自噬在胰腺星状细胞激活及纤维化中的作用

胰腺纤维化是各种致病因子导致的胰腺组织损伤后的修复反应,其特点是反复发作的炎症、坏死引起的以胶原纤维增生为主的细胞外基质过度沉积。胰腺星状细胞(pancreatic stellate cells,PSCs)过度激活、合成和分泌大量的细胞外基质成分是贯穿胰腺纤维化的核心事件。近年研究发现,PSCs自噬参与其激活过程,随着PSCs由静息态向激活态的转变,自噬逐渐增强;自噬抑制剂能同时抑制PSCs自噬和激活。因此,深入研究PSCs自噬及其调控机制可能为胰腺纤维化的治疗提供新靶点和新策略。该文就自噬在PSCs激活及胰腺纤维化中的作用研究进展进行了综述。     

瘦素在胆道闭锁患儿肝纤维化中的表达及其临床意义

目的:研究瘦素与胆道闭锁患儿肝纤维化的关系及其在胆道闭锁肝纤维化中的作用,为胆道闭锁患儿的治疗提供依据。方法:回顾性分析2019年8月至2021年8月在天津市儿童医院普外科手术的胆道闭锁或先天性胆总管扩张症(CBD)患儿的临床资料。共纳入31例患儿,其中男性14例,女性17例,中位年龄[60(30,63)]d。胆道闭锁...     

肝纤维化时星状细胞激活的分子生物学机制

本文就肝纤维化时星状细胞激活的分子生物学机制作一综述.肝纤维化时星状细胞初始活化和持续活化主要表现在肝星状细胞的表型改变上.肝纤维化时星状细胞的活化涉及多基因、多因子的表达变化.     

特异性siRNA抑制肝星状细胞Smad2表达

目的构建Smad2特异性siRNA表达克隆,探讨其在肝星状细胞(HSCs)中是否能够有效地抑制Smad2的表达。方法利用生物软件进行Smad2mRNA二级结构模拟,选择合适的靶位点,使用pBSKU6载体表达Smad2特异性siRNA,在体外转染HSC鄄T6细胞株,应用RT鄄PCR和Western鄄blot技术检测Smad2mRNA和蛋白的表达情况。结果成功构建siRNA表达克隆,转染后Smad2在基因和蛋白的表达水平均受到了明显抑制。在构建成功的4个克隆中,CR4抑制mRNA表达最为显著,达90%以上;抑制蛋白表达率也达到80%左右。同时CR1和CR4能有效地下调HSC分泌Ⅲ型胶原。结论在体外实验中,载体表达的特异性siRNA可有效地抑制HSCs中Smad2的表达,并可对激活的HSCs分泌细胞外基质产生有益的影响。     

肝星状细胞在肝脏疾病中的作用

肝星状细胞（HSC）是肝脏内重要的非实质细胞之一,可分泌、释放多种胶原纤维和细胞骨架蛋白参与肝脏疾病的病理生理过程。正常状态下,HSC通过调节细胞外基质蛋白的合成和降解维持肝脏正常的组织结构;肝脏损伤时,HSC被激活,活化的HSC导致细胞外基质的增加是肝纤维化形成并最终导致肝硬化、肝衰竭的主要原因。因此,深入研究HSC在肝脏疾病发生与发展中的作用和机制,并研究与HSC相关的治疗策略,对于提高患者生存率具有一定意义。     

细胞自噬与门静脉栓塞后肝再生

肝癌是我国常见的恶性肿瘤之一,手术切除是目前治疗肝癌最有效的方法,但术后剩余肝脏体积(FLR)不足是限制肝切除手术的重要因素。术前门静脉栓塞术(PVE)可以使栓塞侧肝脏萎缩和对侧肝脏增生,使术后FLR增大,扩大了手术指征。但PVE的应用目前还存在一些问题:PVE后肿瘤的生长速度可能加快,肝再生的速度短时间内难以达到手术的标准等。细胞自噬是真核细胞自身保护机制,对于细胞生长、分化和细胞内稳态的调节至关重要。近年来研究表明,自噬密切调节PV E后肝再生;对自噬与PVE后肝再生机制将加深对PVE后肝再生机制全面了解,为解决PVE中遇到的问题提供新的机会和途径。     

胆道闭锁肝纤维化无创性诊断的研究进展

胆道闭锁是儿童进行肝移植的最常见的胆源性肝脏疾病,进行性肝纤维化是本病的显著特点,即使进行了肝门空肠R-Y吻合术,大多数患儿术后仍不可避免的出现肝纤维化的进行性加重,直至发展成为肝硬化、肝衰竭,并出现一系列并发症.因此肝纤维化评估是胆道闭锁患儿术后随访的重要内容,准确了解肝纤维化程度,对胆道闭锁患儿的病情评估有着重要的意义.肝组织活检是判断肝纤维化分级的金标准,但是其本身存在许多问题.运用多项无创性指标对患儿肝纤维化程度进行评估成为目前胆道闭锁预后研究的热点,本文对主要应用于胆道闭锁患儿肝纤维化的若干无创性诊断进行系统性分析,并分别从影像学和血清学两个方面进行综述.     

