FK1706 enhances the recovery of erectile function following bilateral cavernous nerve crush injury in the rat

IntroductionAdvances in neurobiology have led to a surge of clinical interest in the development of protective and regenerative neuromodulatory strategies, as surgical therapies for prostate cancer often result in neuronal damage and debilitating loss of sexual function.AimTo investigate the dose-dependent efficacy of FK1706, a nonimmunosuppressant immunophilin ligand, for the recovery of erectile function following bilateral cavernous nerve crush injury in the rat.Main Outcome MeasuresRecovery of erectile function was assessed by cavernous nerve electrostimulation and reported as maximal increase of intracavernous pressure (ICP) and area under the curve (AUC). Changes in animal weights, percentage completion of treatment course, and survival were compared between groups.MethodsThirty-five Sprague–Dawley male rats were randomly divided into five equal groups: seven animals received a sham operation, whereas 28 animals underwent bilateral cavernous nerve crush injury, followed by subcutaneous injection of vehicle alone (1.0 mL/kg), or low (0.1 mg/kg), medium (0.32 mg/kg), or high dose (1.0 mg/kg) FK1706 5 days per week for 8 weeks.ResultsErectile dysfunction did not occur in the sham group (mean maximal ICP increase of 100.8 ± 6.3 cmH₂O), whereas nerve injury and vehicle treatment produced a significant reduction in ICP response to 34.4 ± 12.8 cmH₂O. The mean ICP increase for high-dose FK106 treatment was 73.9 ± 6.3 cmH₂O (P < 0.01 vs. vehicle) compared with 58.3 ± 7.4 cmH₂O and 56.9 ± 8.3 for low and medium doses (P > 0.05). Similar stepwise findings were observed using AUC data. No significant maximal aortic blood pressure or weight differences occurred between groups and all animals completed treatment.ConclusionHigh-dose subcutaneous FK1706 therapy promoted recovery of erectile function following bilateral cavernous nerve crush injury in the rat. No significant differences between groups were observed for changes in weight, and the 8-week treatment course was completed for all animals. Bella AJ, Hayashi N, Carrion RE, Price R, and Lue TF. FK1706 enhances the recovery of erectile function following bilateral cavernous nerve crush injury in the rat.     

亚低温对新生大鼠缺氧缺血性脑损伤p-ERK/p38MAPK的影响

目的 探讨丝裂原活化蛋白激酶(mitogen-activated protein kinases,MAPK)信号转导通路p-ERK/p38部分活化在亚低温治疗新生大鼠缺氧缺血性脑损伤(hypoxic-ischemic brain damage,HIBD)中的作用及意义.方法 实验分模型组和对照组.采用7日龄Sprague-Dawley(SD)清洁级大鼠,建立新生大鼠HIBD标准模型,随机分为常温缺氧缺血组(10只、肛温=37℃)和亚低温缺氧缺血组(10只、肛温=33℃);对照组为假手术动物,也分为常温组和亚低温组(各8只,余同上).取背侧海马冠状平面采用脱氧核糖核苷酸末端转移酶介导的原位缺口末端标记(TUNEL)结合苏木素-伊红(HE)、神经元Nissl染色和电镜观察凋亡小体检测脑细胞凋亡情况.采用Western印迹检测MAPK成分磷酸化p42/44ERK(简称p-ERK1/2)、磷酸化p-38 MAPK(简称p-p38)的变化时程及其意义.结果 72 h亚低温缺氧缺血组脑细胞凋亡发生率(6.4±1.7)%,与常温缺氧缺血组(25.3±1.5)%比较P＜0.01,差异有统计学意义.而细胞坏死发生率无明显改善.HIBD后MAPK信号转导系统活化,p-ERK1/2水平在IN组结扎侧大脑组织逐渐降低,而p-p38水平却逐渐升高,二者作用相拮抗.亚低温治疗可逆转二者的反向变化. 结论 亚低温治疗可通过p-ERK/p38MAPK信号转导通路抑制HIBD后的细胞凋亡.     

FUT8 drives the proliferation and invasion of trophoblastic cells via IGF-1/IGF-1R signaling pathway

