COMPARATIVE STUDY OF PHOSPHOTYROSYL PROTEIN PHOSPHATASE OF MICE NORMAL LIVER, REGENERATING LIVER, AND MICE H22A HEPATOMA ASCITES CELL

In regenerating liver of mice, marked increase of the activity of phosphotyrosyl protein phosphatase (PTPP) in cytosol was observed. The PTPP activity varied with time and reached the highest level between 24 to 48 hours after partial hepatectomy. In H22a cells the PTPP activity found in every subcellular fraction was lower than that of the normal liver. The PTPP activity was mostly concentrated in lysosomes of normal liver, but mainly distributed in nucleus, cytosol and microsome of regenerating liver. In H22a cells PTPP activity seemed distribute evenly. Five similar major PTPP peaks (I-V) were obtained on DEAE cellulose chromatography of cytosols from all three of liver cells studied. However, two additional PTPP peaks, a and b, were also obtained from cytosol of liver.

Anti-tumor Effect and Mechanism of Pratia Extract on H22 Tumor-bearing Mice

[Objectives]This study was conducted to investigate the inhibitory effect of pratia extract on H22 tumor-bearing mice and the effects on immune organs.[Methods]With the application of H22 liver tumor-bearing mice as an animal model,the animals were divided into such three Pratia extract groups as the high,medium and low dose groups(400,200 and 100 mg/kg)and cyclophosphamide CTX group(20 mg/kg).15 d after the administration,the animals were killed by cervical dislocation,and the tumors,thymuses and spleens were taken and weighed,followed by the calculation of the tumor inhibitory rate and the thymus and spleen index,and the serum tumor necrosis factor-α(TNF-α)and interleukin-2(IL-2)levels were determined by ELISA assay.[Results]The inhibitory rates were 54.1,32.6 and 8.2%,respectively,and there were significant differences from the model group(P<0.05);and the spleen index of the tumor-bearing mice was reduced,while the thymus index was improved.The serological results showed that the drug-administrated groups significantly improved the IL-2 levels in the tumor-bearing mice,but had no effects on TNF-α.[Conclusions]Pratia extract has an antitumor effect on H22 tumor-bearing mice,and show certain dose-effect relationship,and its mechanism may be related to enhancing the immune function in tumor-bearing mice by regulating IL-2.

肝癌H-22细胞膜抗原肽提取物对小鼠免疫功能的影响及其抑瘤作用

目的 应用小鼠肝癌H 2 2细胞膜相关抗原肽 (TAP)提取物免疫小鼠 ,观察其对小鼠自身移植肿瘤生长及免疫学参数的影响 ,为制备肿瘤疫苗提供实验依据。方法 采用微酸洗脱法制备分子量≤ 30 0 0Da的细胞膜TAP提取物 ,皮下免疫小鼠 ,检测胸腺细胞增殖反应、T细胞亚组百分数的变化 ,脾细胞ConA反应性、IL 2和IFN γ活性、CTL杀伤活性的变化及抑瘤效应。结果 TAP提取物免疫后 ,移植肿瘤的发生率降低 (P <0 .0 1) ,平均出现时间延迟 (P <0 .0 0 1) ,生长速度减慢 ;同时 ,脾细胞ConA反应性 (P <0 .0 1)、IFN γ(P <0 .0 5 )和IL 2分泌活性和CTL杀伤活性 (P <0 .0 5 )不同程度增强 ;胸腺细胞3 H TdR自发掺入率 (P <0 .0 5 )及CD4 + 和CD8+ T细胞百分数 (P <0 .0 5 )也有不同程度增高。结论 小鼠肝癌H 2 2细胞膜TAP提取物具有免疫原性 ,能有效激发免疫系统功能 ,抑制自身移植肿瘤的生长。

