熊去氧胆酸选择性诱导人肝肿瘤细胞凋亡及抑制增殖的实验研究

目的探讨熊去氧胆酸(UDCA)诱导肝肿瘤细胞株凋亡并抑制其增殖的作用和机制。方法用噻唑蓝法、流式细胞术、TUNEL法、Wright-Giemsa染色法、电镜及免疫细胞化学等方法，观察UDCA对肝肿瘤细胞株HepG2、BEL7402和正常人肝细胞株L-02的生长活力、细胞凋亡、细胞周期及Bax／bcl-2基因表达的影响。结果UDCA对HepG2、BEL7402细胞株具有显著的抑制生长、诱导凋亡、阻滞细胞周期于S期、降低bcl-2和提升Bax表达的作用。结论UDCA对HepG2、BEL7402细胞株有显著的抑制增殖及诱导凋亡作用，该作用可能与UDCA阻滞细胞周期、降低bcl-2和提升Bax的表达有关。     

Protective effects of ursodeoxycholic acid on chenodeoxycholic acid-induced liver injury in hamsters

AIM To investigate the effects of ursodeoxycholic acid (UDCA) on chenodeoxycholic acid (CDCA)-induced liver injury in hamsters, and to elucidate a correlation between liver injury and bile acid profiles in the liver.METHODS Liver injury was induced in hamsters by administration of 0.5% (w/w) CDCA in their feed for 7 d.UDCA (50 mg/kg and 150 mg/kg) was administered for the last 3 d of the experiment.RESULTS At the end of the experiment, serum alanine aminotransferase (ALT) increased more than 10 times and the presence of liver injury was confirmed histologically. Marked increase in bile acids was observed in the liver. The amount of total bile acids increased approximately three-fold and was accompanied by the increase in hydrophobic bile acids, CDCA and lithocholic acid (LCA). UDCA (50 mg/kg and 150 mg/kg) improved liver histology, with a significant decrease (679.3 ±77.5 U/L vs 333.6 ± 50.4 U/L and 254.3 ± 35.5 U/L, respectively, P ＜ 0.01) in serum ALT level. UDCA decreased the concentrations of the hydrophobic bile acids, and as a result, a decrease in the total bile acid level in the liver was achieved.CONCLUSION The results show that UDCA improves oral CDCA-induced liver damage in hamsters. The protective effects of UDCA appear to result from a decrease in the concentration of hydrophobic bile acids, CDCA and LCA, which accumulate and show the cytotoxicity in the liver.     

叶酸对胃癌细胞凋亡的影响

目的胃癌的发生与发展中有细胞凋亡的变化,叶酸抗肿瘤的机理涉及到维持ＤＮＡ甲基化水平等,其与凋亡的关系尚未明了。方法以高低两种浓度的叶酸干预培养的人胃癌ＭＫＮ－４５和ＭＫＮ－２８细胞系７２小时后,以原位ＤＮＡ断端切口标记法和ＤＮＡ琼脂糖凝胶电泳检测凋亡形态学改变和ＤＮＡ断裂情况。结果未加干预者两种细胞系的凋亡指数均低于５％,不同分化细胞系间比较差异无显著性;低浓度叶酸可诱导ＭＫＮ－４５细胞凋亡,而高浓度者使ＭＫＮ－２８细胞凋亡率增加。结论胃癌细胞自然凋亡率较低。叶酸干预可影响凋亡的发生。     

雌激素受体α基因多态性对熊去氧胆酸治疗原发性胆汁性肝硬化疗效的影响

目的 探讨雌激素受体α(ERα)基因多态性对熊去氧胆酸(UDCA)治疗女性原发性胆汁性肝硬化(PBC)疗效的影响.方法 采用聚合酶链反应-限制性片段长度多态性方法 检测65例女性PBC患者ERα基因1号内含子PvuⅡ和XbaⅠ酶切位点多态性.患者每日均予UDCA(13～15 mg/kg,分3次口服)治疗.随访患者临床症状和血清总胆红素(TBil)、丙氨酸转氨酶(ALT)、碱性磷酸酶(ALP)、γ-谷氨酰转肽酶(GGT)变化情况,共随访24个月.结果 共60例患者获完整随访资料.XX和PP型患者4项指标均缓慢下降,但至随访结束时未达治疗有效标准.Xx型患者4项指标亦缓慢下降,至随访结束时达治疗有效标准.Pp、pp、xx型患者4项指标均快速下降至治疗有效标准.UDCA治疗PBC有效率75%(45/60).PP、Pp、pp型患者和XX、Xx、xx型患者在UDCA治疗有效组和无效组中的分布差异均有统计学意义(x2=12.13、P=0.003和x2=9.95、P=0.007).Pp、pp、Xx、xx型患者有效率[分别为82.61%(19/23)、80.65%(25/31)、9/14、83.33%(35/42)]高于PP、XX型者(分别为1/6和1/4).结论 PBC患者ERα基因多态性影响UDCA的疗效.     

