白藜芦醇抑制胶质瘤细胞生长与诱导细胞凋亡实验研究

目的探讨白藜芦醇(Res)对脑胶质瘤细胞(C6细胞株)和正常成纤维细胞(3T3细胞株)体外增殖的影响,进而观察Res诱导C6和3T3细胞系的凋亡作用.方法用不同浓度的Res处理C6和3T3细胞,以四甲基偶氮唑蓝(MTT)比色法检测Res对C6和3T3细胞的增殖作用,通过HE染色、扫描电子显微镜(SEM)、原位末端标记法(TUNEL)和流式细胞仪Annexin Ⅴ荧光染色,观察细胞的形态学结构改变,并定量检测细胞凋亡.结果 Res抑制了C6细胞的生长与增殖( P < 0.01 ),呈浓度依赖性反应;Res能明显诱导C6细胞凋亡,经210 μmol/L、120 μmol/L Res处理24 h后及对照组的C6细胞,其凋亡率分别为29.7%、14.6%及2.1%;相应的3T3细胞凋亡率分别为4.3%、3.5%、2.6%. 结论 Res能通过诱导C6细胞凋亡而抑制其生长与增殖,但对3T3细胞无此作用.本研究为临床应用Res治疗脑胶质细胞瘤提供了实验依据.     

吡咯烷二硫氨基甲酸酯对大鼠重症急性胰腺炎的影响

目的　探讨核转录因子 (NF κB)抑制剂吡咯烷二硫氨基甲酸酯 (PDTC)对大鼠重症急性胰腺炎 (SAP)的影响。方法　将 3 6只Wistar大鼠随机分组 :假手术组 (n =6) ,假手术 静脉注射组 (n =6) ,SAP组 (n =12 )和试验组 (n =12 )。各组于造模 6h后 ,测定血清淀粉酶及脂肪酶含量 ,取胰腺组织进行病理学评分 ,采用免疫组织化学法检测NF κB激活水平及胰腺细胞凋亡情况。结果　SAP组和试验组的胰腺细胞内NF κB呈激活状态 ,存在胰腺细胞凋亡 ,与前两组差异有显著性 (P <0 .0 1) ,试验组大鼠的胰腺组织病理学评分、血清淀粉酶及脂肪酶含量、NF κB激活及细胞凋亡水平与SAP组差异存在显著性 (P <0 .0 5 )。结论　在SAP发病机制中 ,NF κB是多种炎症介质的始动因子 ,PDTC可以有效抑制胰腺细胞中NF κB的激活 ,促进胰腺细胞凋亡 ,减少胰酶释放 ,减轻胰腺组织的病理损害。     

Chronic Alcohol Ingestion Enhances Tumor Necrosis Factor-α Expression and Salivary Gland Apoptosis

We investigated the extent of induction in sublingual salivary gland cells apoptosis and tumor necrosis factor-α (TNF-α) expression with chronic ethanol ingestion. The experiments were conducted on rats pair-fed for 8 weeks with alcohol-containing and control liquid diet. The animals were killed, their sublingual glands dissected, and the glandular tissue used for quantitization of TNF-α expression and the assays of acinar cells apoptosis employing sandwich enzyme immunoassay for histone-associated DNA fragments. The mean value for TNF-α in sublingual gland of the control group was 22.3 pg/mg of protein and showed a 1.6-fold increase in the chronic ethanol diet group to 36.5 pg/mg of protein. In comparison with the controls, the sublingual gland of the chronic ethanol diet group also exhibited a 3.4-fold enhancement in acinar cell apoptosis. These findings suggest that chronic ethanol ingestion causes the enhancement in TNF-α expression and leads to the induction in salivary gland acinar cells apoptosis. Thus, the diminished secretion of saliva in alcoholics may be a direct result of increased salivary gland apoptosis.     

单侧睾丸扭转引发对侧睾丸生精损伤免疫机制的实验研究

目的探讨单侧睾丸扭转对侧睾丸生精功能损伤的机理。方法雄性SD大鼠24只,随机分为3组,建立睾丸扭转动物模型。第1组为假手术组,第2、3组扭转左侧睾丸720°并固定维持,分别于12 h和24 h后复位固定。术后1个月留取对侧睾丸称重。应用流式细胞术(FCM)检测各组生精细胞凋亡和各级生精细胞计数,免疫组化SABC法检测抗精子抗体免疫复合物(IgG)在睾丸组织中的沉积。结果第1、2、3组对侧睾丸重量分别为(1555.73±72.34)、(1184.20±101.02)、(783.60±117.93)mg,凋亡细胞数依次为53.25±8.61、1622.00±129.31、3401.25±179.75,抗精子抗体阳性染色率为0、0.55±0.02、0.69±0.03,组间两两比较差异均有统计学意义(P<0.01)。与第1组相比,第2、3组对侧睾丸单倍体、二倍体和四倍体细胞群计数明显减少,同时可见抗精子抗体免疫复合物在睾丸组织中明显沉积,其差异均有统计学意义(P<0.01)。第2、3组间(除四倍体细胞群计数外)差异亦有统计学意义(P<0.01)。结论单侧睾丸扭转导致对侧睾丸生精细胞广泛凋亡,自体免疫反应可能在对侧睾丸生精功能损伤中起重要作用。     

Dendritic Cells Coinjected with Tumor Cells Treated with an Anticancer Drug to Induce Tumor Rejection

