Restoring the Treg cell to Th17 cell ratio may alleviate HBV-related acute-on-chronic liver failure

AIM: To investigate the role of T helper 17 cells (Th17) and regulatory T cells (Treg) in hepatitis B virus (HBV)-related acute-on-chronic liver failure (ACLF).METHODS: We enrolled 79 patients with HBV infection into the study, 50 patients with HBV-related ACLF and 29 patients with chronic hepatitis B (CHB), from the First Affiliated Hospital of Medical College from January 2009 to June 2012. The ACLF patients were diagnosed according to the criteria recommended by The 19th Conference of the Asian Pacific Association for the Study of the Liver in 2009. Twenty healthy individuals with a similar gender and age structures to the two patient groups were also included as the normal controls (NC). Of the 50 ACLF patients, 28 were subsequently classified as non-survivors: 19 patients died from multiorgan failure, 3 underwent liver transplantation, and 6 discontinued therapy during follow-up because of financial reasons. The remaining 22 ACLF patients whose liver and anticoagulation function recovered to nearly normal levels within the next 6 mo were classified as survivors. The number of circulating Treg and Th17 cells was determined upon diagnosis and during the 8th week of follow-up through flow cytometry. RESULTS: The percentage of circulating Treg cells in the ACLF group was significantly higher than that in the CHB group (5.50% ± 1.15% vs 3.30% ± 1.13%, P < 0.01). The percentages of circulating Th17 cells in the ACLF and the CHB groups were significantly higher than that in the NC group (6.32% ± 2.22% vs 1.56% ± 0.44%, P < 0.01; 3.53% ± 1.65% vs 1.56% ± 0.44%, P < 0.01). No significant difference in Treg cell to Th17 cell ratio was observed between the ACLF group and the CHB group (0.98 ± 0.44 vs 1.12 ± 0.64, P = 0.991), whereas those in the two HBV infection groups were significantly lower than that in the NC group (1.85 ± 1.22; both P < 0.01). The percentage of Treg cells in the survivors during the 8th week of follow-up was significantly lower than that during peak ACLF severity [total bili     

Correlation between anti-fibrotic effect of baicalin and serum cytokines in rat hepatic fibrosis

AIM： To investigate the correlation between the antifibrotic effect of baicalin and serum cytokine production in rat hepatic fibrosis, METHODS： Forty male Sprague-Dawley rats were divided randomly into four groups： normal control group, model group, baicalin-treated group, and colchicine-treated group. Except for the normal control group, all rats in the other groups were administered with carbon tetrachloride to induce hepatic fibrosis. At the same time, the last two groups were also treated with baicalin or colchicine. At the end of the 8 wk, all animals were sacrificed. Serum alanine aminotransferase （ALl＇）, aspartate aminotransferase （AST）, transforming growth factor （TGF）-β1, tumor necrosis factor （TNF）-α, interleukin （IL）-6 and IL-10 were measured. Liver index, hepatic hydroxyproline content and the degree of liver fibrosis were also evaluated. RESULTS： The levels of ALT, AST and liver index in the baicalin-treated group were markedly lower than those in the model group （ALT： 143.88 ± 14.55 U/L vs 193.58± 24.35 U/L; AST： 263.66 ± 44.23 U/L vs 404.37± 68.29 U/L; liver index： 0.033 ± 0.005 vs 0.049± 0.009, P 〈 0.01）. Baicalin therapy also significantly attenuated the degree of hepatic fibrosis, collagen area and collagen area percentage in liver tissue （P 〈 0.01）. Furthermore, the levels of serum TGF-β1, TNF-α and IL-6 were strikingly reduced in the baicalin-treated group compared with the model group, while the production of IL-10 was up-regulated： （TGF-β1：260.21 ± 31.01 pg/mL vs 375.49 ± 57.47 pg/mL; TNF-α： 193.40±15.18 pg/mL vs 260.04 ± 37.70 pg/mL; IL-α：339.87 ± 72.95 pg/mL vs 606.47 ± 130.73 pg/mL; IL-10：506.22 ± 112.07 pg/mL vs 316.95 ± 62.74 pg/mL, P 〈 0.01）. CONCLUSION： Baicalin shows certain therapeutic effects on hepatic fibrosis, probably by immunoregulating the imbalance between profibrotic and antifibrotic cytokines.     

