糖尿病大鼠膀胱病理变化及胰岛素治疗的影响

目的观察糖尿病大鼠膀胱逼尿肌形态学表现和生理变化以探讨糖尿病大鼠膀胱收缩功能减退的病理基础以及胰岛素治疗对其的影响。方法6周和10周正常组鼠、糖尿病观察组鼠、糖尿病胰岛素治疗组鼠各7只,观察测量其膀胱残余尿量、压力-容量曲线并留取逼尿肌标本,进行光镜和电镜病理检查。结果与对照组比较,糖尿病观察组膀胱残余尿明显增多,压力峰值明显降低,而胰岛素治疗组曲线基本与对照组相同;光镜下可见逼尿肌肌束排列紊乱松散,胶原纤维减少;电镜下可见逼尿肌细胞间胶原纤维和中间连接减少,线粒体空泡变性。而胰岛素治疗组与对照组比较,光镜及电镜下表现均无差异。结论糖尿病大鼠早期即可出现逼尿肌形态学改变以致收缩功能下降,早期给予胰岛素治疗可以预防或减轻上述变化。     

热休克蛋白20在大鼠急性尿潴留膀胱中的表达及意义

目的研究热休克蛋白20（Hsp20）在大鼠急性尿潴留（AUR）模型膀胱中的表达及其与逼尿肌收缩功能的关系。方法通过结扎尿道口的方法建立大鼠急性尿潴留动物模型,利用充盈性膀胱测压的方法检测膀胱收缩功能,采用免疫荧光染色的方法分别检测急性尿潴留膀胱排空后不同时相点（0 h、1 h、6 h、12 h、24 h、3 d、5 d、7 d）大鼠膀胱逼尿肌Hsp20的表达情况,采用western blot的方法比较尿潴留后不同时相点Hsp20含量的差异。结果 AUR后1h、6h、12h、24h组大鼠逼尿肌收缩力呈时间依赖性递减,均较0h组显著下降（P〈0.05）;3d、5d、7d组最大逼尿肌压较1h、6h、12h和24h组显著回升,但仍低于0h组和对照组（P〈0.05）;AUR各组Hsp20表达明显高于对照组,梗阻后24 h内Hsp20表达呈逐步上升趋势,而24 h至7 d则是逐渐降低的,其差异具有统计学意义（P〈0.05）。结论 AUR后膀胱逼尿肌中Hsp20表达的上调很可能对逼尿肌细胞有一定的保护作用,但AUR后Hsp20的高表达可能也是逼尿肌细胞收缩功能受到抑制的重要因素之一。     

心肌营养素1诱导骨髓间充质干细胞分化为心肌样细胞

目的: 探索心肌营养素1(CT-1)对小型猪骨髓间充质干细胞(MSCs)向心肌样细胞分化的影响。方法: 获取、扩增稳定的成体西藏小型猪MSCs,鉴定成脂、成骨潜能。分4组诱导向心肌样细胞分化,包括空白对照组、5-氮杂胞苷(5-Aza)组、CT-1组、5-Aza和CT-1合用组。取诱导后4周的细胞加肌动蛋白(α-actin)和心肌肌钙蛋白T(cTnT)抗体,进行免疫荧光染色,最后计数红色荧光阳性染色率。结果: 分组诱导分化结果,合用组的诱导分化心肌样细胞α-actin阳性率为29.90%±4.76%,明显大于5-Aza组(17.73%±2.35%,P<0.01)、CT-1组(6.63%±0.55%,P<0.01)和空白对照组(1.62%±0.09%,P<0.01);5-Aza组明显大于CT-1组(P<0.01)和空白对照组(P<0.01);CT-1组明显大于空白对照组(P<0.05)。cTnT红色荧光染色结果显示:合用组的诱导分化心肌样细胞cTnT阳性率为36.50%±4.09%,明显大于5-Aza组(14.37%±1.65%,P<0.01)、CT-1组(7.50%±0.61%,P<0.01)和空白对照组(1.12%±0.23%,P<0.01);5-Aza组明显大于CT-1组(P<0.01)和空白对照组(P<0.01);CT-1组明显大于空白对照组(P<0.01)。结论: 适宜浓度的5-Aza(10 μmol/L)和CT-1(0.1 μg/L)能够在体外诱导小型藏猪骨髓MSCs转化为心肌样细胞,诱导后的细胞获得部分心肌细胞特异性蛋白的表达。CT-1与5-Aza联合可明显提高诱导率。     

