Determinants of benzo[a]pyrene diol epoxide adducts to albumin in workers exposed to polycyclic aromatic hydrocarbons

The present study was undertaken among 260 subjects [133 controls and 127 persons exposed to polycyclic aromatic hydrocarbons (PAHs)] from two steel foundries and a graphite electrode producing plant in order to investigate the relationship between the benzo[a]-pyrene-diol epoxide adducts to albumin (BPDE-alb) and the intensity of exposure to PAHs. Blood samples were collected from each subject and BPDE-alb adduct determination was performed using a high-pressure liquid chromatography technique with fluorescence detection. Exposure to PAHs was assessed by measuring airborne concentration of 13 PAHs including benzo[a]pyrene (BaP) using personal air sampling and 1-hydroxypyrene excretion (1-HOP) in postshift urine. Significantly higher BPDE-alb adduct levels were observed in exposed workers compared to controls but wide interindividual variation was observed between subjects with the same level of exposure. BPDE-alb adduct level was weakly but significantly associated with the airborne concentrations of total PAHs (r = 0.35, P = 0.0001) and BaP (r = 0.30, P = 0.0001), and urinary 1-HOP excretion (r = 0.29, P = 0.0001). Alcohol and dietary habits, place of residence, and renal and hepatic status were not found to influence the concentration of BPDE-alb adducts significantly. However, for the same level of exposure to BaP, smokers had a higher probability of having an elevated BPDE-alb adduct level than nonsmokers. As long as BPDE-protein adduct levels have not been demonstrated to be better related to the risk of adverse genotoxic effects caused by PAHs in target organs than the 1-HOP level in urine, the latter remains the most practical biological parameter for assessing the risk of exposure to PAHs.     

DNA adduct formation of benzo[a]pyrene in white blood cells of workers exposed to polycyclic aromatic hydrocarbons

The major DNA adducts of anti-benzo[a]pyrene diolepoxide (BPDE) were determined by high performance liquid chromatography with fluorescence detection (HPLC-FLD) in white blood cells (WBC) of workers exposed to benzo[a]pyrene (B[a]P). In addition, ambient concentrations of B[a]P at the workplace were determined by personal air sampling. Workers in a refractory setting were examined before (n=26) and 3 months after (n = 33) changing the production material (binding pitch). Furthermore, 9 coke oven workers were examined. The change in the production process in the refractory setting led to a decrease in the median of ambient B[a]P concentrations (0.14 to <0.07 microg/m3). The median of BPDE-DNA adduct levels in WBC also decreased from 0.9 adducts/10(8) nucleotides before changing the production material to <0.5 adducts/10(8) nucleotides 3 months afterwards. The B[a]P concentrations at the workplace for the coke oven workers were found to be significantly higher than in the refractory setting. However, BPDE-DNA adduct concentrations in coke oven workers and refractory setting workers showed no significant difference, which was probably due to the low number of studied subjects in the coke-oven setting. No significant differences could be observed for BPDE-DNA adduct levels between current smokers (n=21) and non-smokers (n=14; p = 0.93) from both plants. In addition, no correlation between B[a]P concentrations in the air and DNA adduct levels in refractory workers and in coke oven workers could be found (r = -0.03, p = 0.87). Because of the missing correlation between personal air sampling and BPDE-DNA adduct levels in WBC, the results may indicate that their formation is either influenced by other routes of exposure to B[a]P (e.g., skin absorption, dietary habits) or interindividual differences in their formation and repair.     

Induction and localization of hepatic CYP4501A in flounder and rainbow trout exposed to benzo[a]pyrene

The hepatic detoxification system in Baltic flounder and rainbow trout was characterized under experimental conditions. Fish were exposed to benzo[a]pyrene (BaP, 10 and 50mg/kg, ip) or vehicle for 2, 5, and 10 days (in rainbow trout also for 20 days) and then sacrificed. Control fish were sampled at days 0 and 10 (flounder) or day 20 (rainbow trout). The hepatic distribution of CYP1A was analyzed immunohistochemically and microsomal ethoxyresorufin O-deethylase activity was determined spectrophotometrically. The kinetics of the CYP1A responses (EROD) was similar in both species, while a species-specific difference in the magnitude of the response was observed. CYP1A was demonstrated in the hepatocytes in both fish species 2 days after BaP administration and throughout the experiment. In rainbow trout a CYP1A response in the vascular endothelium of liver parenchyma was detected 2 days postadministration, while the corresponding reaction in flounder was seen 5 days postadministration. Thus, our results confirm previous reports that the CYP1A response is species specific. Furthermore, the induction of hepatic CYP1A in Baltic flounder reflects pathophysiological effects induced by polycyclic aromatic hydrocarbon compounds and, consequently, is a parameter useful when monitoring the anthropogenic effects on the Baltic Sea environment.     

