辣椒素对肝星状细胞增殖凋亡的影响

目的 探讨辣椒素(capsaicin)对大鼠肝星状细胞(hepatic stellate cell,HSC) T6细胞系生长的影响及其机制.方法 用CCK-8(Cell Counting Kit-8)法检测终浓度50、100、150、200 μmol/L的辣椒素对大鼠肝星状细胞T6细胞株生长的影响;采用流式细胞仪检测辣椒素对大鼠肝星状细胞T6细胞凋亡率的影响;Western blotting法检测Bcl-2、Bax、细胞色素酶c(Cyt c)蛋白的表达.结果 辣椒素能显著抑制大鼠肝星状细胞T6细胞株增殖,呈时间浓度依赖性(P<0.05).辣椒素作用24 h后T6细胞凋亡率与对照组相比明显升高(P<0.05),与对照组相比,辣椒素作用组Bcl-2蛋白表达量明显下降,bax蛋白的表达明显上升,Cyt c蛋白的表达明显上升,Bcl-2/Bax灰度值明显下降(P<0.05).结论 辣椒素抑制T6细胞增殖并诱导其凋亡,其机制可能是与下调Bcl-2蛋白的表达,上调Bax蛋白的表达,促进Cyt c蛋白的释放有关.     

肝癌切除联合门静脉、肝动脉置泵化疗的临床观察

目的 探讨肝癌切除联合门静脉、肝动脉置泵化疗治疗肝癌的临床疗效及其应用价值. 方法 1998年3月至2002年3月采用肝癌切除63例,随机分为2组,Ⅰ组24例仅行肝癌切除,Ⅱ组39例肝癌切除时联合门静脉、肝动脉置泵化疗,58例获随访. 结果 5例手术后3个月内死于肝肾功能衰竭,53例术后恢复良好.术后1,2,3年复发率和生存率据统计学检验,Ⅱ组的手术后复发率明显低于Ⅰ组(P<0.05),Ⅱ组的手术后生存率明显高于Ⅰ组(P<0.01). 结论 肝癌切除联合门静脉、肝动脉置泵化疗,可以降低术后复发率,提高生存率.     

Aurora A激酶抑制剂Alisertib诱导结直肠癌细胞自噬的作用及与p53表达间的关系

目的 探讨Alisertib（ALS）对人结直肠癌Caco-2和HT29细胞的诱导自噬的作用及与p53 表达状态间的关系。方法 本研究采用人结直肠癌Caco-2和HT29细胞进行实验。实验组用0.1、1、5 μmol/L ALS而对照组用同浓度的DMSO处理细胞。流式细胞术检测细胞的自噬率,Wester Blotting法检测细胞自噬过程中关键调节分子的蛋白表达。结果与对照组比较,0.1、1 μmol/L Alisertib组Caco-2细胞自噬率增加,分别为26.4%和26.8%（P<0.01）。1、5 μmol/L ALS组HT29细胞自噬率增加,分别为36.8%和42.6%（P<0.01）。Caco-2细胞不表达p53蛋白,而HT29细胞表达p53蛋白。与对照组比较,1、5μmol/L Alisertib引起Caco-2细胞的p-PI3K/PI3K、p-Akt/Akt和p-p38/p38的比值分别降低为对照组的62.6%、45.2%,30.1%、38.2%和49.0%、30.4%（P<0.01）,LC3-Ⅱ/LC3-I的比值升高32.6%、46.8%（P<0.01）。与对照组比较,1、5 μmol/L Alisertib引起HT29细胞的p-PI3K/PI3K、p-Akt/Akt的比值分别降低为对照组的70.9%、66.3%和85.2%、27.2%（P<0.01）,p-p38/p38和LC3-II/LC3-I的比值升高2.1倍、2.0倍和78.5%、84.8%（P<0.05）。与5 μmol/L Alisertib组比较,10 μmol/L SB202190+5 μmol/L Alisertib引起HT29细胞凋亡率升高64.7%（P<0.001）,而Caco-2细胞凋亡率无明显变化。结论 Alisertib抑制了PI3K/Akt信号通路,诱导了Caco-2和HT29细胞发生细胞自噬。Alisertib对p38 MAPK信号通路的影响不同,激活了HT29而抑制了Caco-2细胞的p38 MAPK信号通路,可能与p53蛋白是否表达有关。     

