胆汁酸对胆管癌细胞株QBC939中IL-6表达与细胞活力的影响

目的 探讨不同胆汁酸对胆管癌细胞株QBC939中IL-6的表达和细胞活力的影响.方法 分别使用不同浓度的游离胆汁酸以及其对应的甘氨酸结合型胆汁酸作用于胆管癌细胞株QBC939,药物浓度分别为:胆酸(CA)800 μmol/L、脱氧胆酸(DCA)100 μmol/L、鹅脱氧胆酸(CDCA)100 μmol/L、甘氨胆酸(GCA)1200 μmol/L、甘氨脱氧月胆酸(GDCA)200 μmol/L以及甘氨鹅脱氧胆酸(GCDCA)300 μmol/L.以MTT法检测不同胆汁酸刺激24、48、72 h后胆管癌细胞活力的变化,并以ELISA法检测肿瘤细胞中IL-6表达的改变.结果 DCA、CDCA、GCDCA作用48 h后,肿瘤细胞活力比值(A处理组/A对照组)分别为0.61、0.58和1.26;作用72 h后,CA、DCA、CDCA、GCA、GDCA和GCDCA各组肿瘤细胞活力比值分别为0.48、0.50、0.42、1.29、1.30和1.41;与对照组相比,以上各组细胞活力变化均具有统计学意义(P＜0.05).对照组肿瘤细胞培养48和72 h后,IL-6表达量分别为(198±32)ng/L和(323±34)ng/L,CA、DCA、CDCA、GCA、GDCA和GCDCA作用48 h后IL-6的表达量分别为(106±33)ng/L、(88±29)ng/L、(116±54)ng/L、(413±21)ng/L、(587±32)ng/L和(366±30)ng/L,作用72 h后IL-6表达量分别为(123±66)ng/L、(45±21)ng/L、(74±45)ng/L、(792±13)ng/L、(1310±22)ng/L和(845±18)ng/L;与对照组相比,以上各组IL-6表达量变化均具有统计学意义(P＜0.05).结论 游离胆汁酸CA、DCA和CDCA能减少胆管癌细胞IL-6的表达,并抑制细胞活力;而结合胆汁酸GCA、GDCA和GCDCA能增加胆管癌细胞IL-6的表达,并促进细胞活力.胆汁酸能通过IL-6途径改变胆管癌细胞活力.     

胆囊结石患者肝细胞胆汁酸分泌的变化

目的: 研究胆汁酸代谢异常与胆囊结石形成的关系.方法: 利用反相高效液相色谱(RP-HPLC)技术和体外原代培养肝细胞的方法,观察肝脏分泌机制与胆囊结石患者胆汁酸的成份变化及亲水-疏水平衡之间的关系.结果: 胆囊结石患者的甘氨鹅脱氧胆酸(GCDCA)和甘氨胆酸(GCA)的比例均较非结石组明显降低(P<0.05),而甘氨脱氧胆酸(GDCA)的比例明显上升(P<0.01).胆囊结石患者的胆汁及肝细胞培养上清液中胆汁酸HI值均较非结石患者显著上升.结论: 胆囊结石患者肝细胞胆汁酸分泌处于病理状态,胆汁中结合CDCA和结合CA比例下降,而结合DCA比例上升很可能为肝脏分泌胆汁机制受损所致,而胆汁中亲水-疏水平衡被打破是肝脏分泌机制紊乱的一个重要表现.     

胆囊切除术对二甲基肼诱发小鼠大肠癌的影响

目的 探讨胆囊切除术对大肠癌发生的影响及作用机理。方法 胆囊切除术后以二甲基肼诱发小鼠大肠癌，观察大肠肿瘤发生情况；流式细胞术测定大肠粘膜上皮细胞S期百分比；高效液相色谱仪检测手术前后鼠粪胆酸、鹅脱氧胆酸、脱氧胆酸和石胆酸含量。结果 实验组荷结节鼠大肠平均荷结节数、受检鼠大肠腺瘤发生率及S期百分比明显高于对照组（P＜0．05）；胆囊切除术后鼠粪石胆酸含量较术前和对照组明显增加（P＝0．00）。结论 胆囊切除后二甲基肼诱发的小鼠大肠肿瘤发生数量有所增加，并且大肠粘膜上皮细胞增殖旺盛，提示胆囊切除术对二甲基肼诱发小鼠大肠肿瘤有协同促进作用，可能与术后肠道内次级胆汁酸－石胆酸含量增加有关。     

