胆囊黏膜ABCG5和ABCG8与胆囊胆固醇息肉关系的研究

目的:研究胆囊黏膜ABCG5和ABCG8的基因表达与胆囊胆固醇息肉发病的关系.方法:收集63例因胆囊疾患而行腹腔镜胆囊切除术患者的胆石、胆汁及部分胆囊黏膜组织,其中胆囊胆固醇息肉32例,胆囊胆固醇结石18例,对照13例(胆囊腺瘤6例,非胆固醇胆囊结石7例),分别测定胆石胆固醇含量,胆汁胆固醇、胆汁酸、磷脂的浓度,实时定量PCR检测胆囊黏膜ABCG5和ABCG8的mRNA表达.结果:胆固醇息肉组胆汁胆固醇饱和指数较对照组明显增高(P<0.01),结石组胆囊黏膜ABCG5和ABCG8的表达量较息肉组和对照组显著增高,息肉组和对照组ABCG5和ABCG8的表达量差异无统计学意义(P>0.05).结论:胆囊黏膜ABCG5/ABCG8 mRNA的相对低水平表达,可能是导致胆囊胆固醇息肉发病的重要因素之一.     

胆囊黏膜ABCG5和ABCG8基因在胆固醇结石病中的作用

目的探讨ABCG5和ABCG8基因在胆囊黏膜中的表达与胆固醇结石病的关系。方法采集28例胆囊胆固醇结石病患者和10例无胆石对照的胆囊黏膜和胆汁、胆石。测定胆汁胆固醇、胆汁酸和磷脂含量;Real-time PCR测定胆囊黏膜中ABCG5和ABCG8的表达量。结果胆石组中胆固醇摩尔百分比和饱和指数均较对照组显著升高(P<0.01);ABCG5和ABCG8的表达呈显著正相关(r=0.55,P<0.01),胆石组中ABCG5和ABCG8的mRNA表达分别是对照组的2.8倍(P<0.01)和1.9倍(P<0.05)。结论胆固醇结石病患者的胆囊黏膜ABCG5和ABCG8基因表达增高,限制了胆囊上皮对胆固醇的吸收,使胆汁过饱和状态得以维持。     

胆囊黏膜ABCA1的表达和胆囊胆固醇息肉发病关系的研究

目的：研究ABCA1在胆囊黏膜的表达及其表达与胆囊胆固醇息肉发病的关系。方法：收集因胆囊疾患而行腹腔镜胆囊切除术病人的胆石、胆汁、胆囊黏膜及胆囊壁全层组织共计42例,其中胆囊胆固醇息肉15例,胆囊胆固醇结石15例,对照组12例（胆囊腺瘤5例,非胆固醇胆囊结石7例）。分别测定胆石胆固醇含量、胆汁胆固醇、胆汁酸、磷脂的浓度;实时PCR定量检测胆囊黏膜ABCA1、LXRα、RXRα的mRNA表达：胆囊壁全层组织做病理切片;免疫组化显示ABCAl蛋白在胆囊黏膜的表达．结果：胆固醇息肉组胆汁胆固醇饱和指数为1．0±0．2,较对照组胆固醇饱和指数0．6±0．3明显增高,其差别有统计学显著性（P〈0．01）。免疫组化显示ABCA1在胆囊黏膜上皮细胞有明显表达。息肉组、结石组和对照组胆囊黏膜ABCAI、LXRa、RXRα mRNA相对表达量比较,各组之间差异无统计学意义。结论：胆囊黏膜ABCA1的表达可能并不是导致胆囊胆固醇息肉发病的重要原因。     

胆囊胆固醇沉积症病人胆囊黏膜脂肪酸合成关键基因表达的研究

目的:研究胆囊胆固醇沉积症病人胆囊黏膜内与脂肪酸合成相关的基因表达。方法:采集38例因胆囊结石病行腹腔镜胆囊切除术病人的胆囊黏膜组织,其中23例伴有胆固醇沉积症(弥漫性12例,息肉样11例),15例无胆固醇沉积症。采用定量PCR方法测定胆囊黏膜中脂肪酸合成途径关键基因的mRNA表达。结果:无论是弥漫性还是息肉样胆固醇沉积症病人,其胆囊黏膜中乙酰辅酶A羧化酶表达分别较无胆固醇沉积症病人增加34%和32%(P<0.05),硬脂酰辅酶A去饱和酶1的表达分别升高8.35倍和5.27倍(P<0.05),脂肪酸合成酶的表达也有增高趋势,但是无统计学差异。转录因子甾醇反应元件结合蛋白1c在两组病人的胆囊黏膜表达也增加88%和132%。结论:胆囊黏膜脂肪酸合成增加可能是胆囊胆固醇沉积症形成的原因之一。     

