小柴胡汤防治肝病机制研究进展

小柴胡汤是<伤寒论>中著名方剂.许多实验研究和临床实践已经表明,他可以通过多个途径阻断由肝炎至肝纤维化再至肝癌的发展过程,对肝病防治具有重要意义.小柴胡汤防治肝病的机制包括抑制肝炎病毒的复制、抗肝炎病毒活性、保护肝细胞膜、防治肝损伤、多途径抑制肝纤维化、免疫调节和抗肿瘤作用等.     

小柴胡汤对大鼠胰腺纤维化的治疗作用

背景：目前尚无防治胰腺纤维化的有效方法，日本曾报道小柴胡汤对胰腺星状细胞（PSCs）活性有一定抑制作用。从而可能抑制胰腺纤维化。目的：探讨中药小柴胡汤抑制大鼠胰腺纤维化的作用机制，探索慢性胰腺炎的治疗方法。方法：予雄性Sprague—Dawley大鼠胰管内注射含2％三硝基苯磺酸（TNBS）的10％乙醇PBS以诱发慢性胰腺炎，治疗组每日喂食50g/L小柴胡汤（0．8g／ks），持续8周，其余各组不作处理。分别于2、4、6、8周时处死大鼠，比较各组大鼠胰腺的组织病理学改变。检测胰腺组织内超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH—Px）活性、丙二醛（MDA）含量以及α-平滑肌肌动蛋白（SMA）、结蛋白（desmin）、胶原（Col）Ⅰ、ColⅢ、转化生长因子（TGF）-β1和纤维连接蛋白（FN）的表达。结果：TNBS诱导4周后，治疗组胰腺纤维化程度和腺体破环程度均显著低于非治疗组（P〈0．001），治疗时间越长，作用越明显；同时胰腺组织内SOD、GSH．Px活性显著高于、MDA含量显著低于非治疗组，α-SMA、结蛋白、ColⅠ、ColⅢ、TGF—β1和FN的表达亦较非治疗组有不同程度的减少。结论：早期应用小柴胡汤并不能完全抑制胰腺纤维化的发生，但在一定程度上抑制了纤维化的进展，4周后疗效显著。小柴胡汤治疗TNBS诱导的大鼠慢性胰腺炎的机制可能为抗氧化、抑制PSCs增生、活化和减少TGF—β1的分泌。     

化积汤对肝星状细胞增殖与凋亡的影响

体外培养肝星状细胞株HSC-T6,MTT法检测细胞增殖,流式细胞仪检测细胞凋亡的改变。发现化积扬可呈剂量依赖性抑制肝星状细胞增殖;化积汤培养后。细胞周期分析发现S期细胞减少,G2／M期细胞显著增加。提示化积汤可以显著抑制肝星状细胞增殖并诱导其凋亡。     

小柴胡汤治疗肝纤维化的研究进展

肝纤维化的产生是多种类型细胞、氧化压力、细胞因子和生长因子等一系列复杂作用的结果。其中参与的细胞主要有:肝星状细胞(HSC)、肝窦内皮细胞(SEC)和枯否氏细胞(KC);生长因子有:转化生长因子-β_1(TGF-β_1)、胰岛素样生长因子-1(IGF-1)、血小板源性生长因子(PDGF)等;其他细胞因子有:肿瘤坏死因子(TNF)、一氧化氮(NO)、内皮素-1(ET-1)等。其中,KC和HSC的激活是肝纤维化发生过程     

激活素A对人肝星状细胞系LX-2细胞增殖的影响

目的观察激活素A对人肝星状细胞(hepatic stellate cell,HSC)系LX-2细胞增殖的影响。方法体外培养人HSC LX-2。设正常对照组和激活素A干预组,其中激活素A干预组分为9个浓度梯度,药物终浓度分别为1、2.5、5、10、20、40、80、160、320μg/L,采用CCK-8法检测激活素A对细胞增殖的影响;再选取25、50、100、200、300μg/L5个浓度分别作用24、48、72 h,观察激活素A对细胞增殖的影响。结果激活素A可刺激LX-2细胞增殖,1～2.5μg/L浓度范围内作用轻微(P>0.05),而80～320μg/L的激活素A作用明显增强(P<0.01),且在细胞指数生长期对刺激细胞增殖有时间浓度依赖性。结论激活素A可通过刺激HSC增殖,参与肝纤维化的形成。     