柴芩四君子汤对胆囊切除术后患者肠道菌群的影响

目的探讨柴芩四君子汤对胆囊切除术后患者肠道菌群的影响。方法:选取2017 年10 月—2019 年9 月在天津市南开医院诊断为胆囊结石并行胆囊切除术的患者40 例,采用随机数字表法分为对照组19 例、柴芩四君子汤组21 例。柴芩四君子汤组自术后第1 天开始口服柴芩四君子汤,1 剂/d,早晚分服,连服14 d。分别于术前1 d 和14 d 收集患者粪便,采用16S rDNA 扩增子测序(16S rDNA Amplicon Sequencing)对肠道菌群进行测序。结果:柴芩四君子汤组与对照组比较,术前1 d,ACE、Chao1、香农和辛普森指数差异无统计学意义(P ＞ 0.05);术后14 d,ACE、Chao1 指数差异无统计学意义(P ＞ 0.05),柴芩四君子汤组香农和辛普森指数低于对照组(P =0.001、0.002)。术后14 d,与对照组比较,柴芩四君子汤组中产气柯林斯氏菌减少(P =0.001),Dorea_longicatena 减少(P =0.015),布劳特氏菌_obeum 减少(P =0.001),克里斯滕森菌科减少(P =0.017),氨基酸球菌科增加(P =0.033),不明瘤胃球菌属和罕见小球菌属减少(P =0.016、0.010)。结论:柴芩四君子汤可以调节胆囊切除术后放线菌门和厚壁菌门中的部分菌群,调节肠道菌群。     

胆囊结石患者肝细胞胆汁酸分泌与代谢

目的 :研究胆汁酸代谢异常与胆囊结石形成的关系。 方法 :利用反相高效液相色谱 (RP -HPLC)技术和体外原代培养肝细胞的方法 ,观察肝脏分泌机制与胆囊结石患者胆汁酸的成分变化及亲水一疏水平衡之间的关系。 结果 :胆囊结石患者的结合鹅脱氧胆酸和结合胆酸的比例均较非结石组明显降低 (P <0 0 5 ) ,而结合脱氧胆酸的比例明显上升 (P <0 0 1)。胆囊结石患者的胆汁及肝细胞培养上清液中胆汁酸极性指数值均较非结石患者显著上升。 结论 :胆囊结石患者肝细胞胆汁酸分泌处于病理状态 ,胆汁中结合鹅脱氧胆酸和结合CA比例下降而结合脱氧胆酸比例上升很可能为肝脏分泌胆汁机制受损所致 ,而胆汁中亲水一疏水平衡被打破是肝脏分泌机制紊乱的一个重要表现。     

胆酸对肝硬化大鼠肝大部分切除术后肝脏功能的影响

目的探讨胆酸对肝硬化大鼠肝切除术后肝脏功能的影响。方法24只肝硬化大鼠肝切除术后随机分为胆酸组和对照组喂养1周,每组12只。对照组给以标准饲料喂养,胆酸组给以含0.2%胆酸的饲料喂养。1周后检测大鼠胆汁分泌速度及胆汁总胆汁酸（TBA）含量,检测肝功能、有丝分裂指数（MI）、增殖细胞核抗原（PCNA）标记指数、细胞核DNA含量。结果胆酸组胆汁分泌速度、胆汁TBA含量、血清ALB含量、MI、PCNA标记指数、细胞核DNA含量均高于对照组（P〈0.01）,血清谷草转氨酶（AST）、谷丙转氨酶（ALT）含量均低于对照组（P〈0.01）。结论胆酸可以增加肝硬化大鼠肝切除术后胆汁分泌速度及胆汁TBA含量,促进大鼠肝功能恢复和肝再生。     

