| 48 | 0 | 73 |
| 下载次数 | 被引频次 | 阅读次数 |
目的 探讨高血压情况下抑制整合素α5β1下调阴茎海绵体组织中Rho A/ROCK信号蛋白的表达，并影响大鼠勃起功能的相关机制。方法 将15只12周龄自发性高血压大鼠(SHR)随机分为SHR组、SHR+LV-empty组、SHR+LV-siRNA组，每组5只。将15只12周龄正常血压Wistar-Kyoto(WKY)大鼠随机分为WKY组、WKY+LV-empty组、WKY+LV-siRNA组，每组5只。SHR组和WKY组阴茎海绵体中部注射20μL生理盐水; SHR+LV-empty组和WKY+LVempty组阴茎海绵体中部注射20μL空慢病毒载体(滴度1×10~8TU/m L); SHR+LV-siRNA组和WKY+LV-siRNA组阴茎海绵体中部注射20μL携带干扰整合素α5β1的慢病毒(滴度1×10~8TU/m L)。注射1周后检测各组大鼠的体重、血清睾酮水平、阴茎海绵体内最大压(ICPmax)与平均动脉压(MAP)比值、阴茎海绵体中的整合素α5β1、FAK、ROCK1、ROCK2、e NOS的表达水平和NO含量。结果 各组大鼠体重、血清睾酮水平相比差异无统计学意义(P > 0.05)。与WKY组相比，SHR组3、5 V刺激下ICPmax/MAP水平显著降低(P<0.05)，整合素α5β1、FAK、ROCK1、ROCK2的表达显著上调(P<0.05)，而e NOS表达及NO含量显著降低(P<0.05)。整合素α5β1干扰慢病毒成功转染后，与WKY+LV-empty组相比，WKY+LV-siRNA组中3、5 V刺激下ICPmax/MAP水平、阴茎海绵体组织中e NOS表达及NO含量的差异无统计学意义(P > 0.05)，而整合素α5β1、FAK、ROCK1、ROCK2的表达显著降低(P<0.05);与SHR+LV-empty组相比，SHR+LVsiRNA组3、5 V刺激下ICPmax/MAP水平显著升高(P<0.05)，阴茎海绵体组织中e NOS表达及NO含量的差异无统计学意义(P > 0.05)，整合素α5β1、FAK、ROCK1、ROCK2表达显著降低(P<0.05)。结论 高血压上调阴茎海绵体组织内整合素α5β1的表达，激活Rho A/ROCK信号通路，导致阴茎勃起功能障碍。抑制SHR阴茎海绵体组织中整合素α5β1表达，下调Rho A/ROCK信号通路，可改善SHR勃起功能。
Abstract：Objective To explore the mechanism by which inhibition of integrin α5β1 in the context of hypertension downregulates the expression of RAS homolog gene family members( Rho A)/Rho-associated kinase( ROCK) signaling proteins in the penile corpus cavernosum tissue,and affects the erectile function of rats. Methods Fifteen 12-week-old spontaneously hypertensive rats( SHR) were randomly divided into the SHR group,SHR + LV-empty group( transfected with empty lentiviral vector) and SHR + LV-siRNA group( transfected with lentivirus carrying siRNA targeting integrin α5β1),with 5 rats in each group. Fifteen 12-week-old normotensive Wistar-Kyoto( WKY) rats were randomly divided into the WKY rat group,WKY + LVempty group( transfected with empty lentiviral vector) and WKY + LV-siRNA group( transfected with lentivirus carrying siRNA targeting integrin α5β1),with 5 rats in each group. In the WKY group and SHR group,20 μL of normal saline was injected into the middle part of the penile corpus cavernosum. While in the SHR + LV-empty group and WKY + LV-empty group,20 μL of empty lentiviral vector( titer 1 × 108 TU/m L) was injected into the middle part of the penile corpus cavernosum. In the SHR +LV-siRNA group and WKY + LV-siRNA group,20 μL of lentivirus carrying integrin α5β1 interference( titer 1 × 108 TU/m L)was injected into the middle part of the penile corpus cavernosum. One week after transfection,the body weight,levels of serum testosterone,ratio of the maximum intracavernous pressure to the mean arterial pressure( ICPmax/MAP) of rats in each group,the expression of integrin α5β1,FAK,ROCK1,ROCK2,and e NOS in penile corpus cavernosum,as well as the content of nitric oxide( NO) were measured. Results There was no statistically significant difference in body weight and serum testosterone levels among the rats in each group( P > 0. 05). Compared with the WKY group,the ICPmax/MAP in the SHR group was significantly lower( P < 0. 05),and the expressions of integrin α5β1,FAK,ROCK1,and ROCK2 in the penile corpus cavernosum of the SHR group were significantly higher( P < 0. 05),while the expression of e NOS and the content of NO decreased significantly( P < 0. 