邻苯二甲酸(2-乙基己基)酯低剂量暴露对雄性小鼠生殖发育的影响

目的观察邻苯二甲酸(2-乙基己基)酯(DEHP)低剂量暴露对雄性小鼠的生殖发育毒性作用。方法将清洁级ICR小鼠随机分为4个DEHP染毒组(1/120LD50,1/60LD50,1/30LD50,1/15LD50,即250、500、1000、2000mg/kg组),阳性对照组(环磷酰胺,40mg/kg)和阴性对照组(玉米油);DEHP染毒组和阴性对照组灌胃染毒30d,阳性对照组灌胃5d,每天1次。采用一般生殖毒性试验和显性致死突变试验的方法,研究DEHP低剂量暴露对雄性小鼠生殖细胞的损伤作用和生育能力的影响。结果一般生殖毒性试验结果显示,低剂量DEHP染毒雄性小鼠睾丸、附睾重量降低(r睾丸=-0.578,P<0.01;r附睾=-0.661,P<0.01),精子总数减少(r=-0.608,P<0.01),精子畸形率增高(r=0.836,P<0.01),精子活动度降低(P<0.05)。显性致死突变试验研究结果显示,250mg/kgDEHP染毒组小鼠受孕率降低,但与阴性对照组比较,差异无统计学意义(P>0.05);DEHP染毒组子代小鼠平均着床数减少(r=-0.892,P<0.01),与阴性对照组比较,差异均具有统计学意义(P<0.01);平均早期死亡胚胎数随染毒剂量增加而增加(r=0.996,P<0.01),250mg/kg染毒组与阴性对照组比较,差异无统计学意义(P>0.05)。结论DEHP低剂量暴露对雄性小鼠具有生殖毒性作用,可降低小鼠受孕率,使子代小鼠平均着床数减少,平均早期死亡胚胎数增加。     

Estrogenic activities of di-2-ethylhexyl phthalate

Phthalate esters are widespread in the environment. They have been described as being one of the most abundant man-made environmental contaminants that may be adverse to human health. Particularly, di-2-ethylhexyl phthalate (DEHP) has been shown to cause reproductive and developmental toxicity and is suspected to be an endocrine disruptor. The primary objective of this study is to determine the estrogenic activity of DEHP. Estrogenic activities of DEHP were studied by in vitro assays of human breast cancer MCF-7 cell proliferation. Estrogen-dependent MCF-7 cells were grown in RPMI1640 medium containing 10% fetal bovine serum. Five days before the addition of the test compounds, the cells were washed by phosphate balanced solution (PBS), and the medium was substituted with a phenol red-free RPMI1640 medium containing 5% dextral charcoalstripped Fetal Bovine Serum (FBS). Fresh medium was added to the respective test compounds and the control cell received only the vehicle (ethanol). The proliferation of MCF-7 cell was analyzed by the MTT assay, growth curves, mitotic index and colony forming efficiency. Compared with the ethanol control cells, the proliferation of tested cells treated with DEHP, like estradiol, was significantly enhanced and the activity of the cell proliferation reached the maximum at 1 × 10⁻³ mol/L DEHP. The relative proliferative potency of DEHP was 0.000 001 with a relative proliferative effect of 97.32%. During the log phase, the mitotic index of the tested cells treated with DEHP and estradiol was significantly increased. The cell cloning efficiency was enhanced, which was treated by 10⁻³ mol/L DEHP only for 48 hours. The results show a time-dependent and dose-dependent model. Di-2-ethylhexyl phthalate enhanced the proliferation of human breast cancer MCF-7 cells in vitro and might demonstrate an estrogenic activity.     

邻苯二甲酸二丁酯对小鼠卵巢功能的影响

目的 探讨邻苯二甲酸二丁酯(DBP)对小鼠卵巢功能的影响.方法 将雌性昆明小鼠按体重随机分为高(400mg/kg)、中(40mg/kg)、低(4mg/kg)DBP染毒组和1个溶剂对照组(二甲基亚砜),每组12只.每天腹腔注射1次[0.005ml/(g·d)],连续染毒14 d.阴道脱落细胞涂片法观察动情周期变化,分别于终止染毒第1、14天处死,分离卵巢和子宫,计算脏器系数,并观察卵巢病理变化.结果 各染毒组小鼠食欲降低,活动减少,精神萎靡不振,被毛光泽度下降,部分出现腹泻,体重下降;对照组小鼠食欲正常,精神较好,被毛有光泽.不同组别小鼠染毒前、后体重比较,差异均无统计学意义(P＞0.05).中、高剂量组小鼠动情后期、间期和动情周期时间长于对照组,高剂量组小鼠动情后期、间期和动情周期时间长于低剂量组,差异均有统计学意义(P＜0.05).随着染毒剂量的增加,小鼠动情周期时间逐渐延长,呈剂量-反应关系(rs=0.677,P＜0.001),并主要表现为动情后期和动情间期的延长.终止染毒第1、14天,高、中、低剂量组小鼠卵巢脏器系数与对照组相比,差异均无统计学意义(P＞0.05).终止染毒第1天,高、中、低剂量组小鼠子宫脏器系数高于对照组,终止染毒第14天,中、高剂量组小鼠子宫脏器系数高于对照组,差异均有统计学意义(P＜0.05).随染毒剂量的增高,子宫脏器系数有升高的趋势,呈剂量-反应关系(rs=0.564,P=0.010).终止染毒第1天,高剂量组成熟卵泡数低于对照组,中、高剂量组闭锁卵泡数高于对照组,差异均有统计学意义(P＜0.01);终止染毒第14天,高剂量组闭锁卵泡数高于对照组,差异有统计学意义(P《0.05).结论 邻苯二甲酸二丁酯可影响小鼠卵巢的内分泌功能及卵泡的发育、成熟,在体内可能具有类雌激素作用.     

