消癃通闭对小鼠前列腺上皮细胞DNA合成的影响

去势昆明雄性小鼠24只,皮下注的丙酸睾丸素5mg/kg·d~(-1),同时将其分为4组,分别给予高剂量消癃通闭、低剂量消癃通闭、雌二醇及睾丸素。于15天、30天时分别腹腔注射~3H TdR(20uci/g),24小时后处死,取前列腺右头叶和腹叶称重,制成石腊切片进行放射自显影,观察前列腺上皮细胞核上标记银盐颗粒。结果结示~3H TdR标记呈不均一性,腺管末梢标记明显。单纯给予丙酸睾丸素组被标记的腺上皮明显高于其它组。用消癃通闭15天后标记细胞减少,但30天后4组间无明显差异。提示消癃通闭对小鼠前列腺的生长、DNA合成有抑制作用。前列腺内DNA合成局限于腺管末梢,前列腺发育到一定程度,其合成也相对停止。     

消癃通闭对大鼠前列腺上皮细胞分裂周期的影响

目的：进一步了解中药消癃通闭对前列腺上皮细胞增殖的影响。方法：用去势Ｗｉｓｔａｒ大鼠，每天皮下注射丙酸睾酮，消癃通闭每天灌胃，于２０ｄ时断头处死，取前列腺各叶称重及测体积，取相同部位腹叶，采用流式细胞技术对大鼠前列腺上皮细胞分裂周期进行定量检测。结果：消癃通闭高剂量组的前列腺体积、腺重量及腺重与体重的比值均明显小于睾酮组，Ｐ＜００１；前列腺上皮细胞分裂Ｇ２＋Ｍ期所占比例亦明显长于睾酮组，Ｐ＜０００５。结论：消癃通闭可抑制前列腺上皮细胞的有丝分裂，从而抑制了前列腺细胞的生长     

消癃通闭对大鼠前列腺组织碱性成纤细胞生长因子表达的影响

为了解消癃通闭对前列腺组织bFGF表达的影响,以去势Wistar大鼠,经皮下每天注射睾酮5mg/kg,来维持其性腺的发育,同时给予中药消癃通闭药粉灌胃,20天时断髓处死。取相同部位前列腺腹叶,以免疫组化定量技术对前列腺组织bFGF进行测试。结果显示,消癃通闭可明显抑制腺组织中的bFGF的表达,与睾酮组相比,P<0.01,并发现bFGF的表达主要位于腺上皮细胞的胞浆,而胞核及间质部分表达较少。     

“消癃通闭”对大鼠前列腺平滑肌α1肾上腺素受体的拮抗作用

采用大鼠离体前列腺平滑肌收缩功能实验方法,研究了中药复方”消癃通闭”对α_1-AR的拮抗作用。结果表明:“消癃通闭”使NE介导的大鼠前列腺平滑肌收缩效应曲线平行右移。“消癃通闭”70.0g/L时,使NE的PD_2值由对照时5.1±0.15减少到4.7±0.02,两组间有明显差异(P<0.01)。“消癃通闭”拮抗的PKB值为44.0±14.0gq“消癃通闭”170g/L时,使NE的PD_2值由对照时5.4±0.16,减少到4.6±0.22,两组间有显著性差异(P<0.01)。消癃通闭的拮抗PKB值为30.0±8.0g/L。两种浓度“消癃通闭”间拮抗的PKB值间无显著性差别。提示“消癃通闭”对α_1-AR有拮抗效应。     

中药"消癃通闭"对大鼠前列腺上皮细胞凋亡的影响

目的：研究中药“消癃通闭”对前列腺上皮细胞凋亡的影响。方法：将大鼠分为消癃通闭组，保列治组，去势组、空白组，采用免疫组化原位末端标记染色法，因实验后第7d,14d,21d时，检测各组大前列腺上皮细胞的凋亡百分率。结果：消癃通闭组及保列治组在实验第7d时，凋亡百分率达到高峰，分别为9．27％、5．65％，第14d时稍减少，第21d锂有所恢复，去势组在第7d时凋亡即出现，到第14d时凋亡百分率达到高峰，与其它组相比差异显著。结论：中药消癃通闭可在较短时间促进大鼠前列腺上皮细胞凋亡，为该药治疗前列腺增生提供了理论依据。     

消癃通闭对小鼠前列腺上皮细胞DNA合成的影响

为了解消癃通闭对前列腺上皮细胞增生的影响，用去势昆明种雄性小鼠，皮下注射丙酸睾酮每天５ｍｇ／ｋｇ，以维持其副性腺的生长发育。消癃通闭灌胃１５天、３０天时，分别经腹注射３ＨＴｄＲ（７４ｋＢｑ／ｇ），２４小时后断髓处死，取前列腺右头叶和腹叶制片，观察前列腺上皮细胞核上标记银盐颗粒。结果显示：３ＨＴｄＲ标记呈不均一性，腺管末梢标记明显。单纯给予丙酸睾酮组标记的腺上皮明显高于其它组，用消癃通闭１５天后标记细胞减少（Ｐ＜００１），但给药３０天后组间标记细胞数无显著性差异。本研究表明：消癃通闭对小鼠前列腺的生长、再生、ＤＮＡ合成有抑制作用，同时说明了前列腺内ＤＮＡ合成局限于腺管末梢，末梢ＤＮＡ的合成待小鼠成熟后，即前列腺发育到一定限度，其合成也就相对停止。     