BCAT1对HSC-LX2人肝星状细胞纤维化相关基因表达的影响

[摘要] 目的 探讨支链氨基酸转氨酶1（BCAT1）对HSC-LX2人肝星状细胞纤维化相关基因表达的影响。方法 构建携带BCAT1基因的慢病毒载体,将病毒转染HSC-LX2细胞,运用qRT-PCR和Western blotting检测转染BCAT1过表达病毒后的HSC-LX2细胞中BCAT1、ACTA2、TIMP1、MMP2和COL1A1基因的表达情况。结果 携带BCAT1基因的慢病毒转染HSC-LX2细胞后,实验组（转染p-BCAT1）HSC-LX2细胞和对照组（转染Vec）HSC-LX2细胞相比,促进纤维化的ACTA2、TIMP1、COL1A1基因的mRNA及蛋白表达均升高,抗纤维化的MMP2基因的mRNA及蛋白表达均下降,两者的差异均具有统计学意义（P < 0.05）。结论 BCAT1增强HSC-LX2人肝星状细胞中ACTA2、TIMP1、COL1A1促纤维化相关基因的表达,抑制抗纤维化相关基因MMP2的表达。     

大鼠肝星状细胞和库普弗细胞的分离培养及鉴定

目的 应用改良的方法同步分离、培养和鉴定原代大鼠肝星状细胞(HSC)和库普弗细胞(KC).方法 采用Ⅳ型胶原酶肝脏灌注消化及Percoll密度梯度离心体外同步分离、纯化及培养HSC和KC,并用光学显微镜、免疫组织化学染色、电子显微镜等方法进行鉴定.结果 每只大鼠HSC和KC的细胞获得率分别为(1.8～2.9)×107和(0.8～1.4)×107,平均纯度分别为94.7%±1.7%和95.2%±1.5%,活力分别为95.5%±1.3%和96.4%±2.4%.结论 Ⅳ型胶原酶肝脏灌注消化及Percoll密度梯度离心体外同步分离培养HSC和KC方法简单,细胞获得率、活力和纯度高.     

肝星状细胞、相关因子与肝纤维化关系的研究进展

目前多数认为肝纤维化的形成机制是致病因子造成肝细胞损伤，引起肝Kupffer细胞(KC)、血小板、肝窦内皮细胞和肝细胞激活，分泌多种细胞因子(cytokine)，与某些化学递质共同作用于肝星状细胞(hepatic stellate cell，HSC)，使其激活，转化为肌成纤维细胞(myofibroblast，MFB)，通过旁分泌与自分泌作用，使HSC增殖，合成大量的细胞外基质(extracellular matrix，ECM)，以致其在肝内大量沉积，肝纤维化逐渐形成。即表明，HSC活化是各种病因肝纤维化发生的共同中心环节。     

尿CMV快速培养在胆道闭锁患儿诊治中的作用

目的调查我科收治的胆道闭锁(biliary atresia,BA)患儿巨细胞病毒(cy-tomegalovirus,CMV)感染率并探讨尿CMV快速培养在BA患儿诊治中的作用。方法对21例BA患儿血清CMV-IgM、CMV-IgG及尿CMV快速培养结果进行总结,同时采用Masson染色法检测患儿肝纤维化程度;根据尿CMV检测结果进行分组,比较两组患儿肝功能、肝纤维化程度及胆汁排出率的差别。结果BA患儿尿CMV快速培养阳性率为57.1%,高于血清CMV-IgM检测结果;尿CMV阳性组患儿肝功能及肝纤维化程度明显高于尿CMV阴性组,而术后胆汁排出率低于尿CMV阴性组。结论BA发生与CMV感染相关;尿CMV快速培养可用于诊断BA患儿CMV活动性感染;伴有CMV活动性感染的BA患儿肝损伤及肝纤维化程度均较严重,应尽早手术。     

The effect of hepatic portal dissection on the portal vein structure in biliary atresia

Superior mesenteric portograms were performed on 30 patients with biliary atresia (BA) at the time of initial portoenterostomy in 20, and at the stoma closure operation in 10. A withered-branch-shaped irregularity of the intrahepatic portal vein (PV) and collateral vessels were seen in 2 of 11 patients with portal pressures (PP) of 200 to 300 mmH₂O; in 1 of 2 patients with PPs of over 300 mmH₂O at the initial operation; and in 3 jaundice-free patients with PPs of 285, 320, and 305 mmH₂O, respectively, at the stoma closure operation. Collaterals were the only abnormalities seen in two additional jaundice-free patients with PPs of 370 and 183 mmH₂O, respectively. No anatomic changes in the extrahepatic PV at the porta hepatis were found on the portograms of either group of patients. Thus, we conclude that portal dissection itself does not affect the PV structure anatomically, a finding which has important implications in determining whether or not portoenterostomy adversely affects potential liver transplantation.     

