过氧化物酶体增殖物活化受体γ配体15d-PGJ2对肝星状细胞增殖活化的影响

目的观察过氧化物酶体增殖物活化受体γ(PPARγ)的天然配体15d-PGJ2对HSC增殖及活化的影响,以探讨PPARγ在HSC活化过程中的作用。方法采用MTT法和RT-PCR方法观察5μmol/L及10μmol/L 15d-PGJ2对体外培养的HSC自发活化及血小板衍生生长因子(PDGF)引起的HSC增殖及活化的影响。结果以5μmol/L 15d-PGJ2处理原代HSC 3 d后,可明显抑制HSC活化标志物α-平滑肌肌动蛋白的表达,而PPARγ的表达较未处理组明显增高(0.64±0.03对比0.09±0.01,t=36.0517,P<0.01);15d-PGJ2可剂量依赖性地抑制PDGF引起的HSC增殖;经5μmol/L和10μmol/L 15d-PGJ2预处理后再用PDGF干预,则PPARγ的表达较单用PDGF干预组明显增高(分别为0.03±0.02对比0.60±0.03,t=42.6616,P<0.01;以及0.03±0.02对比0.69±0.04,t=33.83,P<0.01),而HSC的活化指标α-平滑肌肌动蛋白、α1(I)型胶原及单核细胞趋化蛋白-1的表达则受抑制。结论激活PPARγ可调控HSC的促纤维化和促炎症作用,促进PPARγ的表达可能成为抗肝纤维化的新手段。     

金雀异黄酮对成骨细胞增殖的影响及其可能的分子生物学机制

目的观察金雀异黄酮（GS）对绝经期骨质疏松患者骨膜成骨细胞增殖的影响及其分子生物学机制。方法用噻唑蓝比色法和流式细胞术分别检测GS对成骨细胞增殖活力和细胞周期分布的影响；Western—blot技术分析GS对细胞周期蛋白D和E表达的影响。结果GS可显著提高成骨细胞增殖活力，与溶剂对照组比较，10^-8mol／L、10^-7mol／L和10^-6mol／L GS时，细胞增殖率分别为137％、155％和161％；提高S期和M／G2期细胞分布比例，并伴有G0／G1期细胞比例降低；它莫西芬可抑制10^-7mol／L和10^-6mol／L GS促成骨细胞增殖效应。Western—blot的检测结果显示，10^-8mol／L和10^-7mol／L GS对成骨细胞处理24h，提高细胞周期蛋白D和E蛋白质的表达（P〈0．05）。结论由于雌激素受体拮抗剂它莫西芬可抑制GS对成骨细胞的促增殖作用，表明金雀异黄酮可通过激活雌激素受体途径，表现出雌激素效应，并促进骨组织代谢中的骨形成作用。     

牛磺酸通过调控细胞周期蛋白抑制肝星状细胞增殖

目的进一步研究牛磺酸对肝星状细胞(HSC)增殖抑制作用的机制。方法用四甲基偶氮唑盐法检测细胞增殖;流式细胞仪测定细胞周期;免疫细胞化学和实时荧光定量PCR测定细胞周期调控蛋白Cyclin D1和P21waf1表达。结果牛磺酸对HSC增殖具有抑制作用,在浓度为5、10、20,30、40、50 mmol/L 作用48h时的抑制率分别为6.7%、14.4%、23.3%、32.2%、36.7%和45.6%,t值为2.939～6.369,P<0.05～0.01。流式细胞仪检测发现牛磺酸可阻滞HSC由G0/G1期向S期转换,使G0/G1期细胞增多,S期细胞减少。G0/G1期、S期细胞,牛磺酸浓度为40 mmol/L时,分别为(68.2±1.4)%和(26.2±1.3)%,与对照组分别为(56.2±1.7)%和(38.5±0.8)%,差异有统计学意义,t≥5.422,P<0.01。牛磺酸可抑制Cyclin D1表达、促进P21waf1表达,用免疫细胞化学染色结合数码图像分析系统软件分析发现牛磺酸浓度在40 mmol/L时HSC的Cyclin D1表达的平均吸光度为0.13±0.02,P21waf1为0.19±0.02,对照组分别为0.18±0.02和0.14±0.01,差异有统计学意义,t=6.689和t=6.528,P<0.01。实时荧光定量PCR检测也发现经40 mmol/L牛磺酸处理的HSC的Cyclin D1 mRNA表达量(拷贝数与106磷酸甘油醛脱氢酶比值)降低为5776.7±3345.0,对照组为18 400.6±1374.8,而P21waf1 mRNA表达量(拷贝/106磷酸甘油醛脱氢酶)增多为44 866.7±3910.7,对照组为16 933.3±960.9。结论牛磺酸通过抑制Cyclin D1表达、促进P21waf1表达,使HSC阻滞于G0/G1期,而抑制HSC增殖。     