消癃通闭对大鼠前列腺组织碱性成纤细胞生长因子表达的影响

目的：了解中药消癃通闭对前列腺组织碱性成纤维细胞生长因子（ＢａｓｉｃＦｉｂｒｏｂｌａｓｔＧｒａｏｔｈＦａｃｔｏｒ,ｂＦＧＦ）表达的影响。方法：以去势Ｗｉｓｔａｒ大鼠,经皮下每天注射睾酮,同时给予中药消癃通闭药粉灌胃,２０ｄ时断髓处死。取相同部位前列腺腹叶,以免疫组化定量技术对前列腺组织ｂＦＧＦ进行测试。结果：消癃通闭高剂量组的ｂＦＧＦ表达明显低于睾酮组,Ｐ＜００１;并发现ｂＦＧＦ的表达主要位于腺上皮细胞的胞浆,而胞核及间质部分表达较少。结论：中药消癃通闭治疗前列腺增生的机理之一是该药明显抑制了前列腺组织中的ｂＦＧＦ的表达,进而抑制了前列腺细胞的生长     

消癃通闭对大鼠前列腺上皮细胞凋亡的影响

目的研究中药消癃通闭对前列腺上皮细胞凋亡的影响。方法将大鼠分为消癃通闭组、保列治组、去势组、空白组,采用免疫组化原位末端标记染色法,分别在实验后第7、14、21天时,检测各组大鼠前列腺上皮细胞的凋亡百分率。结果消癃通闭组及保列治组在实验第7天时,凋亡百分率达到高峰,分别为9．27％、5．65％,第14天时稍减少,第21天时有所恢复。去势组在第7天时凋亡即出现,到第14天时凋亡百分率达到高峰,与其它组相比差异显。结论中药消癃通闭可在较短时间促进大鼠前列腺上皮细胞凋亡,为该药治疗前列腺增生提供了理论依据。     

中药"消癃通闭"对前列腺平滑肌中NOS神经的影响

目的　 探讨一氧化氮 (NOS)与前列腺增生 (BPH)发病的关系以及中药消癃通闭对前列腺平滑肌中NOS神经的影响。 方法　 去势雄性大鼠 ,皮下注射丙酸睾酮 5mg/ (kg·d) ,分别给予消癃通闭、保列治灌胃 ,2 1d后断髓处死 ,应用NADPH组化染色结合形态学定量分析方法 ,检测各组前列腺中NOS神经含量。 结果　NOS神经纤维主要分布在前列腺平滑肌细胞周围 ,消癃通闭高剂量组在治疗 3周后 ,前列腺组织中NOS神经的长度密度 (Lv)相对值为 (113.4 9± 2 3.30 )× 10 -3 ,与其它 3组相比均有显著性差异 (P <0 .0 1)。 结论　 中药消癃通闭可增加模型BPH大鼠前列腺中NOS神经含量 ,为该药治疗前列腺增生改善临床症状提供了科学依据     

中药消癃通闭对犬前列腺组织中T、E_2、DHT、AR和5α-还原酶的影响

取前列腺增生老龄犬,经口喂给消癃通闭药粉32天后,用RIA技术测定前列腺组织中睾酮(T)、雌二醇(E_2)、双氢睾酮(DHT),并与血清中浓度对照;用Scatchard Plots方法及双复管单点法测定雄激素受体(AR);用酶学法测定5α—还原酶Ⅰ、Ⅱ型。结果发现:正常犬微粒体中5α—还原酶Ⅰ型的活性为97.145±45.729,Ⅱ型的活性为15.745±15.093(单位均为DHT Pmol/mg Pro—tein·30min~(-1),下同)。予消癃通闭lg/kg·d~(-1)组犬的测定值分别为92.454±57.703和11.328±10.060;给予消癃通闭2g/kg·d~(-1)组犬的测定值分别为42.837±31.909和9.288±11.209。各组的前列腺组织中AR的测定值无明显差异。提示消癃通闭治疗前列腺增生的机理为抑制前列腺组织中5α—还原酶的活性。     

他莫昔酚对大鼠前列腺增生动物模型的抑制作用及机理研究

目的 :探讨他莫昔酚抑制前列腺增生的作用机理。　方法 :将Wistar成年大鼠肌肉注射丙酸睾丸酮 ,同时以他莫昔酚常规灌胃。于给药 7、15、30d时分批处死大鼠 ,称取前列腺重量 ,标本做常规病理切片 ,电镜观察前列腺组织细胞结构变化。　结果 :给药 7d时单用睾酮组前列腺指数高于对照组 ,也高于灌胃组。剩余大鼠分 2组 ,其中 1组继续单用他莫昔酚灌胃 ,于第 15、30d时分组处死剩余大鼠。光镜染色观察及电镜扫描均证实他莫昔酚组前列腺上皮细胞及基质的增殖被抑制。　结论 :他莫昔酚通过对雌激素的拮抗作用 ,阻断雌激素在前列腺增生中的作用 ,从而抑制前列腺增生。     

