柞蚕雄蛾提取液逆转大鼠放射性免疫抑制的作用机制研究

目的探讨柞蚕雄蛾提取液对放疗引起的免疫抑制的调节作用及其机制。方法50只Wistar雄性大鼠随机分为对照组、单纯照射组和柞蚕雄蛾提取液大、中、小剂量治疗组,于治疗后第14天观察各组胸腺、脾脏指数变化,流式细胞术检测大鼠外周血细胞中淋巴细胞亚型改变,免疫组化检测大鼠脾脏中Th1类细胞因子IL-2、IFN-γ和Th2类细胞因子IL-4、IL-10表达变化。结果单纯照射组与对照组相比,胸腺、脾脏指数下降,外周血细胞中CD3~ 、CD4~ 淋巴细胞明显减少,脾脏中IL-4、IL-10表达增加,差异有极显著意义(P＜0．01),IL-2、IFN-γ表达元明显改变。大剂量治疗组与单纯照射组相比胸腺、脾脏指数增加,外周血细胞中CD3~ 、CD4~ 淋巴细胞升高,差异有极显著意义(P＜0．01),脾脏中IL-2、IFN-γ表达增加,IL-4、IL-10表达下降(P＜0．01)。结论柞蚕雄蛾提取液可以有效降低放疗引起的免疫抑制,促进T细胞增殖,诱导Th1类细胞因子的表达,促进Th2类细胞因子向Th1类细胞因子漂移。     

阿米福汀对大鼠肺放射性肺损伤保护作用的实验研究

背景与目的:放射性肺损伤是胸部肿瘤放射治疗后的常见并发症,在放射治疗中如何保护肺组织免受或少受放射性损伤显得非常重要。本研究采用阿米福汀作为放射保护剂,探讨其对采用胸壁切线放射治疗技术照射大鼠肺组织放射性损伤的保护作用。方法:将35只雌性Fish-344大白鼠编号后按随机数字法分为常规照射部分和单次照射部分。常规照射部分:20只给予照射5周,总剂量为50 Gy,共25次。随机分为2组,用药照射组10只(随机照射左侧、右侧各5只);对照照射组10只(随机照射左侧、右侧各5只)。单次照射部分:15只右侧胸部切线单次照射20 Gy,随机分为用药照射组5只,对照照射组5只,空白对照组5只。用药照射组照射前30 min内腹腔注射阿米福汀(140 mg/kg),对照照射组用相同剂量照射。采用60Co治疗机,前后野半野对穿照射。照射结束后记录呼吸频率、体质量,尾静脉取血测转化生长因子-β1(transforming growth factor-β,TGF-β),取大鼠照射侧肺及对照组同侧肺进行TGF-β1免疫组织化学染色。结果:常规照射部分用药照射组在放射后4周呼吸频率达到最高,对照照射组在放射后2周达到最高,差异有统计学意义(P<0.05)。单次照射部分对照照射组升高较用药照射组差异有统计学意义(P<0.05)。常规照射部分及单次照射部分中用药照射组与单纯照射组各段时间内的血清TGF-β1差异无统计学意义(P>0.05)。在常规照射部分,肺组织HE染色和马松染色显示,与正常肺泡壁比较,单纯照射组的肺泡壁明显增厚,其肺泡间质大部分为增生纤维组织和弹力纤维组织。而在照射加药组虽也有肺泡壁增厚和纤维化,但与单纯照射组相比较不甚明显,可见大部分肺泡结构存在,肺泡壁的纤维化增生较单纯照射组减轻。免疫组织化学TGF-β1染色显示正常组织为阴性或弱阳性的表达,单纯照射组的肺组织TGF-β1染色表达多为阳性或中强阳性,照射加药组表达为弱阳性或阳性。在常规照射期间,用药照射组在放疗开始后体质量明显下降,而对照照射组体质量下降不明显,差异有统计学意义(P<0.05),在放疗结束后两组之间的体质量平均值逐渐接近。单次照射部分3组之间体质量差异无统计学意义(P>0.05)。结论:本研究显示,在单次大剂量照射部分和常规照射部分,阿米福汀可以改善大鼠受到照射后引起的肺部症状,改善了照射后肺部肺泡增生和纤维化程度。但是常规照射中连续应用阿米福汀会带来一定的不良反应,在实际应用中造成了大鼠体质量减轻。     