I_κB kinase-beta inhibitor attenuates hepatic fibrosis in mice

AIM: To investigate the anti-fibrosis effect of IκB kinase-beta inhibitor (IKK2 inhibitor IMD0354) in liver fibrosis. METHODS: Twenty male C57BL6 mice were divided into four groups. Five high-fat fed mice were injected with lipopolysaccharide (LPS, 10 mg/kg) intraperitoneally and five high-fat fed mice were without LPS injection to build models of liver injury, and the intervention group (five mice) was injected intraperitoneally with IKK2 inhibitor (IMD 30 mg/kg for 14 d), while the remaining five mice rec...     

肝脏前体细胞移植对CCl4诱导的肝纤维化自发逆转大鼠肝组织学的影响*

目的 研究肝脏前体细胞（HPCs）移植对纤维化自发逆转大鼠肝脏组织学的影响。方法 采用四氯化碳（CCl4）腹腔注射制备大鼠肝纤维化模型,取14只模型大鼠,随机分为自发逆转组7只和HPCs移植组7只,后者采用经脾内注射HPCs。取肝组织行苏木精-伊红和天狼猩红染色,观察组织病理学变化;采用免疫组织化学法检测肝组织OV6和α-平滑肌肌动蛋白（α-SMA）表达;采用Western blot法测定α-SMA、基质金属蛋白酶组织抑制剂（TIMP）-1蛋白表达;采用ELISA法测定血清I型胶原和白蛋白水平。结果 在CCl4腹腔注射8 w后动物肝组织OV6阳性细胞数量达到高峰,12 w时开始下降;与自发逆转组比,HPCs移植组肝组织纤维化和炎症程度减轻,胶原面积显著降低【（3.14±0.99）对 （5.05±0.89）,P<0.05】;自发逆转组肝组织α-SMA和TIMP-1蛋白相对表达量分别为（1.22±0.10）和（0.43±0.09）,均显著高于HPCs移植组的【（0.31±0.08）和（0.25±0.032）,P<0.05】;自发逆转组血清I型胶原水平为（41.33±8.08） ng/ml,显著高于HPCs移植组的【（31.12±10.08）ng/ml,P<0.05】,自发逆转组白蛋白为（33.18±4.69） g/L,显著低于HPCs移植组的【（48.30±2.15）g/L,P<0.05】。结论 肝脏前体细胞移植能加速肝纤维化的逆转,改善肝功能。     