视黄醇类核内受体-α基因慢病毒载体的构建及其对肝星状细胞的影响

目的:通过视黄醇类核内受体-α(retinoid X receptor-alpha,RXR-α)基因慢病毒表达载体的构建,及其对肝星状细胞(hepatic stellate cells,HSC)活化、增殖影响的研究,探讨RXR-α基因在肝纤维化中的作用。方法:①构建RXR-α基因慢病毒表达载体;②分离和体外培养大鼠HSC;③将自发活化的HSC分成3组,即正常对照组、阴性对照组和RXR-α载体组;分别将空白慢病毒载体和RXR-α慢病毒载体转染自体活化的HSC,采用Western印迹法检测各组细胞中RXR-α、α-SMA和Ⅰ型胶原蛋白表达变化,用MTT法检测各组HSC培养24、48、72及96 h后的增殖情况。结果:正常对照组及阴性对照组HSC中几乎没有RXR-α蛋白的表达,而RXR-α载体转染组的HSC中出现RXR-α蛋白的明显表达。与正常对照组及阴性对照组相比,RXR-α载体组转染的HSC中α-SMA蛋白表达量显著下降(t=3.767,P0.01和t=8.491,P0.05),Ⅰ型胶原蛋白表达也显著降低(t分别为10.449和10.756,P值均0.01),同时HSC的增殖能力也显著下降(t分别为4.381和1.778,P值均0.05)。而阴性对照组与正常对照组相比,α-SMA和Ⅰ型胶原蛋白表达量和HSC增殖能力无明显变化(P0.05)。结论:RXR-α基因在HSC内的增强表达能显著抑制HSC的活化和增殖,具有潜在的抑制肝纤维化的作用。     

siRNA干扰ADAMTS2基因对肝星状细胞的影响

目的 探讨siRNA干扰大鼠含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶2(ADAMTS2)基因后对大鼠肝星状细胞(HSC)生物学特性的影响及其机制,评估ADAMTS2基因作为一种新的靶向基因在抗肝纤维化治疗中的应用前景.方法 设计并化学合成3对针对大鼠ADAMTS2 mRNA 2237、2597及690位点的siRNAs.用筛选出的抑制效率最高的siRNA体外转染HSC-T6,应用实时PCR、Westem blot及MTT等方法研究沉默ADAMTS2基因对大鼠HSC活化、增殖等生物学特性及对基因ADAMTS2、α-SMA、COL1α1、COL(I)、TGF-β1、MMP-2和TIMP-3表达的影响.结果 在相同注射剂量(50 nmol/L)及时间(48 h)下,2237位点的siRNA-ADAMTS2具有最高的抑制效率;能显著抑制ADAMTS2基因mRNA和蛋白表达(抑制率80％以上);显著抑制COL(I)、α-SMA和TGF-β1在HSC中的表达.HSC的增殖能力显著降低.结论 化学合成的siRNA-ADAMTS2干扰载体能有效抑制ADAMTS2基因在大鼠HSC-T6细胞系中的表达,抑制HSC的活化和增殖,同时显著降低COL(I)、α-SMA和TGF-β1的表达,具有潜在的阻止甚至逆转肝纤维化的作用.ADAMTS2可能是抗肝纤维化治疗的有效靶位,抑制ADAMTS2在肝脏的表达可能是一种有效的抗肝纤维化治疗策略.     