Effects and mechanisms of silibinin on human hepatoma cell lines

AIM To investigate in vitro effects and mechanisms of silibinin on hepatocellular carcinoma (HCC) cell growth. METHODS Human HCC cell lines were treated with different doses of silibinin. The effects of silibinin on HCC cell growth and proliferation, apoptosis, cell cycle progression, histone acetylation, and other related signal transductions were systematically examined. RESULTS We demonstrated that silibinin significantly reduced the growth of HUH7, HepG2, Hep3B, and PLC/PRF/5 human hepatoma cells. Silibinin-reduced HuH7 cell growth was associated with significantly upregulated p21/CDK4 and p27/CDK4 complexes, downregulated Rb-phosphorylation and E2F1/DP1 complex. Silibinin promoted apoptosis of HuH7 cells that was associated with down-regulated survivin and upregulated activated caspase-3 and -9. Silibinin's anti angiogenic effects were indicated by down-regulated metalloproteinase-2 (MMP2) and CD34. We found that silibinin-reduced growth of HuH7 cells was associated with increased activity of phosphatase and tensin homolog deleted on chromosome ten (PTEN) and decreased p-Akt production, indicating the role of PTEN/ PI3K/Akt pathway in silibinin-mediated anti-HCC effects. We also demonstrated that silibinin increased acetylation of histone H3 and H4 (AC-H3 and AC-H4), indicating a possible role of altered histone acetylation in silibininreduced HCC cell proliferation.CONCLUSION Our results defined silibinin's in vitro anti-HCC effects and possible mechanisms, and provided a rationale to further test silibinin for HCC chemoprevention.     

中等剂量熊去氧胆酸治疗原发性胆汁性肝硬化的系统评价

目的评价长期应用中等剂量熊去氧胆酸治疗原发性胆汁性肝硬化的疗效及安全性。方法对全世界关于中等剂量(13～15mg·kg-1·g-1)熊去氧胆酸与安慰剂对照治疗原发性胆汁性肝硬化的随机对照试验进行系统评价。结果共纳入7项随机对照试验,累计1038例患者。熊去氧胆酸能显著改善患者的肝功能生化检测指标,但不能改善疲劳和瘙痒等症状。病程Ⅰ至Ⅱ期的患者治疗2年后肝脏组织学显著好于对照组(P=0.03),但分析所有患者时差异无统计学意义(P=0.08)。荟萃分析显示治疗组与对照组间死亡率(OR0.99,95%可信区间0.62～1.58)、肝病相关死亡率(1.05,0.53～2.05)、肝移植率(0.87,0.53～1.41)、死亡和(或)肝移植率(0.92,0.64～1.31)和肝功能失代偿率(0.94,0.60～1.49)差异无统计学意义。结论熊去氧胆酸能有效改善肝功能,但不能改善症状,也无足够证据支持熊去氧胆酸能延长患者的生存期。早期患者及早并长期应用熊去氧胆酸可能延缓肝脏组织学进展。     