05). After successful transfection with integrin α5β1 interfering lentivirus,compared with the WKY + LVempty group,there were no statistically significant differences in ICPmax/MAP,e NOS expression in the penile corpus cavernosum,and NO content in the WKY + LV-siRNA group( P > 0. 05),but the expressions of integrin α5β1,FAK,ROCK1,and ROCK2 were significantly decreased( P < 0. 05). Compared with the SHR + LV-empty group,the ICPmax/MAP in the SHR +LV-siRNA group was significantly increased( P < 0. 05). There were no statistically significant differences in e NOS expression in the penile corpus cavernosum and NO content( P > 0. 05). While the expressions of integrin α5β1,FAK,ROCK1,and ROCK2 were significantly decreased( P < 0. 05). Conclusion Hypertension upregulates the expression of integrin α5β1 in the penile corpus cavernosum,activates the Rho A/ROCK signaling pathway,and leads to penile erectile dysfunction. Inhibiting the expression of integrin α5β1 in penile cavernous tissue of SHRs downregulates the Rho A/ROCK signaling pathway and improves the erectile function of SHRs.
[1]Muneer A,Kalsi J,Nazareth I,et al. Erectile dysfunction[J]. BMJ,2014,348:g129.
[2]Shamloul R,Ghanem H. Erectile dysfunction[J]. Lancet,2013,381(9861):153-165.
[3]Vlachopoulos C,Jackson G,Stefanadis C,et al. Erectile dysfunction in the cardiovascular patient[J]. Eur Heart J,2013,34(27):2034-2046.
[4]Wang XY,Huang W,Zhang Y. Relation between hypertension and erectile dysfunction:A meta-analysisof cross-section studies[J]. Int J Impot Res,2018 ,30(3):141-146.
[5]de Simone G,Mancusi C. Erectile dysfunction and arterial hypertension:Still looking for a scapegoat[J]. Eur J Intern Med,2020,81:22-23.
[6]刘继红，栾阳．男性勃起功能障碍的分子生物学研究进展[J]．中华男科学杂志，2015,21(2):99-106.
[7]Sauzeau V,Loirand G. In vitro and in vivo approaches to assess Rho kinase activity[J]. Methods Mol Biol,2017,1527:213-218.
[8]Liu S,Calderwood DA,Ginsberg MH. Integrin cytoplasmic domainbinding proteins[J]. J Cell Sci,2000,113(Pt 20):3563-71.
[9]Hood JD,Cheresh DA. Role of integrins in cell invasion and migration[J]. Nat Rev Cancer,2002,2(2):91-100.
[10]van der Flier A,Sonnenberg A. Function and interactions of integrins[J]. Cell Tissue Res,2001,305(3):285-98.
[11]Miyamoto S,Teramoto H,Coso OA,et al. Integrin function:Molecular hierarchies of cytoskeletal and signaling molecules[J]. J Cell Biol,1995,131(3):791-805.
[12]Jambusaria A,Hong Z,Zhang L,et al. Endothelial heterogeneity across distinct vascularbeds during homeostasis and inflammation[J]. Elife,2020,9:e51413.
[13]Kalucka J,de Rooij LPMH,Goveia J,et al. Single-cell transcriptome atlas of murine endothelial cells[J]. Cell,2020,180:764-779. e20.
[14]Aman J,Margadant C. Integrin-dependent cell-matrix adhesion in endothelial health and disease[J]. Circ Res,2023,132(3):355-378.
[15]Zoungas S,Asmar RP. Arterial stiffness and cardiovascular outcome[J]. Clin Exp Pharmacol Physiol,2007,34(7):647-651.
[16]Failer T,Amponsah-Offeh M,Neuwirth A,et al. Developmental endothelial locus-1 protects from hypertension-induced cardiovascular remodeling via immunomodulation[J]. J Clin Invest,2022,132(6):e126155.
[17]De Leonardis F,Colalillo G,Finazzi AgròE,et al. Endothelial dysfunction,erectile deficit and cardiovascular disease:An overview of the pathogenetic links[J]. Biomedicines,2022,10(8):1848.
[18]Lv PC,Jiang AQ,Zhang WM,et al. FAK inhibitors in cancer,a patent review[J]. Expert Opin Ther Pat,2018,28(2):139-145.