大麻素受体2介导的N-花生四烯酸氨基乙醇对肝星状细胞增殖和活化的影响

目的 观察内源性大麻素N-花生四烯酸氨基乙醇(AEA)及大麻素受体(CBR)2对肝星状细胞(HSC)增殖活化的影响,以探讨内源性大麻素及其受体系统在肝纤维化发展中的作用.方法 采用免疫荧光观察血小板衍生生长因子(PDGF)刺激前后HSC中CBR1和CBR2的表达.Western blot、PCR法观察不同浓度AEA及CBR2拮抗剂AM630对PDGF刺激下HSC增殖及活化的影响,同时用四甲基偶氮唑盐、流式细胞仪分析AEA对HSC活力及凋亡的影响.结果 HSC中CBR2的表达较CBR1高(F=116.797,P<0.01),且PDGF刺激后CBR2的表达明显增强(F=7.878,P<0.05).AEA可剂量依赖地抑制HSC的增殖,在浓度为10,20、50μmol/L时抑制率分别为7.12%±0.34%、12.52%±0.78%、80.13%±1.57%,差异有统计学意义(F=533.41,P<0.01);但对HSC凋亡的影响不明显.同时AEA可抑制HSC的活化指标α-平滑肌肌动蛋白、转化生长因子β1、Ⅰ型胶原、Ⅲ型胶原及基质金属蛋白酶抑制因子等的表达,但这种抑制作用在给予CBR2拮抗剂AM630后明显减弱,差异有统计学意义(P<0.05).结论 CBR2在AEA引起的HSC增殖及活化抑制中起关键作用,AEA和CBR2可望成为肝纤维治疗的新靶点.     

Blocking effects of genistein on cell proliferation and possible mechanism in human gastric carcinoma

AIM: To study the blocking effects of genistein on cell proliferation cycle in human gastric carcinoma cells (SGC-7901) and the possible mechanism. METHODS: MTT assay was applied in the detection of the inhibitory effects of genistein on cell proliferation. Flow cytometry was used to analyze the cell cycle distribution. Immunocytochemical technique and Western blotting were performed to detect the protein expression of cyclin D_1, cyclin B_1 and p21~(waf1/cip1). RESULTS: Genistein significantly inhibited the growth and proliferation of human gastric carcinoma cells (SGC-7901). Seven days after treatment with different concentrations of genistein (2.5, 5.0, 10.0, 20.0 μg/mL), the growth inhibitory rates were 11.2%, 28.8%, 55.3%, 84.7% respectively and cell cycles were arrested at the G(2)/ M phase. Genistein decreased cyclin D_1 protein expression and enhanced cyclin B_1 and p21~(waf1/cip1) protein expression in a concentration-dependent manner. CONCLUSION: The growth and proliferation of SGC-7901 cells can be inhibited by genistein via blocking the cell cycle, with reduced expression of cyclin D_1 and enhanced expression of cyclin B_1 and p21~(waf1/cip1) protein in the concentration range of 0-20 μg/mL.     