慢性丙型肝炎和丙型肝炎肝硬化患者外周血Treg和Th17水平变化临床意义探讨*

目的 探讨外周血Treg和Th17细胞比率检测在评估HCV感染患者发生肝硬化的临床意义。方法 2017年3月~2019年1月我院收治的丙型肝炎肝硬化患者35例、慢性丙型肝炎（CHC）患者31例和同期接受健康体检者66名,采用ELISA法检测血清白细胞介素-6（IL-6）、IL-17和转化生长因子-β（TGF-β）水平,使用流式细胞仪检测外周血Treg和Th17细胞比率。绘制ROC曲线观察各指标预测肝硬化的发生。结果 丙型肝炎肝硬化患者血清IL-6、IL-17和TGF-β水平分别为（8.08±2.71） ng/L、（17.01±4.55） ng/L和（5.20±1.35） ng/L,显著高于健康人[分别为（6.64±2.33） ng/L、（12.76±3.46） ng/L和（4.19±1.07） ng/L,P<0.05]或CHC 患者[分别为（7.20±2.49） ng/L、（14.12±3.87） ng/L和（4.41±1.20） ng/L,P<0.05],血清高水平的TGF-β能预测肝硬化的发生（AUC=0.963,P=0.000）;肝硬化患者外周血Treg细胞比率为（6.27±1.02）%,显著高于CHC患者或健康人[分别为（5.75±0.84） %和（5.37±0.99） %,P<0.05],而Th17/Treg比值为（0.19±0.05）,显著低于CHC患者或健康人[分别为（0.23±0.06）和（0.22±0.05）,P<0.05];ROC曲线分析显示外周血Treg细胞比率或Th17/Treg细胞比值可有效预测肝硬化[AUC=0.820和AUC=0.607,P=0.000,P=0.041],而TH17细胞对肝硬化无预测价值。结论 丙型肝炎肝硬化患者血清TGF-β水平和外周血Treg细胞比率有明显的变化,监测它们的变化可能有助于在慢性丙型肝炎病毒感染者中早期发现肝硬化,做到早诊断、早治疗,对改善预后有重要的临床意义。     

Glycyrrhizic acid inhibits apoptosis and fibrosis in carbontetrachloride-induced rat liver injury

AIM:To investigate anti-apoptotic effects of glycyrrhizic acid(GA) against fibrosis in carbon tetrachloride(CCl4)-induced liver injury and its contributing factors.METHODS:Liver fibrosis was induced by administration of CCl4 for 8 wk.Pathological changes in the liver of rats were examined by hematoxylin-eosin staining.Collagen fibers were detected by Sirius red staining.Hepatocyte apoptosis was determined by TUNEL assay and the expression levels of cleaved caspase-3,Bax,α-SMA,connective tissue growth factor(CTGF),matrix metalloproteinase(MMP) 2 and MMP9 proteins were evaluated by western blot analysis,and α-SMA m RNA,collagen type Ⅰ and Ⅲ m RNA were estimated by real-time PCR.RESULTS:Treatment with GA significantly improved the pathological changes in the liver and markedly decreased the positive area of Sirius red compared with rats in the CCl4-treated group.TUNEL assay showed that GA significantly reduced the number of TUNEL-positive cells compared with the CCl4-treated group.The expression levels of cleaved caspase-3,Bax,α-SMA,CTGF,MMP2 and MMP9 proteins,and α-SMA m RNA,collagen type Ⅰ and Ⅲ m RNA were also significantly reduced by GA compared with the CCl4-treated group(P 0.05).CONCLUSION:GA treatment can ameliorate CCl4-induced liver fibrosis by inhibiting hepatocyte apoptosis and hepatic stellate cell activation.     