胆固醇结石病人肝脏胆小管侧膜ATP基因表达差异的研究

目的:本研究旨在测定比较胆石病人与对照组肝脏胆小管侧膜转运蛋白表达差异,以探讨胆石病发生的分子生物学机制。方法:研究包括20例胆囊胆固醇结石病人和11例无胆石症的对照。测定血清胆固醇和甘油三酯、胆汁胆固醇、胆汁酸和磷脂含量,采用Carey表计算胆汁胆固醇饱和指数。实时定量PCR法测定肝脏胆小管侧膜转运蛋白(ABCG5、ABCG8、ABCBll和ABCB4)mRNA的表达量。结果:胆石组血清胆固醇低于对照组(P<0.05)。胆石组胆汁胆固醇摩尔百分比和胆固醇饱和指数较对照组显著升高(P<0.01)。胆石组肝脏胆小管侧膜胆固醇转运蛋白ABCG5和ABCG8表达高于对照组,且后者差异具有统计学显著性(ABCG5:31.44±3.17Vs25.72±3.27,ABCG8:27.53±3.06vs17.81±2.23)。ABCBll和ABCB4表达在两组间差异无显著性。结论:本研究显示,胆石病主要病理生理异常为胆汁胆固醇过饱和,与肝脏胆小管侧膜胆固醇转运蛋白ABCG5和ABCG8的mRNA表达增加有关。     

胆固醇结石病人肝脏脂质代谢异常的分子生物学研究

目的:研究导致胆石病人胆汁胆固醇过饱和的肝脏胆固醇和胆汁酸代谢途径中的分子生物学改变。方法:收集22例胆石病人和13例无胆石病的对照病人肝脏活检组织、胆囊胆汁和血浆。采用实时定量PCR检测肝脏基因表达,采用Western印迹法测定蛋白含量。结果:胆石病人较对照组ABCG5/ABCG8和LXRα基因的mRNA表达水平分别增加51%、59%和102%。肝脏SRBI的mRNA和蛋白含量均增加。结论:胆石病人ABCG5/ABCG8基因表达上调,可能与LXRα表达增加促进相关,这些异常是导致胆汁胆固醇过饱和的原因。此外,胆汁中过多的胆固醇可能来源于经肝脏高密度脂蛋白受体SRBI的摄取,而不是由于肝脏合成和酯化的异常。     

胆囊胆固醇息肉和胆囊结石发病关系的研究

目的 探讨胆囊胆固醇息肉和胆囊结石的发生关系.方法 选择2008年4～9月上海交通大学医学院附属瑞金医院外科微创中心行腹腔镜胆囊切除术病人62例,分别收集病人的胆石、胆汁及胆囊壁全层组织,其中胆固醇息肉31例,胆固醇胆囊结石18例,对照组13例.入院后上午餐前及餐后1h行B超胆囊三径测量,计算胆囊排空率对比显示胆囊功能.测定结石胆固醇含量及胆汁胆固醇、磷脂、胆汁酸浓度,胆囊壁组织做病理检查.结果 胆固醇息肉组病人平均胆囊排空率为(47.3±18.6)%.较健康成人平均胆囊排空率(71.7±8.1)%明显降低,其差异有统计学意义(P<0.01),而胆固醇息肉组和胆固醇结石组病人平均胆囊排空率(47.6±23.7)%比较,差异无统计学意义.胆固醇息肉组胆汁胆固醇饱和指数为(1.0±0.2),较对照组胆固醇饱和指教(0.6±0.3)明显增高,其差异有统计学意义(P<0.01),而胆固醇息肉组和胆固醇结石组胆固醇饱和指数(1.0±0.2)比较,差异无统计学意义.31例胆固醇息肉病人中,合并胆囊结石13例,结石发病率为41.9%.结论 胆囊胆固醇息肉的存在可以促使胆囊结石形成.     