转化生长因子β1对不同活化状态肝星状细胞增殖与Ⅰ型胶原表达的影响

目的 观察不同活化状态肝星状细胞(HSC)对外源性转化生长因子-β_1(TGF-β_1)旁分泌刺激的生物学效应作用。方法 原代分离培养大鼠HSC,无包被塑料培养皿上分别培养1、4、7d,细胞处于静止、中间活化与完全活化状态,继以10～500 pmol/L TGF-β_1温育细胞24h,~3H—TdR掺入法测定细胞增殖,western blot法检测细胞α-平滑肌肌动蛋白(α-SMA)与Ⅰ型胶原蛋白表达沉积,~3H-脯氨酸掺入与胶原酶消化法测定细胞总胶原的分泌量。100pmol/L TGF-β_1温育细胞15～90min,northern blot法检测细胞Ⅰ型前胶原mRNA的表达水平。结果 TGF-β_1浓度依赖性抑制培养1d HSC的细胞增殖,10～500 pmol/L TGF-β_1浓度组细胞内~3H—TdR掺入率分别为对照组的52.8％～16.8％,与对照组比较,q值为5.44～10.37,P<0.01。但TGF-β_1对培养4d与7d的细胞增殖无影响。随细胞活化,HSC基础性α-SMA、Ⅰ型胶原蛋白与mRNA水平明显增加,而TGF-β_1刺激各培养时间HSC以上蛋白与基因的表达。培养1、4、7d HSC基础水平与TGF-β_1刺激的总胶原分泌量分别为(804±274)dpm/孔与(1 200±708)dpm/孔;(2 966±1 701)dpm/孔与(6 160±1 123)dpm/孔;(2 580±767)dpm/孔与(4 583±1 467)dpm/孔,后2组组内比较,t值分别为3.84与2.96,P<0.01或P<0.05。以培养4d HSC     

4′,5,7-三羟基异黄酮抗骨髓瘤细胞增殖机制的研究

目的：为了研究4′,5,7-三羟基异黄酮（genistein,Gen）是否能抑制人多发性骨髓瘤（multipule myeloma,MM）细胞株XG1的增殖,研究Gen处理骨髓瘤细胞后B细胞淋巴瘤/白血病-2基因（bcl-2）、bcl-xl、细胞周期蛋白D1（cyclin D1）、细胞间黏附因子1（ICMA-1）基因表达变化及骨髓瘤细胞中核转录因子κB（NF-κB）表达的变化。方法：利用体外培养人MM细胞株XG1,依据剂量梯度分别给予Gen,用噻唑蓝染色法（MTT）观察不同药物浓度的Gen对MM细胞的增殖影响;5、10、15μg/mLGen与溶剂对照组分别作用XG1细胞48h后,半定量逆转录聚合酶链反应（RT-PCR）法检测XG1细胞bcl-2、bcl-xl、细胞周期蛋白D1、ICMA-1基因表达;应用5μg/mLGen处理XG1细胞,用免疫组织化学法观察Gen处理前后细胞内NF-κB的表达情况。结果：Gen作用XG1细胞48h,随浓度增加,细胞增殖逐渐下降;5、10、15μg/mLGen处理组mRNA的表达均低于溶剂对照组（P〈0.05）,伴随Gen处理浓度逐渐增加,bcl-2、bcl-xl、细胞周期蛋白D1、ICMA-1基因mRNA的表达逐渐下降;Gen能够使NF-κB在XG1细胞内重新分布,胞浆内出现NF-κB表达,胞核内NF-κB表达下降。结论：Gen可能通过下调骨髓瘤细胞核中NF-κB的表达来抑制bcl-2、bcl-xl、细胞周期蛋白D1、ICMA-1基因mRNA的表达,进而抑制骨髓瘤细胞的增殖、黏附、转移。     

Suppressive effects of 171β-estradiol on hepatic fibrosis in CCI4-induced rat model