Synthesis and cytotoxic activity of benzo[a]acronycine and benzo[b]acronycine substituted on the A ring

The impact of substitutions at position 10 in the A ring of the cytotoxic benzo[a]acronycine and benzo[b]acronycine series has been explored. 10-Bromobenzo[a] and 10-bromobenzo[b]acronycine were prepared in 12% and 15% yield respectively from commercially available chemicals. Their 1,2-dihydro-1,2-dihydroxy diesters were synthesized. The different derivatives were tested against two cell lines KB-3-1 and L1210. Their cytotoxic activities were found in the same range of magnitude as their non-substituted counterparts. These structure-activity relationships permitted to conclude that the introduction of a substituent at position 10 maintains the activity in both the benzo[a] and [b]acronycine series and open the way to further pharmacomodulations.     

Modulation of carcinogen bioavailability by immunisation with benzo[a]pyrene-conjugate vaccines

Benzo[a]pyrene (B[a]P) conjugate vaccines based on ovalbumin, tetanus toxoid and diphtheria toxoid (DT) as carrier proteins were developed to investigate the effect of specific antibodies on the bioavailability of this ubiquitous carcinogen and its metabolites. After metabolic activation of this prototype carcinogen, B[a]P forms DNA adducts which initiate chemical carcinogenesis. B[a]P-DT conjugate induced the most robust immune response. The antibodies reacted not only with B[a]P but also with the proximate carcinogen 7,8-diol-B[a]P. Antibodies modulated the bioavailability of B[a]P and its metabolic activation in a dose-dependent manner by sequestration in the blood. Our results showed that this immune prophylactic strategy influences the pharmacokinetic of B[a]P and further studies to investigate their effects on chemical carcinogenesis are warranted.     

Synthesis and anticonvulsant activity of some new thiazolo[3,2-a][1,3]diazepine, benzo[d]thiazolo[5,2-a][12,6]diazepine and benzo[d]oxazolo[5,2-a][12,6]diazepine analogues

A new series of 6,7-dihydro-thiazolo[3,2-a][1,3]diazepines (9–12), benzo[d]thiazolo[5,2-a][12,6]diazepines (19–21) and benzo[d]oxazolo[5,2-a][12,6]diazepine (24) analogues were synthesized and evaluated for their anticonvulsant activity. Compounds (E)-2-bromo-6,7-dihydro-thiazolo[3,2-a][1,3]diazepine-8(5H)-thione (12), 3-chloro-benzo[d]thiazolo[5,2-a][12,6]diazepin-10-one (20), and 4-chloro-benzo[d]oxazolo[5,2-a][12,6] diazepin-10-one (24) showed 100% protection against PTZ- and bicuculline-induced seizures; 70%, 33%, 70% protection against MES-induced tonic extension; and 70%, 66%, 100% protection against picrotoxin-induced convulsions, respectively. Compounds 12, 20, and 24 proved to act as GABA_A receptor agonists, with ED₅₀ values of 252, 380, 251 mg/kg; TD₅₀ values of 398, 417, 355 mg/kg; PI values of 1.58, 1.09, 1.41; LD₅₀ values of 380, 617, 537 mg/kg and TI values of 1.51, 1.62, 2.14, respectively.     