阿米福汀对放射性肺损伤保护作用的实验研究

目的用荷瘤和无瘤大鼠的放射性肺损伤模式评价阿米福汀(Amifostine,WR-2721)的放射性肺损伤保护作用.方法选150～160 g的Fisher-344雌性大鼠,在右后胸壁种植R3230AC人乳腺癌细胞株,肿瘤长至直径为1.0～1.5 cm时和无肿瘤大鼠随机进入实验组.用4 MVX射线、剂量DT35 Gy 5分次5 d照射右全肺,每次照射前30 min腹腔内注射Amifostine(150 mg/kg).照射后观测呼吸频率、肿瘤大小和转化生长因子(TGF-β1)水平.当出现呼吸困难伴体重下降时,终止观察,否则在6个月后终止观察.肺组织进行羟脯胺酸和TGF-β1表达等生物学检测.结果治疗后第3 d体重下降明显,照射加Amifostine(15%)和单用Amifostine(11%)与单纯放射(7%)间有明显的差别.肿瘤生长延迟和肿瘤生长情况及因肿瘤肺转移死亡贡献生存率,用药与未用药者相似.无瘤大鼠单纯照射组的平均呼吸频率增加早(第9周开始)、幅度高(125～127次/min),照射加药组在第12周开始,115～118次/min(P<0.001).单纯照射组羟脯胺酸含量明显高于照射加amifostine组(P=0.042)、单用药组和对照组(P<0.01).血浆TGF-β1含量在照射后1～3个月增加、2个月达高峰,单放组的TGF-β1含量[(5.32±1.21) ng/mL]明显高于用药组[(2.80±0.23)     

MnTBAP对大鼠急性放射性肺损伤保护作用的实验研究

目的:观察锰四(4-苯甲酸)卟啉[MnTBAP]对大鼠急性放射性肺损伤的保护作用。方法:200～230g SD雄性大鼠分为正常对照组、照射损伤组和MnTBAP治疗组。用6 MV X射线、单次照射全肺28Gy,建立大鼠肺损伤模型。损伤组腹腔注射生理盐水(1.5mL/kg),MnTBAP组腹腔注射MnTBAP(10mg/kg)。经1、4、8和12周,采用ELISA法检测血清转化生长因子β1(TGF-β1)水平;肺组织作病理检查和生化检测超氧化物歧化酶(SOD)、脂质过氧化产物丙二醛(MDA)、羟脯氨酸(Hyp)、抗超氧阴离子自由基(ASAR)。结果:病理检查显示,MnTBAP组肺组织损伤程度较损伤组明显减轻;生化检测结果显示,经1～12周,与照射损伤组相比较,MnTBAP治疗后SOD含量显著增加(P值均<0.001),ASAR含量显著增加(P值均<0.001),MDA含量显著减少(1周,P<0.001;4～12周,P值均<0.05),TGF-β1含量显著减少(P值均<0.001),Hyp含量1～4周没有变化,8～12周明显降低(8周,P<0.05;12周,P<0.01)。结论:MnTBAP具有抗氧化清除自由基功能,对放射性肺损伤具有保护作用,可能是潜在的放射性损伤保护剂。     