大鼠Walker-256、小鼠H22肿瘤模型建立及应用

目的研究大鼠Walker-256及小鼠H22两种肿瘤模型的成瘤性和稳定性,并探讨其应用。方法建立大鼠Walker-256及小鼠H22两种皮下肿瘤模型,1周后观察其出瘤率和肿瘤大小,观察并绘制肿瘤生长曲线,实验结束后进行病理学检测。结果1周后两种鼠肿瘤模型出瘤率分别为98.57%、97.14%(P>0.05);肿瘤直径分别为(9.61±2.90)mm、(8.25±1.89)mm(P>0.05);大鼠Walker-256模型有肿瘤自然消退现象,而小鼠H22模型肿瘤生长稳定且少数出现肺转移;病理学检测支持上述现象。结论小鼠H22皮下肿瘤模型较大鼠Walker-256模型稳定,可用于抗肿瘤疗效评价。

华蟾素对小鼠H_(22)肝癌移植瘤作用的研究

目的 :观察华蟾素对小鼠移植肝肿瘤H2 2 生长的影响 ,初步探讨华蟾素对肝肿瘤细胞的作用机制。方法 :以瘤株 (H2 2 )接种于小鼠 ,制备肝癌模型 ,设正常对照组、荷瘤模型组、5 -FU组、华蟾素组。各组小鼠采血计白细胞数 ,切除肿瘤、胸腺及脾脏 ,称取重量 ,计算肿瘤抑制率 ;光、电镜下观察肿瘤组织结构。结果 :华蟾素和 5 -FU组小鼠肿瘤抑制率分别为 3 0 .99%和 74.2 8% ,均明显高于荷瘤模型组 (P <0 .0 1)。华蟾素组白细胞数与正常组比较无明显差异 (P>0 .0 5 ) ,显著高于 5 -FU组 (P <0 .0 1)。电镜观察华蟾素组有典型的细胞凋亡形态学特征。结论 :华蟾素通过提高机体免疫功能 。

小鼠肝癌H_(22)细胞LDL受体活性的研究

目的 :观察小鼠肝癌H2 2 细胞LDLR活性。方法 :对小鼠肝癌H2 2 细胞和正常肝细胞进行受体结合的Scatchard分析和单点分析。结果 :①H2 2 细胞对LDL的亲和性与正常肝细胞相同 ,但H2 2 细胞Bmax明显增多 ;②H2 2 细胞对LDL的非特异性结合、内移和降解能力与正常肝细胞相同。结论 :小鼠肝癌H2 2 细胞LDLR活性增高 ,其对LDL的内移和降解功能未变。

人参皂甙Rh_2注射液对荷肝癌(H_(22))小鼠免疫功能的影响

本研究以荷肝癌 (H2 2 )小鼠为实验模型 ,观察人参皂甙Rh2 注射液对荷瘤小鼠腹腔巨噬细胞吞噬功能、血清溶血素、NK细胞毒活性和IL - 2免疫指标的影响。结果表明 :人参皂甙Rh2 注射液能提高荷瘤小鼠腹腔巨噬细胞吞噬功能、增加血清溶血素抗体生成能力、促进小鼠脾NK细胞杀伤活性和IL - 2活性。人参皂甙Rh2注射液明显提高荷瘤小鼠免疫功能。

Possible mechanisms associated with immune escape and apoptosis on anti-hepatocellular carcinoma effect of Mu Ji Fang granules