溃疡性结肠炎患者肿瘤坏死因子α与T细胞亚群的变化及其相关性研究

目的和方法：探讨溃疡性结肠炎（UC）之肿瘤坏死因子α与T细胞亚群的变化及其相关性,测定25例溃疡性结肠炎患者外周血单个核细胞（PBMC）经培养于体外自发和经LPS诱生的TNFα含量及其外周血T细胞亚群百分率和比值。结果：①UC组与对照组自发产生的TNFα含量间无明显差异（P>0.05）。LPS诱生后TNFα含量UC组明显低于对照组（P<0.05）。②UC组与对照组CD3细胞百分率无明显差异（68.86%±4.10% vs 71.10%±5.04%,P>0.05）。UC组CD4细胞显著低于对照组（32.72%±6.06% vs 41.15%± 7.26%,P<0.01）,而其CD8细胞则高于对照组（24.96%±4.02% vs 21.88%±4.17%,P<0.05）,使得其CD4/CD8的比值明显低于对照组（P<0.01）。③两组诱生后的TNFα含量与其CD4、CD8、CD4 /CD8之间均无相关性。结论：UC患者TNFα的诱生能力低下,T细胞亚群数量及比值异常,TNFα诱生水平与T细胞亚群的变化间无相关性。     

SGK1抑制剂EMD638683对膀胱出口梗阻小鼠肾组织细胞焦亡的作用及机制研究

目的:确定小鼠膀胱出口梗阻(bladder outlet obstruction,BOO)模型引起下尿路梗阻介导的肾脏炎性损伤;研究血清和糖皮质激素调节激酶1(serum and glucocorticoid-regulated kinase 1,SGK1)抑制剂EMD638683抗梗阻性肾病炎症的作用机制,探讨SGK1与NLRP3-caspase-1-IL-1β/细胞焦亡(pyroptosis)通路诱导细胞损伤的关系。方法:构建小鼠BOO模型,HE染色观察肾组织病理变化及炎症细胞浸润,PAS染色观察肾脏的组织病变,Masson染色法检测肾小管的胶原沉积,免疫组化法和Western blot法检测NLRP3、caspase-1、F4/80、gasdermin D-N末端片段(GSDMD-N)、IL-1β和SGK1的表达。醛固酮(aldosterone,ALD)处理小鼠肾小管上皮细胞(mouse renal tubular epithelial cells,mRTECs),EMD638683进行干预,Western blot法检测NLRP3、caspase-1、IL-1β、SGK1及GSDMD-N的表达水平;ELISA法检测各组细胞上清液IL-1β含量。结果:梗阻3周的小鼠肾脏中HE染色观察到炎症细胞浸润,PAS染色出现肾脏组织病变,Masson染色检测到肾小管间质有胶原沉积。与正常组小鼠比较,梗阻3周小鼠组NLRP3、caspase-1、F4/80、GSDMD-N、IL-1β和SGK1含量显著升高(P<0.05或P<0.01)。细胞实验中,与正常组比较,ALD组NLRP3、caspase-1、IL-1β、SGK1及GSDMD-N含量显著升高(P<0.05或P<0.01);与ALD组相比,ALD加EMD638683组NLRP3、caspase-1、IL-1β、SGK1及GSDMD-N含量显著减少(P<0.05或P<0.01)。结论:成功构建小鼠BOO模型引起下尿路梗阻介导的肾脏炎性损伤;EMD638683通过抑制NLRP3-caspase-1-pyroptosis通路阻断细胞焦亡,从而发挥抗炎作用。     