邻苯二甲酸二丁酯和苯并[a]芘联合染毒对大鼠睾丸支持细胞的影响

目的 探讨邻苯二甲酸二丁酯和苯并[a]芘单独或联合染毒对大鼠睾丸支持细胞增殖及乳酸分泌的影响.方法 分离纯化大鼠睾丸支持细胞,油红O染色鉴定纯度大于90%后,以0.1、1、10、100、500 μg/ml 邻苯二甲酸二丁酯(dibutyl phthalate,DBP)和0.01、0.1、1、10、100 μg/ml 苯并[a]芘[benzo(a)pyrene,BaP]单独或依次联合染毒,用MTT法检测对支持细胞增殖的影响,并测定培养液中的乳酸含量.结果 DBP染毒后100 μg/ml 吸光度值[D(490)]显著高于对照组(P＜0.01).BaP染毒各组的D(490)值与对照组比较,无统计学差异.DBP 100 μg/ml BaP 10 μg/ml联合染毒组D(490)值显著上升(P＜0.01),而在500 μg/ml DBP 50 μg/ml BaP联合染毒组D(490)值下降(P＜0.05).DBP 100 μg/ml组、 100 μg/ml DBP 10 μg/ml BaP组和500 μg/ml DBP 50 μg/ml DBP组培养液中的乳酸显著增加(P＜0.01),DBP 500 μg/ml组和BaP 50 μg/ml组培养液中的乳酸含量也增高(P＜0.05).结论 一定剂量DBP可刺激支持细胞增殖并增加乳酸分泌,BaP虽不抑制支持细胞的增殖,但可使乳酸分泌增加.二者联合作用,在一定剂量下以DBP的效应为主,但在高剂量可抑制细胞增殖,呈现一定的协同作用.     

Colon delivery of prednisolone based on chitosan coated polysaccharide tablets

Colon drug delivery is advantageous in the treatment of colonic disease and oral delivery of drugs unstable or suceptible to enzymatic degradation in upper GI tract. In this study, multilayer coated system that is resistant to gastric and small intestinal conditions but can be easily degraded by colonic bacterial enzymes was designed to achieve effective colon delivery of prednisolone. Variously coated tablets containing prednisolone were fabricated using chitosan and cellulose acetate phthalate (CAP) as coating materials. Release aspects of prednisolone in simulated gastrointestinal fluid and rat colonic extracts (CERM) were investigated. Also, colonic bacterial degradation study of chitosan was performed in CERM. From these results, a three layer (CAP/Chitosan/CAP) coated system exhibited gastric and small intestinal resistance to the release of prednisolone in vitro most effectively. The rapid increase of prednisolone in CERM was revealed as due to the degradation of the chitosan membrane by bacterial enzymes. The designed system could be used potentially used as a carrier for colon delivery of prednisolone by regulating drug release in stomach and the small intestine.     