苯并[a]芘亚慢性染毒对大鼠海马胆碱能系统的影响

目的 通过观察亚慢性染毒苯并[a]芘(B[a]P)对大鼠神经行为及海马组织中乙酰胆碱(Ach)含量、胆碱酯酶(AChE)活力和乙酰胆碱受体α7亚型(nAChRα7)mRNA和蛋白表达的影响,探讨B[a]P的神经毒性机制.方法 选择60只健康雄性SD大鼠,随机分为空白对照组,溶剂对照组,1.0、2.5、6.25 mg/kg B[a]P染毒组,隔日腹腔注射连续染毒90 d.Morris水迷宫和跳台试验检测大鼠学习记忆能力.碱性羟胺法检测海马组织Ach含量,DNTB法检测AChE活力.荧光定量PCR和Western-blot法分别检测海马组织nAChRα7 mRNA和蛋白表达水平.结果 Morris水迷宫、跳台试验结果显示,2.5、6.25 mg/kg B[a]P组大鼠学习记忆能力比空白和溶剂对照组大鼠明显下降,差异有统计学意义(P＜0.05).2.5、6.25 mg/kgB[a]P组大鼠海马组织Ach含量明显低于空白、溶剂对照和1.0 mg/kg B[a]P组,差异有统计学意义(P＜0.05),6.25 mg/kg B[a]P组AChE活力明显低于空白对照、溶剂对照和1.0 mg/kg B[a]P组,差异有统计学意义(P＜0.05).各组之间nAChRα7 mRNA和蛋白表达水平的差异无统计学意义(P＞0.05).大鼠海马组织Ach含量与大鼠平均逃避潜伏期和总路程呈负相关(相关系数r分别为-0.567和-0.503,P＜0.01),与平台象限滞留时间呈正相关(r=0.800,P＜0.01).结论 亚慢性染毒B[a]P可损伤大鼠学习记忆功能,其机制与大鼠海马组织Ach含量下降有关     

French firefighters exposure to Benzo[a]pyrene after simulated structure fires

Introduction

Firefighters have occupational exposure to toxic compounds during firefighting, but not only. Surface contamination of equipment has never been studied in French firefighters.

Elimination of 1-hydroxypyrene after human volunteer exposure to polycyclic aromatic hydrocarbons

The aim of this study was to estimate the kinetics of 1-hydroxypyrene (1-HP) elimination after inhalation exposure to polycyclic aromatic hydrocarbons (PAHs). Samples of inhaled and exhaled air were collected on glass fiber filters backed with tubes filled with Amberlit XAD-2 resin. The filters were extracted by cyclohexane and Amberlit – by acetonitrile. Extracts for the determination of pyrene and benzo[a]pyrene (B[a]P) concentrations were analyzed by high-performance liquid chromatography (HPLC). 1-Hydroxypyrene in urine was determined after its preconcentration on a C-18 column (solid phase extraction method) using the same analytical technique. Five male volunteers were exposed for 6 h (two times, with a 1-month interval) to a PAH mixture at an aluminium plant. The volunteers were breathing at rest through facial mask equipped with a 1000-ml compensation container which allows collection of the exhaled air. Inhaled air samples were collected in the breathing zone of each volunteer. Urine samples were collected until the 71st hour after the onset of exposure. The average respiratory retention of pyrene was found to be 61%. The 1-HP elimination process could be described by one-compartment model with the half-live of 9.8 hour (95% CI 7.9–11.7 h). The simulation of 1-HP elimination in urine during a working week (4 days) indicates that the balance between absorption and elimination is achieved at the end of the second day. Received: 29 July 1996 / Accepted: 21 February 1997     

Skin permeation of polycyclic aromatic hydrocarbons: A solvent-based in vitro approach to assess dermal exposures against benzo[a]pyrene and dibenzopyrenes

Consumer products with high contents of polycyclic aromatic hydrocarbons (PAHs) were repeatedly identified by market surveillance authorities. Since several of the individual compounds have been identified as genotoxic carcinogens, there might be health risks associated with the usage of these items. It therefore becomes reasonable to argue to reduce PAH contents in consumer products to a level as low as possible. This study presents data on the migration of PAHs from consumer products into aqueous sweat simulant or aqueous ethanol and on its combined migration and penetration into human skin. Product specimens were either submerged in simulant, or placed directly on test skins in Franz cell chambers to simulate dermal contacts. Migration of hexacyclic dibenzopyrenes became detectable by using ethanolic simulant, but not in aqueous sweat simulant. Similarly, migration of the pentacyclic model carcinogen benzo[a]pyrene (B[a]P) into aqueous sweat simulant was significantly lower when compared with human skin or skin models. The results point to a gross underestimation (about two orders of magnitude) when using aqueous sweat simulant instead of human skin for assessing PAH migration. On the other side, the usage of 20% ethanol as simulant revealed good agreement to the actual exposure of human skin against B[a]P migrating out of contaminated products. Our results underline that aqueous sweat simulant is not suitable to study dermal migration of highly lipophilic compounds.     