AIM: To investigate the pathway via which 17β-estradio(β-Est) exerts suppressive effects on rat hepatic fibrosis.METHODS: In vivo study was done in CCI4-induced female hepatofibrotic rats. Fibrosis-suppressive effect of β-Est(20 μg/kg&#183;d) was evaluated in intact and ovariectomized rat models. Six weeks after the treatment, all the rats were sacrificed and specimens of serum or liver tissue were collected for the studies. Serum liver enzymes,fibrosis markers and estradiol levels were determined by standard enzymatic methods, ELISA and RIA, respectively.Degrees of fibrosis and areas of hepatic stellate cells(HSCs) positive for alpha-smooth muscle actin (a-SMA) in the liver were determined by van Gieson (VG) stain and immunohistochemistry. In vitro studies, HSCs were isolated by a combination of pronase-collagenase perfusion and density gradient centrifugation. First-passage HSCs were randomly divided into 10 groups, and different concentrations of β-Est, 2-hydroxyestradiol (2OHE) or 2-methoxyestradiol(2MeOE) were separately added to the cell groups. After incubation for 72 h, the degree of cell proliferation, collagen production, ~-SMA or estrogen receptor (ER) expression was determined by MTT assay, ELISA and immunohistochemistry,respectively.RESULTS: β-Est treatment reduced aspartate aminotransfer-ase (AST), alanine aminotransferase (ALT), hyaluronic acid(HA) and type IV collagen (C IV) in sera, suppressed hepatic collagen content, decreased the areas of HSCs positive for α-SMA significantly in both intact and ovariectomized female hepatofibrotic rats. There was a negative correlation between the percentage of fibrotic area of liver tissue and the serum estradiol level; the calculated correlation coefficient was -0.57 (P&lt;0.01). β-Est and its metabolites concentration-dependently (10^-9 mol/L-10^-7 mol/L) inhibited HSC proliferation and collagen synthesis. At the concentration of 10^-7 mol/L, they could inhibit α-SMA expression. The order of potency was 2MeOE&gt;2OHE&gt;β-Est.CONCLUSION: β-Est may suppress hepatic fibrosis probably via its biologically active metabolites.     

二丙酸倍氯米松和布地奈德对白细胞介素13激活的肺成纤维细胞的影响

目的观察吸入型糖皮质激素二丙酸倍氯米松（BDP）和布地奈德（BUD）对白细胞介素13（IL-13）激活的肺成纤维细胞的影响。方法IL-13加入培养的肺成纤维细胞，采用噻唑蓝法、免疫组织化学法、免疫印迹法、逆转录聚合酶链反应、ELISA等方法检测细胞增殖、α-肌动蛋白（α-SMA）表达及分泌白细胞介素6（IL-6）、嗜酸粒细胞趋化因子（eotaxin）的变化，观察BDP和BUD的作用。结果加入20ng／ml的IL-13后，成纤维细胞分泌IL-6明显增加[（20±2）ng／L和（140±8）ng／L]，eotaxin也明显增加[（64±25）ng／L和（334±51）ng／L]。10^-8mol／L～10^-5mol／L的BDP组和BUD组IL-6表达水平[（112±3）ng／L～（53±2）ng／L和（55±14）ng／L～（32±6）ng／L]比IL-13组明显下降，IL-6 mRNA表达比IL-13组明显减少，细胞上清液中eotaxin水平[（297±59）～（226±43）ng／L和（287±59）～（183±43）ng／L]比IL-13组明显降低。IL-13诱导成纤维细胞表达α-SMA mRNA及蛋白明显增加，并促进成纤维细胞增殖（1．5±0．2）倍。BDP组和BUD组对IL-13诱导的α-SMA mRNA及蛋白的表达无显著影响，并明显具有协同IL-13促进成纤维细胞增殖的作用。结论BDP和BUD对IL-13刺激成纤维细胞的作用具有多重调节效应。BDP和BUD抑制IL-13诱导成纤维细胞合成、释放炎症介质IL-6和eotaxin的作用，有利于改善气道上皮下纤维化，但对于IL-13促进成纤维细胞转化为肌成纤维细胞及其增殖方面起负面影响。     