家族性胆固醇结石患者肝组织CYP7A1 mRNA的表达

目的:研究家族遗传性胆囊胆固醇结石患者、散发性胆固醇结石患者和非结石患者肝组织CYP7A1基因mRNA的表达改变及其与胆汁成石性变化的关系.方法:应用逆转录-聚合酶链反应(RT-PCR)技术,研究了28例具有家族遗传性胆囊胆固醇结石患者、30例散发性结石患者和32例非结石患者肝组织CYP7A1 mRNA的表达变化;并采用生物化学技术行胆汁脂质分析,计算成石指数.结果:家族遗传性和散发性胆固醇结石组CYP7A1 mRNA表达水平较非胆固醇结石组降低,差异有显著性;在家族遗传性和散发性胆固醇结石组两组间无统计学差别;各组肝组织CYP7A1 mRNA表达与胆汁成石指数(LI)呈负相关.结论:胆囊胆固醇结石患者肝组织CYP7A1 mRNA水平降低,因而CYP7A1 mRNA表达下调可能是胆囊胆固醇结石形成的重要原因之一;CYP7A1是决定胆汁成石性和胆囊胆固醇结石发生的一个重要基因.     

胆结石病人肝脏核受体LRH-1及其调控基因表达的研究

目的 研究胆囊胆固醇结石病人肝脏的肝脏受体类似物1(Liver receptor homolog 1,LRH-1)及其调控的三磷酸腺苷结合盒(ABC)转运体家族成员abcg5/8的表达,探讨胆固醇结石病发病的机制.方法 27例胆囊胆固醇结石病人,男6例,女21例;年龄平均52岁.10例无胆石症的胆囊息肉病人为对照,男6例,女4例;平均年龄48岁.测定胆汁脂类成分和计算胆汁胆固醇饱和指数.实时定量PCR法测定肝脏LRH-1及abcg5/8 mRNA的表达量.结果 胆石组LRH-1表达高于对照组(14.18±1.80vs7.22±2.22,P<0.05),胆石组肝脏abcg基因mRNA表达量均较对照组增高(abcg5:49.34±3.68 vs 33.48±2.77,P<0.05)abcg8:38.93±5.70 vs 18.70±3.42,P<0.05).胆石组胆汁呈胆固醇过饱和(1.17±0.02).结论 该研究结果提示人类肝脏LRH-1及其调控的abcg5/8的表达增高与胆囊胆固醇结石形成有关.     

胆囊黏膜G蛋白偶联胆汁酸受体1表达与胆囊结石致石胆汁关系的研究

目的 研究胆囊结石患者胆囊黏膜G蛋白偶联胆汁酸受体1 (GPBAR1)表达与致石胆汁形成的关系.方法 收集34例胆囊结石病和15例无胆石对照患者的胆囊黏膜、囊壁、胆汁、静脉血.HE常规染色病理检查胆囊壁,免疫组化染色法检测胆囊壁GPBAR1、黏蛋白1(MUC1)和黏蛋白5AC(MUC5AC)的表达水平;RT-PCR反应检测胆囊黏膜GPBAR1、MUC1和MUC5AC的基因表达水平;检测术前血清和胆囊胆汁的主要脂质成分等.结果 胆囊结石组胆囊HE染色均呈慢性炎症表现;胆囊结石组GPBAR1、MUC5AC的蛋白和mRNA表达均较对照组明显升高(61.34±8.06比43.05±7.83,P＜0.01; 52.11±9.62比45.05±9.27,P＜0.05;0.87±0.07比0.80±0.09,P＜0.05;1.04±0.22比0.8±0.17,P＜0.01).胆囊结石组血清总胆固醇水平和胆汁总胆固醇浓度、总胆固醇摩尔百分比、胆固醇饱和指数、黏蛋白浓度均较对照组明显升高(5.07±1.64比3.62±1.42,P＜0.01;17.23±3.67比12.47±2.31,P＜0.01;7.47±0.65比5.05±0.24,P＜0.01;1.03±0.58比0.69±0.38,P＜0.01;92.02±20.89比76.36±19.71,P＜0.05);而胆汁总胆汁酸浓度、胆汁酸摩尔百分比较对照组明显降低(162.68±20.19比180.21±26.05,P＜0.05; 71.28±1.84比73.29±0.96,P＜0.01).胆囊结石组胆囊GPBAR1 mRNA表达与胆汁总胆汁酸水平负相关(γ=-0.341,P＜0.05),GPBAR1表达与胆汁总胆汁酸水平和总脂质水平均负相关(γ=-0.365,P＜0.05;γ=-0.403,P＜0.05).结论 GPBAR1在胆囊结石患者胆囊黏膜表达增强,介导胆汁酸吸收,可能与致石胆汁形成有关.     