慢性丙型肝炎和肝硬化患者外周血Th17和Treg细胞及其相关细胞因子水平变化*

目的 调查慢性丙型肝炎（CHC）和丙型肝炎肝硬化（LC）患者外周血辅助性T淋巴细胞17（Th17）、调节性T淋巴细胞（Treg）及其相关细胞因子水平的变化。方法 2014年10月~2016年10月本院收治的CHC患者33例,丙型肝炎肝硬化患者27例和健康体检者30例。给予CHC患者α-干扰素联合利巴韦林(PR)方案治疗12 m。采用ELISA法检测白介素-6 （IL-6）、IL-10、IL-17和转化生长因子-β（TGF-β）,使用流式细胞仪检测外周血Th17和CD4⁺CD25⁺Foxp3⁺Treg细胞数。结果 LC和CHC患者外周血Th17和Treg细胞比率和Th17/Treg比值分别为（4.6±0.7）%、（2.7±0.5）%和（1.6±0.4）,和（5.8±0.9）%、（3.1±0.6）%和（1.9±0.5）,均显著高于健康人[（2.3±0.4）%、（1.7±0.4）%和（1.4±0.2）,P<0.05],LC患者外周血Th17和Treg细胞比率和Th17/Treg比值显著高于CHC患者 （P<0.05）;LC和CHC患者血清IL-6、IL-10、IL-17和TGF-β水平分别为（9.9±2.5）ng/L、（18.0±3.2） ng/L、（15.3±2.8） ng/L和（5.5±0.8） ng/L,和（14.1±3.0） ng/L、（21.4±4.0） ng/L、（23.3±3.9）ng/L和（7.2±1.2）ng/L,显著高于健康人[（6.7±1.1） ng/L、（14.2±2.9） ng/L、（5.3±0.6） ng/L和（4.3±1.0） ng/L,P<0.05],LC患者血清IL-6、IL-10、IL-17和TGF-β水平显著高于CHC患者（P<0.05）;CHC患者治疗后外周血Th17和Treg细胞比率及血清TGF-β、IL-10和IL-17水平分别为（3.4±0.6）%、（1.9±0.4）%、（4.7±0.6） ng/L、（15.1±2.5） ng/L和（11.5±2.1） ng/L,均显著低于治疗前,差异有统计学意义（t=7.477、t=7.177、t=4.596、t=4.102、t=6.237,P<0.05）。结论 CHC和丙型肝炎肝硬化患者外周血Th17和Treg细胞比率增高,相关细胞因子水平也升高,可能参与了丙型肝炎病毒感染的发病过程,在抗病毒治疗后血清细胞因子水平降低,可能有助于对疗效的观察。     

Oxymatrine liposome attenuates hepatic fibrosis via targeting hepatic stellate cells

AIM: To investigate the potential mechanism of Arg-Gly-Asp (RGD) peptide-labeled liposome loading oxymatrine (OM) therapy in CCl₄-induced hepatic fibrosis in rats.METHODS: We constructed a rat model of CCl₄-induced hepatic fibrosis and treated the rats with different formulations of OM. To evaluate the antifibrotic effect of OM, we detected levels of alkaline phosphatase, hepatic histopathology (hematoxylin and eosin stain and Masson staining) and fibrosis-related gene expression of matrix metallopeptidase (MMP)-2, tissue inhibitor of metalloproteinase (TIMP)-1 as well as type I procollagen via quantitative real-time polymerase chain reaction. To detect cell viability and apoptosis of hepatic stellate cells (HSCs), we performed 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-diphenytetrazoliumromide assay and flow cytometry. To reinforce the combination of oxymatrine with HSCs, we constructed fluorescein-isothiocyanate-conjugated Arg-Gly-Asp peptide-labeled liposomes loading OM, and its targeting of HSCs was examined by fluorescent microscopy.RESULTS: OM attenuated CCl₄-induced hepatic fibrosis, as defined by reducing serum alkaline phosphatase (344.47 ± 27.52 U/L vs 550.69 ± 43.78 U/L, P < 0.05), attenuating liver injury and improving collagen deposits (2.36% ± 0.09% vs 7.70% ± 0.60%, P < 0.05) and downregulating fibrosis-related gene expression, that is, MMP-2, TIMP-1 and type I procollagen (P < 0.05). OM inhibited cell viability and induced apoptosis of HSCs in vitro. RGD promoted OM targeting of HSCs and enhanced the therapeutic effect of OM in terms of serum alkaline phosphatase (272.51 ± 19.55 U/L vs 344.47 ± 27.52 U/L, P < 0.05), liver injury, collagen deposits (0.26% ± 0.09% vs 2.36% ± 0.09%, P < 0.05) and downregulating fibrosis-related gene expression, that is, MMP-2, TIMP-1 and type I procollagen (P < 0.05). Moreover, in vitro assay demonstrated that RGD enhanced the effect of OM on HSC viability and apoptosis.CONCLUSION: OM attenuated hepatic fibrosis by inhibiting viability and inducing apoptosis of HSCs. The RGD-labeled formulation enhanced the targeting efficiency for HSCs and the therapeutic effect.     