Exposure to polycyclic aromatic hydrocarbons in occupational versus urban environmental air

To evaluate the balance between occupational and environmental exposure to suspended particulate matter (SPM) and polycyclic aromatic hydrocarbons (PAHs), comparison measurements were performed in a coal-fired power plant and the urban atmosphere from the town nearby. Methods: The analysis of SPM for PAH content was done according to a high-performance liquid chromatography (HPLC)-based method. The microscopic assessment was performed using scanning electron microscopy (SEM) by silver coverage of the samples derived by air filter. Results: Contrary to expectations, the results showed low levels of particle-bound PAHs in the occupational environment (<1 ng benzo(a)pyrene/m³ air) and high levels in urban air (range 80–1250 ng benzo(a)pyrene/m³). The SPM collected from the power plant exhibited non-respirable characteristics (particles larger than 10 μm), whereas urban SPM almost exclusively contained respirable airborne particles (<3 μm). Conclusions: The PAH burden, combined with the enhanced probability of respiratory absorption, confers a much greater hazard potential to the urban SPM. Under these conditions, in areas or countries in which old technologies remain in use, occupational exposure to SPM containing PAHs might represent a severe underestimation of the total risk as it does not take into account the background air pollution. Received: 6 December 1997 / Accepted: 12 June 1998     

苯并[a]芘、铅染毒小鼠神经毒性及脑组织细胞凋亡的研究

目的研究苯并[a]芘(BaP)、铅及其联合作用引起的小鼠神经毒性及脑组织细胞凋亡.方法将80只昆明小鼠随机分为10组,即:①未处理对照组;②溶剂(植物油)对照组;③低浓度铅(5.4 mg/L,饮水)染毒组;④高浓度铅(54 mg/L,饮水)染毒组;⑤低剂量BaP(0.5 mg/kg体重,每周4次腹腔注射)染毒组;⑥高剂量BaP(5 mg/kg体重,每周4次腹腔注射)染毒组;⑦低浓度铅 低剂量BaP联合染毒组;⑧低浓度铅 高剂量BaP联合染毒组;⑨高浓度铅 低剂量BaP联合染毒组;⑩高浓度铅 高剂量BaP联合染毒组.实验中观察记录一般情况,处理8周后测脑脏器系数,TUNEL法检测小鼠脑组织细胞凋亡率并对凋亡率与脑脏器系数进行相关分析.结果①高浓度BaP单独染毒组(0.98%)及各联合染毒组(0.95%、0.93%、0.92%、0.90%)小鼠脑组织脏器系数低于对照组(1.08%、1.08%, P＜0.05～P＜0.000 1).②除低剂量铅单独染毒组外的其余各染毒组小鼠脑组织细胞凋亡率(11.91%、18.09%、44.62%、20.12%、60.84%、52.28%、90.17%)显著高于对照组(5.89%、5.87%, P＜0.000 1). ③细胞凋亡率与小鼠脑组织脏器系数间存在负相关关系(r=-0.827 1,P＜0.05).结论①BaP、铅对小鼠均有一定的中枢毒性,两者联合作用可使毒性增强;②细胞凋亡可能是BaP引起中枢神经系统毒性的机制之一.     

Effects of tributyltin, benzo[a]pyrene, and their mixture on antioxidant defense systems in Sebastiscus marmoratus

It has been reported that there is a metabolic interaction between tributyltin (TBT), an organometal used as an antifouling biocide, and benzo[a]pyrene (BaP), a widespread carcinogenic polycyclic aromatic hydrocarbon. This study was therefore designed to examine the potential in vivo influence of TBT, BaP, and their mixture on hepatic antioxidant defense systems of Sebastiscus marmoratus, which were given a single ip injection of TBT (0.5, 1, 5, and 10mg/kg), BaP (0.5, 1, 5, and 10mg/kg), or both in combination (0.5, 1, 5, and 10mg/kg); control fish received olive oil vehicle only. Samples were collected for biochemical analysis after injection for 7 days. Cotreatment with BaP caused a significant inhibition of TBT-mediated malondialdehyde contents elevation. Cotreatment with TBT decreased BaP-mediated glutathione peroxidase activity induction. Cotreatment with TBT and BaP did not significantly alter the reduced glutathione levels, which were significantly induced by TBT or BaP alone. TBT-induced suppression of BaP bioactivation or BaP-induced stimulation of the phase II metabolism of TBT and its biliary excretion, both of which have been reported previously, could explain the observed antagonism. The results suggest that combined exposure of TBT and BaP increases the vulnerability of the fish to oxidative stress. BaP cotreatment decreased the induction of glutathione S-transferase (GST) activity by the lower dose of TBT, while cotreatment with TBT and BaP at the highest dose (10mg/kg) resulted in inhibition of the GST activity by 4.8-fold. The results suggest that these biomarkers should be interpreted with caution in biomonitoring studies. Combined effects of TBT and BaP exposure at environmental levels on these biomarkers should be further researched.     