Evidence for segmental bile drainage by hepatic portoenterostomy for biliary atresia: Cholangiographic, hepatic venographic, and histologic evaluation of the liver taken at liver transplantation

Background

The result of hepatic portoenterostomy for biliary atresia (BA) has improved, but there are some patients who experience worsened liver function in the long term after one decrease in jaundice owing to portoenterostomy. However, the cause of the liver dysfunction in the long term has not been clearly ascertained.

Methods

Five patients (5 to 28 years of age) with BA underwent liver transplantation (LT) because of liver dysfunction after successful portoenterostomy. To clarify the cause of liver dysfunction occurring in the long term, the authors performed a cholangiogram, hepatic venogram, and macroscopic/microscopic examination of the liver just after LT.

Results

(1) Macroscopically, the liver could be divided into 3 areas, the hypertrophic, atrophic, and intermediate, with findings between those of the hypertrophic and atrophic areas. (2) The divided areas clearly corresponded to the liver segments. Segment IV was the hypertrophic area in all patients, but segments VI and VII were the atrophic areas in 4 of the 5 patients. (3) Based on the cholangiographic and microscopic findings, the hypertrophic area had near-normal structure with bile ducts. The atrophic area had severe fibrosis and contained only a few bile ducts in the intralobular spaces of liver.

Conclusions

It seems that segmental bile drainage must have been established by hepatic portoenterostomy in some patients and that some postoperative patients might have worsened liver function in the long-term follow-up period accompanied with progression of fibrosis and impaired bile drainage. These pathologic changes occur in each liver segment.     

Effects of the herbal medicine Inchinko-to on liver function in postoperative patients with biliary atresia--a pilot study

Background/Purpose:

A continuation of liver fibrosis after undergoing successful Kasai operation has become the important clinical issue in the long-term follow-up of patients with biliary atresia (BA). The aim of this study is to evaluate the efficacy of the herbal medicine Inchinko-to (TJ-135) on the treatment of liver fibrosis in patients with BA without jaundice, especially from the viewpoint of the long-term effects of TJ-135.

Methods:

Six postoperative patients with BA ranging between 3 and 13 years of age with normal serum total bilirubin levels (total bilirubin < 1.0 mg/dL [17 μmol/L]) received TJ-135 from 2 to 4 years. The liver enzyme (glutamic oxaloacetic transaminase [GOT], glutamic pyruvic transaminase [GPT], gamma glutamyl transpeptidase[γ-GTP]transpeptidase[γ-GTP] levels and hyaluronic acid (HA) levels were compared before and after the administration of TJ-135. The monthly collected data were averaged on a 1-year basis. The record of one postoperative patient with BA and a normal serum total bilirubin level was incorporated as a control. This patient showed portal hypertension and did not receive TJ-135.

Results:

Five of the six patients who showed abnormal values for liver enzymes, exhibited a significant decrease in serum GOT, γ-GTP, or GPT levels after a 1 to 3-year administration of TJ-135, and the improvement in these parameters persisted thereafter. Furthermore, one patient who had an abnormally high value of HA also showed a significant decrease in the serum level of HA. In the remaining patient with normal liver enzyme values, no significant change was observed during the administration of TJ-135. The control patient exhibited a chronological decrease in the serum GOT and GPT levels by 5 years of age, but the serum γ-GTP and HA levels remained stable throughout the postoperative period.

Conclusions:

The long-term effectiveness of TJ-135 was only found in those patients with abnormal liver enzyme levels and HA, thereby suggesting that TJ-135 has a protective and antifibrotic effect on the liver.     