地尔硫卓对增殖血管平滑肌细胞的原癌基因表达的影响

目的观察地尔硫卓（Dil）对增殖血管平滑肌细胞（VSMC）的原癌基因c-myc、c-fos、c-jun和ras mRNA表达的影响。方法将组织贴块法培养的大鼠胸主动脉VSMC随机分为5组,即空白组、模型组和Dil1、2、3组（浓度分别为10^-5、10^-6、10^-7mol/L）,应用MTT检测增殖能力,用流式细胞术检测VSMC的增殖指数,用rt-PCR检测c-myc、c-fos、c-jun和ras mRNA的表达。结果与模型组比较各浓度Dil都能抑制VSMC增殖,PI值均显著下降（各组PI值分别为21.53±1.72、28.63±0.96、15.95±0.37、19.28±0.94、20.33±0.67;P〈0.05）;Dil1、2组c-myc、c-fos、c-jun mRNA的表达显著减少（模型组和Dil1、2组的c-myc/GAPDH值分别为2.454±0.03、1.509±0.05、1.660±0.04,c-fos/GAPDH值分别为0.0046±0.0004、0.0023±0.0003、0.0038±0.0005,c-jun/GAPDH值分别为1.950±0.03、1.077±0.03、1.725±0.03;P〈0.05）,rasmRNA的表达显著增加（模型组和Dil1、2组的ras/GAPDH值分别为1.941±0.03、3.811±0.02、2.501±0.02;P〈0.05）。结论Dil抑制VSMC增殖机制可能与下调原癌基因c-myc、c-fos、c-jun的表达和上调ras的表达有关。     

生长因子对HSC-T6细胞c-fos和c-jun基因表达的影响

目的 了解血小板衍生生长因子(PDGF)和转化生长因子β(TGF β)分别对肝星状细胞c～fos和c-jun基因表达的影响.方法 在培养的肝星状细胞系HSC-T16细胞中分别加入不同浓度的PDGF(终浓度分别为8、40、200 ng/ml)和TGF β(终浓度分别为0.2,1.0、5.0 ng/ml);于8、24,48、72h四个时间点分别收集细胞,提取细胞总RNA;用逆转录定量PCR法测定c～los和c-jun的基因表达水平.结果 PDGF处理的3组HSC-T6细胞8、24、48、72h时,c-fos基因表达水平均明显高于对照组,并呈剂量依赖性l培养8 h时达到表达最高峰,对照组,PDGF 8 ng/ml组,PDGF 40 ng/ml组,PDGF 200ng/ml组c-fos表达分别为0.63±0.13,1.13±0.19、1.75±0.20、2.40±0.23,3组间差异有统计学意义(F=7.03,P<0.01).TGF β处理的3组HSC-T6细胞8、24,48、72 h时,c-iun基因表达水平均明显高于对照组,并呈剂量依赖性,培养8 h时达到表达最高峰,对照组,TGF β 0.2ng/ml组、TGF β 1.0ng/ml组、TGF β 5.0ng/ml组cjun基因表达分别为0.93±0.13、1.69±0.26、2.34±0.30、2.96士0.37; 3组间差异有统计学意义(F=6.34,P<0.01).结论 PDGF和TGF β分别对肝星状细胞c-los和c-jun基因表达有明显的上调作用.     

细胞外信号调节激酶信号通路对乙醛刺激的大鼠肝星状细胞周期的影响

目的 观察特异性丝裂原细胞外信号反应激酶1(MEK 1)阻断剂(PD98059)对乙醛刺激的大鼠肝星状细胞(HSC)增殖及细胞周期的影响,并探讨其作用机制。 方法 用不同浓度的PD98059对乙醛刺激的HSC进行处理;以四甲基偶氮唑蓝法检测细胞增殖,流式细胞仪检测细胞周期,逆转录聚合酶链反应方法检测HSC内细胞周期蛋白-D1(Cyclin D1)mRNA和细胞周期蛋白依赖性激酶(CDK4)mRNA的表达。 结果 20、50、100μmol/L的PD98059均能显著且剂量依赖性地抑制乙醛刺激的HSC增殖,3组A值分别为0.109±0.020、0.081±0.010、0.056±0.020,与乙醛组A值0.146±0.030相比较,F=31.385,P<0.05;20、50、100 μmol/L的PD98059可显著抑制乙醛刺激的HSC由G1期进入S期,G0/G1期细胞百分比逐渐升高,3组G0/G1期细胞百分比分别为(61.9±6.3)％、(64.1±3.3)％、(70.9±4.8)％,与乙醛组(55.2±4.4)％相比较,F=16.402,P<0.05;50、100μmol/L的PD98059能显著抑制乙醛刺激的HSC内Cylin D1 mRNA表达,2组平均光强度比值分别为0.56±0.04,0.46±0.03,与乙醛组0.65±0.07相比较,F=68.758,P<0.05;50、100μmol/L的PD98059能显著抑制乙醛刺激的HSC内CDK4 mRNA表达,2组平均光强度比值分别为0.39±0.07,0.33±0.05,与乙醛组0.50±0.06相比较,F=29.406,P<0.05。 结     