改良小柴胡汤对慢性肝病患者肝纤维化标志物的作用

应用改良小柴胡汤制剂对８１例慢性肝病患者进行治疗学研究．旨在观察其对血清肝纤维化标志物的影响。结果显示改良小柴胡汤治疗可使血清透明质酸（ＨＡ）和Ⅲ型前胶原肽（ＰⅢＰ）由治疗前的２４５．８９±１４８．４９ ｎｇ／ｍｌ和２６１．８９±１３４．４２μｇ／Ｌ分别降至１２５．４３±８５．４６ ｎｇ／ｍｌ和１４４ ０４±８３．９９μｇ／Ｌ（Ｐ＝０．０００），均优于对照组；治疗组的ＨＡ和ＰⅢＰ复常率分别为３４．３８％（２２／６４ ）和５０．６８％（３７／７３）。提示改良小柴胡汤制剂对肝纤维化过程具有一定的抑制作用，长期应用将有助于延缓肝炎肝硬变的发生。     

益气软肝汤对血吸虫病兔肝纤维化的防治作用

目的:观察益气软肝汤的抗肝纤维化作用。方法:用日本血吸虫尾蚴感染新西兰免,制成肝纤维化模型,病兔均予吡喹酮顿服杀虫治疗,继之中药组开始服用益气软肝汤治疗肝纤维化,并与γ-干扰素组及生理盐水组比较,观察活化的肝星状细胞(HSC)被α-SMA染色后的阳性表达情况,以及肝胶原沉积情况,并判断肝纤维化程度。结果:中药组、干扰素组的肝纤维化程度及α-SMA阳性细胞表达情况均显著轻于生理盐水组(P<0.05),3组的抗纤效果以干扰素组为最佳,中药组次之,生理盐水组无效(各组间比较P<0.05)。结论:益气软肝汤具有较好的抗血吸虫病肝纤维化作用。     

γ干扰素与小柴胡汤联合抗肝纤维化的研究

目的:寻找抗肝纤维化的有效方法。方法:用γ干扰素(IFN-γ)和小柴胡汤联合治疗慢性乙型肝炎患者60例,并与单纯应用IFN-γ治疗慢性乙型肝炎50例比较,观察治疗前后患者透明质酸(HA)、Ⅳ型胶原(Ⅳ-C)、层粘蛋白(LN)、肝功能等指标变化。结果:IFN-γ联合小柴胡汤组患者血清肝圩堆化指标、ALT、TBil下降明显,A/G比值上升显著,与治疗前、单用IFN-γ组比较有显著性差异。结论:IFN-γ和小柴胡汤联合应用可有效改善患者肝功能,减轻和抑制肝纤维化的发生。     

氧化苦参碱对大鼠肝星状细胞增殖的影响

目的探讨氧化苦参碱对大鼠肝星状细胞增殖的影响及其抗肝纤维化的机理.方法用链酶蛋白酶和胶原酶原位灌流消化正常大鼠肝脏,Nycodenz密度梯度离心分离肝星状细胞,以MTT比色法观察氧化苦参碱对肝星状细胞毒性作用,3H-TdR法观察氧化苦参碱对肝星状细胞增殖的效应.结果氧化苦参碱浓度＞10-5mol/1时对肝星状细胞增殖有抑制作用(P＜0.05).结论氧化苦参碱可抑制肝星状细胞的增殖,有抗肝纤维化的作用.     