原发性胆汁性肝硬化患者对熊去氧胆酸应答欠佳的影响因素

目的 观察对熊去氧胆酸应答欠佳的原发性胆汁性肝硬化(PBC)患者的临床特点.方法 回顾性研究经熊去氧胆酸治疗1年以上的38例PBC患者,分析其在确诊时的性别、年龄及就诊时主要主诉;治疗前及治疗1年后的肝功能生化指标(血清ALT、AST、ALP、Y-谷氨酰转肽酶、总胆红素、白蛋白和球蛋白等);治疗前免疫学指标(IgG、IgM、血清抗核抗体和抗线粒体抗体).计量资料比较采用独立样本均数的t检验或近似法t'检验,计数资料的比较用Fisher's Exact Test.结果 38例PBC患者中,应答欠佳组患者17例,主要就诊原因为出现肝病相关症状,包括皮肤、巩膜黄染(41.1%),乏力、纳差(23.5%),下肢水肿、腹胀(11.7%)等.较好应答组患者21例,主要就诊原因为体检肝功能异常(33.3%).实验室检查结果显示,应答欠佳组与较好应答组相比,基线总胆红素[(40.15±25.08)μmol/L比(18.07±8.73)μmol/L]、碱性磷酸酶[(498.60±277.50)U/L比(313.70±182.10)U/L]、ALT[(105.60±69.22)U/L比(65.84±37.11)U/L]、AST[(113.80±54.24)U/L比(62.52±26.22)U/L]、球蛋白[(40.64±8.82)g/L比(33.06±5.01)g/L]和IgG[(19.52±4.44)g/L比(15.18±4.06)g/L]均较高,差异有统计学意义(t值分别为3.428、2.315、2.180、3.532、2.386和3.021,P值均＜0.05).结论 PBC患者中,因肝病相关症状就诊者、肝脏生物化学指标明显异常者以及自身免疫特征较多者,可能对熊去氧胆酸的应答欠佳.

Abstract：

Objective To observe the characteristics of primary biliary cirrhosis (PBC) with a suboptimal biochemical response to ursodeoxycholic acid.Methods A total of 38 Chinse PBC patients (5male patients,33 female patients,average age 55 years old) with treatment of ursodeoxycholic acid in our hospital from January 1999 to January 2009 were erolled and studied retrospectively.Results 17 suboptimal biochemical responders mainly presented with liver diseases related symptoms including jaundice (41.1%),fatigue,anorexia (23.5%),edema and abdominal distension (11.7%).21 good biochemical responders mainly presented with abnormal liver function tests without symptoms.The suboptimal biochemical responders had significantly higher baseline levels of total serum bilirubin,alkaline phosphatase,alanine aminotransferase,aspartate aminotransferase,immunoglobulin G and globulin as compared to the good biochemical responsers.There were no differences in gender,age and the dose of UDCA.Conclusion PBC patients with liver diseases related symptoms,marked abnormal liver tests and characteristics of autoimmune hepatitis may have a suboptimal biochemical response to ursodeoxycholic acid treatment.     

脂氧合酶抑制剂对HepG2细胞增殖及凋亡的影响

目的探讨脂氧合酶抑制剂对HepG2细胞增殖及凋亡的影响及其相关机制。方法应用不同浓度脂氧合酶抑制剂去甲二氢愈创木酸（NDGA）处理HepG2细胞，利用MTY比色法、生长曲线、细胞克隆形成观察NDGA对HepG2细胞的增殖作用，H-E染色、流式细胞术观察细胞凋亡并行细胞周期分析，细胞免疫化学或Western印迹技术测定Caspase-3、增殖细胞核抗原（PCNA）、野生型（wt）P53、c-erbB-2蛋白表达水平变化。结果NDGA对HepG2细胞具有明显生长抑制作用，不同浓度NDGA（50、100、150、200μmol／L）处理HepG2细胞72h的抑制率分别为27．34％、41．76％、57．08％和69．17％，其IC50值为109．13μmol／L；细胞克隆形成率分别为21．63％、17．33％、12．53％、7．97％，明显低于对照组的31．70％。同时NDGA可诱导HepG2细胞凋亡，与细胞周期被阻滞于G1／S期有关。细胞免疫化学、Western印迹分析显示NDGA对HepG2细胞的PCNA、c-erbB-2蛋白表达有抑制作用，并可促进Caspase-3、wtP53蛋白表达。结论NDGA对HepG2细胞有较强的细胞毒作用且以剂量、时间依赖方式抑制细胞增殖，并诱导凋亡，使细胞周期阻滞于G1／S期。此作用与改变某些原癌基因、抑癌基因的蛋白表达水平有关。     

熊去氧胆酸及普瑞博思治疗碱性反流性胃炎的实验研究

目的：为研究碱性反流性胃炎的机理和治疗，在建立大鼠碱性反流性胃炎模型的基础上观察熊去氧胆酸及西沙必利的治疗作用。方法：正常对照组１０只，行幽门弹簧扩张手术３５只中手术对照组１０只，熊去氧胆酸治疗组１５只，普瑞博思治疗组１０只。８周后，测胃液ｐＨ、甘氨胆酸、磷脂、氨基己糖含量、血清胃泌素及胃粘膜病理检查。结果：手术对照组胃液甘氨胆酸及ｐＨ值升高，胃粘膜病理改变明显；熊去氧胆酸及普瑞博思治疗组，大鼠胃液甘氨胆酸明显降低，胃粘膜病理改变轻，结论：①胃幽门弹簧扩张术能够建立实验性碱性反流性胃炎模型；②熊去氧胆酸及普瑞博思对大鼠碱性反流性胃炎均有较好的疗效。     