胆囊癌中TβR Ⅰ与细胞增殖及细胞周期的关系

目的 探讨T13R Ⅰ与胆囊癌细胞增殖和细胞周期的关系,分析其在胆囊癌发病中的机制,评价其对胆囊癌预后的价值.方法 用免疫组化法对原发性胆囊癌及胆囊腺瘤、慢性胆囊炎行转化生长因子βⅠ型受体(transforming growth factor β receptor type Ⅰ,TβR Ⅰ)、增殖细胞核抗原(proliferative cell nuclear antigen,PCNA)、细胞周期素E(Cychn E)进行检测,对胆囊癌患者进行随访.结果 胆囊癌中TβR Ⅰ阳性表达率34.29%,显著低于胆囊腺瘤、胆囊炎(P均小于0.05).伴淋巴结和远处转移者TβR Ⅰ阳性表达率明显低于无转移者(P<0.05),TβR Ⅰ阳性表达率与PCNA LI呈负相关(r=0.4024,p=0.0166).TβR Ⅰ表达阳性者预后比阴性者好(P<0.05).结论 TβR Ⅰ与胆囊癌细胞增殖和细胞周期关系密切,是判断胆囊癌发生发展的重要生物学标志.TβR Ⅰ低表达而导致TGF-β负性生长调控作用的逃逸,可能是胆囊癌变过程中的重要机制.TβR Ⅰ是判断胆囊癌预后的良好指标.     

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。     

瘦素受体在小鼠日本血吸虫病肝纤维化组织中的动态表达及作用

目的 观察小鼠感染日本血吸虫后不同时期肝脏瘦素长受体(OB-Rb)mRNA和蛋白动态表达,探讨瘦素及其受体在日本血吸虫病肝纤维化发生发展中的作用.方法 日本血吸虫尾蚴皮肤敷贴法感染小鼠构建日本血吸虫病肝纤维化模型,于感染后2、4、8、12、16、20、24周取肝脏标本.HE和VG染色对肝组织进行病理学检查,免疫组织化学SP法动态观察OB-Rb及肝星状细胞(HSC)标记物α-平滑肌动蛋白(α-SMA)表达;逆转录-聚合酶链反应(RT-PCR)检测OB-Rb mRNA动态表达.结果 α-SMA和OB-Rb蛋白阳性表达随病程进展进行性增加,两者共表达于胶原纤维沉积处;模型组OB-Rb mRNA在感染4周开始表达,随病程进展逐渐增高,持续至24周.相关分析表明,α-SMA蛋白表达与胶原含量(r=0.956,P《0.01)及肝纤维化程度(r=0.804,P《0.01)均呈正相关;OB-Rb蛋白和mRNA表达与胶原含量(r=0.965,P《0.01;r=0.945,P《0.01)及肝纤维化程度(r=0.823,P《0.01;r=0.880,P《0.01)均呈正相关.结论 瘦素、HSC和日本血吸虫病肝纤维化之间有着极为密切的关系,瘦素可能是日本血吸虫病肝纤维化形成过程中一个新的促肝纤维化因子.     

miR-21调节缺血缺氧诱导的大鼠肝卵圆细胞自噬的研究

目的探讨miR-21在缺血缺氧诱导的大鼠肝卵圆细胞自噬中的作用。方法体外培养肝卵圆细胞,慢病毒转染肝卵圆细胞构建稳定的细胞株,分别提取各组细胞RNA逆转录后行SYBR Green实时荧光定量PCR,检测各组miR-21的表达,以转染好的稳定的细胞建立缺血缺氧模型,共分为3组:miR-21空载体慢病毒转染HOC自噬组,miR-21增强慢病毒载体转染HOC自噬组,miR-21-inhibition慢病毒载体转染HOC自噬组。Hoechst 33258染色观察细胞凋亡;应用丹(磺)酰戊二胺(Monodansylcadaverine,MDC)染色荧光定位法、观察各组细胞的自噬;免疫印迹法(Western blotting)检测各组细胞LC3-Ⅱ/Ⅰ蛋白表达情况。结果与空载体组比,增强组miR-21基因水平表达增强,而MDC染色减弱(自噬减少),蛋白LC3-Ⅱ/LC3-Ⅰ的比值减少;而抑制组miR-21基因水平表达减少,MDC染色增强(自噬增强),蛋白LC3-Ⅱ/LC3-Ⅰ的比值增多。结论抑制miR-21过表达可以增强缺血缺氧引起的肝卵圆细胞的自噬,有利于肝卵圆细胞在缺血缺氧微环境中稳定细胞内环境,维持细胞的存活。     