Purpose: We examined whether bone marrow-derived dendritic cells (DCs) could induce antitumor immunity when a chemotherapeutic drug was added. Methods: CT26 (a murine colon cancer cell line syngeneic with BALB/c) and CT26-bearing mice were treated with mitomycin C (MMC) intraperitoneally (i.p.). Next, mice immunized with a coinjection of DCs and MMC-treated CT26 (i.p.) were given an intradermal inoculation of CT26. Finally, CT26-bearing mice were treated with MMC (i.p.) with or without DCs, given peritumorally. Results: Although the inoculated tumor was not rejected in the control mice, CT26 was rejected in 50% of the mice injected with MMC alone. Apoptosis was observed in the MMC-treated CT26 cells in vitro and in vivo. Immunization with DCs and apoptotic CT26 cells, but not with apoptotic CT26 alone, gave protection against tumor challenge in 7 of 13 mice. A significantly higher level of cytotoxic T-cell activity and interferon-γ production was seen in the protected mice. When MMC (i.p.) treatment was followed by peritumoral DC injection in the CT26-bearing mice, remarkable therapeutic effects were observed. Conclusion: DCs can collaborate with chemotherapy-induced apoptotic tumor cells and elicit improved antitumor immunity, probably through the acquisition of tumor-associated antigens from apoptotic tumor cells. Received: January 7, 2002 / Accepted: September 3, 2002 Acknowledgments. We thank Dr. Kazuo Kinoshita for his useful advice on using flow cytometry. This research was partly supported by the Ministry of Education, Culture, Sports, Science and Technology (No. 11671160). Reprint requests to: S. Yamasaki     

Studies of neurotrophin biology in the developing trigeminal system

The accessibility of the primary sensory neurons of the trigeminal system at stages throughout their development in avian and mammalian embryos and the ease with which these neurons can be studied in vivo has facilitated investigation of several fundamental aspects of neurotrophin biology. Studies of the timing and sequence of action of neurotrophins and the expression of neurotrophins and their receptors in this well characterised neuronal system have led to a detailed understanding of the functions of neurotrophins in neuronal development. The concepts of neurotrophin independent survival, neurotrophin switching and neurotrophin cooperativity have largely arisen from work on the trigeminal system. Moreover, in vitro studies of trigeminal neurons provided some of the first evidence that the neurotrophin requirements of sensory neurons are related to sensory modality. The developing trigeminal system has been studied most extensively in mice and chickens, each of which has particular advantages for understanding different aspects of neurotrophin biology. In this review, I will outline these advantages and describe some of the main findings that have arisen from this work.     

治疗性高碳酸血症对大鼠移植肺细胞凋亡的影响

目的探讨治疗性高碳酸血症对移植肺细胞凋亡的影响及其可能的机制。方法26只Wistar大鼠,供体鼠和受体鼠各13只,肺移植成功后随机分为2组：对照组（n=6）和治疗组（n= 7）。对照组再灌注期间吸入50%N2-50%O2混合气体;治疗组再灌注期间吸入40%N2-60%O2- 100%CO2混合气体,调节吸入二氧化碳和氧气的流量,维持吸入氧浓度50%,动脉血二氧化碳分压（PaCO2）80～100 mm Hg。记录受体鼠机械通气15 min时平均动脉压（MAP）、PaCO2和动脉血氧分压（PaO2）作为基础值,再灌注期间每15分钟记录1次,直至实验结束。再灌注90 min时放血处死大鼠,采用考马斯亮蓝法测定支气管肺泡灌洗液（BALF）蛋白浓度;取左肺组织,计算湿/干重比;采用免疫组化法测定Bax、Bcl-2的表达;采用TUNEL法检测细胞凋亡情况。结果与基础值比较,再灌注期间对照组MAP和PaO2降低（P〈0.05）;与对照组比较,再灌注期间治疗组MAP、PaCO2和PaO2升高,BALF蛋白浓度、肺组织W/D、Bax表达和细胞凋亡指数降低（P〈0.05）,Bcl-2表达差异无统计学意义（P〉0.05）。结论治疗性高碳酸血症可抑制大鼠移植肺细胞凋亡,其机制与下调Bax蛋白表达有关。     

山莨菪碱、旋覆花素、苦碟子对过度训练致急性心肌损伤大鼠的影响

目的探讨山莨菪碱、旋覆花素、苦碟子对过度训练致急性心肌损伤大鼠的影响。方法雄性Wistar大鼠80只,体重200～220 g,随机分为5组:正常对照组(C组,n=8)、力竭对照组(E组, n=24)、山莨菪碱组(A组,n=16)、旋覆花素组(IB组,n=16)、苦碟子组(IS组,n=16)。采用游泳致力竭建立过度训练动物模型。A组于力竭前即刻腹腔注射山莨菪碱10 mg/kg,IB组于力竭前24 h和力竭前即刻口腔灌入旋覆花素25 ml/kg,IS组于力竭前即刻腹腔注射苦碟子20 ml/kg。取标本检测大鼠血清磷酸肌酸激酶(CK)及其同工酶(CK-MB)的活性,光镜观察心肌组织形态学,采用流式细胞术及TUNEL法检测大鼠心肌细胞凋亡,并计算心肌细胞凋亡率。结果与C组比较,E组、A组、IB组和IS组血清CK及CK-MB升高,心肌细胞凋亡率升高(P<0.05);与E组比较,A组、IB组及IS组大鼠血清CK及CK-MB及心肌细胞凋亡率均降低(P<0.05)。光镜下,E组的心肌细胞核深染,有固缩现象;而A组、IB组及IS组的心肌心肌组织学变化均未见异常。结论山莨菪碱、旋覆花素、苦碟子预先给药通过抑制心肌细胞的凋亡,可减轻过度训练致大鼠的心肌损伤。