瘦素对血脂及胆汁成分的调节在胆囊胆固醇结石形成中的作用

目的:分析胆囊胆固醇结石患者瘦素与血脂及胆囊内胆汁成分的关系,探讨瘦素在胆囊胆固醇结石形成中的作用。方法:选择胆囊胆固醇结石接受腹腔镜胆囊切除术的患者30例(结石组)与同期因胆囊息肉行腹腔镜胆囊切除术的患者22例(息肉组),检测患者血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL)、高密度脂蛋白(HDL)、瘦素、胆囊胆汁中的TC与总胆汁酸(TBA)水平,以及胆囊壁组织瘦素受体mRNA水平。结果:与息肉组比较,结石组血清TC、TG、LDL、瘦素水平以及胆囊内胆汁TC/TBA含量比率与胆囊组织瘦素受体mRNA水平均明显升高,而血清HDL明显降低(均P0.05)。结石组的血清瘦素水平与血清TG、TC及胆汁TC均呈正相关(r=0.633,P=0.002;r=0.224,P=0.025;r=0.384,P=0.000),与HDL和TBA呈负相关(r=-0.205,P=0.014;r=-0.548,P=0.024)呈负相关,而息肉组血清瘦素与以上指标间均无关(均P0.05)。结论:瘦素参与了胆囊胆固醇结石的形成,瘦素及其受体水平的升高可能与胆囊胆固醇结石患者胆固醇代谢异常、胆囊胆汁成分失调密切相关。     

临床预测胆石化学类型研究的进展

随着非手术疗法胆石症的进展,治疗前确定胆石化学类型变得十分重要。口服鹅去氧胆酸(CDCA)/熊去氧胆酸(UDCA)溶石,胆囊/胆道内灌注甲基叔丁基醚溶石都仅适用于胆固醇类结石;而有些溶石剂只适用于胆色素结石。经皮经肝穿刺胆囊/胆道插管溶石是一种侵入性治疗,治疗前必需对胆石性质了解清楚。为提高体外冲击波碎石的疗效,在碎石前后辅以CDCA/CDCA口服溶石,也只对胆固醇类结石起作用。治疗失败的主要原因是治疗前未能确切了解胆结石的化学类型。随着医学影象技术的发展,很多学者正在各领域寻求一种准确而又简便的方法,以便在溶石、碎石治疗前能判断患者结石化学类型,选择病例,以提高疗效。兹分别综述如下:     

胆道狭窄致胆色素结石时胆汁酸成分的变化

目的 研究胆汁中胆汁酸的变化与胆色素结石形成的关系。方法 结扎豚鼠的胆总管造成狭窄，一周后测定胆汁中胆汁酸及其它成分并行病理研究。结果 狭窄组(S组)成石率达91．3%，均为胆色素结石（PS)，胆汁中总胆红素（TBr）、非结合胆红素（Br），钙离子(JCE）、糖蛋白(GP）均较对照组（C组）明显升高。总胆汁酸（TBA）也明显增高。S组胆汁中牛磺鹅脱氧胆酸(TCDCA）明显升高，而甘氨鹅脱氧胆酸（GCDCA）明显降低，G/T值显著下降。超微病理研究表明，S组肝细胞内线粒体明显肿胀，滑面内质网显著减少。结论 胆汁中G/T的下降与肝脏病变均有利于PS形成。     

胆固醇结石差异lncRNA表达基因的分析及其功能研究

目的探索胆固醇结石形成相关的差异表达lncRNA基因,并通过生物信息学方法分析差异lncRNA基因的生物学功能。方法将24只C57BL/6小鼠随机分为胆固醇结石组16只和空白对照组8只。胆固醇结石组小鼠饲喂致石饲料5周,空白对照组饲喂常规饲料。5周后,将胆固醇结石组小鼠随机分为模型对照组8只和熊去氧胆酸组8只。之后空白对照组饲喂常规饲料,模型对照组饲喂致石饲料,熊去氧胆酸组饲喂致石饲料并给予熊去氧胆酸灌胃。2周后处死小鼠,收集3组小鼠的肝脏组织及胆汁样本,观察胆囊大体样本并对小鼠胆汁进行脂质分析。同时对3组小鼠的肝脏组织进行lnc RNA表达谱测定,并分析差异表达基因,包括Cis-/Trans-靶基因的基因本体(GO)和京都基因与基因组百科全书(KEGG)Pathway分析。结果 16只胆固醇成石小鼠模型构建成功。模型对照组、熊脱氧胆酸组及空白对照组胆囊胆汁总胆固醇含量的差异有统计学意义(P0.05),模型对照组较空白对照组和熊脱氧胆酸组明显升高(P0.05),但熊脱氧胆酸组与空白对照组比较差异无统计学意义(P=0.59);而3组的胆囊胆汁总胆汁酸、总胆红素及直接胆红素水平比较差异均无统计学意义(P0.05)。3组小鼠肝脏组织的差异lncRNA基因分析结果显示,模型对照组与熊脱氧胆酸组共有49种差异lncRNA基因。将差异基因分别进行GO和KEGG pathway分析后发现,模型对照组与空白对照组的差异基因富集得到88个GO term和18条pathway,熊脱氧胆酸组与空白组的差异基因富集得到205个GO term和20条pathway。结论熊脱氧胆酸对胆囊结石有治疗作用;差异表达lncRNA基因在胆固醇结石形成及熊脱氧胆酸的预防结石形成中起到重要调控作用,为进一步探讨lncRNA的具体调控机制奠定了基础。     