小RNA干扰DDR2基因表达对肝星状细胞的影响

目的 探讨siRNA干扰大鼠盘状结构域受体2(discoidin domain receptor2,DDB2)基因对肝星状细胞(hepatic stellate cell,HSC)的影响,评估DDR2在肝纤维化发生中的作用.方法 (1)化学合成3对针对DDR2基因的siRNAs,转染肝星状细胞株(HSC-T6),筛选抑制效率最高的siRNA用于干扰实验;(2)将肝星状细胞株HSC-T6分为3组:正常组(normal,N组)、阴性对照组(negative control,NC组)和干扰组(siRNA-DDR2,SI组).将筛选的抑制效率最高的siRNA以脂质体法转染HSC-T6细胞株,以RT-PCR检测其DDR2,α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)和I型胶原(collagen-Ⅰ)的mRNA表达,Western blot检测DDR2蛋白表达变化;同时四唑盐(MTT)法检测细胞增殖情况.结果 (1)868位点的siRNA抑制效率最高;(2)与正常组相比,siRNA-DDR2转染组DDR2和α-SMA的mRNA表达均显著下降(分别t=6.43,t=7.34,均P<0.01),DDR2蛋白的表达亦显著降低(t=4.49,P<0.01),同时HSC的增殖能力也显著降低(t=18.32,P<0.01);相反,阴性对照组与正常组相比,DDR2、α-SMA的mRNA表达以及DDR2蛋白和HSC的增殖能力则无明显改变.结论 siRNA-DDR2能显著抑制HSC的活化与增殖,进而抑制纤维化发生,具有潜在的抗纤维化作用.     

Oridonin inhibits hepatic stellate cell proliferation and fibrogenesis

Background

Liver fibrosis is a common response to liver injury and, in severe cases, leads to cirrhosis. The hepatic stellate cells (HSCs) become activated after liver injury and play a significant role in fibrogenesis. The activated HSC is characterized by increased proliferation, overexpression of α smooth muscle actin, and excessive production of extracellular matrix (ECM) proteins. Oridonin, a naturally occurring diterpenoid, has been shown to induce apoptosis in liver and gastric cancer cells. However, its effects on the HSC are unknown.

Methods

We tested the effects of oridonin on the activated human and rat HSC lines LX-2 and HSC-T6, and the human hepatocyte cell line C3A. Transforming growth factor β1 (TGF-β1) was used to stimulate LX-2 cells.

Results

Oridonin significantly inhibited LX-2 and HSC-T6 proliferation. In contrast, oridonin had no antiproliferative effect on C3A cells at our tested range. Oridonin induced apoptosis and S-phase arrest in LX-2 cells. These findings were associated with an increase in p53, p21, p16, and cleaved Poly (ADP-ribose) Polymerase (PARP), and with a decrease in Cyclin-dependent kinase 4 (Cdk4). Oridonin markedly decreased expression of α smooth muscle actin and ECM protein type I collagen and fibronectin, blocked TGF-β1-induced Smad2/3 phosphorylation and type I collagen expression.

Conclusions

Oridonin induces apoptosis and cell cycle arrest involving the p53-p21 pathway in HSC and appears to be nontoxic to hepatocytes. In addition, oridonin suppressed endogenous and TGF-β1-induced ECM proteins. Thus, oridonin may act as a novel agent to prevent hepatic fibrosis.     

Hepatic stellate cell activation and hepatic fibrosis induced by ischemia/reperfusion injury

Objective

Warm ischemia causes severe allograft damage in liver transplantation. However, the long-term effects of ischemia/reperfusion injury (IRI) on fibrosis have not been fully elucidated. In this study, we used a partial warm hepatic ischemia mouse model to monitor fibrosis in the ischemic liver.