A dose-up of ursodeoxycholic acid decreases transaminases in hepatitis C patients

AIM： To examine whether a dose-up to 900 mg of ursodeoxycholic acid （UDCA） decreases transaminases in hepatitis C patients. METHODS： From January to December 2007, patients with chronic hepatitis C or compensated liver cirrhosis with hepatitis C virus （HCV） （43-80 years old） showing positive serum HCV-RNA who had already taken 600 mg/d of UDCA were recruited into this study. Blood parameters were examined at 4, 8 and 24 wk after increasing the dose of oral UDCA from 600 to 900 mg/d. RESULTS： Serum alanine aminotransferase （ALT）, aspartate aminotransferase （AST）, and gamma- glutamyl transpeptidase （GGT） levels were significantly decreased following the administration of 900 mg/d as compared to 600 mg/d. The decrease in ALT from immediately before the dose-up of UDCA to 8 wk after the dose-up was 14.3 IU/L, while that for AST was 10.5 IU/L and for GGT was 9.8 IU/L. Platelet count tended to increase after the dose-up of UDCA, although it did not show a statistically significant level （P = 0.05）. Minor adverse events were observed in 3 cases, although no drop-outs from the study occurred.CONCLUSION： Oral administration of 900 mg/d of UDCA was more effective than 600 mg/d for reducing ALT, AST, and GGT levels in patients with HCV-related chronic liver disease.     

熊去氧胆酸防治结肠肿瘤的研究进展

熊去氧胆酸(UDCA)是一种存在于人胆汁中的游离胆汁酸,为天然亲水二羟胆盐,临床上用于治疗多种肝胆疾病,如原发性胆汁性肝硬化、胆固醇结石、胆囊炎、胆管炎、慢性活动性肝炎及胆汁淤积性肝病等.近年发现UDCA和结肠肿瘤的关系密切,本文综述UDCA对防治结肠肿瘤作用的研究进展.     

Efficacy of ursodeoxycholic acid for Japanese patients with autoimmune hepatitis

Purpose

This study aimed to investigate the efficacy of ursodeoxycholic acid (UDCA) for Japanese patients with autoimmune hepatitis (AIH).

Methods

One hundred forty-seven patients were investigated.

Results

As initial treatment, 25 patients received UDCA (300–600 mg/day) monotherapy (UDCA group), 40 received a combination of prednisolone (PSL) (≥20 mg/day) and UDCA (combination group), 68 received PSL monotherapy (PSL group), and 14 received other treatments. During the follow-up, in the UDCA group, PSL was added to 8 of 12 patients failing to achieve the normalization of serum transaminase levels with UDCA monotherapy. Cumulative incidence of the normalization of serum transaminase levels was 64% in the UDCA group, 95% in the combination group, and 94% in the PSL group (log-rank test, P = 0.0001). UDCA group required longest periods until the normalization of serum transaminase levels. Eleven patients, who achieved persistent normalization of serum transaminase levels with UDCA monotherapy, did not reach liver failure or develop hepatocellular carcinoma for 49.7 (range = 13.4–137.3) months. Meanwhile, during the taper of PSL, doses of PSL at the initial relapse were lower in patients treated with PSL and UDCA than in those treated with PSL monotherapy, and initial relapse occurred earlier in patients treated with PSL monotherapy.

Conclusions

UDCA monotherapy is effective for some Japanese AIH patients; however, UDCA monotherapy for patients with either high-grade inflammatory activity or poor residual capacity of liver function is not recommended because they may reach liver failure before achievement of remission. Meanwhile, additional use of UDCA during the taper of corticosteroids may be effective for the prevention of early relapse.     