川芎嗪逆转肿瘤多药耐药性及其机制的研究

目的:探讨川芎嗪(tetrameth-ylpyrazine,TMP)逆转人乳腺癌MCF-7/ADM细胞对多柔比星(adriamycin,ADM)的耐药及其P-糖蛋白(P-glycopro-tein,P-gp)表达的影响。方法:MTT法测定细胞的药敏性,荧光分光光度法检测细胞内多柔比星浓度的变化,流式细胞术检测耐药细胞凋亡率的变化,流式细胞术观测细胞P-gp的表达。结果:非细胞毒性剂量(320mg/L)川芎嗪能显著降低MCF-7/ADM的IC50(P=0·0031),逆转倍数为2·13倍;且能明显增加耐药细胞ADM的浓度(P=0·0042)和凋亡率(P=0·0026);320mg/L川芎嗪使耐药细胞的P-gp表达率由(90·6±0·41)%降低至(69·1±1·65)%。结论:川芎嗪具有部分逆转人乳腺癌MCF-7/ADM细胞对多柔比星的耐药性,其逆转机制可能与抑制该细胞P-gp的表达有关。     

养阴抗毒胶囊Ⅱ对放射损伤大鼠防护效应的实验研究

目的:研究养阴抗毒胶囊Ⅱ对重度造血型放射损伤大鼠的防护作用。方法:以6GyX射线照射30只SD大鼠造成重度造血型放射损伤模型。SD大鼠随机分成3组,即:正常对照组(A组);照射对照组(B组);照射中药组(C组)。外周血红、白细胞计数;检测血浆总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)、过氧化氢酶(Cat)、丙二醛(MDA);骨髓切片组织病理学检测;观察胸腺指数和脾脏指数的变化。结果:与B组相比,C组的WBC、T-AOC、Cat、SOD水平显著上升(P<0.05),MDA水平下降(P<0.01);胸腺指数和脾脏指数增大,具有显著差异(P<0.01)。光镜下B组大鼠骨髓切片可见满视野大量脂肪细胞,细胞肿胀变性,细胞间联系不清,细胞核固缩、浓染;C组可见少量细胞变性。结论:养阴抗毒胶囊Ⅱ对辐射损伤大鼠具有防护效应。     

斯普林对小鼠免疫功能影响的研究

目的探讨斯普林对小鼠脾细胞增殖转化能力、体内细胞因子分泌的影响和小鼠免疫功能的作用。方法采用MTT比色法测定脾细胞的增殖程度,细胞因子生物学活性测定法检测小鼠体内白细胞介素2(interleukin2,IL2)和IFN的产生情况。结果应用斯普林注射液的实验组小鼠分为3.50和1.75mg/(kg·d)免疫剂量,在淋巴细胞转化试验中,两个剂量的实验组小鼠吸光度值(OD490)分别为0.63±0.14和0.56±0.09,刺激指数分别为1.93±0.28和1.76±0.16;与对照组小鼠比较差异有统计学意义,t=3.543,P=0.008;t=3.332,P=0.011。实验组小鼠脾细胞上清中IL2和IFN细胞因子的分泌量均较对照组增高,两者间差异有统计学意义,t=4.229,P=0.003;t=3.140,P=0.013。结论斯普林可明显增强小鼠的免疫功能,具有较强的抗肿瘤作用。     

小牛血去蛋白提取物对急性放射性肠炎大鼠小肠黏膜的修复作用及凋亡相关基因的影响

目的 探讨小牛血去蛋白提取物(商品名爱维治)对急性放射性肠炎大鼠小肠黏膜的修复作用及对肠上皮细胞bcl-2、bax基因蛋白表达的影响.方法 以高能X线直线加速器给予实验大鼠全腹照射(9.0 Gy),建立辐射损伤模型.实验大鼠随机分成正常对照组、模型对照组、爱维治低、中、高剂量组.造模后连续4 d腹腔注射给药,取相应部佗的小肠制成病理切片,图像分析仪测定相关形态学指标,用免疫组化方法 检测小肠黏膜上皮细胞中凋亡相关蛋白bcl-2、bax的表达.结果 爱维治中、高剂量组小肠绒毛高度、隐窝深度、黏膜厚度和全层厚度分别为(254.66±26.71)μm、(166.47±25.31)μm、(510.44±30.27)μm、(610.38±37.56)μm和(261.71±30.12)μm、(165.41±19.89)μm、(511.71±29.64)μm、(608.98±34.23)μm,较模型对照组明显改善(P＜0.05).爱维治中、高剂量组bax的表达量分别为(24.54±8.59)%和(23.24±9.10)%,低于模型对照组(P＜0.05);爱维治中、高剂量组bcl-2的表达量分别为(55.54±8.59)%和(52.21±8.32)%,高于模型对照组(P＜0.05);爱维治中、高剂量组bcl-2/bax的比值分别为2.2632和2.1275,高于模型对照组(0.3425,P＜0.01).结论 爱维治通过促进抑凋亡蛋白bcl-2的表达,抑制促凋亡蛋白bax的表达,减少肠黏膜细胞凋亡,加速急性放射性肠炎受损肠黏膜的修复.     