IntroductionTrophoblast proliferation and invasion are essential for embryo implantation and placentation. Protein glycosylation is one of the most common and vital post-translational modifications, regulates protein physical and biochemical properties. FUT8 is the only known fucosyltransferase responsible for catalyzing α1,6-fucosylation in mammals, and α1,6-fucosylated glycoproteins are found to participate in various physiopathological processes. However, whether FUT8/α1,6-fucosylation modulates the functions of trophoblastic cells remains elusive.MethodsFUT8 in human placenta villi during 6-8 gestational weeks and trophoblastic cells were detected by Western blot and immunofluorescent staining. α1,6-fucosylation in tissues or cells were measured by Lectin LCA (Lens culinaris) fluorescent staining and Lectin blot. FUT8 expression was down-regulated by siRNA transfection in JAR and JEG-3 cells, and cell viability, motility and invasiveness ability were detected by the functional experiments. α1,6-fucosylation of insulin-like growth factor receptor (IGF-1R) was examined by immunoprecipitation, and the amount of phosphorylated IGF-1R was detected in FUT8 down-regulated JAR cells.ResultsHuman placenta villi and trophoblastic cells expressed FUT8/α1,6-fucosylation. Knockdown FUT8 by siRNA transfection suppressed the proliferation, epithelial-mesenchymal transition, migration and invasion of JAR and JEG-3 cells. Furthermore, we found that FUT8 modified the α1,6-fucosylation of IGF-1R, and regulated IGF-1 dependent activation of IGF-1R, MAPK and PI3K/Akt signaling pathways in JAR cells.ConclusionsOur results implicate a critical role for FUT8 in maintaining the normal functions of trophoblastic cells, suggesting manipulating FUT8 may be an effective approach in pregnancy.     

Egr1信号途径参与NDRG1表达调节的机制研究

目的:研究人宫颈癌细胞中早期生长反应基因1(Egr1)信号途径参与N-myc下游调节基因1(NDRG1)表达调节的机制。方法:以人宫颈癌细胞株SiHa为研究对象,利用siRNA干扰技术使Egr1基因沉默,用RT-PCR和W estern blot检测转染前后Egr1表达的变化。通过RT-PCR和W estern blot检测细胞在低氧条件下,以及转染siRNA-Egr1后在低氧或低氧模拟物(DFX)条件下,Egr1与NDRG1表达的变化。结果:siRNA能有效地抑制Egr1基因表达(P0.001)。低氧条件下,Egr1、NDRG1的蛋白水平均明显升高(P0.001)。转染siRNA-Egr1 48h后,细胞在低氧条件下作用1h,Egr1 mRNA表达显著下降(P0.05)。细胞转染siRNA-Egr1 48h后,在低氧及DFX条件下作用24h,结果显示DFX使NDRG1 mRNA的表达显著高于低氧条件(P0.05)。同时也显示siRNA-Egr1能够使NDRG1 mRNA水平明显减少(P0.05)。结论:体内环境因素如低氧、DFX等上调NDRG1基因的表达,通过Egr1信号途径参与NDRG1在宫颈癌细胞中表达的调节。     

过表达NDRG1通过调控Wnt/β-catenin信号通路促进滋养层细胞上皮-间质转化

目的:探讨N-myc下游调节基因1(NDRG1)过表达对滋养层细胞上皮-间质转化(EMT)的影响以及对Wnt/β-catenin信号通路的作用。方法:采用携带NDRG1基因的慢病毒感染人胎盘滋养层细胞系HTR-8/SVneo,建立NDRG1基因稳定过表达的HTR-8/SVneo细胞株。qRT-PCR和Western blot法检测慢病毒感染后细胞中NDRG1基因mRNA和蛋白的表达水平,Transwell小室法观察细胞侵袭和迁移能力,免疫荧光实验检测β-catenin蛋白核转位情况,Western blot法分析EMT相关蛋白、Wnt/β-catenin信号通路相关蛋白及基质金属蛋白酶-2(MMP-2)和MMP-9等蛋白表达。结果:慢病毒介导NDRG1过表达后,HTR-8/SVneo细胞中NDRG1 mRNA和蛋白表达水平显著上调。NDRG1过表达可显著增强细胞的侵袭和迁移能力,并上调MMP-2、MMP-9、波形蛋白(Vimentin)、神经性钙黏附蛋白(N-cadherin)、β-catenin、Snail1和环氧化酶-2(COX-2)等蛋白的表达水平,下调上皮钙黏蛋白(E-cadherin)和糖原合成酶激酶3β(GSK3β)磷酸化水平,同时促进β-catenin蛋白由细胞质向细胞核转位。结论:NDRG1过表达可促进HTR-8/SVneo细胞EMT过程,提高细胞侵袭和迁移能力,其机制可能与激活Wnt/β-catenin信号通路转导相关。     