熊去氧胆酸抗免疫性大鼠肝纤维化作用的研究

目的　研究熊去氧胆酸 (UDCA)对免疫性大鼠肝纤维化的影响 ,并探讨其作用机制。方法　 3 6只SD大鼠随机分为 3组。 2 4只大鼠腹腔内注射猪血清 (每周 2次 ,共 8周 )诱导建立肝纤维化模型 ;其中 12只予以UDCA[15mg/(kg·d]干预 (UDCA组 ) :余 12只为模型组。对照组 12只大鼠未予任何处理。 8周后处死动物。观察肝组织纤维化程度、胶原表达及血清透明质酸 (HA) ;羟脯氨酸 (Hyp)水平的变化 ;免疫组化染色观察α 平滑肌肌动蛋白 (α SMA)、转化生长因子 β1(TGF β1)及核因子κB(NFκBp65 )表达的情况 ;各组结果分别进行比较。结果　模型组大鼠肝内胶原面积明显高于对照组 ( 11 1± 2 0 9vs 0 73± 0 15 ,P <0 0 5 ) ,血清HA、Hyp及α SMA、TGF β1,NFκBp65在肝内的表达亦显著增高 (P <0 0 5 )。与模型组相比较 ,UDCA组肝内胶原面积无明显变化 ( 9 49±1 3 1vs 11 1± 2 0 9,P >0 0 5 ) ,其余指标在两组间均无显著性差异 (P >0 0 5 )。结论　一般剂量的UDCA对大鼠免疫性肝纤维化无显著改善作用     

维甲酸对人肝癌细胞凋亡的影响及临床意义

目的：观察全反式维甲酸（ATRA）对肝癌细胞凋亡的作用并探讨ATR的作用机制。方法：在肝癌细胞株SMMC－7721细胞加入ATRA,使终浓度为0．5μg/ml,对照组加等剂量的0．02％DMSO,培养4d,ATRA对SMMC－7721细胞的影响通过苏木精－伊红染色,在光镜下进行细胞形态学观察;通过细胞计数绘制生长曲线及计算细胞生长抑制率,流式细胞术分析不同DNA含量的细胞分布,计算细胞凋亡百分率,凋亡相关基因Fas,p53,Bcl-2蛋白表达的检测亦采用流式细胞仪检测。结果：苏木精－伊红染色可见较多细胞核碎裂,胞浆浓缩,染色质深染,聚集于核膜下,部分细胞膜突起呈小泡样,小泡脱落形成凋亡小体,而对照组无明显的形态学改变,作用4d生细胞生长抑制率为47．5％,与对照组比较,差异显著（P＜0．05）,流式细胞仪分析在G1期前出现亚二倍体凋亡峰;凋亡细胞百分比为16．0％,与对照的6．9％比较有显著性差异（P＜0．01）。观察发现,ATRA对SMMC－771细胞Fax,P53的表达明显增加,并下调Bcl-2的表达,结论：0．5μg/ml的ATRA对肝癌细胞的生长有显著抑制效果,并可见癌细胞凋亡形态改变,ATRA促进SMMC－721肝癌细胞凋亡的机制之一可能是通过调控Fas,P53及Bcl-2的表达。     

鹅脱氧胆酸衍生物HS-1200诱导人肝癌细胞株凋亡的作用及机制

鹅脱氧胆酸衍生物HS-1200可有效地抑制肝肿瘤细胞株BEL7402的生长,其作用随药物浓度提高和作用时间延长而增强,呈一定的剂量、时间依赖性;鹅脱氧胆酸衍生物HS-1200对人肝肿瘤细胞株BEL7402有显著的生长抑制作用,该生长抑制作用与HS-1200诱导肝肿瘤细胞凋亡有关;HS-1200对正常肝细胞株无生长抑制及诱导凋亡的作用.HS-1200诱导凋亡的机制可能是HS-1200提升Bax而降低Bcl-2的表达,从而使线粒体膜通透性增高,使细胞色素C由线粒体释放入胞质,活化easpase-9,活化的caspase-9切割pro-caspase-3生成caspase-3,从而诱导凋亡;但上述凋亡过程caspase-8特异性抑制剂未改变HS-1200诱导的凋亡,因而HS-1200诱导肝肿瘤细胞凋亡途径为内源性凋亡途径.     