酒精性肝病患者血浆白细胞介素-21水平及重组IL-21对LX-2肝星状细胞增殖和活化的影响

目的：分析酒精性肝病患者血浆白细胞介素-21（IL-21）水平及重组IL-21体外对LX-2肝星状细胞增殖和活化的影响。方法采用ELISA法检测17例酒精性肝炎、51例酒精性肝硬化患者和20例健康人血浆IL-21水平;体外培养LX-2肝星状细胞,以IL-21（1ng/ml或10ng/ml）处理24 h或48 h,检测LX-2细胞增殖及α-平滑肌肌动蛋白表达。结果与健康对照人群比,酒精性肝炎和酒精性肝硬化患者血浆IL-21水平均显著升高（P＆lt;0.05）,但酒精性肝炎和肝硬化患者之间无显著性差异,不同Child分级的肝硬化患者之间也无显著性差异;在IL-21作用24～48 h后,LX-2细胞增殖水平与对照组比无显著性差异,但α-平滑肌肌动蛋白表达均较对照组显著升高。结论血浆IL-21可能通过促进肝星状细胞活化参与了酒精性肝病患者肝纤维化的发生和发展。     

Xiaochaihu decoction for nonalcoholic fatty liver disease: A protocol for a systematic review and meta-analysis

Background:Nonalcoholic fatty liver disease (NAFLD) is a clinicopathologic syndrome characterized by excessive deposition of fat in hepatocytes except for alcohol and other specific hepatic factors. Xiaochaihu decoction (XD) has been widely used to treat NAFLD in China. However, there is no systematic review found. In order to evaluate the efficacy and safety of XD in the treatment of NAFLS, we need to conduct a meta-analysis and systematic evaluation.Methods:There are enrolled randomized controlled trials (RCTs) evaluating the effectiveness and safety of XD in the treatment of NAFLD. Data come mainly from 4 Chinese databases (CNKI, CBM, Wanfang, and VIP Database) and 4 English databases (Pubmed, Embase, Cochrane Library, and Web of science). The enrollment of RCTs is from the starting date of database establishment till September 30, 2021. The work such as selection of literature, data collection, quality evaluation of included literature, and assessment of publication bias will be conducted by 2 independent researchers. Meta-analysis will be performed by RevMan 5.0 software.Results:This study will provide high-quality evidence for the effectiveness and safety of XD in the treatment of NAFLD.Conclusion:The results of the study will help us determine whether XD can effectively treat NAFLD.Ethics and dissemination:This study does not require ethical approval. We will disseminate our findings by publishing results in a peer-reviewed journal.OSF registration number:DOI 10.17605/OSF.IO/A5XEM     

加味小柴胡汤对肝癌患者栓塞化疗后脂质过氧化反应的影响

[目的]探讨原发性肝癌患者介入栓塞化疗(TACE)导致肝损害等不良反应的过氧化损伤机制,观察加味小柴胡汤的抗氧化作用.[方法]将56例肝癌患者随机分2组(TACE后均行水化治疗),各28例.对照组口服肌苷、肝泰乐、胃复安、维生素B6等;观察组给予加味小柴胡汤.观测TACE前、后2、7、14、21、28 d肝功能及过氧化指标的动态变化.[结果]TACE后2 d肝功能异常加重,过氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性、谷胱甘肽(GSH)水平明显降低,丙二醛(MDA)水平增加,以对照组变化显著.观察组肝功能及氧化指标恢复较快,均优于对照组(P＜0.01).[结论]肝癌TACE引起的不良反应及肝损害,与化疗药诱致的脂质过氧化反应有关.加味小柴胡汤有明显的抗氧化作用.     

加味小柴胡汤对乙型肝炎病毒YMDD变异率的影响

[目的]探讨加味小柴胡汤对乙型肝炎病毒（HBV）-YMDD变异率的影响。[方法]120例患者随机分成2组,治疗组以加味小柴胡汤加拉米夫定、对照组单用拉米夫定治疗,分别观察其对乙肝病毒标志物的变化、肝功能以及对HBV-YMDD变异率的影响,观察期为1年。[结果]治疗组治疗后nBeng、HBeAb、HBV-DNA阴转率分别为54．3％、42．8％、40．0％,对照组分别为26．7％、17．0％、13．3％,2组比较,其差异有统计学意义（P〈0．05）;在对肝功能恢复方面,治疗组优于对照组,差异有统计学意义（P〈0.05）;治疗组治疗后的6个月和12个月HBV-YMDD变异率分别为3．38％、10．17％,均较对照组（13．11％、21．31％）有所降低,2组比较差异均有统计学意义（P〈0．05）。[结论]加味小柴胡汤具有一定的抗HBV作用,并可降低HBV-YMDD的变异率。     