胆囊结石病人小肠胆固醇吸收相关基因表达的初步研究

目的 该研究通过分析胆石病人小肠胆固醇吸收相关基因的表达情况探讨小肠胆固醇吸收的遗传凶素在胆石病发生中的作用.方法 研究包括10例胆囊结石病人和7例无胆石症的对照.实时定量PCR法测定近段空肠黏膜胆固醇吸收相关基因mRNA的表达量.结果 胆石组小肠胆固醇吸收相关基因的mRNA表达量与对照组没有统计学差异(NPCIL1:1.15±0.60 vs 0.80±0.29;ABCG5:3.11±2.70 vs 1.92±1.38;ABCG8:3.65±2.08 vs 2.85±1.33;ACAT2:0.30±0.25vs 0.20±0.16;MTTP:12.35±8.10 vs 8.22±5.74,P>0.05).结论 胆石病人小肠胆固醇吸收相关基因表达差异不是胆石病发生的主要遗传因素.     

High glucose increases mesangial lipid accumulation via impaired cholesterol transporters

Diabetes, whether it occurs before or after transplantation, plays an important role to decrease graft function and survival. In addition renal lipid accumulation has been suggested to play a role in the development and progression of chronic renal allograft rejection. Intracellular lipid accumulation is governed by a balance between the influx and efflux of lipid. Cholesterol transporters, such as scavenger receptor (SR)-A1, CD36, and ATP binding cassette (ABC) A1 and G1 (ABCG1), coordinate to regulate cellular lipid status. Therefore, in the present study, we examined whether high glucose caused lipid accumulation in mesangial cells as a result of altered cholesterol transporters. Mouse mesangial cells were stimulated with 30 mmol/L D-glucose (high glucose); 100 μmol/L oleic acid (OA) used as a positive control. Cellular lipid accumulation was measured by Oil Red O staining. Protein and mRNA expression of cholesterol influx (SR-A1 and CD36) and efflux (ABCA1 and ABCG1) transporters were evaluated using Western blot analysis and real-time quantitative polymerase chain reaction, respectively. High glucose was shown to significantly increase lipid accumulation in mesangial cells at 24 hours as was observed for OA. SR-A1 and CD36 mRNA expression levels were 1.5-fold and 3.5-fold higher, respectively, in high glucose-stimulated than control mesangial cell, whereas ABCG1 mRNA expression decreased to 60% of controls; however, there was no decrease in ABCA1 mRNA. Altered protein expression of each transporter in mesangial cells cultured under conditions of high glucose concentrations was consistent with mRNA expression. Osmotic control using mannitol did not significantly affect any of the measured parameters in the present study. These results demonstrated that high glucose, in itself, can induce mesangial lipid accumulation; this effect may be associated with an impaired balance between the influx and efflux of cholesterol.     

氟尿嘧啶对人结肠癌SW480细胞ABCG2表达的影响

目的观察氟尿嘧啶（5-FU）对人结肠癌SW480细胞ABCG2表达的影响。方法用不同药物浓度的5-FU处理SW480细胞,用CCK8法检测5-FU在SW480中的IC50,流式细胞仪检测SW480细胞ABCG2的阳性表达率．RT—PCR检测ABCG2的mRNA在SW480细胞中的表达差异。结果5-FU对SW480细胞的IC50随着药物浓度的增加而升高（P〈0．05）。流式细胞仪检测发现,正常SW480细胞（A组）中ABCG2阳性表达率为（6．26±0．86）％;在药物处理48h后即刻检测时（B组）的阳性表达率下降至（3．43±1．18）％（P〈0．05）;在药物处理48h后的第2代细胞检测时（c组）则升高至（12．91±3．42）％（P〈0.05）。3组ABCG2mRNA表达趋势与流式细胞仪检测结果的趋势一致。结论不同浓度的5-FU可以影响人结肠癌SW480细胞ABCG2的表达。     

三磷酸腺苷结合盒转运体成员2在人脑胶质瘤中的表达及其与神经巢蛋白表达的关系

目的 探讨三磷酸腺苷结合盒转运体成员2(ABCG2)在人脑胶质瘤组织中的表达及其与胶质瘤分级和神经巢蛋白(nestin)表达的关系.方法 实时荧光定量聚合酶链反应(PCR)检测ABCG2在52例不同病理级别胶质瘤中的mRNA表达;免疫组织化学SABC法检测ABCG2和nestin的蛋白表达.结果 ABCG2在脑胶质瘤中的mRNA表达与正常脑组织比较呈过表达(P<0.05),且随着病理级别的升高而增加(P<0.05);免疫组织化学显示ABCG2在52例胶质瘤中的阳性表达率为32.7%(17/52),并与病理级别明显相关(x2=4.62,P<0.05),主要呈亲血管分布;ABCG2的表达与nestin的表达明显相关(x2=7.60,P<0.05).结论 ABCG2在人脑胶质瘤中的表达随着病理级别的升高而逐渐增加,且与nestin的表达呈正相关;脑肿瘤干细胞可能与神经干细胞具有一定的同源性.     

