹腔内注射麻醉各组大鼠，仔细分离并暴露出大鼠右侧颈总动脉，用充满肝素生理盐水的26G针头穿刺成功后将其连接于压力换能器，连续监测大鼠颈动脉平均动脉压(MAP)的变化。沿下腹正中线打开腹腔，于前列腺两侧叶后方找到星状的海绵体盆神经节作为电刺激点，以充满肝素生理盐水的24G针头插入大鼠阴茎海绵体固定并连接到压力换能器，测定大鼠海绵体内压(ICP)。给予海绵体盆神经节不同强度的电刺激（刺激强度分别为0V、3V5V，波幅为5ms，刺激频率12Hz，持续刺激时间45s，刺激时间间隔3min）。采用压力换能器记录ICPmax/MAP值。实验大鼠勃起功能测定完毕后，取阴茎海绵体标本并采血，检测两组大鼠血清睾酮水平及血清S1P水平。将阴茎海绵体标本分为两部分，一部分立即用于免疫组化分析，另一部分于-80℃超低温冰箱保存，供Western-blot实验分析。免疫组化和Western-blot印迹方法分别检测S1P1-3、P-eNOS（Ser-1177）、eNOS、ROCK1、ROCK2在大鼠阴茎海绵体中的表达。结果：血清睾酮水平SHR组（430.89±98.67）ng/dl与对照组（492.93±54.23）ng/dl无显著差异。0V电压时SHR组ICPmax/MAP比值（0.109±0.014）较对照组（0.155±0.011）显著降低（P0.05）；3V、5V电刺激海绵体盆神经节后SHR组ICPmax/MAP比值（0.190±0.013、0.219±0.014）均较对照组（0.393±0.018、0.524±0.019）极显著降低（P0.01）。血清S1P水平SHR组（4.156±0.362） ng/ml与对照组（4.549±0.876）ng/ml无显著差异。免疫组化结果显示S1P1主要表达在血管内皮细胞胞膜，，S1P2主要表达在海绵体平滑肌细胞和血管内皮细胞，S1P3主要表达在血管内皮细胞胞膜和海绵体平滑肌细胞，P-eNOS、eNOS主要表达于血管内皮细胞和海绵窦腔内，ROCK1、ROCK2主要表达于海绵体血管平滑肌细胞胞浆中，阳性颗粒呈棕黄色染色。免疫组化数据分析采用积分光密度值(IOD)表示，显示S1P1、P-eNOS、eNOS在SHR组表达显著低于对照组（P 0.05），S1P2、S1P3、ROCK1、ROCK2在SHR组表达显著高于对照组（P 0.05）。Western blot印迹显示S1P1、P-eNOS、eNOS在SHR阴茎海绵体中表达的相对光密度值较对照组显著降低（P 0.05），S1P2、S1P3、ROCK1、ROCK2在SHR阴茎海绵体中表达的相对光密度值较对照组显著升高（P 0.05）。结论：高血压并发ED可能与阴茎海绵体内S1P1表达下调并抑制eNOS/NO信号通路，S1P2、S1P3表达升高并激活RhoA/Rho激酶信号通路有关。
[Abstract]:Objective : To study the relationship between hypertension and erectile dysfunction ( ED ) . The serum testosterone levels and serum S1P levels were measured by pressure transducer . The serum testosterone levels and serum S1P levels were measured by immunohistochemistry and Western - blot . Results : Serum testosterone level in SHR group ( 430 . 89 卤 98.67 ) ng / dl was significantly lower than that of control group ( 492.93 卤 54.23 ) ng / dl .
Immunohistochemical analysis showed that the expression of S1P1 , P - eNOS and eNOS in SHR group was significantly lower than that of the control group ( P 0.05 ) , and the expression of S1P1 , P - eNOS and eNOS in SHR group was significantly lower than that of the control group ( P 0.05 ) . Western blot analysis showed that the relative optical density of S1P1 , P - eNOS and eNOS was significantly lower than that of the control group ( P 0.05 ) , and the relative optical density values of S1P1 , S1P2 , S1P2 , S1P2 , S1P2 , S1P3 , respectively were significantly higher than those in the control group ( P 0.05 ) . Conclusion : The expression of S1P1 and the expression of eNOS / NO signaling pathway , S1P2 , S1P3 in the cavernous body of the penile cavernous body may be regulated by the ED and the activation of the signal pathway of RhoA / rho kinase .
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R544.1;R698

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