卡托普利通过抑制CCL-2表达缓解大鼠急性放射性肺损伤的机制研究

目的 研究卡托普利对大鼠急性放射性肺损伤的抑制作用及可能机制。方法 将64只雌性Wistar大鼠随机分为对照组、照射组和照射+卡托普利低/高剂量组。除对照组外,其余各组均给予右肺单次20 Gy照射建立大鼠急性放射性肺损伤模型。于第1、2、4、8周处死大鼠HE染色观察大鼠肺组织变化,RT-qPCR、蛋白印迹法分别检测肺组织中CCL-2mRNA水平及蛋白含量,免疫组化检测各组大鼠肺组织中巨噬细胞(CD₆₈)数量。采用单因素方差分析。结果 卡托普利能够减轻急性放射性肺损伤大鼠肺组织炎症反应(P<0.05),抑制单核细胞在损伤肺组织中聚集(P<0.05),降低CCL-2在大鼠肺组织中的含量(P<0.05)。结论 卡托普利可能通过降低CCL-2的表达,从而抑制单核细胞在急性放射性肺损伤大鼠肺组织中的聚集,进而减轻炎症反应。     

柞蚕雄蛾浓缩液影响小鼠免疫功能的实验研究

目的 :研究柞蚕雄蛾浓缩液对小鼠细胞免疫 ,体液免疫及小鼠单核吞噬系统功能的影响。方法 :每项实验均40只小鼠随机分为生理盐水组和高、中、低 3个剂量的柞蚕雄蛾浓缩液给药组 ,经口灌胃给药连续 15d。分别测定计算代表细胞免疫的足垫肿胀反应值FSR(mm) ,代表体液免疫反应的半数溶血值 (HC50 )值 ,反映小鼠单核吞噬系统对异物胶体炭粒清除功能的吞噬指数K及校正吞噬指数α。结果 :2 4h及 48h足垫肿胀反应值在给药各组均较对照组增高 ,经方差分析差异有统计学意义 ( 2 4hP <0 0 1,48hP <0 0 0 1) ;HC50 值 ,高剂量组与对照组相比有统计学意义 ,P <0 0 1,且高剂量组分别与低剂量组和中剂量组相比有统计学意义 ,P <0 0 1;吞噬指数K及校正吞噬指数α虽有一定增高趋势 ,但经方差分析各组之间差异无统计学意义 ,P >0 0 5。结论 :柞蚕雄蛾浓缩液可显著增强小鼠细胞免疫功能 ,对小鼠体液免疫反应有一定增强作用 ,对小鼠单核吞噬系统功能无显著影响     

柞蚕雄蛾提取液对放疗后大鼠脾细胞中Th1/Th2类细胞因子的影响

目的：探讨柞蚕雄蛾提取液对放疗后大鼠脾细胞表达Th1类细胞因子IL-2、IFN-g和Th2类细胞因子IL-4、IL-10的影响,分析其作用机制。方法：将30只Wistar雄性大鼠随机分为对照组、单纯放疗组和药物治疗组,于放疗后第14天观察各组大鼠的胸腺和脾脏指数变化,用免疫组化法检测大鼠细胞表达IL-2、IFN-g和L-4、IL-10水平的变化。结果：单纯放疗组与对照组大鼠相比,其胸腺和脾脏指数明显下降,脾细胞表达IL-4和IL-10的水平增加,差异有统计学意义,P〈0.05。但IL-2和IFN-g的表达无明显变化,P〉0.05。药物治疗组与单纯放疗组相比大鼠的胸腺、脾脏指数明显增加,脾细胞表达IL-2和INF-g的水平增加,IL-4和IL-10的水平下降,P〈0.05。结论：柞蚕雄蛾提取液可以有效降低放疗引起的免疫抑制,该作用可能与其诱导Th1类细胞因子的表达以及促进Th2样细胞因子向Th1类细胞因子漂移有关。     