阻断丝裂原活化蛋白激酶信号通路对子宫内膜癌生长抑制作用的研究

目的:研究丝裂原活化蛋白激酶(MAPK/ERK)信号通路抑制剂(PD98059)对ER阳性表达的Ishikawa和ER低表达的HEC-1A人子宫内膜癌细胞系的裸鼠体内抗增殖作用,探讨阻断MAPK/ERK信号传导通路治疗子宫内膜癌的可行性。方法:体外培养人子宫内膜癌Ishikawa、HEC-1A细胞,将对数生长期的两种细胞分别接种于裸鼠背部皮下,建立子宫内膜癌裸鼠移植瘤模型。将两种细胞系成瘤阳性裸鼠随机分为对照组和实验组,每组6只,分别腹腔注射生理盐水和PD98059,每周2次,共3周,观察各组裸鼠移植瘤的生长情况,建立移植瘤生长曲线,计算抑瘤率。实验结束时取肿瘤组织切片HE染色,原位末端标记(TUNEL)法检测组织细胞的凋亡,Western blot检测各组织中ERK1/2的活化。结果:(1)PD98059能有效抑制人子宫内膜癌Ishikawa、HEC-1A细胞裸鼠皮下移植瘤的生长;(2)HE染色可见实验组瘤细胞核浆比相对较小,血管密度降低;TUNEL染色可见两种细胞移植瘤中实验组细胞凋亡率较对照组明显增加;(3)Western blot检测结果:PD98059能有效降低裸鼠移植瘤组织中p-ERK表达。结论:PD98059通过阻断MAPK/ERK信号传导通路增加Ishikawa和HEC-1A细胞裸鼠移植瘤组织中细胞凋亡,抑制肿瘤生长,抗肿瘤效果明显,可成为治疗子宫内膜癌的有效方法。     

Beneficial treatment effects of dietary nitrate supplementation on testicular injury in streptozotocin-induced diabetic male rats

Research questionDo low doses of dietary nitrate help to attenuate the progression of diabetic reproductive disorders in streptozotocin-induced diabetic male rats?DesignFifty male Wistar rats were divided into five groups: controls receiving distilled water; controls receiving 100 mg/l nitrate in distilled water; diabetic rats receiving distilled water; diabetic rats receiving insulin 2–4 U/day of neutral protamine hagedorn insulin; and diabetic rats receiving 100 mg/l nitrate in distilled water. Diabetes was induced by 45 mg/kg streptozotocin. Nitrate and insulin treatment were started 4 weeks after diabetes induction for 8 weeks. Serum insulin, nitrogen oxide, stereology of testis, apoptosis, sperm parameters, and mRNA expression of Pdcd4, Pacs2, p53 and miR-449a were assessed at the end of the study.ResultsBlood glucose, apoptotic index of seminiferous tubules and expression of p53, Pdcd4, and Pacs2 mRNA were significantly higher in the diabetic rats (P < 0.001). Decreased body weight, serum insulin and nitrogen oxide level, and miR-449a were observed in the diabetic group (P < 0.01 for insulin; P < 0.001 for others). Most sperm parameters and stereological results differed between diabetic and control rats; nitrate recovered almost all these alterations, including dead spermatozoa, sperm motility grade, sperm deformity index, spermatozoa with damaged DNA, malformations in abnormal spermatozoa, total volume of seminiferous tubule, germinal epithelium, capsule, lumen, interstitial tissue, seminiferous tubule diameter, germinal epithelium height, the number of spermatogenic, Sertoli and Leydig cells.ConclusionsTreatment with sodium nitrate could modulate apoptosis, which is a major cause of diabetic testicular disorder. These experiments suggest that nitric oxide plays an important role in the function of the reproductive system.     

调节性T细胞和Notch1信号通路在原因不明复发性自然流产中的作用

目的探讨调节性T细胞(Treg)和Notch1信号通路在原因不明复发性自然流产(URSA)中的作用。方法流式细胞仪检测URSA患者(URSA组)及正常妊娠妇女(对照组)蜕膜CD4~+CD25~+T细胞Treg表达比例,real time RT-PCR及Western blotting检测蜕膜中CD4~+T细胞中Notch1信号通路和叉头转录因子家族3(Foxp3)表达情况。结果 URSA组CD4~+CD25~+T细胞/淋巴细胞、CD4~+Foxp3~+T细胞/淋巴细胞和CD4~+Foxp3~+T细胞/CD4~+T细胞比例均低于对照组(P0.05)。URSA组CD4~+T细胞中Notch1-Ic、RBPJκ、Foxp3 m RNA及蛋白表达均低于对照组。结论 URSA患者蜕膜CD4~+T细胞中Notch1信号通路和Foxp3表达下调,CD4~+CD25~+T细胞表达比例下降,提示URSA患者Notch1信号通路和Foxp3表达下调可能阻碍CD4~+T细胞转化为CD4~+CD25~+T细胞,进而诱发免疫排斥,诱导流产。     

Protective role of SIRT1-mediated Sonic Hedgehog signaling pathway in the preeclampsia rat models

Journal of Assisted Reproduction and Genetics - To explore the role of silent information regulator 1 (SIRT1)-mediated Sonic Hedgehog (SHH) pathway in reduced uterine perfusion pressure (RUPP)...