HBV特异性CTLs表面CD244和PD-1共表达与慢性乙型肝炎严重程度的相关性

 目的：探讨慢性乙型肝炎（CHB）患者外周血乙型肝炎病毒（HBV）特异性细胞毒性T淋巴细胞（CTLs）表面CD244与程序性死亡蛋白1（PD-1) 的共表达模式及其与疾病进展的关系。方法：采集81例CHB患者和14例健康者外周血,分离外周血单个核细胞（PBMC）;利用主要组织相容性复合体I（MHC-I）-肽五聚体技术标定HBV表位特异性CTLs;荧光抗体标记细胞表面CD244与PD-1分子,流式细胞术检测。结果：CHB患者CD244与PD-1在HBV特异性CTLs表面共表达较总CD8⁺ T细胞表面明显升高（67.48%±17.16% vs 14.01%±7.97%,P<0.01）;HBV感染的不同临床类型中,轻中度的CHB患者HBV特异性CTLs表面CD244与PD-1共表达水平较免疫耐受者明显升高(72.05%±16.86% vs 53.11%±18.05%,P<0.05),肝衰竭组CD244与PD-1共表达水平较轻中度CHB下调（63.11%±13.87% vs 72.05%±16.86%,P<0.05）并伴随干扰素γ（IFN-γ）分泌功能的恢复（30.95%±20.29% vs 13.63%±10.46%,P<0.01）。结论：CHB患者HBV特异性CTLs细胞表面高共表达CD244与PD-1分子,CD244与PD-1共表达在CHB不同临床类型中有差异,并与IFN-γ分泌水平呈负相关,提示CD244可能与CHB患者HBV特异性CTLs功能低下及CHB疾病进展相关。     

热休克蛋白27在大鼠急性尿潴留膀胱中的表达及意义

目的研究热休克蛋白27（HSP27）在大鼠急性尿潴留（AUR）模型膀胱中的表达及其与逼尿肌收缩功能的关系。方法通过尿道口钳夹的方法建立大鼠急性尿潴留动物模型,利用充盈性膀胱测压的方法检测膀胱收缩功能,采用免疫荧光染色的方法分别检测急性尿潴留膀胱排空后不同时相点（0、2、4、6h）大鼠膀胱逼尿肌HSP27的表达情况,采用Western-blot的方法比较尿潴留后不同时相点HSP27含量的差异。结果 AUR后2、4、6h大鼠逼尿肌收缩力呈时间依赖性递减,均较0 h组显著下降（P〈0.05）;其膀胱组织中HSP27的含量亦显著低于0h组,各组的HSP27表达均高于对照组（P〈0.05）。结论 HSP27可能在急性尿潴留致逼尿肌收缩功能障碍发病过程中起到重要作用。     

豚鼠膀胱中二聚体Cajal间质细胞的分布特点及意义

目的　观察二聚体Cajal间质细胞(interstitial cells of Cajal,ICC)在豚鼠膀胱不同组织及部位的分布特点,并探讨其意义。 方法　电子显微镜下观察豚鼠膀胱壁组织切片黏膜层,黏膜下层,肌层内二聚体ICC分布情况;对膀胱组织切片进行免疫荧光染色,用c-Kit抗体标记ICC,激光共聚焦显微镜下观察二聚体ICC在膀胱顶部、体部、颈部的分布特点。 结果　在电子显微镜下见二聚体ICC主要分布于黏膜下层,而肌层主要以单体ICC为主。免疫荧光染色发现每高倍镜视野下膀胱顶部二聚体ICC平均数量为（3.47±0.53）个,体部和颈部为（1.57±0.45）个和(0.49±0.19)个。膀胱顶部二聚体ICC数量明显高于体部和颈部（P<0.01）。 结论　二聚体ICC主要分布于豚鼠膀胱顶部的黏膜下层,可能是感受黏膜张力刺激,引发顶部膀胱自发兴奋的起搏细胞。     