Enhanced CD3+/CD4+ T cell activities and modulation of Th1/Th2 lineage development in irradiated rats due to treatment with the male zooid of antheraea pernyi extracts

Objective  Cancer patients undergoing large dose radiotherapy exhibit multifaceted defects in their immune capacity that are likely to contribute to an increased susceptibility to infections and disease progression. The immune impairment may also constitute a barrier to effective immunotherapeutic interventions. Here, we evaluate whether supplementation with the male zooid of Antheraea pernyi extracts could enhance immune function in irradiated rats. Methods  Fifty male Wistar rats were randomly divided into a control group, a simple radiation group and a treatment group. The mice in the simple irradiation and treatment groups were given whole-chest irradiation with 6Gy. In the treatment group, the male zooid of Antheraea pernyi extracts was gavaged at the doses of 16.53mg/kg (large dose group), 2.62mg/kg (medium dose group), and 0.564mg/kg (small dose group) once a day for 14 days. The thymus and spleen indices were calculated. T cell subsets in peripheral blood were determined by flow cytometry and the expressions of IL-2, IFN-γ, IL-4 and IL-10 in sera were determined by ELISA on the 15th day. Results  The thymus index and spleen index of the high dose treatment group were statistically lower than that of the control group and higher than that of the radiation group (P<0.01). CD3+ and CD4+ T cells in the peripheral blood were increased in the high dose treatment group and decreased in the radiation group (P<0.01). Expression of IL-2 and INF-γ in the radiation group was lower than that in control, and significantly increased during therapy. The production of IL-4 and IL-10 could be induced by radiation and was inhibited in the high dose treatment group (P<0.01). Conclusion  Our data indicate that the male zooid of Antheraea pernyi extracts may be administrated to improve immune function in irradiated rats and reverse the radial immune inhibition of rats by stimulating the proliferation of Th cells and inducing the differentiation of Th2 to Th1. This work was supported by the grants from the National Natural Science foundation of China (No. 30472260), the Administration of Traditional Chinese Medicine of Shandong Province, China(No. 2005-109),, and the Health Protection Committee of Shandong Province, China(No. 2006059).     

卵巢癌化疗耐药及逆转耐药基因水平研究进展

卵巢癌细胞对化疗产生耐药的机制主要有肿瘤细胞内有效药物浓度降低、DNA损伤修复功能异常和细胞凋亡调控异常3个方面.逆转卵巢癌细胞耐药主要包括反义基因治疗、RNA干预、化疗药物的联合应用等.     

冷冻治疗抑制大鼠C6脑胶质瘤增殖的实验研究

目的：研究冷冻治疗对大鼠C6脑胶质瘤的增殖抑制作用及其机制。方法：建立大鼠C6脑胶质瘤模型，冷冻治疗后免疫组化检测CyclinD1蛋白的表达，流式细胞仪检测细胞周期的变化，磁共振检测肿瘤体积变化。结果：与对照组比较，冷冻治疗后CyclinD1蛋白的表达下调，细胞周期出现G1期阻滞，肿瘤体积缩小。结论：冷冻治疗可抑制大鼠C6脑胶质瘤的增殖，其机制与下调CyclinD1蛋白的表达后，肿瘤细胞周期发生G1期阻滞有关。     

Experimental study on immune serum therapy of cancer

The effect of specific antisera to alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA) are described as one of the immune serum therapy of cancer. This fundamental approach especially in case of specific antisera to AFP was developed to confirm the effect of purified antibodies to AFP and its mechanism in experimental systems. Bi-specific monoclonal antibody to hepatoma membrane antigen is also added as one of the models for application of the immune serum therapy of cancer in rats.     