Antifibrotic effect of N-acetyl-seryl-aspartyl-lysyl-proline on bile duct ligation induced liver fibrosis in rats

AIM:To investigate the preventive effect of N-acetyl-seryl-aspartyl-lysyl-proline (AcSDKP) on bile duct ligation (BDL)induced liver fibrosis in rats. METHODS:Liver fibrosis in rats was induced by BDL and AcSDKP was infused subcutaneously for 2 wkvia a osmotic minipump (Alzet 2ML4) immediately after BDL operation. After scarifying, serum and liver specimens were collected. Hematoxylin and eosin staining, Sirius red staining, enzyme linked immunosorbent assay, Western blot or real-time polymerase chain reaction were used to determinate liver functions, histological alterations, collagen deposition, mRNA expression of markers for fibroblasts, transforming growth factor-β1 (TGF-β1) and bone morphogenetic protein-7 (BMP-7). RESULTS:When compared to model rats, chronic exogenous AcSDKP infusion suppressed profibrogenicTGF-β1 signaling, α-smooth muscle actin positivity (α-SMA), fibroblast specific protein-1 (FSP-1) staining and collagen gene expression. Col Ⅰ, Col Ⅲ, matrix metalloproteinase-2, tissue inhibitors of metallopro-teinase-1 and tissue inhibitors of metalloproteinase-2 mRNA expressions were all significantly downregulated by AcSDKP infusion (2.02 ± 1.10vs 14.16 ± 6.50, 2.02 ± 0.45vs 10.00 ± 3.35, 2.91 ± 0.30vs 7.83 ± 1.10, 4.64 ± 1.25 vs 18.52 ± 7.61, 0.46 ± 0.16 vs 0.34 ± 0.12, respectively, P 0.05). Chronic exogenous AcSDKP infusion attenuated BDL-induced liver injury, inflammation and fibrosis. BDL caused a remarkable increase in alanine transaminase, aspartate transaminase, total bilirubin, and prothrombin time, all of which were reduced by AcSDKP infusion. Mast cells, collagen accumulation, α-SMA, TGF-β1, FSP-1 and BMP-7 increased. The histological appearance of liver specimens was also improved. CONCLUSION:Infusion of exogenous AcSDKP attenu-ated BDL-induced fibrosis in the rat liver. Preservation of AcSDKP may be a useful therapeutic approach in the management of liver fibrosis.     

Hepatoprotective effect of Cichorium intybus L.,a traditional Uighur medicine,against carbon tetrachloride-induced hepatic fibrosis in rats

AIM:To investigate the hepatoprotective effect of a Cichorium intybus L.extract(CIE)on CCl4-induced hepatic fibrosis in rats.METHODS:Seventy-two male Wistar albino rats were randomly divided into six groups of twelve rats each.The normal control group was allowed free access to food and water.Liver injury was performed in the remaining five groups with an i.p.injection of a 1.0mL/kg CCl4 and olive oil(2:3 v/v)mixture,twice weekly for 8 weeks.All rats,with the exception of the injury model group,were intragastrically(i.g.,)administered quantum satis(q.s.)dosages[CIE group:6,18,and 54 mg/kg,respectively;Fu Fang Bie Jia Ruan Gan Pian(FFBJRGP)group:780 mg/kg].The oral administration of different drugs was performed on the day before CCl4 administration and subsequently once per day for8 wk.The serum levels of aspartate aminotransferase(AST),alanine aminotransferase(ALT),hexadecenoic acid(HA),laminin(LN),hydroxyproline(Hyp),and glutathione(GSH),malondialdehyde(MDA)and superoxide dismutase(SOD)in the rat livers were measured.Histopathological changes in the liver were assessed for each group using HE staining and a Masson Trichrome examination.The expression of transforming growth factor-β1(TGF-β1)andα-smooth muscle actin(α-SMA)was examined by immunohistochemical analysis.RESULTS:CIE at oral doses of 6,18,and 54 g/kg per day showed a significant hepatoprotective effect,especially at a dose of 54 g/kg per day.CIE doses reduced the levels of AST(149.04±34.44,P<0.01),ALT(100.72±27.19,P<0.01),HA(548.50±65.09,P<0.01),LN(28.69±3.32,P<0.01)and Hyp(263.33±75.82,P<0.01).With regards to hepatoprotective activity,the CIE dose of 54 g/kg per day produced the largest significant effect by increasing GSH(3.11±0.81),SOD(269.98±33.77,P<0.01)and reducing MDA(2.76±0.51,P<0.01)levels in the liver.The expressions of TGF-β1 andα-SMA were measured by immunohistology and found to be significantly reduced by CIE in a dose-dependent manner.     