BACKGROUND Hepatocellular carcinoma(HCC)is one of the most common digestive system cancers with high mortality rates worldwide.The main ingredients in Mu Ji Fang Granules(MJF)are alkaloids,flavonoids,and polysaccharides.MJF has been used in the clinical treatment of hepatitis,cirrhosis and HCC for more than 30 years.Few previous studies have focused on the mechanism of MJF on tumor immunology in the treatment of HCC.AIM To explore the mechanism of action of MJF on tumor immunology in the treatment of HCC.METHODS The absorbable ingredients of MJF were identified using Molecule Network related to High Performance Liquid Chromatography-Electron Spray Ionization-Time of Flight-Mass Spectrometry,and hub potential anti-HCC targets were screened using network pharmacology and pathway enrichment analysis.Forty male mice were randomly divided into the Blank,Model,and MJF groups(1.8,5.4,and 10.8 g/kg/d)following 7 d of oral administration.Average body weight gain,spleen and thymus indices were calculated,tumor tissues were stained with hematoxylin and eosin,and Interferon gamma(IFN-γ),Tumor necrosis factorα(TNF-α),Interleukin-2,aspartate aminotransferase,alanine aminotransferase,alpha-fetoprotein(AFP),Fas,and FasL were measured by Enzyme-linked Immunosorbent Assay.Relevant mRNA expression of Bax and Bcl2 was evaluated by Real Time Quantitative PCR(RTqPCR)and protein expression of Transforming growth factorβ1(TGF-β1)and Mothers against decapentaplegic homolog(SMAD)4 was assessed by Western blotting.The HepG2 cell line was treated with 10 mg/mL,20 mg/mL,30 mg/mL,40 mg/mL of MJF,and another 3 groups were treated with TGF-β1 inhibitor(LY364947)and different doses of MJF.Relevant mRNA expression of TNF-α,IFN-γ,Bax and Bcl2 was evaluated by RT-qPCR and protein expression of TGF-β1,SMAD2,p-SMAD2,SMAD4,and SMAD7 was assessed by Western blotting.RESULTS It was shown that MJF improved body weight gain and tumor inhibition rate in H22 tumorbearing mice,protected immune organs and liver function,reduced the HCC indicator AFP,affected immunity and apoptosis,and up-regulated the TGF-β1/SMAD signaling pathway,by increasing the relative expression of TGF-β1,SMAD2,p-SMAD2 and SMAD4 and decreasing SMAD7,reducing immune factors TNF-αand IFN-γ,decreasing apoptosis cytokines Fas,FasL and Bcl2/Bax,and inhibiting the effect of LY364947 in HepG2 cells.CONCLUSION MJF inhibits HCC by activating the TGF-β1/SMAD signaling pathway,and affecting immune and apoptotic cytokines,which may be due to MJF adjusting immune escape and apoptosis.

Anti-tumor activities of macromolecular fractions of fresh gecko vivo and their induction of Bel-7402 cell differentiation

Objective:To investigate the anti-tumor effect of macromolecular fractions of fresh gecko (M-AG) in vivo and their differentiation-inducing activity in Bel-7402 cells in vitro.Methods:An H22 hepatocarcinoma-bearing mouse model was used to evaluate the anti-tumor activity of M-AG samples.The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was applied to analyze cell viability.Cell morphology was observed by phase contrast microscopy.The quantity of the alpha-fetoprotein was detected by a radioimmunoassay.Chromatometry was used to assay the albumin quantity.Activities of alkaline phosphatase and γ-glutamyl trans-peptidase were measured by biochemical methods.Finally,western blotting was applied to assess proteins in the mitogen-activated protein kinase (MAPK) signaling pathway.Results:Macromolecular fractions of fresh gecko exerted a significant anti-tumor effect in mice.The inhibition rate of tumor growth was 63% in the moderate M-AG dose group.Cells treated with M-AG displayed a differentiated state.The treatment lowered alphafetoprotein secretion and significantly decreased the activities of γ-glutamyl trans-peptidase and alkaline phosphatase in Bel-7402 cells.In contrast,M-AG increased the amount of albumin in the cell culture medium.All biochemical indices demonstrated that M-AG induced Bel7402 cell differentiation.Western blotting showed no changes in the quantities of extracellular signal-regulated kinase (ERK) 1/2,p38MAPK,or c-Jun N-terminal protein kinase 1/2.However,M-AG significantly activated the phosphorylation of ERK1/2 in a dose-dependent manner.In addition,M-AG had no significant influence on the expression of nuclear factor-kappa B.Conclusion:Macromolecular fractions of fresh gecko has an anti-tumor activity in H22 hepatocarcinoma-bearing mice in vivo and inhibits Bel-7402 cell proliferation in vitro by inducing cell differentiation related to activation of ERK1/2.