Materials and Methods

Male BALB/c mice were divided into ischemic and sham groups (n = 30/group). Via a midline laparotomy, an atraumatic clip was used to interrupt the arterial and the portal venous blood supply to the left liver lobe. After 90 minutes of partial hepatic ischemia, the clip was removed initiating hepatic reperfusion. Samples from normal, sham, and ischemic liver tissues were collected at intervals of 1, 5, 10, 15, 20, or 30 days after operation (n = 5 for each time point) for hematoxylin-eosin (H&E), Mallory's trichrome, and alpha-smooth muscle actin (α-SMA) immunohistochemical stains for fibrosis and activation of hepatic stellate cell (HSCs).

Results

IRI caused significant HSC activation in the ischemic liver tissues. Mallory's trichrome stain demonstrated that IRI caused hepatic parenchymal fibrosis near portal tracts and central veins. With prolonged reperfusion time hepatic parenchymal fibrosis was aggravated, showing the same pattern of HSC activation. IRI also caused increased portal tract fibrosis in ischemic liver tissues, especially around biliary tracts.

Conclusions

Hepatic IRI caused HSC activation, increasing hepatic parenchymal and portal tract fibrosis in ischemic liver tissues.     

Protective effect of emodin on renal interstitial fibrosis in mice of unilateral ureteral obstruction via PI3K/Akt/mTOR signaling pathway

Objective To investigate the effect and mechanism of emodin (EM) in renal interstitial fibrosis of unilateral ureteral obstruction (UUO) mice. Methods Male C57BL/6J mice were randomly divided into 4 groups, including sham operation group (n=8), UUO operation group (n=8), UUO operation+losartan (LST) group (n=8) and UUO operation+EM group (n=8). The mice in each group were ingested the suspensions by gavage for 14 days after surgery. Mice in UUO+LST and UUO+EM groups were given 10 mg?kg-1?d-1 LST and 20 mg?kg-1?d-1 EM, respectively. LST and EM were mixed with 0.5% sodium carboxymethyl cellulose. Mice in sham group and UUO group were given 0.5% sodium carboxymethyl cellulose. The mice were sacrificed at the 14th day. Interstitial fibrosis was observed by HE, Masson and PAS stain. Real-time PCR was used to detect LC3, Beclin-1 and mTOR mRNA. Protein expressions of TGF-β1, α-SMA, E-cadherin, LC3, Beclin-1, PI3K, p-Akt and mTOR were detected by Western blotting. The autophagy was observed with transmission electron microscopy in the renal tissue. Results Compared with sham mice, UUO mice at the 14th day displayed obvious renal fibrosis. Meanwhile, UUO mice had increased expressions of TGF-β1 and α-SMA (all P＜0.01), and decreased expressions of E-cadherin (P＜0.01). Their renal expressions of PI3K, p-Akt and mTOR were also raised (all P＜0.01). Compared with those in UUO group, in UUO+LST group and UUO+EM group, expressions of autophagy protein LC3 and Beclin-1 were increased (all P＜0.01), and the number of autophagic was increased. Additionally, expressions of TGF-β1 and α-SMA were reduced in UUO+LST group and UUO+EM group (all P＜0.01), while the expression of E-cadherin was increased by emodin treatment (P＜0.05). And expressions of PI3K, p-Akt and mTOR were decreased in UUO+LST group and UUO+EM group (all P＜0.05), meanwhile renal tissue fibrosis significantly reduced. Conclusions Emodin can promote autophagy, ameliorate renal interstitial fibrosis and protect renal function through PI3K/Akt/mTOR signaling pathway.     