Hepatocyte nuclear factor 4α induces a tendency of differentiation and activation of rat hepatic stellate cells

AIM: To investigate the effect of hepatocyte nuclear factor 4α(HNF4α) on the differentiation and transformation of hepatic stellate cells(HSCs).METHODS: By constructing the recombinant adenovirus vector expressing HNF4α and HNF4αshRNA vector, and manipulating HNF4α expression in HSC-T6 cells, we explored the influence of HNF4α and its induction capacity in the differentiation of rat HSCs into hepatocytes.RESULTS: With increased expression of HNF4αmediated by AdHNF4α, the relative expression of Nanog was downregulated in HSC-T6 cells(98.33 ±12.33 vs 41.33 ± 5.67, P 0.001). Consequently, the expression of G-P-6 and PEPCK was upregulated(G-P-6:14.34 ± 3.33 vs 42.53 ± 5.87, P 0.01; PEPCK: 10.10± 4.67 vs 56.56 ± 5.25, P 0.001), the expression of AFP and ALB was positive, and the expression of Nanog, Type Ⅰ collagen, α-SMA, and TIMP-1 was significantly decreased. HNF4α also downregulated vimentin expression and enhanced E-cadherin expression. The ultrastructure of HNF4α-induced cells had more mitochondria and ribosomes compared with the parental cells. After silencing HNF4α expression,EPCK, E-cadherin, AFP, and ALB were downregulated and α-SMA and vimentin were upregulated.CONCLUSION: HNF4α can induce a tendency of differentiation of HSCs into hepatocyte-like cells. These findings may provide an effective way for the treatmentof liver diseases.     

氧化苯砷体外对大鼠原代肝星状细胞活化的阻抑作用研究*

目的 探讨氧化苯砷(PAO)对肝星状细胞（HSC）活化的影响。方法 采用密度梯度离心法提取SD大鼠原代HSC,在倒置显微镜下观察HSC形态的改变;以25、50、100、150和200 nmol/L浓度PAO处理活化的HSC-T6细胞,以四甲基偶氮唑盐（MTT）法评估PAO的细胞毒性;分别以25、50、100 nmol/L浓度的PAO处理离体培养4 d的HSC 72 h,并设对照组,采用Western blot和Real-time PCR法检测各组细胞α-SMA和I型胶原mRNA和蛋白表达。结果 原代HSC离体培养过程中α-SMA表达量逐渐升高,与培养1 d时（0.762±0.062）比,培养4 d时其α-SMA蛋白表达【(1.51±0.045),P<0.05】 显著升高;PAO在25~100 nmo/L浓度范围内对活化的HSC无明显的细胞毒性,但能浓度依赖性抑制活化的HSC α-SMA 和I型胶原mRNA水平和蛋白表达。结论 离体培养4 d时,HSC呈初始活化状态。PAO在25~100 nmol/L范围可浓度依赖性地阻抑离体培养的HSC的自发激活,提示PAO有潜力成为一类新型的抗肝纤维化药物。     