斑蝥素衍生物与铂络合物抗癌活性的实验研究

目的：研究４种斑蝥素衍生物与铂络合物（斑铂，Ｄｐｔ）对实验动物的抗癌作用，力图开发一种新型的铂族金属抗癌药。方法：以Ｓ１８０肉瘤、Ｈ２２肝癌实体瘤及其腹水瘤小鼠为实验动物，通过腹腔或静脉给予不同剂量的４种化合物Ｄｐｔ１－１５、Ｄｐｔ５－１０、Ｄｐｔ１２－３、Ｄｐｔ６－２，分别观察药物对小鼠瘤重及存活天数的影响，以顺铂为阳性对照，生理盐水为阴性对照。所有数据进行ｔ检验。结果：４种斑铂（Ｄｐｔ１－１５、Ｄｐｔ５－１０、Ｄｐｔ１２－３、Ｄｐｔ６－２）均有抗癌活性。其中Ｄｐｔ５－１０、Ｄｐｔ１－１５对小鼠Ｓ１８０肉瘤及Ｈ２２实体瘤的抑制率与顺铂相似；Ｄｐｔ５－１０对Ｈ２２腹水瘤小鼠生命延长率与顺铂相似；Ｄｐｔ１－１５的有效剂量毒性较大。结论：斑铂是有效的新型抗癌化合物，其中Ｄｐｔ５－１０抗癌作用较好，应进一步改进药物溶解性，并从与顺铂抗药性不同方面开发利用。

“江边一碗水”总木脂素的抗肿瘤作用及一般毒性的研究

为评价"江边一碗水"总木脂素对H22荷瘤小鼠的肿瘤抑制作用和相关毒性,用SPF级雄性小鼠腋下接种肝癌H22瘤株建立了移植性肝癌H22小鼠模型,观察了不同剂量的"江边一碗水"总木脂素给药10 d后对荷瘤小鼠的肿瘤抑制率、体重、免疫器官指数及血尿常规、肝肾功等的影响.结果表明:50,100,200 mg/kg"江边一碗水"总木脂素的抑瘤率分别为44.9%,54.8%和62.1%."江边一碗水"总木脂素对H22小鼠体重、免疫器官指数、造血系统、肝肾功能等生理生化指标无明显影响.故"江边一碗水"总木脂素具有较显著的抗肿瘤作用和较低的毒性,是其抗肿瘤的药效物质基础.

蜜煮川乌对H_(22)荷瘤小鼠免疫功能影响的实验研究

目的 研究蜜煮川乌饮片对H2 2 荷瘤小鼠免疫功能的影响。方法 利用H2 2 荷瘤小鼠为动物模型 ,检测不同剂量蜜煮川乌饮片治疗后小鼠脾脏中T细胞、B细胞增殖和腹腔巨噬细胞吞噬活性。结果 中剂量蜜煮川乌能促进H2 2 荷瘤小鼠T细胞增殖、抑制B细胞增殖、增强腹腔巨噬细胞的吞噬活性。结论 中剂量蜜煮川乌对H2 2 荷瘤小鼠的免疫功能有影响。

克癌力胶囊抗肿瘤作用的实验研究

克癌力胶囊（３０、１０、５ｇ生药／ｋｇＰ．０）能明显抑制小鼠肿瘤Ｕ１４、Ｈ２２和ＦＣ的生长；与环磷酰胺合用对小鼠Ｕ１４肿瘤的治疗有明显的增效作用；能明显提高环磷酰胺中毒小鼠的白细胞、淋巴细胞和骨髓有核细胞数量。

