一种新型嘧啶基取代芳基苯酸衍生物对人多囊肾囊肿衬里上皮细胞增殖抑制的实验研究

目的:观察新型PPARγ激动剂DH9对人多囊肾囊肿衬里上皮细胞(WT9-12)增殖的影响。方法:予以不同浓度的DH9及罗格列酮作用WT9-12细胞24、48、72、96 h,MTT法检测细胞增殖情况;流式细胞术观察细胞周期的改变;AnnexinV+PI双染色流式细胞术测定细胞凋亡率;Western blotting分析cyclinA、P21CIP/WAF1、Bax和Bcl-2的表达。结果:DH9抑制细胞增殖作用明显优于罗格列酮(P<0.05),并呈量-效和时-效关系。比较加入PPAR-γ抑制剂GW9662前后细胞增殖抑制作用差异无统计学意义(P>0.05)。DH9可增加P21CIP/WAF1蛋白合成,减少CyclinA蛋白合成,使细胞阻滞在S期。DH9作用72 h后Bcl-2/Bax的比值分别为较对照组(2.19±0.08)显著减少(P<0.05),具有促进细胞凋亡的作用。结论:DH9抑制WT9-12细胞增殖活性明显优于罗格列酮,且这种增殖抑制作用与DH9作用在细胞周期的不同调节点,促进WT9-12细胞凋亡有关。     

罗格列酮对多囊肾囊肿衬里上皮细胞p38促分裂原活化蛋白激酶信号通路的影响

Objective To investigate the effect of rosiglitazone on p38 mitogen-activated protein kinase (p38MAPK) pathway in polycystic kidney cyst-lining epithelial cells. Methods The cyst-lining epithelial cells (PKD cells) from human polycystic kidney were treated with rosiglitazone (10 μmol/L), peroxisome proliferator-activated receptor-γ (PPARγ) inhibitor GW9662 (10 μmol/L), rosiglitazone (10 μmol/L) +GW9662 (10 μmol/L), p38MAPK specific inhibitor SB203580 (10 μmol/L), SB203580 (10 μmol/L)+ rosiglitazone(10 μmol/L) for 2 hours followed by epidermal growth factor (EGF) stimulation. Protein expressions of p38, phuspho-p38 (p-p38) and proliferating cell nuclear antigen (PCNA) were detected by Western blot. p38 mRNA was examined by RT-PCR. Expression of c-fos and c-jun was observed by immunocytochemistry. Results (1) EGF markedly up-regulated the expressions of p38, p-p38, PCNA, c-fos anti c-jun compared with control group (P<0.01). (2) Compared with EGF treated group, rosiglitazone significantly reduced p38 activation and mRNA expression (P<0.01, respectively). Rosiglitazone, rosiglitazone+SB203580 could significantly down-regulated p-p38, PCNA, c-fos and c-jun expression (P<0.01, respectively) with no significant difference between these two groups. (3) GW9662 partially reversed the reduction effect of rosiglitazone. Conclusions Rosiglitazone can inhibit proliferation of autosomal dominant polycystic kidney disease cyst-lining epithelial cells partially through down-regulating p38 activation and reducing c-fos, c-jun and PCNA expression. The above effect of rosiglitazone is in part PPARγ-independcnt.     

罗格列酮对多囊肾囊肿衬里上皮细胞p38促分裂原活化蛋白激酶信号通路的影响

Objective To investigate the effect of rosiglitazone on p38 mitogen-activated protein kinase (p38MAPK) pathway in polycystic kidney cyst-lining epithelial cells. Methods The cyst-lining epithelial cells (PKD cells) from human polycystic kidney were treated with rosiglitazone (10 μmol/L), peroxisome proliferator-activated receptor-γ (PPARγ) inhibitor GW9662 (10 μmol/L), rosiglitazone (10 μmol/L) +GW9662 (10 μmol/L), p38MAPK specific inhibitor SB203580 (10 μmol/L), SB203580 (10 μmol/L)+ rosiglitazone(10 μmol/L) for 2 hours followed by epidermal growth factor (EGF) stimulation. Protein expressions of p38, phuspho-p38 (p-p38) and proliferating cell nuclear antigen (PCNA) were detected by Western blot. p38 mRNA was examined by RT-PCR. Expression of c-fos and c-jun was observed by immunocytochemistry. Results (1) EGF markedly up-regulated the expressions of p38, p-p38, PCNA, c-fos anti c-jun compared with control group (P<0.01). (2) Compared with EGF treated group, rosiglitazone significantly reduced p38 activation and mRNA expression (P<0.01, respectively). Rosiglitazone, rosiglitazone+SB203580 could significantly down-regulated p-p38, PCNA, c-fos and c-jun expression (P<0.01, respectively) with no significant difference between these two groups. (3) GW9662 partially reversed the reduction effect of rosiglitazone. Conclusions Rosiglitazone can inhibit proliferation of autosomal dominant polycystic kidney disease cyst-lining epithelial cells partially through down-regulating p38 activation and reducing c-fos, c-jun and PCNA expression. The above effect of rosiglitazone is in part PPARγ-independcnt.     