表没食子儿茶素没食子酸酯对人胃癌细胞株MKN45诱导凋亡作用的研究

目的了解表没食子儿茶素没食子酸酯（EGCG）是否诱导人胃癌细胞株MKN45凋亡及其凋亡信号传导途径，为其临床应用提供进一步的理论依据。方法用四甲基偶氮唑蓝（MTT）比色法检测EGCG对MKN45细胞生长的抑制作用；膜连蛋白V-异硫氰基荧光素（Annexin V-FITE）＋碘化丙啶（PI）方法测定EGCG作用后MKN45细胞的凋亡率；酶联免疫吸附测定（ELISA）检测EGCG对MKN45细胞内castmse-3活性的影响；Rhodamine123染色检测EGCG作用后MKN45细胞内线粒体膜电位的改变情况。结果EGCG作用后MKN45细胞发生凋亡，随着EGCG作用时间的延长及浓度的增加．凋亡率增加。在EGCG作用8h后MKN45细胞内的caspase-3的活性开始升高，并于作用12h后活性明显升高。线粒体的膜电位于EGCG作用4h后就开始明显下降，并与时间和浓度正相关。结论EGCG可以诱导人胃癌细胞株MKN45细胞凋亡，且与作用时间及浓度正相关。EGCG对MKN45细胞凋亡的诱导是通过线粒体途径发生的．     

奥曲肽与胰岛素对人肝癌细胞影响的体外实验研究

目的 旨在探讨胰岛素能否促进人肝癌细胞增殖,而奥曲肽对其诱导的增殖活动有无抑制作用。方法 将奥曲肽与胰岛素直接作用于体外培养的人原发性肝癌细胞株（ＢＥＬ－７４０２）,运用四氧唑盐、直接细胞计数及流式细胞仪测定等方法观察奥曲肽与胰岛素对ＢＥＬ－７４０２细胞株活细胞数及其比值、细胞总数、细胞周期及增殖指数（ＰＩ）的影响。结果 胰岛素（０～５ｕｇ／ｍｌ）使ＢＥＬ－７４０２细胞株的细胞总数、活细胞数及其比值增加,呈明显的量效关系。而奥曲肽（０～２ｕｇ／ｍｌ）则使ＢＥＬ－７４０２基础的及胰岛素刺激的细胞总数、活细胞数及其比值降低,亦呈明显的量效关系。另外,５ｕｇ／ｍｌ的胰岛素增加细胞的ＰＩ值,而１ｕｇ／ｍｌ的奥典肽降低其ＰＩ值（Ｐ〈０．０５）。并对细胞周期同步化的ＢＥＬ－７４０２细胞株产生Ｓ期阻滞作用（Ｐ〈０．０５）。结     

熊去氧胆酸治疗原发性胆汁性胆管炎患者血清甲状腺激素水平变化*

目的 探讨应用熊去氧胆酸（UDCA）治疗原发性胆汁性胆管炎患者血清甲状腺激素水平的变化。方法 2016年1月~2017年6月我院治疗的原发性胆汁性胆管炎患者70例和原发性胆汁性肝硬化患者95例,另选20例健康人作为对照组。给予患者UDCA治疗3月,检测治疗前后血清甲状腺激素水平变化。结果 在治疗3个月末,两组患者肝功能指标得到改善,除胆管炎患者血清ALP和GGT水平仍显著高于肝硬化患者外,其他肝功能指标比较,差异无统计学意义（P>0.05）;治疗前,胆管炎患者血清总三碘甲状腺原氨酸（TT3）、总甲状腺素（TT4）、游离三碘甲状腺原氨酸（FT3）和游离甲状腺素（FT4）水平分别为1.6±0.2 nmol/L、3.2±0.4 pmol/L、95.6±28.7 nmol/L和10.3±3.6 pmol/L,肝硬化患者分别为1.1±0.2 nmol/L、3.0±0.3 pmol/L、88.2±26.7 nmol/L和9.7±2.5 pmol/L,均显著低于健康人【分别为（2.5±0.5 nmol/L、4.7±0.8 pmol/L、115.7±31.6 nmol/L和13.8±4.1 pmol/L,P<0.05】,两组患者血清促甲状腺素（TSH）水平与健康人比,差异无统计学意义【分别为（2.9±0.5 mIU/L和 2.9±0.7 mIU/L对 3.3±0.6 mIU/L,P>0.05】; 在治疗3个月末,胆管炎患者血清TT3、FT3、TT4和FT4水平分别为2.4±0.6 nmol/L、4.5±0.8 pmol/L、108.2±36.2 nmol/L和12.9±3.5 pmol/L,较治疗前显著上升,已与健康人无统计学差异（P>0.05）,而肝硬化患者血清TT3、FT3、TT4和FT4水平分别为1.2±0.3 nmol/L、3.4±0.4 pmol/L、93.7±29.8 nmol/L和9.9±3.1 pmol/L,无明显改善,仍显著低于健康人（P<0.05）。结论 原发性胆汁性胆管炎患者经熊去氧胆酸治疗后,血清甲状腺激素水平可获得改善,因此应该早期诊断、早期治疗。     