亚慢性染毒苯并[a]芘对大鼠海马热休克蛋白70表达的影响

目的 研究亚慢性染毒苯并[a]芘(B[a]P)对大鼠海马热休克蛋白70( Hsp70)的影响。方法 选择48只健康雄性SD大鼠,饲养1周后将动物随机分为空白对照组、溶剂对照组及0.5、1.5、4.5、10.0 mg/kg B[a]P染毒组,隔日腹腔注射染毒90d,Morris水迷宫试验进行行为学测试。免疫印迹(Westem-blot)法和反转录-聚合酶链反应(RT-PCR)法分别检测大鼠海马Hsp70蛋白和Hsp70 mRNA的表达水平,免疫组织化学法观察大鼠海马各区Hsp70表达情况,图像分析系统测定Hsp70平均灰度值。结果 Morris水迷宫结果显示,4.5、10.0 mg/kg B[a]P组大鼠平均潜伏期和平均总路程明显高于空白对照组、溶剂对照组、0.5、1.5 mg/kg B[a]P组,差异有统计学意义(P＜0.05);10.0 mg/kg B[a]P组平台象限滞留时间明显低于空白对照组、溶剂对照组和0.5mg/kg B[a]P组,差异有统计学意义(P＜0.05)。B[a]P染毒组大鼠海马hsp70蛋白和hsp70 mRNA的表达水平均随染毒剂量增高而下降。其中,4.5、10.0 mg/kgB[a]P组Hsp70蛋白表达水平明显低于空白对照组、溶剂对照组和0.5mg/kg B[a]P组,差异有统计学意义( P＜0.01,P＜O.05);0.5、4.5、10.0 mg/kg B[a]P组hsp70 mRNA表达水平明显低于空白对照组、溶剂对照组,差异有统计学意义(P＜0.01),10.0 mg/kg B[a]P组hsp70 mRNA表达水平明显低于其他染毒组。免疫组化结果显示,海马CA1、CA2、CA3、DG区Hsp70表达平均灰度值均随着B[a]P染毒剂量的增高而呈下降趋势。Pearson相关性分析结果显示,大鼠海马Hsp70表达水平与平均潜伏期呈负相关（r=-0.428,P＜O.05）;大鼠海马hsp70 mRNA表达水平与平均潜伏期、平均总路程呈负相关(r值分别为-0.677、-0.594,P＜0.01),与平台象限滞留时间呈明显正相关(r=0.597,P＜O.01)。结论 亚慢性染毒B[a]P可抑制大鼠海马Hsp70的表达,与大鼠学习记忆和空间探索能力损害相关,该抑制作用无区域特异性。     

Enhanced Bio-Mineralization by Riboflavin Photosensitization and its Significance to Detoxification of Benzo[a]pyrene

In this study, ¹⁴C-benzo[a]pyrene (BaP) was chosen as a model compound to investigate if photosensitization by riboflavin enhances the subsequent microbial mineralization of polycyclic aromatic hydrocarbons (PAHs) in natural aquatic environments. After photolysis, BaP showed an increased toxicity to human epithelial cell and natural microbial assemblage. However, BaP mineralization rate in a river water sample containing riboflavin is roughly twice of that without riboflavin after the 2-day incubation. Thus, the results imply that microbial assemblage can mineralize BaP photoproducts to carbon dioxide and a combination of riboflavin photosensitization and microbial degradation could lead to complete detoxification of PAHs.     