成纤维细胞生长因子对关节软骨细胞转化生长因子β1作用的免疫组织化学观察

目的观察成纤维细胞生长因子对培养兔关节软骨细胞转化生长因子β１的表达。方法培养１月龄新西兰兔关节软骨细胞，在每次换液时加入成纤维细胞生长因子１００ｎｇ／ｍｌ，铺满后收集细胞，作关节软骨细胞转化生长因子β１常规及电镜免疫组织化学观察。结果光镜下实验组细胞明显呈棕色，电镜下细胞膜表面有明显的胶金颗粒黏附。结论成纤维细胞生长因子能促进关节软骨细胞转化生长因子β１的表达。     

100例小脑镰的形态学研究

调查100例小脑镰,按嵴数可分为单嵴、双嵴、三嵴和四嵴等型,其中以双嵴型为最多见,共有60例。小脑镰的长度平均为41mm,中点宽平均为4 mm,中点高平均为6 mm。通过小脑镰基底部的枕窦共出现66例。     

臀肌注射区的选择与应用

前言肌内注射给药能迅速被机体所吸收,并能使血液中及时地获得有效浓度,因而对于治疗许多疾病是有利的,这些长处都是经口给药所不及的,故为患者所欢迎。虽然注射给药的途径有许多优点,但也要注意防止发生事故,据日     

FGF对关节软骨细胞基质及其TGFβ1表达的作用

目的:观察成纤维细胞生长因子(FGF)对培养兔关节软骨细胞基质及转化生长因子β1(TGFβ1)表达的影响.方法:培养1月龄新西兰兔关节软骨细胞,在每次换液时加入FGF 100 ng/ml,铺满后收集细胞,作关节软骨细胞TGFβ1检测及电镜免疫组织化学观察.采用氯胺T法和Antonopoulos法,作细胞基质的胶原和蛋白多糖含量测定.结果:FGF能明显促进胶原和蛋白多糖的合成,并且与剂量、时间呈正相关(P＜0.01).光镜下实验组细胞明显呈棕色,电镜下细胞膜表面有明显的胶金颗粒粘附.结论:FGF能促进关节软骨细胞TGFβ1的表达及基质合成,推迟了关节软骨细胞的成熟,维持细胞的软骨表现型,这些作用有利于关节软骨损伤后的修复,可能具有一定的临床意义.     

镧示踪法透射电镜显示肾脏细胞糖萼的方法学研究

目的 不同方案灌注小鼠后电镜观察肾脏各种细胞糖萼的显示情况,探讨镧示踪法显示肾脏细胞糖萼的最佳方法.方法 根据灌注方案的不同,将C57/BL6小鼠分为3组:①生理盐水组:小鼠放血后,用生理盐水5 ml自主动脉灌注,取肾脏后用2％硝酸镧一戊二醛固定液固定;②硝酸镧固定液组:用2％硝酸镧一戊二醛固定液5 ml相同方法灌注,取肾脏后固定方法同前;③硝酸镧溶液组:用2％硝酸镧溶液和2％硝酸镧一戊二醛固定液5 ml先后进行灌注,取肾脏后固定方法同前.各组均应用透射电镜观察肾脏的肾间质血管内皮细胞、肾小球内皮细胞、足细胞和肾间质细胞连接的糖萼的显示情况,计算各组不同肾脏细胞糖萼的显示率,并进行比较.结果 镧示踪法透射电镜显示的小鼠肾脏细胞的正常糖萼表现为附着于细胞表面的一层电子致密物层.生理盐水组的肾间质血管内皮细胞、肾小球内皮细胞糖萼层的显示率均为0％;足细胞糖萼的显示率为20％;肾间质细胞连接的糖萼显示率(92％)较高.硝酸镧固定液组的肾间质血管内皮细胞和肾小球内皮细胞显示率较低(分别为16％和12％);足细胞糖萼和细胞连接的糖萼层显示率相对较高(分别为84％和96％).硝酸镧溶液组的肾脏各种细胞糖萼均能清晰显示,肾间质血管内皮细胞、肾小球内皮细胞、足细胞和细胞连接糖萼的显示率分别为100％、96％、92％和100％.与生理盐水组和硝酸镧固定液组比较,硝酸镧溶液组的肾间质血管内皮细胞、肾小球内皮细胞糖萼的显示率均显著增高(P＜0.01).硝酸镧固定液组和硝酸镧溶液组的足细胞糖萼的显示率均较生理盐水组显著增高(P＜0.01).各组的肾间质细胞连接糖萼的显示率无统计学差异(P＞0.05).结论 2％硝酸镧溶液和2％硝酸镧一戊二醛固定液先后灌注小鼠,取肾脏用相同固定液固定后行透射电镜检查,能清晰显示肾脏各种细胞糖萼层,尤其是肾脏血管内皮细胞糖萼层,这为肾脏糖萼的相关研究提供一种可靠的实验方法.