亮氨酸脑啡肽抑制血管平滑肌细胞增殖和c-fos基因表达

为探讨脑啡肽与去甲肾上腺素一起参与血管运动的局部调节机制，用MTT法、氚标胸腺嘧啶脱氧核苷掺入法和斑点杂交技术发现亮氨酸脑啡肽可抑制SD大鼠胸主动脉平滑肌细胞的增殖、DNA合成和c-fos基因的表达；阿片受体阻断剂纳洛酮无上述抑制作用，但可拮抗亮氨酸脑啡肽的上述抑制作用；上述作用还可被去甲肾上腺素所拮抗。结果提示，脑啡肽通过影响血管平滑肌细胞功能来参与血管运动的局部调节，这种调节可能是通过阿片受体介导的。     

Intracellular pathways mediating Na+/H+ exchange activation by platelet-derived growth factor in rat hepatic stellate cells

BACKGROUND & AIMS: The Na+/H+ exchanger is the main intracellular pH regulator in hepatic stellate cells (HSCs), and its activity is increased by platelet-derived growth factor (PDGF). Amiloride, an Na+/H+ exchange inhibitor, reduces PDGF-induced HSC proliferation, suggesting that the Na+/H+ exchanger plays a role in regulating HSC proliferative response. The aim of this study was to characterize the intracellular pathways mediating activation of the Na+/H+ exchanger by PDGF in HSCs. METHODS: The activity of the Na+/H+ exchanger and HSC proliferation rate were evaluated under control condition and after incubation with PDGF in the absence or presence of specific inhibitors of the main intracellular pathways of signal transduction. Na+/H+ exchange protein expression was evaluated by means of Western blot. RESULTS: PDGF induced a significant increase in the activity of the Na+/H+ exchanger without modifying protein expression. Inhibition of the calcium/calmodulin- and protein kinase C-dependent pathways resulted in a significant inhibition of both Na+/H+ exchange activity and of PDGF-induced HSC proliferation. The involvement of the two pathways was confirmed by showing that incubation of HSCs with both phorbol-12-myristate-13-acetate, a potent protein kinase C activator, and thapsigargin, which increases intracellular calcium levels, significantly increased both the Na+/H+ exchanger activity and HSC proliferation rate. Inhibition of the protein kinase A pathway did not modify either PDGF-induced Na+/H+ exchange activation or PDGF-induced HSC proliferation. On the contrary, inhibition of the mitogen-activated protein kinase- and of phosphatidylinositol 3-kinase-dependent pathways significantly reduced PDGF-induced HSC proliferation without affecting the activity of the Na+/H+ exchanger. CONCLUSIONS: Activation of the Na+/H+ exchanger by PDGF in HSCs is mediated by calcium/calmodulin- and protein kinase C-dependent pathways. PDGF-induced HSC proliferation is mediated by Na+/H+ exchange-dependent and -independent pathways.     

Selective Na+/H+ exchange inhibition by cariporide reduces liver fibrosis in the rat