KN-93, a specific inhibitor of CaMKⅡ inhibits human hepatic stellate cell proliferation in vitro

AIM: To investigate the effects of KN-93, a CaMKⅡ selective inhibitor on cell proliferation and the expression of p53 or p21 protein in human hepatic stellate cells. METHODS: Human hepatic stellate cells (LX-2) were incubated with various concentrations (0-50 mmol/L) of KN-93 or its inactive derivative, KN-92. Cell proliferation was measured by CCK-8 assay, and the expression of two cell cycle regulators, p53 and p21, was determined by SDS-PAGE and Western blotting. RESULTS: KN-93 (5-50 mmol/L) decreased the proliferation of human hepatic stellate cells in a dose- dependent manner from 81.76% (81.76% ± 2.58% vs 96.63% ± 2.69%, P < 0.05) to 27.15% (27.15% ± 2.86% vs 96.59% ± 2.44%, P < 0.01) after 24 h treatment. Incubation of 10 mmol/L KN-93 induced the cell growth reduction in a time-dependent manner from 78.27% at 8 h to 11.48% at 48 h. However, KN-92, an inactive derivative of KN-93, did not inhibit cell proliferation effectively. Moreover, analysis of cell cycle regulator expression revealed that KN-93 rather than KN-92 reduced the expression of p53 and p21.     

姜黄素对肝星状细胞增殖及分泌细胞外基质的影响

观察姜黄素对大鼠肝星状细胞 (HSC)增殖及分泌细胞外基质的影响。用不同浓度的姜黄素处理大鼠肝星状细胞 ;以MTT比色法测定细胞的增殖 ;放射免疫法测定透明质酸和层粘连蛋白 ;酶联免疫吸附法 (ELISA)测定Ⅰ型胶原的水平。姜黄素可剂量依赖性地抑制HSC增殖 ,不同程度抑制Ⅰ型胶原、透明质酸和层粘连蛋白分泌。姜黄素可显著抑制HSC的增殖及分泌细胞外基质。     

KN-93, a specific inhibitor of CaMK Ⅱ inhibits human hepatic stellate cell proliferation in vitro

AIM: To investigate the effects of KN-93, a CaMKⅡ selective inhibitor on cell proliferation and the expression of p53 or p21 protein in human hepatic stellate ceils.METHODS: Human hepatic stellate cells (LX-2) were incubated with various concentrations (0-50 μmol/L) of KN-93 or its inactive derivative, KN-92. Cell proliferation was measured by CCK-8 assay, and the expression of two cell cycle regulators, p53 and p21, was determined by SDS-PAGE and Western blotting.RESULTS: KN-93 (5-50 μmol/L) decreased the proliferation of human hepatic stellate cells in a dosedependent manner from 81.76% (81.76% + 2.58% vs 96.63% + 2.69%, P ＜ 0.05) to 27.15% (27.15% + 2.86% vs 96.59% + 2.44%, P ＜ 0.01) after 24 h treatment.Incubation of 10 μmol/L KN-93 induced the cell growth reduction in a time-dependent manner from 78.27% at 8 h to 11.48% at 48 h. However, KN-92, an inactive derivative of KN-93, did not inhibit cell proliferation effectively. Moreover, analysis of cell cycle regulator expression revealed that KN-93 rather than KN-92 reduced the expression of p53 and p21.CONCLUSION: KN-93 has potent inhibitory effect on proliferation of LX-2 cells by modulating the expression of two special cell cycle regulators, p53 and p21.     