雷公藤内酯醇对急性T淋巴细胞白血病Jurkat细胞系APC基因去甲基化

 目的　以抑癌基因即腺瘤性结肠息肉病相关基因（APC）存在异常甲基化的急性T淋巴细胞白血病Jurkat细胞为研究对象,探究雷公藤内酯醇（TPL）对APC基因的影响,并对其机制进行初步探讨。方法　运用四甲基偶氮唑盐（MTT）法等检测TPL对Jurkat细胞株增生的影响。巢式甲基特异性PCR（n-MSP）检测TPL对Jurkat细胞APC基因甲基化模式的影响。半定量RT-PCR检测经TPL作用后Jurkat细胞APC基因、甲基转移酶DNMT3A、DNMT3B mRNA的表达。Western blotting检测TPL作用后APC蛋白的表达。结果　TPL对Jurkat细胞生长有明显的抑制作用,并有时间及剂量依赖性,48 h IC50为19.7 ng／ml;TPL可逆转APC基因的高甲基化;TPL能够诱导Jurkat细胞APC基因mRNA的表达,具剂量依赖性;TPL能够诱导Jurkat细胞APC蛋白重新表达,亦有剂量依赖性。结论　小剂量TPL可明显抑制Jurkat细胞的增生,其可能机制为通过诱导Jurkat细胞中异常甲基化的APC基因去甲基化,使APC基因恢复表达。     

Promotion of experimental breast carcinoma in rats treated with extract of liver from rats with portacaval shunt.

Liver homogenates or extracts of liver homogenates from rats in which portacaval shunt had been performed were found to have a significant growth-promoting effect on 7,12 dimethyl (a) benzanthracene (DMBA)-produced breast carcinoma in rats. Tumor potentiation was manifested by increased incidence of animals developing tumors, increased number of tumors, and increased tumor size, when compared with animals receiving injections of shunted or control liver. These observations suggest the existence of a tumor-stimulating factor in liver from which portal blood has been completely and chronically diverted by portacaval shunt. The demonstration of tumor growth stimulating factor(s) present in shunted liver, together with previously reported observations of the modification of the growth of various types of tumors in animals with a portacaval shunt, suggests that the liver is capable of playing an important role in tumor-host interactions. The portacaval shunt appears to be useful as a technique in elucidating ways that liver function may influence tumor growth.     

Experimental induction of neoplasia in the accessory sex organs of male Lobund-Wistar rats

Experimental induction of neoplasia in the urogenital tract was studied in male Lobund-Wistar rats. Animals were given single 30.0-mg/kg i.v. injections of N-nitroso-N-methylurea (NMU) followed 7 days later by s.c. implantation of a 2.0-cm Silastic capsule containing testosterone propionate (TP). Additional rats were given the NMU or TP treatments individually. Control animals were given a single i.v. injection of saline followed by implantation of an empty Silastic capsule. The Silastic implants for each group were replaced every 2 months. This hormone treatment regimen produced significantly (P less than 0.05) elevated serum testosterone concentrations relative to control for 42 days following implantation. Animals were killed at 92, 177, 259, 361, or 427 days post-NMU injection. A high treatment-related incidence of adenocarcinoma occurred in the dorsal and lateral prostatic lobes of animals given the combined NMU-TP treatment. In addition, a few animals had adenocarcinomas of the coagulating gland or the seminal vesicle. The estimated probability of neoplasia in the accessory sex organs by 427 days after initiation of the NMU-TP treatment was 68%, with no occurrence before 9 months. The NMU-TP treatment was also associated with an incidence of focal dysplasia in the accessory sex organs, particularly in the coagulating gland. These findings indicate that NMU-TP treatment of Lobund-Wistar rats can provide a useful experimental system to study the biochemical and molecular events involved in the induction of accessory sex organ neoplasia.