糖尿病大鼠膀胱肾上腺素能α_1受体表达改变

目的研究糖尿病大鼠膀胱结构功能的改变及其肾上腺素能α1受体及其亚型的表达改变,并探讨其意义。方法采用1%链脲佐菌素(STZ)建立糖尿病大鼠的模型,实验分为正常对照组和糖尿病组。分别于2周、4周、6周进行在体充盈性膀胱测压评估膀胱功能,处死动物后取膀胱组织标本常规HE染色观察膀胱逼尿肌病理组织学变化,并采用Western blot与免疫荧光检测α1受体亚型(α1A受体与α1D受体)的表达改变。结果糖尿病大鼠体重减轻,尿量和膀胱湿重增加;充盈性膀胱测压检查结果显示糖尿病大鼠膀胱容量、顺应性、逼尿肌压力上升,残余尿量明显增加,排尿效率明显降低;常规HE染色光镜下观察可见糖尿病大鼠膀胱逼尿肌肌束进行性结构松散、紊乱,甚至出现肌束断裂。Western blott结果显示α1A受体及α1D受体的表达在逼尿肌中表达量随病程进行性下降,糖尿病组与正常对照组间对比差异有统计学意义。免疫荧光结果显示糖尿病膀胱实验组α1A受体和α1D受体灰度值均较相应对照组减少,且随病程进行性减少。结论糖尿病大鼠早期即可出现膀胱逼尿肌形态学和功能的改变,糖尿病膀胱中的α1-AR表达存在负相关关系,提示糖尿病可引起膀胱部位α1-AR表达减少。α1-AR表达减少可能是引起糖尿病膀胱顺应性增加,残余尿量增加的影响因素之一。     

豚鼠糖尿病性膀胱病逼尿肌中c-kit mRNA和蛋白的表达变化及意义

目的: 检测豚鼠糖尿病性膀胱病(DCP)逼尿肌中酪氨酸蛋白激酶生长因子受体(c-kit)mRNA和蛋白的表达水平,探讨c-kit表达与DCP的关系及其发病机制。方法: 60只豚鼠随机分成对照组20只,实验组40只,实验组采用链脲佐菌素(STZ)建立糖尿病豚鼠模型,2组豚鼠饲养10周后运用尿动力学检测并将实验组筛选出DCP组和糖尿病性非膀胱病(NDCP)组,应用RT-PCR、激光共聚焦显微镜技术分别检测各组膀胱组织c-kit mRNA和蛋白的表达。结果: DCP组c-kit mRNA表达显著低于对照组(P<0.01)及NDCP组(P<0.05),相对平均吸光度值分别为4.65±0.47、5.66±0.54、5.54±1.28。糖尿病性膀胱病组c-kit蛋白表达明显低于对照组(P<0.01)与NDCP组(P<0.01),c-kit蛋白荧光值分别为548.69±48.51、844.67±59.24、856.52±53.03。结论: 豚鼠DCP逼尿肌中c-kit mRNA、c-kit蛋白表达减少,导致c-kit信号通路异常,从而使Cajal样细胞在膀胱中的功能减弱而导致逼尿肌功能障碍发生DCP,因此c-kit表达异常可能是DCP的发病机制之一。     

A gap junction blocker inhibits isolated whole bladder activity in normal rats and rats with partial bladder outlet obstruction

We examined the effect of 18alpha-glycyrrhetinic acid (18alpha-GA), a gap junction blocker, or propiverine hydrochloride on the activity of isolated whole bladders obtained from intact rats and rats with partial bladder outlet obstruction (BOO). Thirty-two female Sprague-Dawley rats were divided into an intact group and a BOO group. The whole bladder was harvested from each rat and isovolumetric cystometry was performed in Krebs solution. Changes of bladder activity were recorded after addition of 18alpha-GA or propiverine hydrochloride to the perfusate. Propiverine hydrochloride inhibited the amplitude and duration of contraction in both intact and BOO groups. Propiverine hydrochloride also reduced the baseline bladder pressure in the BOO group, but not in the intact group. In contrast, 18alpha-GA inhibited the amplitude and duration of bladder contraction, and also reduced the baseline pressure, in both intact and BOO groups. BOO bladders showed inhibition of the amplitude and duration of bladder contraction at lower concentrations of 18alpha-GA than intact bladders. A gap junction blocker suppressed the in vitro activity of BOO bladders more effectively than that of intact bladders. Therefore, inhibition of intercellular communication in the bladder via gap junctions may be useful for treating detrusor overactivity, as well as propiverine hydrochloride.     