间充质干细胞外泌体对体外肝星状细胞增殖和活化的影响

目的 探讨间充质干细胞来源的外泌体(MSCs-Exo)对人肝星状细胞系LX-2细胞增殖及活化的影响。方法 取MSCs-Exo,在透射电子显微镜下观察。取LX-2细胞,分为对照组、5 μg/L、10 μg/L和20 μg/L转化生长因子-β1(TGF-β1)处理组,采用RT-PCR和Western blot法检测细胞α-SMA和COL1A1 mRNA及其蛋白表达。取TGF-β1诱导活化的LX-2细胞,以50 μg/mL、100 μg/mL和200 μg/mL MSCs-Exo干预24 h,采用MTT法检测细胞增殖。结果 经电镜观察及粒径分析表明,实验所用MSCs-Exo符合外泌体的形态特征;实验所用MSCs-Exo阳性标志蛋白CD9和CD63阳性,而GM130蛋白阴性;经TGF-β1诱导活化的LX-2细胞α-SMA和COL1A1 mRNA水平及其蛋白表达均显著升高;当TGF-β1诱导浓度为20 μg/L时,细胞α-SMA和COL1A1 mRNA及其蛋白表达量最高;与对照组比,MSCs-Exo-100组和MSCs-Exo-200处理细胞增殖活力显著降低(均P<0.0001);50 μg/mL、100 μg/mL和200 μg/mL MSCs-Exo作用TGF-β1诱导活化的LX-2细胞α-SMA mRNA水平和COL1A1 mRNA水平较对照组显著降低(P<0.01),细胞α-SMA和COL1A1蛋白水平也较对照组显著降低(P<0.0001);MSCs-Exo-100和MSCs-Exo-200处理的LX-2细胞增殖活力较对照细胞显著降低(均P<0.0001)。结论 MSCs-Exo可以抑制TGF-β1诱导的LX-2细胞增殖和活化,其应用研究值得期待。     

Characteristics of the cell proliferation profile of activated rat hepatic stellate cells in vitro in contrast to their fibrogenesis activity

Purpose. Alterations in the kinetics of hepatic stellate cells (HSCs) after the cells are activated once have not been well documented. We investigated the characteristic profiles of cell proliferation of once-activated HSCs in contrast to the in fibrogenesis activity. Methods. HSCs from male Wistar rats were submitted to primary culture for 14 days and to secondary culture for 7 days. The potential for cell proliferation was evaluated by the number of the cells in G2/M phase, based on flow cytometric analysis of the cell cycle. The fibrogenesis activity was assessed by Northern blot analysis of the expression of type I and type III procollagen mRNA. Results. The number of HSCs in G2/M phase was maintained at a low level in primary culture after 6 days, while a significantly (P < 0.05) elevated number of HSCs in G2/M phase was observed on days 3 to 4. In secondary culture, the number of HSCs in G2/M phase was also consecutively maintained at a decreased level. By contrast, HSCs showed progressively increased type I and type III procollagen mRNA expression during the experimental periods of primary culture. Conclusions. These results clearly demonstrated consecutively decreased proliferative activity, evaluated by the potential for cell mitogenesis, in once-activated HSCs, in contrast to their progressively increased fibrogenesis activity. Received: April 12, 2000 / Accepted: December 22, 2000     

小柴胡片联合干扰素治疗慢性乙型肝炎肝纤维化的临床研究

[目的]观察小柴胡片联合干扰素（IFN）治疗慢性乙型肝炎（CHB）肝纤维化的临床疗效。[方法]选择CHB患者137例，随机分为2组，对照组（64例）用IFN 300万U隔日1次肌内注射，治疗组（73例）加用小柴胡片，疗程均为6个月，观察2组治疗前、后肝纤维化及影像学指标的变化。[结果]2组疗程结束后肝功能和肝纤维化指标均有明显改善，治疗组优于对照组，且能改善影像学指标（P〈0．05，〈0．01）。[结论]小柴胡片与IFN联合应用对CHB具有较好的临床疗效，在改善肝纤维化方面两药合用具有协同作用。