转入人肿瘤坏死因子α基因的鱼腥藻IB02(+)的细胞毒活性及其体内抗肿瘤药效学研究

目的测定转入人肿瘤坏死因子α(hTNFα)基因的鱼腥藻IB02(+)的抗肿瘤药效。方法在相同的培养条件下通过100L光生物反应器培养4组转入hTNFα基因鱼腥藻IB02(+),分别命名为批次1、2、3、4,经过反复冻融和真空冷冻干燥处理,得到转入hTNFα基因鱼腥藻IB02(+)粗提物干粉样品。采用结晶紫染色法检测转入hTNFα基因鱼腥藻IB02(+)粗提物干粉样品的细胞毒活性,并通过测定不同批次间的细胞毒活性差异检测其稳定性。采用转入人肿瘤坏死因子α(hTNFα)基因鱼腥藻IB02(+)对小鼠肝癌细胞H-22的抑制来测定体内抗肿瘤药效。结果转入hTNFα基因鱼腥藻IB02(+)的细胞毒活性约为8000U/mg。4种不同批次的转入hTNFα基因蓝藻IB02(+)细胞毒活性相当,hTNFα蛋白性质比较稳定。小鼠1次口服最大耐受量为16.0g/kg,在观察期内所有受试小鼠无一死亡,生长发育正常,主要脏器未见异常,未显示明显毒性。药效学研究表明,口服6g/kg转入hTNFα基因蓝藻IB02(+)对小鼠肝癌细胞H-22的抑制率平均值(37.7±3.4)%,最高值为41.5%(P<0.001)。结论转入hTNFα基因鱼腥藻IB02(+)抗肿瘤作用明显,无明显的毒副作用且药效学性质稳定。

小鼠肝癌H_(22)细胞LDLR的分析研究

目的 :观察分析小鼠肝癌 H2 2 细胞及正常肝细胞低密度脂蛋白受体 (L DL R)对低密度脂蛋白 (L DL)的结合。方法 :用 1 2 5 I标记 L DL ,对小鼠肝癌 H2 2 细胞及正常肝细胞 L DL R进行受体饱和分析。结果 :H2 2 细胞及正常肝细胞主要通过其 L DL R结合 L DL。 H2 2 细胞及正常肝细胞 L DL R对 L DL的亲和性和特异性相同 ,但 H2 2 细胞Bmax明显增多。结论 :小鼠肝癌 H2 2 细胞 L DL R数目增多 。

维生素E促进小鼠H-22移植瘤的作用及机制

[目的]利用H-22荷瘤小鼠研究维生素E(vitamin E,VE)对肿瘤发生发展的作用及机制。[方法]建立H-22小鼠荷瘤模型,计算肿瘤指数与肝脏指数,利用试剂盒检测血糖、血清低密度脂蛋白(low density lipoprotein,LDL)、总胆固醇(total cholesterol,TC)以及碱性磷酸酶(alkaline phosphatase,AKP)水平,利用苏木素-伊红染色与Real-time PCR检测VE对肿瘤发展和转移的作用及分子机制。[结果]肿瘤-VE组LDL水平、肿瘤组和肿瘤-VE组TC、AKP水平显著高于对照组(LDL,P <0.05;TC,P <0.01;AKP,P <0.001)。肿瘤组织中肿瘤-VE组小鼠p27K ip1、Stat3以及Cxcl12基因表达水平显著高于肿瘤组(p27Kip1和Stat3,P <0.05,Cxcl12,P <0.01)。肝脏组织中肿瘤-VE组Jak2、Pigf及Cxcl12基因表达水平显著高于肿瘤组(P <0.05),且肿瘤组Jak2、Pigf及Cxcl12基因表达水平显著高于对照组(P <0.05)。[结论] VE处理的H-22荷瘤小鼠肿瘤组织中p27Kip1、Stat3及Cxcl12表达水平显著升高,肝脏组织中Jak2、Pigf及Cxcl12表达水平显著升高,VE可能通过调控细胞周期、肿瘤转移及炎症相关基因的表达促进小鼠H-22肿瘤的发展。