Investigation of early protein changes in the urinary bladder following partial bladder outlet obstruction by proteomic approach

We investigated the pathophysiological mechanism by proteomic approach as a possible tool to detect the marker proteins to develop lower urinary tract symptoms following bladder outlet obstruction (BOO). Rats were randomized into 3 groups; control, sham operation and BOO groups. BOO group was divided into 1, 3, and 5 day-group. Conventional proteomics was performed with high resolution 2-D gel electrophoresis followed by computational image analysis and protein identification using mass spectrometry using rat urinary bladders. A comparison of bladder of BOO group with control bladder showed that three proteins of optineurin, thioredoxin and preprohaptoglobin were over-expressed in the bladder of BOO group. In addition, four proteins, such as peroxiredoxin 2, transgelin, hippocampal cholinergic neurostimulating peptide (HCNP) and beta-galactoside-binding lectin, were under-expressed in the bladder of BOO group. These data supported that downregulation of HCNP might make detrusor muscle be supersensitive to acetylcholine, up-regulation of optineurin means the protection of nerve injury, and down-regulation of transgelin means the decreased contractility of detrusor muscle. Beside these proteins, other proteins are related to oxidative stress or have a nonspecific function in this study. However more information is needed in human bladder tissue for clinical usage.     

The effect of testosterone replacement therapy on bladder functions,histology, apoptosis,and Rho-kinase expression in bladder outlet obstruction and hypogonadism rat model

Background/aimThe effect of testosterone replacement therapy was investigated on bladder functions, histology, apoptosis as well as Rho-kinase expression in the rat bladder outlet obstruction (BOO) and hypogonadism models. Materials and methods30 mature male rats divided into 4 groups: sham group (n = 8), BOO group (n = 8), BOO + orchiectomy group (n = 7), BOO + orchiectomy + testosterone (T) treatment group (n = 7). Cystometric findings, apoptosis index, Rho-kinase (ROCK-2) expression, and smooth muscle/collagen ratio were compared. Results BOO did not change ROCK-2 expression level, compared to sham group (P > 0.05). However, when compared to BOO group (P < 0.01), BOO + orchiectomy led ROCK-2 increase. The testosterone treatment failed to reverse the up-regulation of ROCK-2 induced by orchiectomy although it tended to lower ROCK-2 level. Compared to sham group (P = 0.002), changes in maximal bladder capacity and leak point pressure were higher (P = 0.026, P = 0.001), and bladder compliance was lower in BOO group. Also, the apoptosis index was different between the two groups (P = 0.380). Smooth muscle/collagen ratio was higher in BOO + orchiectomy + T group than in BOO + orchiectomy group (P = 0.010).ConclusionsThe research draws attention to alternating treatment approaches in case of the presence of hypogonadism and BOO.     

肾交感神经去除术对心肌肥厚和心肌纤维化的影响

 目的:观察肾交感神经去除术（renal sympathetic denervation, RDN）对心肌肥厚和心肌纤维化的影响,并探讨其可能机制。方法:选用12周龄的健康SD雄性大鼠60只,随机分为假手术组、假手术+RDN组、主动脉缩窄组、主动脉缩窄+RDN组,8周后用介入生理记录仪检测血流动力学和心功能指标,HE染色、苦味酸-天狼星红染色分别观察心肌肥厚和心肌纤维化情况,放射免疫分析法测量血浆肾上腺素浓度、肾素活性、血管紧张素II浓度及心脏血管紧张素II含量。结果:与主动脉缩窄组相比,RDN可显著改善主动脉缩窄大鼠心脏舒张功能［左室舒张末期压力（LVEDP）：(8.03±1.66) mmHg vs(15.77±2.14) mmHg;等容舒张期左室压力下降最大速率（-dp/dt）：(7 793±587) mmHg/s vs(6 353±475) mmHg/s;P＜0.01］、防止其心肌肥厚和纤维化［左心室重量指数：3.340±0.121 vs4.244±0.102;心肌细胞面积：(332.9±28.9) μm2 vs(401.6±33.2) μm2;胶原容积分数：7.76%±0.85% vs12.48%±1.82%;P＜0.01］。然而,RDN不能降低主动脉缩窄大鼠的血压（P＞0.05）。RDN导致主动脉缩窄大鼠的血浆肾上腺素浓度、肾素活性、血管紧张素II浓度及心脏血管紧张素II含量均明显减少（P＜0.01）。结论: RDN可以通过降低交感和肾素-血管紧张素系统活性直接抑制心肌肥厚和心肌纤维化,从而改善心脏功能。     