[19]Claesson-Welsh L,Dejana E,Mc Donald DM. Permeability of the endothelial barrier:Identifying and reconciling controversies[J].Trends Mol Med,2021,27(4):314-331.
[20]Wang W,Lollis EM,Bordeleau F,et al. Matrix stiffness regulates vascular integrity through focal adhesion kinase activity[J]. FASEB J,2019 ,33(1):1199-1208.
[21]Albarrán-Juárez J,Iring A,Wang S,et al. Piezo1 and Gq/G11promote endothelial inflammation depending on flow pattern and integrin activation[J]. J Exp Med,2018 ,215(10):2655-2672.
[22]Mutlu E,ilhan S,Onat E,et al. The effects of novokinin,an AT2agonist, on blood pressure, vascular responses, and levels of ADMA,NADPH oxidase,and Rho kinase in hypertension induced by NOS inhibition and salt[J]. Turk J Med Sci,2016,46(4):1249-1257.
[23]晏萧，夏纪毅，姜睿．靶向S1P2基因的siRNA有效片段慢病毒载体的筛选[J]．中国病理生理杂志，2017,33(7):1338-1344.
[24]Quinlan DM,Nelson RJ,Partin AW,et al. The rat as a model for the study of penile erection[J]. J Urol,1989,141(3):656-661.
[25]Wang B,Jiang J,Fan Z,et al. Expression of sphingosine 1-phosphate 1-3 on penile cavernous tissue in hypertensive and normotensive rats[J]. Urology,2014 ,84(2):490. e7-13.
[26]Anderle N,Schäfer-Ruoff F,Staebler A,et al. Breast cancer patientderived microtumors resemble tumor heterogeneity and enable proteinbased stratification and functional validation of individualized drug treatment[J]. J Exp Clin Cancer Res,2023,42(1):210.
[27]Li R,Maminishkis A,Zahn G,et al. Integrin alpha5beta1 mediates attachment,migration,and proliferation in human retinal pigment epithelium:relevance for proliferative retinal disease[J]. Invest Ophthalmol Vis Sci,2009,50(12):5988-5996.
[28]Kuramoto K,Liang H,Hong JH,et al. Exercise-activated hepatic autophagy via the FN1-α5β1 integrin pathway drives metabolic benefits of exercise[J]. Cell Metab,2023,35(4):620-632. e5.
[29]徐律韵，朱从健．整合素α5β1与肿瘤发生、发展相关性的研究进展[J]．临床肿瘤学杂志，2014,19(1):86-89.
[30]Li B,He J,Lv H,et al. c-Abl regulates YAPY357 phosphorylation to activate endothelial atherogenic responses to disturbed flow[J]. J Clin Invest,2019,129(3):1167-1179.
[31]Yurdagul A Jr,Green J,Albert P,et al. α5β1 integrin signaling mediates oxidized low-density lipoprotein-induced inflammation and early atherosclerosis[J]. Arterioscler Thromb Vasc Biol,2014,34(7):1362-73.
[32]Lemay SE,Montesinos MS,Grobs Y,et al. Exploring integrinα5β1 as a potential therapeutic target for pulmonary arterial hypertension:Insights from comprehensive multicenter preclinical studies[J]. Circulation,2025,151(16):1162-1183.
[33]Spescha RD,Glanzmann M,Simic B,et al. Adaptor protein p66(Shc)mediates hypertension-associated,cyclic stretch-dependent,endothelial damage[J]. Hypertension,2014,64(2):347-353.
[34]Traish AM,Goldstein I,Kim NN. Testosterone and erectile function:From basic research to a new clinical paradigm for managing men with androgen insufficiency and erectile dysfunction[J]. Eur Urol,2007,52(1):54-70.
[35]Xiong W,Kong X,Jiang J,et al. Low androgen status inhibits erectile function by inducing e NOS uncoupling in rat corpus cavernosum[J]. Andrology,2020,8(6):1875-1883.
[36]Long H,Jiang J,Xia J,et al. Icariin improves SHR erectile function via inhibiting e NOS uncoupling[J]. Andrologia,2018,50(9):e13084.
基本信息:
DOI：10.13263/j.cnki.nja.2026.02.001
中图分类号:R544.1
引用信息:
[1]龚豪,姜睿.整合素α5β1对自发性高血压大鼠勃起功能的影响[J].中华男科学杂志,2026,32(02):97-105.DOI:10.13263/j.cnki.nja.2026.02.001.
基金信息:
泸州市科技计划(2021-JYJ-58)