Human platelets inhibit liver fibrosis in severe combined immunodeficiency mice

AIM:To investigate the role of human platelets in liver fibrosis.METHODS:Severe combined immunodeficiency(SCID)mice were administered CCl4and either phosphate-buffered saline(PBS group)or human platelet transfusions(hPLT group).Concentrations of hepatocyte growth factor(HGF),matrix metallopeptidases(MMP)-9,and transforming growth factor-β(TGF-β)in the liver tissue were compared between the PBS and the hPLT groups by enzyme-linked immunosorbent assay(ELISA)and Western blotting.The effects of a human platelet transfusion on liver fibrosis included the fibrotic area,hydroxyproline content,and-smooth muscle actin(α-SMA)expression,which were evaluated by picrosirius red staining,ELISA,and immunohistochemical staining using an anti-mouse-SMA antibody,respectively.Phosphorylations of mesenchymal-epithelial transition factor(Met)and SMAD3,downstream signals of HGF and TGF-β,were compared between the two groups by Western blotting and were quantified using densitometry.Hepatocyte apoptosis was evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling.Furthermore,the accumulation of human platelets in the liver 2 h after platelet transfusion was compared between normal and fibrotic livers by immunohistochemical staining using an anti-human CD41 antibody.RESULTS:The fibrotic area and hydroxyproline content in the liver were both significantly lower in the hPLT group when compared to the PBS group(fibrotic area,1.7%±0.6%vs 2.5%±0.6%,P=0.03;hydroxyproline content,121±26 ng/g liver vs 156±47 ng/g liver,P=0.04).There was less α-smooth muscle actin staining in the hPLT group than in the PBS group(0.5%±0.1%vs 0.8%±0.3%,P=0.02).Hepatic expression levels of mouse HGF and MMP-9were significantly higher in the hPLT group than in the PBS group(HGF,109±13 ng/g liver vs 88±22 ng/g liver,P=0.03;MMP-9,113%±7%/GAPDH vs 92%±11%/GAPDH,P=0.04).In contrast,the concentration of mouse TGF-β in the liver tissue was significantly lower in the hPLT group than in the PBS group(22±5ng/g liver vs 39±6 ng/g liver,P=0.     

复方鳖甲软肝片对二乙基亚硝胺所致原发性肝癌大鼠肝组织PCNA和α-SMA表达的影响*

目的 研究复方鳖甲软肝片对二乙基亚硝胺（DEN）所致原发性肝癌大鼠肝癌组织增殖细胞核抗原（PCNA）和平滑肌肌动蛋白（α-SMA）表达的影响。方法 将40只SD大鼠随机平均分为对照组、模型组、大剂量和小剂量复方鳖甲软肝片处理组,每组均为10只。应用DEN注射法制备原发性肝癌模型,分别给予复方鳖甲软肝片0.625 g.kg^-1和1.25 g.kg^-1或生理盐水灌胃,连续给药20周。采用免疫组化法检测肝组织PCNA和α-SMA表达。结果 大剂量组肝脏质量、肝脏癌变率、肝脏指数和肝表面癌结节数分别为（17.34±1.78）g、10.0%、（3.10±0.72）和（3.16±0.45）个,显著低于模型组 【（25.59±2.38） g、100.0%、（4.95±0.58）和（14.14±2.37）个,P<0.05】;小剂量组肝脏质量、肝脏癌变率、肝脏指数和肝表面癌结节数分别为（19.89±1.80）g、30.0%、（3.89±0.81）和（7.38±1.01）个,也显著低于模型组（P<0.05）;大剂量组PCNA和α-SMA相对表达量分别为（5.98±1.12）和（7.96±1.37）,小剂量组分别为（9.23±1.34）和（12.34±2.88）,均分别明显低于模型组【（19.59±2.67）和（25.12±3.19）,P<0.05】。结论 复方鳖甲软肝片对DEN所致原发性肝癌大鼠具有较好的防治作用,可能是通过调节肝组织PCNA和α-SMA表达发挥作用的。