Urodynamics and bladder muscarinic receptors in rats with cerebral infarction and bladder outlet obstruction

We characterized muscarinic receptor binding and urodynamic parameters in rats with cerebral infarction and chronic bladder outlet obstruction as models of detrusor overactivity. Bladder weight showed little significant difference between the cerebral-infarcted and sham rats, but the bladder weight was about three times greater in the bladder outlet-obstructed rats. Bladder capacity and voided volume were significantly lower (36.7 and 55.1%, respectively) in the cerebral-infarcted than in the sham rats. Involuntary contractions before micturition were seen in the bladder outlet-obstructed rats but not in sham rats. The bladder outlet-obstructed rats showed significant increases (2.65 and 2.57 times, respectively) in bladder capacity and voided volume, compared with those in sham rats. Bmax values for specific [N-methyl-3H]scopolamine ([3H]NMS) binding in the bladder were significantly (34%) increased in the cerebral-infarcted rats compared with sham rats, whereas Kd was unaffected by infarction. On the other hand, there was little significant change in Kd and Bmax for specific [3H]NMS binding in the bladder-obstructed rats compared with sham rats. In conclusion, the present study shows that cerebral infarction but not bladder outlet obstruction in rats causes up-regulation of bladder muscarinic receptors, and that such regulation of bladder muscarinic receptors may be at least partly associated with the symptoms of detrusor overactivity subsequent to cerebral infarction.     

Treatment for chronic ischaemia-induced bladder detrusor dysfunction using bone marrow mesenchymal stem cells: an experimental study

The mechanism of ischaemia-induced bladder dysfunction is not entirely clear, but is thought to be a result of the ischaemia-related M-receptor hypersensitivity to acetylcholine. In addition to nerve injury, ischaemia may cause bladder detrusor fibrosis and urethra de-epithelialization. Bladder dysfunctions caused by bladder outlet obstruction (BOO) and aging detrusor were considered to be associated with chronic ischaemia. To date, there has been no effective treatment for the histological and functional changes of the bladder caused by bladder ischaemia. The present study evaluated the feasibility and effectiveness of using bone marrow mesenchymal stem cells in the treatment of chronic ischaemia-induced bladder detrusor dysfunction in an experimental model. Bone marrow mesenchymal stem cells from Sprague-Dawley (SD) rats were injected into the common iliac artery of experimental animals, then bilateral iliac arteries were ligated and doxazosin mesylate was intragastrically administered. Eight weeks later, urodynamic examination and intravesical pressure measurements were performed on experimental animals. Histological changes of the taken bladder from sacrificed SD rats were evaluated by immunohistochemistry and trichrome staining and the images captured were analyzed by a software program. The average intravesical pressure and detrusor contraction power of the ischaemia group was 16.21±5.26 and 17.26±5.72; those of the experimental group were 24.02±10.06 and 25.84±11.99; the average intravesical pressure and detrusor contraction power of the control group was 28.56±4.48 and 29.57±5.01. The average intravesical pressure and detrusor contraction power of the ischaemia group were significantly lower than those of the experimental and control group, while no significant difference was shown between the experimental and control groups. 5-Bromo-2'-deoxyuridine (BrdU) staining for the experimental group was positive. The percentage of the smooth muscle content in the bladder wall was (25.21±6.28)%, (49.38±6.32)% and (48.00±17.39)% for ischaemia, experimental and control group, respectively. The percentage of smooth muscle in the bladder wall of the ischaemia group was significantly lower than that of the experimental and control group, while no significant difference was observed between the experimental and control groups. The number of nerve fibers per high power field of the experimental group was significantly higher than that of the ischaemia group, but lower than the control group. In conclusion, the percentage of smooth muscle content and the number of nerve cells per high power field decreases in ischaemia bladder, with detrusor contractility decreased. Injection of stem cell suspension into the common iliac artery in rats with ischaemic bladder, followed by intragastric administration of doxazosin mesylate, makes transplanted stem cells regenerate in the bladder tissue, increases the percentage of smooth muscle content and nerve cells per high power field in the bladder wall, and improve bladder detrusor contraction function.     