狼疮肾炎患者自噬水平及其对足细胞相关蛋白表达水平的影响

目的探讨狼疮肾炎(lupus nephritis,LN)患者自噬水平及其对足细胞相关蛋白表达水平的影响。方法选择2017年5月至2019年5月于榆林市第一医院收治的69例LN患者为LN组,50例系统性红斑狼疮(systemic lupus erythematosus,SLE)患者为SLE组,50例肾切除手术患者为对照组,观察3组肾脏足细胞内自噬体数量,比较3组肾脏组织中自噬相关蛋白、微管相关蛋白轻链3(LC3)、B淋巴细胞瘤蛋白质-2-相互作用蛋白(Beclin1)的表达,以及足细胞相关蛋白,肾病蛋白(Nephrin)、足突蛋白(Podocin)的表达。分离狼疮肾炎患者肾脏足细胞,将足细胞分为自噬抑制组和自噬诱导组,自噬抑制组加入100 nmol/L 3-甲基腺嘌呤(3-MA),自噬诱导组加入100 nmol/L雷帕霉素(RAPA),比较3组足细胞内自噬体数量及LC3、Beclin-1、Podocin、Nephrin等蛋白的表达。结果LN组、SLE组肾脏足细胞内自噬体数量及LC3、Beclin-1、Podocin、Nephrin蛋白表达量显著高于对照组(P<0.05);SLE组肾脏足细胞内自噬体数量及LC3、Beclin-1、Podocin、Nephrin蛋白表达量显著高于LN组(P<0.05);自噬诱导组足细胞内自噬体数量及LC3、Beclin-1、Podocin、Nephrin蛋白表达量显著高于正常对照组、自噬抑制组(P<0.05);正常对照组足细胞内自噬体数量及LC3、Beclin-1、Podocin、Nephrin蛋白表达量显著高于自噬抑制组(P<0.05)。结论LN患者自噬水平呈升高状态,自噬水平升高可能通过上调足细胞相关蛋白Podocin、Nephrin水平而减轻足细胞损伤,抑制LN病情进展。     

抗氧化剂治疗肝纤维化的研究进展

肝纤维化是肝内弥漫性纤维组织沉积,是多种慢性肝损伤的共同后果,持续发展则成为肝硬化甚至肝癌。肝星状细胞（HSC）的活化使细胞外基质（ECM）的产生与降解不平衡,是导致肝纤维化发生发展的重要环节。氧化应激反应与肝脏的纤维化密切相关,作为一种重要的机制参与到HSC的活化和ECM的沉积,因此,抗氧化剂作为一种重要的治疗肝纤维化药物成为研究热点。笔者就近几年抗氧化剂治疗肝纤维化的研究进展进行综述。     

Connective tissue growth factor expression is increased in biliary epithelial cells in biliary atresia

Background/Purpose

Connective tissue growth factor (CTGF) has been implicated in the pathogenesis of hepatic fibrosis and is elevated in the serum of children with biliary atresia (BA). The objective of this study was to evaluate hepatic CTGF messenger RNA (mRNA) expression and its relationship to hepatic histology in children with BA.

Methods

Connective tissue growth factor mRNA expression was evaluated by in situ hybridization in 26 liver biopsies from 11 patients with BA, 11 with other diseases, and 4 autopsy controls. Serial sections were immunostained with cell-specific markers to characterize the cells expressing CTGF. Biopsies were scored for CTGF expression (0-4) and inflammation and fibrosis (1-4).

Results

High levels of CTGF expression were observed in 9 of 11 BA with localization to biliary epithelial cells and vascular endothelial cells. Connective tissue growth factor mRNA expression was correlated with fibrosis in BA and all livers. In the 11 patients with other liver diseases, 7 had CTGF expression limited to hepatic stellate cells and vascular endothelial cells. None of the 4 livers in children without liver disease had significant levels of CTGF.

Conclusions

In BA livers, novel biliary epithelia CTGF mRNA expression is high and correlates with severity of fibrosis. These data support a role for biliary epithelial cell signaling in fibrogenesis.     