孕期暴露苯并[a]芘对子代大鼠神经发育毒性的影响

Objective To study the effects of prenatal exposure to benzo[a]pyrene (B[a]P) on the physical development, early behavioral development, the adaptability to new environment and the learning and memory ability of rat offspring. Methods Pregnant rats were randomly divided into five groups: control group,olive oil group, 3 exposure groups (25, 50 and 100 mg/kg B [a]P). The rats were exposed to B [a]P) by intraperitoneal injection on the 17th-19th days during gestation. The offspring were weighed on postnatal days (PND)1, PND 4, PND 7 and PND 28, the indices of physical development, reflective ability and sensory function were detected for offspring, the Morris water maze and Open-field tests were used to measure the ability of learning and memory and the adaptability to new environment of offspring. Results The time of ear opening in middle and high-dose groups[(4. 1 ±0.4),(5.0±0.4) d] was posterior to that in untreated and solvent groups[(3.3±0.5),(3.4±0.6) d](P＜0.01).The attainment rate (6.5%) of the surface righting reflex test in highdose group on the 4th day was significantly lower than that (36.1%) in untreated group, the attainment rate(50.0%) in high-dose group on PND7 was significantly lower than those (81.3% and 79.3%) in untreated group and solvent group (P＜0.05). Compared to the untreated group, the time of forelimb hanging test in all exposure groups on PND12 and PND14 significantly decreased; compared to the solvent group the time of forelimb hanging test decreased in high-dose group on the 14th day significantly decreased (P＜0.01). The attainment rate (61.9%) of olfactory discrimination in high-dose group on PND 12 was significantly lower than that (94.3%) in untreated group (P＜0.05). The results of morris water maze test showed that the escape latency of different dose groups significantly increased, and the time of spatial probe and the times of traversing flat in high-dose group decreased significantly, as compared to the untreated and solvent groups(P＜0.01). The results of open-field test indicated that the center retention time in middle and high-dose groups significantly prolonged, the times of crossing lattice obviously reduced, and the rearing times decreased in high-dose group,as compared to untreated(P＜0.05).Compared to the solvent group, the times of crossing lattice in all exposure groups reduced significantly(P＜0.01 or P＜0.05). Conclusion The prenatal exposure to B[a]P could inhibit the physical development and early behavioral development, and influence the adaptability to new environment and learning and memory ability for offspring.     

苯并（a）芘染毒大鼠血、肺中SOD、LPO及SOD／LPO变化的研究

用苯并（ａ）芘对大鼠经气管内染毒，在染毒后９０、１８０、２７０、３６０和５４０天时动态观察肺组织和血中超氧化物歧化酶（ＳＯＤ）、脂质过氧化物（ＬＰＯ）及ＳＯＤ／ＬＰＯ比值的变化．结果表明，苯并（ａ）芘可诱发大鼠血和肺组织中自由基反应增强，ＳＯＤ活性降低，脂质过氧化反应增高，ＬＰＯ含量升高，抗氧化能力降低，ＳＯＤ／ＬＰＯ比值降低．体内抗氧化和脂质过氧化平衡失调．     

苯并[a]芘在大鼠脑组织中分布动态观察

目的 观察[¹⁴C]标记苯并[a]芘在大鼠脑内的分布。方法 将100只SD大鼠随机分为对照组和实验组,实验组尾静脉注射[¹⁴C]标记苯并[a]芘3.7×10⁵Bq/kg,对照组注射等量生理盐水;在注射后1,6,12,24和48 h用光镜放射自显影法观察苯并[a]芘在脑内的分布情况。结果 与对照组比较,实验组大鼠脑组织注药后1 h即有苯并[a]芘银粒出现,随着时间的增加,银粒数也逐渐增加,24 h达高峰,48 h明显减少,注射后1,6,12,24,48 h银粒数分别为(17.68±1.79),(22.67±2.15),(25.79±2.55),(32.33±2.78),(18.01±1.68)粒,差异有统计学意义(F=69.67,P<0.01);银粒在脑中分布不均匀,注药后1 h主要集中在海马,6 h主要集中于大脑皮层,12 h则分布于纹状体;各时相脑组织中神经元银粒数明显高于神经胶质细胞(P<0.05)。结论 苯并[a]芘在各脑区的分布不均匀,24 h达到高峰,神经元细胞可能是苯并[a]芘靶细胞。     

磷酸化Rb在苯并[a]芘致皮质神经元凋亡中的作用

目的探讨磷酸化视网膜母细胞瘤蛋白（phospho-retinoblastoma protein,p-Rb）在苯并[a]芘致神经元凋亡过程中的作用。方法用CCK-8试剂检测细胞活性;Annexin—V／PI双染法检测细胞凋亡率,免疫荧光细胞化学染色观察p．Rb表达,免疫印记法（Western-blot）检测p—Rb的表达情况。结果与对照组相比,各染毒组神经元活性降低（P〈0．05）,凋亡率增高（P〈0．05）,p-Rb表达量增高（P〈0．05）,2μmol／L组加入抑制剂后细胞活性增强（P〈0．05）,凋亡率下降（P〈0．05）,p—Rb阳性细胞数减少（P〈0．05）,p-Rb表达量降低（P〈0．05）。结论苯并[a]芘可引起磷酸化Rb水平升高,是苯并[a]芘致神经元凋亡的可能机制。