Bile acids and the increased risk of colorectal tumours after truncal vagotomy

An association between colorectal cancer and previous peptic ulcer surgery is reported. In a prospective screening study, 100 asymptomatic patients (80 men and 20 women) who had undergone truncal vagotomy at least 10 years previously were investigated by barium enema, colonoscopy and gallbladder ultrasonography. Control data were obtained from forensic autopsy subjects. The incidence of neoplasms greater than or equal to 1.0 cm in the vagotomized group was 14 per cent (11 adenomas, 3 carcinomas) and 3 per cent in controls (P = 0.01). Duodenal bile obtained at endoscopy from 21 vagotomized patients with normal gallbladders and from 21 control patients undergoing endoscopy was analysed by high performance liquid chromatography. The mean percentage of cholic (CA), chenodeoxycholic (CDCA), deoxycholic (DCA) and lithocholic (LCA) acids in the bile of vagotomized patients was 32.3, 45.6, 20.7 and 1.4 per cent respectively compared with 45.3, 36.2, 17.9 and 0.7 per cent respectively in controls. The increased proportions of CDCA and LCA and decreased proportions of CA in the duodenal bile of vagotomized patients were significant (P less than 0.001; P = 0.02; P = 0.007). Abnormalities in bile acid metabolism may help to explain the increased risk of colorectal neoplasia 10 years after truncal vagotomy.     

Altered bile acid metabolism in nonobese, spontaneously diabetic (NOD) mice

Cholesterol and bile acid metabolism was examined in nonobese, spontaneously diabetic (NOD) female mice before and after the development of diabetes. After the development of glucosuria, the plasma and liver cholesterol levels, gallbladder bile weight after 5-h fasting, biliary cholesterol, phospholipid and bile acid concentrations, the lithogenic index, the pool size of bile acids, and fecal sterol excretion markedly increased, but fecal bile acid excretion and fractional turnover rates for the cholic acid and chenodeoxycholic acid groups decreased. The distribution percentage of bile acids in the small intestine did not change, but it increased in the gallbladder and decreased in the large intestine. One striking finding was a change in the bile acid composition: increases were recorded in cholic and deoxycholic acids while decreases occurred in bile acids derived from chenodeoxycholic acid, such as beta-muricholic and ursodeoxycholic acids in the bile and alpha-muricholic, beta-muricholic, omega-muricholic, and hyodeoxycholic acids in the feces. Therefore, the cholic acid group/chenodeoxycholic acid group (CA/CDCA) ratio increased in the bile, feces, and small and large intestines after the development of diabetes. These changes were very similar to those observed in alloxan-treated mice, suggesting that the changes found in NOD mice are caused by insulin deficiency.     

Changes in serum bile acid composition in relation to histological findings after liver transplantation in piglets

Changes in the composition of serum bile acids were investigated after orthotopic liver transplantation in piglets. Serum levels of all bile acid components rapidly increased during the anhepatic phase and then quickly decreased, returning to the preoperative state within 6 h of revascularization of the allograft. In the animals in which extrahepatic biliary stenosis was a complication, serum total bile acids (TBA) increased remarkably. A marked increase of hyocholic acid and marked decrease of hyodeoxycholic acid were noted; as percentages of TBA, the average levels per day were 91.3 +/- 1.3 and 5.0 +/- 1.3%, respectively. No change in chenodeoxycholic acid (CDCA) was observed. In the animals suffering acute rejection without extrahepatic biliary stenosis, serum TBA increased slightly and the percentage of CDCA rose, the average level being 29.6 +/- 1.5%. These results suggest that the measurement of the composition of serum bile acids may serve as a useful means of assessing allograft function after liver transplantation.     