The aim of this study was to evaluate the effect of cariporide, a selective Na(+)/H(+) exchange inhibitor, on isolated and cultured hepatic stellate cells (HSCs) and in 2 in vivo models of rat liver fibrosis. Platelet-derived growth factor (PDGF)-induced HSC proliferation, evaluated by measuring the percentage of bromodeoxyuridine-positive cells, was significantly inhibited by cariporide, with a maximal effect at 10 micromol/L. Incubation with cariporide did not inhibit PDGF-induced extracellular-regulated kinase 1/2 (ERK1/2), Akt (a downstream component of the phosphatidylinositol [PI]-3 kinase pathway), and protein kinase C (PKC) activation but reduced PDGF-induced activation of the Na(+)/H(+) exchanger, with a maximal effect at 10 micromol/L. Rats treated with dimethylnitrosamine (DMN; 10 mg/kg) for 1 and 5 weeks received a diet with or without 6 ppm cariporide. Treatment with cariporide reduced the degree of liver injury, as determined by alanine aminotransferase (ALT) values, also when administered after the induction of hepatic damage. This was associated with reduced HSC activation and proliferation and reduced collagen deposition, as determined by morphometric evaluation of alpha-smooth muscle actin (SMA)/proliferating cell nuclear antigen-positive cells and percentage of Sirius red-positive parenchyma, respectively. Moreover, cariporide was also able to reduce alpha(1)I procollagen messenger RNA (mRNA) expression. Similar effects were observed in bile duct-ligated (BDL) rats. In conclusion, selective inhibition of the Na(+)/H(+) exchanger by cariporide may represent an effective therapeutic strategy in the treatment of hepatic fibrosis.     

Effects of Yigan Decoction on proliferation and apoptosis of hepatic stellate cells

AIM：To investigate the effects of Chinese herb Yigan Decoction on proliferation and apoptosis of the hepatic stellate cells(HSC) in vitro .METHODS:The study in vitro was carried out in the Culture of HSC lines.Various comcentration of Yigan Decoction were added and incubated .Cell proliferation was detected with MTT colorimetric assay.Cell apoptosis was detected by electron microscopy,flow cytometry and TUNEL.RESULTS:The proliferation of HSC was inhibited by Yigan Decoction,which depending on dose and time significantly.The HSC proliferation rates of groups at the end concentrations 144 and 72(g.L^-1) were 21.62%and 40.54% respectively,significantly lower than that of normal control group(P&lt;0.01),The HSC proliferation rates of groups at the and concentrations 36,18 and 9(g.L^-1) were 54.05%,45.95%and 51\35% respectively,lower than that of control group(P&lt;0.05).When the end concentration was 4.5g.L^-1,the proliferation rate was 83.78%,which appeared no significant differences compared with control group.At the same concentrations of 18 g.L^-1,the inhibitory effects of Yigan Decoction at 24h,48h and 72h time point were observed,the effects were time-dependent and reached a peak at 72h.Meanwhile,it was showed that the inducing effects of Yigan Decoction on HSC apoptosis were dose dependent and time-dependent,The apoptosis index(AL)was detected by TUNEL.After Yigan Decoction had been incubated for 48h at the end concentration of 18g.L^1,the Al(14.5&#177;31.%),was significantly higher than that of control group(4.3&#177;1.3)^(P&lt;0.01).When visualized under transmission electron microscopy,some apoptotic stellate cells were found ,i.e.dilated endoplasmic reticulum,irregular nuclei,chromatin condensation and heterochromatin ranked along inside of nuclear membrane.By flow cytometry detection,after HSC was treated with Yigan Decoction at different concentrations of 36,18and 9(g.L^-1)for48h,Al(%)were 13.3&#177;3.2,10.7&#177;2.7and 10.1&#177;2.5respectively,which were significantly higher than that of control group(4.1&#177;1.9)(P&lt;0.01),At the same concentration of 18g.L^-1 or 24h,48h and 72h,Al(%)were9.3&#177;1.8,10.7&#177;2.7and 14.6&#177;4.3repectively,which were significantly higher than that of control group(P&lt;0.01).CONCLUSION:Yigan Decoction could significantly inhibit HSC proliferation and increase the apoptosis index of HSC dose-dependently and time-dependently,which may be related to its mechanism of antifibrosis.     

Dynamic expression of extracellular signal-regulated kinase in rat liver tissue during hepatic fibrogenesis

INTRODUCTION Mitogen-activated protein kinase (MAPK) pathway is an important intracellular signal transduction system[1], and extracellular signal-regulated kinase 1 (ERK1) is the critical and classical pathway of MAPK and plays an important role in sever…