Molecular mechanisms of bladder outlet obstruction in transgenic male mice overexpressing aromatase (Cyp19a1)

We investigated the etiology and molecular mechanisms of bladder outlet obstruction (BOO). Transgenic (Tg) male mice overexpressing aromatase (Cyp19a1) under the ubiquitin C promoter in the estrogen-susceptible C57Bl/6J genetic background (AROM+/6J) developed inguinal hernia by 2 months and severe BOO by 9 to 10 months, with 100% penetrance. These mice gradually developed uremia, renal failure, renal retention, and finally died. The BOO bladders were threefold larger than in age-matched wild-type (WT) males and were filled with urine on necropsy. Hypotrophic smooth muscle cells formed the thin detrusor urinae muscle, and collagen III accumulation contributed to the reduced compliance of the bladder. p-AKT and ERα expression were up-regulated and Pten expression was down-regulated in the BOO bladder urothelium. Expression of only ERα in the intradetrusor fibroblasts suggests a specific role of this estrogen receptor form in urothelial proliferation. Inactivation of Pten, which in turn activated the p-AKT pathway, was strictly related to the activation of the ERα pathway in the BOO bladders. Human relevance for these findings was provided by increased expression of p-AKT, PCNA, and ERα and decreased expression of PTEN in severe human BOO samples, compared with subnormal to mild samples. These findings clarify the involvement of estrogen excess and/or imbalance of the androgen/estrogen ratio in the molecular pathogenetic mechanisms of BOO and provide a novel lead into potential treatment strategies for BOO.     

依降钙素干预膝骨关节炎模型大鼠软骨下骨的变化

文题释义： 依降钙素：为人工合成的鳗鱼降钙素多肽衍生物,其主要作用是抑制破骨细胞活性,减少骨的吸收,防止骨钙丢失,同时可降低正常动物和高钙血症动物血清钙,对实验性骨质疏松有改善骨强度、骨皮质厚度、骨钙质含量、骨密度等作用。 p38丝裂原活化蛋白激酶(p38 MAPK)：p38 MAPK是MAPK亚族中的一员,它以转录因子为作用目标,通过对转录因子的磷酸化来调节许多转录因子的活性,进而介导炎症反应和细胞凋亡,参与体内细胞的应激反应。研究认为p38 MAPK是调节膝关节骨关节炎发病机制中炎症产生的关键上游信号,影响骨关节炎的发生与发展,是近年来研究的热点。 背景：骨关节炎存在软骨下骨吸收和骨形成失去平衡,课题组前期研究发现膝骨关节炎大鼠关节软骨改变前软骨下骨就已经发生改变。 目的：通过观察膝骨关节炎大鼠膝关节软骨下骨的变化,探讨依降钙素对软骨下骨p38 MAPK表达及软骨下骨吸收的影响。 方法：雌性3月龄SD大鼠随机分为3组：假手术组(n=6)不切除卵巢,切除同卵巢等体积大小的脂肪组织,切开双侧膝关节,但不损伤交叉韧带;模型组(n=6)切除双侧卵巢后,再切断双侧膝关节前交叉韧带,建立卵巢切除+前交叉韧带切断模型,不进行药物干预;依降钙素组(n=6)建立卵巢切除+前交叉韧带切断模型后,依降钙素5 IU/kg,每周2次肌肉注射。12周后分析软骨下骨的骨密度、软骨下骨p38蛋白表达水平以及血清Ⅰ型胶原交联C-末端肽、Ⅱ型胶原交联C-末端肽、白细胞介素1、白细胞介素6、骨特异性碱性磷酸酶、抗酒石酸酸性磷酸酶5b水平。实验方案经南华大学动物实验伦理委员会批准。 结果与结论：①依降钙素组软骨下骨的骨体积分数、骨小梁数量显著高于模型组(P < 0.05),骨小梁分离度显著低于模型组(P < 0.05);②模型组的骨体积分数、骨小梁数量、骨小梁厚度显著低于假手术组(P < 0.01或< 0.05)、骨小梁分离度显著高于假手术组(P < 0.01);③依降钙素组血清Ⅰ型胶原交联C-末端肽、白细胞介素1、白细胞介素6、抗酒石酸酸性磷酸酶5b显著低于模型组(P < 0.05),模型组Ⅰ型胶原交联C-末端肽、Ⅱ型胶原交联C-末端肽、白细胞介素1、白细胞介素6、抗酒石酸酸性磷酸酶5b明显高于假手术组(P < 0.05);④软骨下骨p38蛋白表达水平依降钙素组显著低于模型组(P < 0.01)、模型组显著高于假手术组(P < 0.01);⑤结果说明,依降钙素可能通过下调p38 MAPK表达,抑制骨关节炎促炎因子分泌及软骨下骨吸收。 ORCID: 0000-0002-2108-9820(伍琦) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