Ursodeoxycholic acid increased bile flow and affects bile composition in the early postoperative phase following liver transplantation

Abstract Orally given ursodeoxycholic acid (UDCA) has beneficial effects on laboratory parameters in different cholestatic conditions. In order to investigate the effect on early graft function after liver transplantation, 33 patients were randomized to receive either UDCA 15 mg/kg per day or placebo from the 1st postoperative day until 3 months after transplantation. All liver grafts produced bile within 24 h after revascularization. In both groups there was an increasing bile flow each day until day 5 after transplantation. This increase was more pronounced in the UDCA group where the flow on day 2 reached a mean value of 183 ± 28 ml/day compared to 106 ± 17 ml/day in the placebo group ( P < 0.05). The average daily volume of bile produced during the first 10 days was also found to be higher in the UDCA group compared to the placebo group (242 ± 20 ml vs 176 ± 18 ml, P < 0.02). In the UDCA group a significant decrease in total bile acid output between the 5th and 10th postoperative days was found, while in the placebo group the amount of bile acids excreted remained stable over time. The composition of bile differed between the two groups with an increase in the portion of UDCA in the UDCA group from the 2nd postoperative day (25 % vs 4.6 %, P < 0.0003). The fraction of UDCA then remained high during the whole study period with a peak at day 3 when 38.1 ± 6.6% of the bile acids consisted of UDCA. In the placebo group, the fraction of UDCA was low from the beginning and diminished further over time. Prophylactic UDCA treatment was found to have a significant positive impact on the ALT level during the 4th and 5th postoperative days, but had no effect on bilirubin or GGT in the early postoperative phase (days 1–10). No differences in cyclosporin requirement were found between the two groups.     

Promoting effect of bile acids on gastric carcinogenesis induced by MNNG in rats

The promoting effect of bile acids on the development of gastric carcinoma was examined in rats treated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). At the first experiment, two hundred and fifteen male Wistar rats were divided into 5 groups; after oral administration MNNG (120 micrograms/ml) for 24 weeks, group 1 received tap water, group 2 administered of chenodeoxycholic acid (CDCA) solution, group 3 had deoxycholic acid (DCA) solution for the next 12 weeks. Group 4 received CDCA solution and group 5 received DCA solution for 36 weeks without MNNG. At the second experiment, fifty one rats were divided into 3 groups; for the first 12 weeks, group 1 received tap water, group 2 CDCA and group 3 DCA. These 3 groups received MNNG for the next 24 weeks followed by tap water for 12 weeks. The incidence of gastric adenocarcinoma in MNNG-treated rats was significantly higher in group 3 (63.6%) as compared with that in group 1 (36.7%) in the first experiment. No carcinoma lesions was found in groups 4 and 5. In the second experiment, no significant changes was observed among 3 groups. Undifferentiated adenocarcinomas were identified in groups 2 and 3, especially treated with MNNG plus bile acids. The result suggested a promoting effect of bile acids, especially DCA, in stomach carcinogenesis.     

The non-operative management of gallstones.

During the last decade the non-surgical management of gallbladder stones has become possible. Oral chenodeoxycholic acid (CDCA) and more recently, ursodeoxycholic acid (UDCA), have proved to be safe and effective. For the treatment of bile duct stones, various methods including flushing, chemical dissolution and mechanical procedures are now available. A combination of these techniques can be used in a logical sequence and should enable the successful treatment of most patients with bile ducts stones.     

THE NON-OPERATIVE MANAGEMENT OF GALLSTONES

During the last decade the non-surgical management of gallbladder stones has become possible. Oral chenodeoxycholic acid (CDCA) and more recently, ursodeoxycholic acid (UDCA), have proved to be safe and effective. For the treatment of bile duct stones, various methods including flushing, chemical dissolution and mechanical procedures are now available. A combination of these techniques can be used in a logical sequence and should enable the successful treatment of most patients with bile ducts stones.