Testosterone induces vascular endothelial growth factor synthesis in the ventral prostate in castrated rats

PURPOSE: Recent studies suggest that the vasculature is important for the control of prostate growth. Castration induces an involution of the prostate gland and its vasculature. Replacement of testosterone stimulates endothelial cell proliferation and normalizes vascular volumes and blood flow several days before organ regrowth. Antiangiogenesis treatment inhibits the growth of prostate tumors. Understanding the regulation of the prostate vasculature may therefore provide important knowledge of the mechanisms responsible for the growth of non-malignant and malignant prostate tissue. Castration induced regression and testosterone stimulated regrowth of the prostatic vasculature have here been used to study the involvement of the angiogenic factor vascular endothelial growth factor (VEGF) and its receptors flt-1 and flk-1/KDR in the regulation of the prostatic vasculature. MATERIALS AND METHODS: VEGF, flt-1, and flk-1/KDR levels were quantified in the rat ventral prostate following castration and testosterone replacement. Methods used were competitive RT-PCR, Western blot and immunohistochemistry. RESULTS: VEGF mRNA and protein levels were significantly decreased by castration and testosterone treatment induced VEGF synthesis in the rat ventral prostate epithelium. Flt-1 and flk-1/KDR receptor levels were unaffected by castration and testosterone treatment. CONCLUSIONS: Castration down regulates VEGF and testosterone induces VEGF synthesis in epithelial cells in the rat ventral prostate.     

Differential effects of growth factor antagonists on neoplastic and normal prostatic cells

Growth factors such as epidermal growth factor and fibroblast growth factor have been suggested to be involved as paracrine or autocrine mediators of androgen action in normal and malignant prostatic cells. Suramin and protamine are potent in vitro growth factor antagonists. To evaluate the effect of growth factor antagonists on prostatic growth, suramin and protamine were administered to castrated rats simultaneously receiving exogenous testosterone replacement in an attempt to block androgen-dependent restoration of the normal rat ventral prostate. Using this prostatic restoration model, there was no statistical difference in the prostate wet weight or total DNA content attained between rats given testosterone alone and those given testosterone in combination with either suramin or protamine. In intact rats, suramin administration caused an 80% reduction in serum testosterone; concomitantly, these rats had a 40% reduction in mean prostate weight. This decrease in size could be blocked with androgen supplementation. To examine the effects of growth factor antagonists on neoplastic prostatic cell growth, rats bearing the androgen-independent AT-2 subline of the Dunning R-3327 tumor were treated with either suramin or protamine. The same dosing regimen found to be ineffective in blocking the restoration of the involuted prostate of castrated rats resulted in a significant reduction in the growth rate of AT-2 tumors. These results suggest that the growth factor antagonists suramin and protamine given in vivo have a differential ability to slow the growth of malignant vs. normal prostatic cells.     

Prostatic development, cytodifferentiation and growth: epithelial-mesenchymal interaction and heterogeneity of prostatic cellular activity

Prostatic growth and hormonal effects on the prostate play a basic role in the pathogenesis of abnormal proliferative diseases (i.e. benign prostatic hyperplasia and prostatic carcinoma). During the embryonic period, the prostate is organized through the budding and branching of the epithelial cords into the urogenital sinus mesenchyme. The urogenital sinus mesenchyme has an inductive potential for the prostatic epithelial development and cytodifferentiation under the influence of androgen and an epithelial-mesenchymal interaction. In this interaction, mesenchymal cells are considered as a mediator of hormonal action on epithelial cells. Postnatal prostatic growth is obtained further ductal branching morphogenesis and regulated by the epithelial-mesenchymal interaction, androgen, and epithelial/mesenchymal ratio. Castration-induced degeneration, and androgen-induced regeneration of the glandular architecture of the mouse prostate were investigated by microdissection techniques that permitted precise quantitation of the numbers of the terminal ductal tips and ductal branch-points. During the first 15 days after birth, active branching morphogenesis occurred as a result of focally high levels of DNA synthesis confined almost exclusively to the distal tips of the branching ducts. Following castration about 35% of the ductal tips and branch-points were lost in distal regions (usually near the capsule). By contrast, in more proximal regions of the prostate the ducts survived in an atrophic condition. The lost distal ductal morphology completely regenerated following administration of testosterone propionate (TP) to the castrated males. Whole-mount autoradiography demonstrated that labelling intensity reached a maximum on the third day of TP treatment in distal ductal areas. Recognition of the mesenchymal-epithelial interaction and heterogeneities in the morphogenesis, androgen dependency, and DNA synthetic activity within the prostatic architecture is fundamental to understanding the mechanism of androgenic regulation of normal or abnormal prostatic growth and development.