粒细胞集落刺激因子改善阿尔茨海默病模型大鼠学习记忆功能及抗炎作用机制

目的 探讨粒细胞集落刺激因子（G-CSF）对阿尔茨海默病（AD）大鼠认知障碍的改善作用及其对脑内炎症反应的影响。 方法 45只雄性SD大鼠,随机分为3组,包括假手术组、模型组、G-CSF组,每组15只。采用β-淀粉样蛋白1-42（Aβ _1-42）复制AD大鼠模型,并给予G-CSF治疗。观察并比较大鼠的学习记忆功能及脑组织病理学变化。采用Western blotting检测核因子κB p65（NF-κB p65）、磷酸化p-38丝裂原活化蛋白激酶（p-p38MAPK）、诱导型一氧化氮合酶（iNOS）、环氧化酶-2（COX-2）、肿瘤坏死因子α（TNF-α）和白细胞介素-1β（IL-1β）蛋白的表达,采用Real-time PCR检测iNOS、COX-2、TNF-α和 IL-1β mRNA水平。 结果 模型组大鼠神经细胞数量明显减少,细胞核固缩明显,核仁不清。G-CSF组大鼠脑组织神经细胞核固缩现象均有显著改善。假手术组、模型组、G-CSF组逃避潜伏期分别为（35.68±6.73）s、（57.92±7.35）s及（40.27±8.91）s;目标象限游泳时间分别为54.72%±4.22%、36.73%±3.21%及44.68%±4.01%;穿过平台次数分别为（8.7±2.1）次、（3.9±1.6）次及（6.5±1.7）次;与模型组相比,假手术组、G-CSF组的逃避潜伏期均显著缩短,目标象限游泳时间显著增加,穿过平台次数显著增加,差异均有统计学意义（P<0.05）。假手术组、模型组、G-CSF组NF-κB p65蛋白表达水平为0.144±0.033、0.502±0.035及0.473±0.061;p-p38 MAPK蛋白水平为0.194±0.021、0.511±0.039及0.266±0.048,与模型组相比,假手术组、G-CSF组NF-κB p65、p-p38MAPK蛋白水平均显著降低,差异均有统计学意义（P<0.05）。与模型组比较,假手术组、G-CSF组iNOS、COX-2、TNF-α 和 IL-1β的蛋白和mRNA 水平显著降低（P<0.05）。 结论 G-CSF具有良好的抑制AD模型大鼠脑神经细胞凋亡,改善大鼠学习记忆功能障碍的作用,其作用可能通过抑制p38MAPK和NF-κB p65的信号通路的过度激活,下调iNOS、COX-2、TNF-α 和 IL-1β等炎性因子的表达,抑制脑内神经炎症状态而实现。