落新妇苷对大鼠肝脏缺血再灌注损伤及自噬的影响

目的探讨落新妇苷对大鼠肝脏缺血再灌注损伤（HIRI）的保护作用及自噬的影响。 方法将54只SD大鼠随机分为假手术组（Sham组）、缺血再灌注组（HIRI组）以及落新妇苷组（40 mg·kg^-1·d^-1,连续7 d）,每组18只。建立大鼠HIRI模型,于再灌注4、8、16 h后取下腔静脉血及肝左外叶组织。全自动生化分析仪检测血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）水平;光学显微镜下观察肝细胞显微结构变化;Western blotting分析肝组织中自噬相关蛋白LC3、Beclin-1的表达;电镜下观察肝脏组织内自噬小体数量情况。 结果同检测时间点比较,Sham组和落新妇苷组的血清ALT和AST水平均低于HIRI组（P<0.05）,且肝细胞肿胀、炎性细胞浸润、汇管区结构损伤明显较轻。落新妇苷组LC3-Ⅱ/ LC3-Ⅰ灰度值比值及Beclin-1蛋白灰度值在术后4、8、16 h明显低于HIRI组（P<0.05）。落新妇苷组肝组织的自噬小体数量较HIRI组有所减少（3.68±0.42 vs 7.12±0.60,t=36.382,P<0.01）。 结论落新妇苷预处理可减轻大鼠HIRI,其作用机制可能与其抑制自噬有关。     

Dynamic changes of autophagy during hypertrophic scar formation and the role of autophagy intervention

BackgroundThe role of autophagy in the formation of hypertrophic scars (HS) remains unclear. This study aimed to explore the role and potential mechanism of autophagy during the development of HS.MethodsRNA and protein expression levels of Beclin-1, p62, and LC3II in normal skin tissues and HS specimens from different patients were examined. Autophagy inducers and inhibitors were used to cure established HS in rabbit ears, and the expression of Beclin-1, p62, and LC3II at the RNA and protein level was determined. Lastly, the effects of autophagy inducers and inhibitors on HS development were analyzed.ResultsCompared to normal skin tissues, the expression of LC3Ⅱ and Beclin-1 was higher (P<0.05), while that of p62 was lower (P<0.05) in HS tissues. In addition, the LC3II/LC3I ratio was increased during HS formation, and the altered expression of the three proteins stabilized after one year. Administration of autophagy inducers enhanced the formation of HS as well as the expression levels of LC3II and Beclin-1 but decreased p62 expression. Meanwhile, administration of autophagy inhibitors increased the expression of LC3II, Beclin-1, and p62, along with reduced HS formation.ConclusionAutophagic activity increased during HS initiation and subsequent stabilization. In addition, autophagy inhibitors were able to inhibit HS formation by suppressing autophagy, whereas autophagy inducers promoted scar hyperplasia by enhancing autophagy.     

Abnormal hepatic expression of fibrillin-1 in children with cholestasis

Fibrillin-1, one of the main constituents of microfibrils, is present in normal adult liver and overexpressed in fibrotic area around cirrhotic nodules and hepatocellular carcinoma. In this work fibrillin-1 expression was studied by immunohistochemistry in liver samples from children with various cholestatic diseases corresponding to paucity of intrahepatic bile ducts, biliary atresia, congenital hepatic fibrosis, Byler's disease, mitochondrial cytopathy, sclerosing cholangitis, or choledochal cyst. As controls, histologically normal liver samples were used. In control liver, as in adult, fibrillin-1 was expressed in vessel walls, sinusoids, and portal connective tissue, particularly at the interface with the limiting hepatocytic plate and close to the basement membrane of bile ducts. In paucity of intrahepatic bile ducts without fibrosis, the fibrillin-1 distribution was similar to controls. In cholestatic diseases associated with severe fibrosis, such as biliary atresia, congenital hepatic fibrosis, Byler's disease, mitochondrial cytopathy, or sclerosing cholangitis, an enhanced deposition of fibrillin-1 was observed in portal connective tissue and fibrous septa. The strong fibrillin-1 expression close to the basement membrane of biliary structures was lost in cholestatic diseases, except biliary atresia. Finally, in normal and pathologic tissues, fibrillin-1 was co-localized with its putative receptor alphaVbeta3 in sinusoids but not around biliary structures.