Beneficial effects of ursodeoxycholic acid via inhibition of airway remodelling,apoptosis of airway epithelial cells,and Th2 immune response in murine model of chronic asthma

Background and aimsIn previous studies, anti-inflammatory, anti-apoptotic and immunomodulatory effects of ursodeoxycholic acid (UDCA) on liver diseases have been shown. In this study, we aimed to investigate the effects of UDCA on airway remodelling, epithelial apoptosis, and T Helper (Th)-2 derived cytokine levels in a murine model of chronic asthma.MethodsTwenty-seven BALB/c mice were divided into five groups; PBS-Control, OVA-Placebo, OVA-50 mg/kg UDCA, OVA-150 mg/kg UDCA, OVA-Dexamethasone. Mice in groups OVA-50 mg/kg UDCA, OVA-150 mg/kg UDCA, OVA-Dexamethasone received the UDCA (50 mg/kg), UDCA (150 mg/kg), and dexamethasone, respectively. Epithelium thickness, sub-epithelial smooth muscle thickness, number of mast and goblet cells of samples isolated from the lung were measured. Immunohistochemical scorings of the lung tissue for matrix metalloproteinase-9 (MMP-9), vascular endothelial growth factor (VEG-F), transforming growth factor-beta (TGF-β), terminal deoxynucleotidyl transferase-mediated dUTP nick endlabeling (TUNEL) and cysteine-dependent aspartate-specific proteases (caspase)-3 were determined. IL-4, IL-5, IL-13, Nitric oxide, ovalbumin-specific immunoglobulin (Ig) E levels were quantified.ResultsThe dose of 150 mg/kg UDCA treatment led to lower epithelial thickness, sub-epithelial smooth muscle thickness, goblet and mast cell numbers compared to placebo. Except for MMP-9 and TUNEL all immunohistochemical scores were similar in both UDCA treated groups and the placebo. All cytokine levels were significantly lower in group IV compared to the placebo.ConclusionsThese findings suggested that the dose of 150 mg/kg UDCA improved all histopathological changes of airway remodelling and its beneficial effects might be related to modulating Th-2 derived cytokines and the inhibition of apoptosis of airway epithelial cells.     

Gallic acid induced apoptotic events in HCT-15 colon cancer cells

AIM: To investigate the inhibitory action of diet-derived phenolic compound gallic acid(GA) against HCT-15 colon cancer cells.METHODS: The antiproliferative effect of GA against colon cancer cells was determined by performing thiazolyl blue tetrazolium bromide(MTT) assay. The colony forming ability of GA treated colon cancer cells was evaluated using the colony forming assay. The cell cycle changes induced by GA in HCT-15 cells were analyzed by propidium iodide staining. Levels of reactive oxygen species(ROS) and mitochondrial membrane potential of HCT-15 exposed to GA was assessed using 2',7'-dichlorfluorescein-diacetate and rhodamine-123 respectively, with the help of flow cytometry. Morphological changes caused by GA treatment in the colon cancer cells were identified by scanning electron microscope and photomicrograph examination. Apoptosis was confirmed using flow cytometric analysis of GA treated HCT-15 cells after staining with Yo-Pro-1.RESULTS: MTT assay results illustrated that GA has an inhibitory effect on HCT-15 cells with IC50 value of 740 μmol/L. A time-dependent inhibition of colony formation was evident with GA treatment. Cell cycle arrest was evident from the accumulation of GA treated HCT-15 cells at sub-G1 phase(0.98 ± 1.03 vs 58.01 ± 2.05)with increasing exposure time. Flow cytometric analysis of GA treated HCT-15 cells depicted early events associated with apoptosis like lipid layer breakage and fall in mitochondrial membrane potential apart from an increase in the generation of ROS which were in a time dependent manner. SEM and photomicrograph images of the GA-treated cells displayed membrane blebbing and cell shrinking characteristics of apoptosis. Further apoptosis confirmation by Yo-Pro-1 staining also showed the time-dependent increase of apoptotic cells after treatment.CONCLUSION: These results show that GA induced ROS dependent apoptosis and inhibited the growth of colon cancer cells.