胆固醇对人半月板纤维软骨细胞影响的初步探讨

目的:探究胆固醇对人半月板纤维软骨细胞基质合成及降解相关基因表达的影响及其机制。方法:获取关节镜手术患者半月板组织并提取纤维软骨细胞,分为对照组（正常细胞不做处理）、阳性对照组（白介素-1 β进行退行性病变造模）和15 μg/mL组（给予15 μg/mL浓度胆固醇处理细胞）、30 μg/mL组（给予30 μ...     

Upregulation of COX-1 and COX-2 in nasal polyps in cystic fibrosis

BACKGROUND: Since abnormalities in prostanoid metabolism occur in the lower airway of patients with cystic fibrosis (CF), it is likely that they could also be detected in the nose. METHODS: The degree of mRNA and protein expression of cyclo-oxygenase (COX) enzymes 1 (COX-1) and 2 (COX-2) was examined using quantitative reverse competitive polymerase chain reaction (RT-PCR) and Western blot analysis in the nasal polyps from 10 patients with CF, nasal polyps from 10 non-CF patients and 11 nasal mucosa specimens. The results are presented as 10(6) cDNA molecules/mug total RNA and the densitometric ratio between protein and beta-actin. RESULTS: COX-1 mRNA levels were significantly higher in CF nasal polyps (median 2.34, 25-75th percentiles 1.6-3.2) than in the nasal mucosa (0.78, 0.11-1.21), while there was no difference with non-CF nasal polyps (1.11, 0.80-3.15). COX-1 protein levels were significantly higher in CF nasal polyps (3.63, 2.71-4.27) than in nasal mucosa (1.55, 0.66-2.33) and non-CF nasal polyps (2.19, 1.72-3.68). COX-2 mRNA was significantly higher in CF nasal polyps (3.34, 2.42-7.05) than in nasal mucosa (1.69, 0.19-3.50). No differences were found in COX-2 mRNA expression between CF and non-CF polyps (1.38, 0.12-6.07). COX-2 protein levels were also significantly higher in CF nasal polyps (0.23, 0.04-0.34) than in non-CF nasal polyps (0.011, 0.009-0.016) or nasal mucosa (0.014, 0.014-0.016). CONCLUSIONS: Upregulation in the expression of COX-1 and COX-2 could explain the high production of prostanoids reported in CF. These findings raise questions regarding the potential use of selective or non-selective COX-2 non-steroidal anti-inflammatory treatment in CF.     

Loss of both ABCA1 and ABCG1 results in increased disturbances in islet sterol homeostasis, inflammation, and impaired β-cell function

Cellular cholesterol homeostasis is important for normal β-cell function. Disruption of cholesterol transport by decreased function of the ATP-binding cassette (ABC) transporter ABCA1 results in impaired insulin secretion. Mice lacking β-cell ABCA1 have increased islet expression of ABCG1, another cholesterol transporter implicated in β-cell function. To determine whether ABCA1 and ABCG1 have complementary roles in β-cells, mice lacking ABCG1 and β-cell ABCA1 were generated and glucose tolerance, islet sterol levels, and β-cell function were assessed. Lack of both ABCG1 and β-cell ABCA1 resulted in increased fasting glucose levels and a greater impairment in glucose tolerance compared with either ABCG1 deletion or loss of ABCA1 in β-cells alone. In addition, glucose-stimulated insulin secretion was decreased and sterol accumulation increased in islets lacking both transporters compared with those isolated from knockout mice with each gene alone. Combined deficiency of ABCA1 and ABCG1 also resulted in significant islet inflammation as indicated by increased expression of interleukin-1β and macrophage infiltration. Thus, lack of both ABCA1 and ABCG1 induces greater defects in β-cell function than deficiency of either transporter individually. These data suggest that ABCA1 and ABCG1 each make complimentary and important contributions to β-cell function by maintaining islet cholesterol homeostasis in vivo.