邻苯二甲酸二(2-乙基己)酯可抑制人原代培养早孕绒毛滋养细胞浸润能力

目的研究邻苯二甲酸二(2-乙基己)酯[di-(2-ethylexyl)phthalate,DEHP]对人原代培养早孕绒毛滋养细胞浸润能力及基质金属蛋白酶(MMP)-2、MMP-9、基质金属蛋白酶抑制因子(TIMP)-1和TIMP-2表达的影响,探讨DEHP对妊娠的影响及其毒性机制。方法2006年12月于北京军区总医院采用不同浓度DEHP预处理人原代培养早孕绒毛滋养细胞,采用Transwell体外浸润实验检测滋养细胞浸润能力的改变。RT-PCR法检测滋养层细胞侵袭性相关基因MMP-2、MMP-9、TIMP-1和TIMP-2的mRNA表达,Western blot法检测MMP-2、MMP-9、TIMP-1和TIMP-2蛋白表达水平。结果DEHP作用后,滋养细胞浸润能力明显下降。当DEEP为50、100μmol/L时,MMP-2、MMP-9、TIMP-1的mRNA表达量分别为0.35±0.07、0.26±0.03、0.97±0.18和0.30±0.10、0.20±0.05、1.02±0.20,与DEEP为0时MMP-2、MMP-9、TIMP-1的mRNA的表达量(0.77±0.15、0.45±0.04、0.80±0.20)比较P<0.05;当DEEP为50、100μmol/L时,MMP-2、MMP-9、TIMP-1的蛋白表达量分别为0.35±0.03、0.23±0.05、0.69±0.08和0.24±0.02、0.17±0.02、0.86±0.10,与DEEP为0时MMP-2、MMP-9、TIMP-1的mR-NA的表达量(0.69±0.04、0.57±0.03、0.24±0.12)比较P<0.05。可见DEHP剂量≥50μmol/L时,滋养细胞MMP-2和MMP-9表达下降、TIMP-1表达增加,且呈剂量依赖性。结论DEHP可通过抑制MMP-2、MMP-9的表达并促进TIMP-1表达,影响滋养细胞浸润能力。     

邻苯二甲酸二丁酯对雌性果蝇生殖能力的双重作用

目的:研究邻苯二甲酸二丁酯(DBP)对雌性果蝇生殖能力的影响。方法:将未交配雌性果蝇随机分为5组,分别给予含DBP0.1%(A组)、0.4%(B组)、1.6%(C组)和6.4%(D组)的培养基喂饲,并以0浓度为对照组(E组),在染毒10d和20d时,分别取存活的实验雌蝇进行交配试验和生育试验。结果:各染毒组雌蝇1h交配率随DBP浓度升高而降低,D组雌蝇的1h交配率和生育率明显低于E组(P<0.05或P<0.01);染毒10d后C组的平均生育数和平均产卵期均明显高于E组(P<0.05),且在连续产卵10d后仍保持较高的生育水平;染毒20d后D组的平均生育数和平均产卵期均明显低于E组(P<0.01)。结论:DBP对雌蝇生殖能力有双重作用,较低浓度可提高雌性果蝇生育力,而较高浓度可使其生育能力降低。     

羟丙基甲基纤维素酖酸酯游离膜的透湿性及附加剂影响

 考察了羟丙基甲基纤维素酞酸酯（HPMCP）的游离薄膜对水蒸汽的穿透性能，并与日本同类产品HP－50、HP－55及国产丙烯酸树酯（Ⅲ）的透湿性作比较，同时考察了几种附加剂对HPMCP游离膜透湿性的影响。由膜的透湿系数K可以看出：HPMCP（K＝0．253）与HP－50（K＝0．254）及HP－55（K＝0．266）的透湿性相近，而大于丙烯酸树酯（Ⅲ）（K＝0．056）。增塑剂邻苯二甲酸二丁酯与蓖麻油对HPMCP膜的透湿性影响均呈现一最小透湿值（在含量为10％时），随后随含量增加透湿值反而增大。含有不溶性填加剂（钛白粉、滑石粉）的HPMCP膜的透湿性均有不同程度的下降。     

Plasticizers in denture soft-lining materials: leaching and biodegradation

The leaching from soft lining materials into an aqueous buffer with the same esterase activity as that of saliva was compared to leaching into buffer without esterase activity. The buffer contained 0.1% non-ionic detergent, giving the medium a capacity to dissolve phthalates to the same degree as saliva. The hypothesis that esterase in the immersion medium will increase the rate of diffusion of plasticizers from denture soft-lining material was confirmed by the results. The average leaching of phthalates from 5.5 g of one of the materials was 4.5 mg kg(-1) d(-1) within the first 2 d and 1.1 mg kg(-1) d(-1) within the first 28 d. These levels may be compared to a LOAEL (lowest observed adverse effect level) of 52 mg kg(-1). It is concluded that an esterase activity, equivalent to that in saliva, in the immersion medium for soft lining materials increased the rate of diffusion of plasticizer from the materials. The measured levels of phthalates leaching from these materials might in vivo only be slightly less than 1/10 of the LOAEL given above.     

The occurrence of phthalic acid esters in various samples of commercially available sodium chloride injections (Indian Pharmacopoeia)

Conclusive evidence for the presence of di(2-ethylhexyl) phthalate (DEHP) in 15 out of 17 commercially available sodium chloride injection (Indian Pharmacopoeia) samples was obtained and concentration levels as high as 11.0 mg/500 ml of saline were detected. The presence of such contaminants in i.v. fluids, avoidable by the institution of appropriate quality control measures before manufacturing and marketing, is a matter of serious concern. Stipulated specifications in different pharmacopoeias are meant only to ensure sterility and pyrogenicity, and it is advisable that cognisance be taken of the presence of toxic contaminants e.g. di(2-ethylhexyl) phthalate.