Doppler ultrasonographic evaluation of hepatic circulation in patients following kasai's operation for biliary atresia

The hepatic circulation of eight children who underwent Kasai's operation for biliary atresia was serially evaluated by Doppler ultrasonography (US). A total of 36 examinations were performed to evaluate the maximal velocities (mv_s) of the main portal vein (MPV), splenic vein (SV), and hepatic artery (HA), and to analyze the spectral waveform and the directions of flow. The mean mv_s-MPV in four patients for whom adequate biliary diversion had been achieved and whose serum bilirubin had fallen to within the normal range (group A), was 19.8±7.5 cm/s. Their MPV waveforms were constant, with blood flowing toward the liver, while their mean mv_s-SV was 12.1±5.8 cm/s. In two other patients with apparent hypersplenism, but whose serum bilirubin levels had fallen to nearly normal (group B), the mean mv_s-MPV was 20.7±10.4 cm/s and the mv_s-SV was 22.4±10.4 cm/s. In contrast, the mv_s-MPV in the two remaining patients, whose serum bilirubin levels had either not fallen to within the normal range, or had fallen initially but increased due to recurrent cholangitis (group C), was 14.2±9.1 cm/s. In these patients, the waveforms were unstable, the MPV flows were occasionally hepatofugal, and their mean mv_s-SV was 18.0±5.8 cm/s. The mv_s-HA were markedly increased in the latter four patients, whose clinical condition had also deteriorated. These observations led to the conclusion that hepatic circulation evaluated by serial Doppler US provides important information about liver status in children who have undergone Kasai's operation for biliary atresia.     

A histopathological study of the remnant of extrahepatic bile duct in so-called uncorrectable biliary atresia

Histopathological study of the remnant of extrahepatic bile ducts in 40 cases of so-called uncorrectable biliary atresia, upon which we operated the last three years, has been performed. The histological findings of the remnant were classified into three types.Only two cases were found to have type 1a ducts in the porta hepatis area, from which we can expect better prognosis postoperatively. We also found that as the patients become older, the size of the duct in the remnant becomes smaller and the hepatic fibrosis becomes more remarkable. Therefore the operation should be performed in the infant with this lesion as young as possible.As for the evaluation of operative results of hepatic portoenterostomy for this lesion, a proper evaluation can be made only in those cases in which a microscopic examination of the remnant of extrahepatic bile duct at the porta hepatis area has been adequately performed.Concerning the pathogenesis of biliary atresia, we presume that congenital abnormalities of bile ducts are a basic factor, and additional nonspecific inflammation and bile stasis complete its pathological condition.     

Technical aspects of hepatic portal dissection in biliary atresia.

During the past 8 yr, 37 patients with a noncorrectable type of biliary atresia have undergone hepatic portoenterostomy or portocholecystostomy at the Kobe Children's Hospital. The hepatic portal dissections employed in this series were classified as "supraportal" (9 procedures), "portal" (25 procedures), and "infra-portal" (3 procedures) based on the level at which the fibrous mass at the porta hepatis was transsected as determined by the operative record and the pathologic findings. Successful biliary drainage was achieved in 19 out of 25 patients (76%) with a "portal" type of dissection, while 1 out of 9 with "supra-portal" and none out of 3 with "infra-portal" type dissections were successful in this respect. Of the 19 patients who achieved significant biliary flow, 8 have lived for 2--7 yr without jaundice and 3 others are jaundice-free for shorter intervals.     

胆道闭锁肝内胆管上皮细胞凋亡与增殖的实验研究

目的　研究胆道闭锁 (biliaryatresia ,BA)肝内胆管上皮细胞凋亡与增殖情况 ,探讨其与BA的关系。方法　应用末端脱氧核苷酸转移酶介导的脱氧三磷酸尿苷缺口末端标记技术(TUNEL法 )和Ki -67免疫组化染色方法 ,观察 3 8例胆道闭锁患儿与 16例正常对照儿童肝内胆管上皮细胞凋亡与增殖情况。结果　胆道闭锁患儿肝内胆管上皮细胞凋亡指数为 (5 1.74±19 .93 ) %明显高于正常对照 (12 .3 4± 19.3 2 ) % (P <0 .0 1) ,增殖指数 (19.0 4± 9.78) %相对正常对照 (12 .68± 15 .5 6) %有所升高 ,但差异无显著性 (P >0 .0 5 )。肝细胞凋亡指数和增殖指数分别与正常对照相比差异无显著性 (P >0 .0 5 )。BA胆管上皮细胞凋亡指数明显高于增殖指数 (P <0 .0 1) ,而肝细胞凋亡指数与增殖指数比较则无统计学差别 (P >0 .0 5 )。正常对照组胆管上皮细胞凋亡指数与增殖指数相比也无统计学意义 (P >0 .0 5 )。结论　胆道闭锁的发生及其进行性发展与肝内胆管上皮细胞凋亡增殖不平衡有关 ,该疾病可能是一种波及整个胆管系统的全胆道病变。     

肝星状细胞的生物学特性及其调节肿瘤微环境的研究进展

肝星状细胞是肝内的非实质性细胞.在消化系肿瘤的发生和肝转移过程中,肝星状细胞发生活化并与肿瘤细胞相互作用,引起基质重建,促进肿瘤的侵袭转移.本文就肝星状细胞的生物学与免疫学特性及其与消化系肿瘤细胞之间相互作用的研究进展作一综述.