戊酸雌二醇、炔雌醇环丙孕酮联合克罗米芬胶囊治疗多囊卵巢综合征80例观察

摘 要 目的：探讨戊酸雌二醇、炔雌醇环丙孕酮片联合克罗米芬胶囊对多囊卵巢综合征不孕症患者的治疗效果。方法: 多囊卵巢综合征不孕患者80例分为观察组和对照组,每组40例。对照组给予炔雌醇环丙孕酮片和克罗米芬胶囊治疗,观察组给予戊酸雌二醇片、炔雌醇环丙孕酮片和克罗米芬胶囊治疗,共4个月经周期。比较两组多囊卵巢综合征不孕患者临床表现,促卵泡素、黄体生成素和睾酮分泌情况,子宫内膜厚度,妊娠率和排卵率。结果: 治疗后,两组多毛、痤疮和月经失调的发生率均低于治疗前（P<0.05）,但两组间比较差异无统计学意义（P>0.05）;两组黄体生成素、睾酮和黄体生成素/促卵泡素水平明显低于治疗前（P<0.05）,但两组促卵泡素水平治疗前后无明显差异（P>0.05）,且两组黄体生成素、睾酮和黄体生成素/促卵泡素水平比较也无明显差异（P>0.05）。月经第5天,两组患者子宫内膜厚度比较差异无统计学意义（P>0.05）,而围排卵期,观察组子宫内膜厚度大于对照组（P<0.05）。观察组妊娠率高于对照组（65.0% vs. 35.0%,P<0.05）,两组排卵率比较差异无统计学意义（P>0.05）。结论:炔雌醇环丙孕酮片和克罗米芬胶囊与戊酸雌二醇、炔雌醇环丙孕酮片联合克罗米芬胶囊治疗均可降低多囊卵巢综合征不孕症患者的多毛、痤疮和月经失调症状,降低血清黄体生成素、睾酮和黄体生成素/促卵泡素。戊酸雌二醇、炔雌醇环丙孕酮片和克罗米芬胶囊治疗还可促进子宫内膜生长和提高妊娠率。     

大气细颗粒物PM2.5诱导肺上皮MLE-12细胞的氧化应激和自噬

目的 研究大气细颗粒物PM_2.5对小鼠肺泡Ⅱ型上皮细胞MLE-12细胞的氧化应激和自噬的影响。方法 用采集和处理的2009年北京市细颗粒物PM_2.5 25,50,100和200 mg·L^-1暴露处理MLE-12细胞24和48 h,用MTT比色法测定细胞存活率,双乙酰基二氯荧光素(DCFH-DA)荧光探针检测细胞内活性氧类(ROS)自由基生成,Western蛋白质印迹法检测微管相关蛋白1轻链3 Ⅰ蛋白(LC3Ⅰ)和LC3Ⅱ表达,激光共聚焦显微镜观察细胞自噬体的形成。结果 与细胞对照组比较,PM_2.5 100和200 mg·L^-1处理24 h组细胞存活率明显降低,分别降低28%和33%(P<0.01);暴露处理48 h,PM_2.5 25~200 mg·L^-1组细胞存活率均明显下降(P<0.01),并呈浓度效应关系(R²=0.4940,P<0.01)。PM_2.5 100和200 mg·L^-1处理MLE-12细胞3 h,胞内ROS水平较细胞对照组显著升高,分别升高27.6%和60.7%(P<0.01)。此外,PM_2.5 100 mg·L^-1处理细胞12,24和48 h及PM_2.5 50,100和200 mg·L^-1处理细胞24 h细胞自噬标志物LC3Ⅱ蛋白表达均明显增强,呈现明显的时间效应(R²=0.9150,P<0.01)和浓度效应(R²=0.6338,P<0.01)关系。PM_2.5 100 mg·L^-1处理24 h细胞内自噬体荧光强度明显升高(P<0.05),细胞核周边有明显的自噬泡环绕。结论 PM_2.5诱导肺泡Ⅱ型上皮MLE-12细胞的氧化应激,并引发细胞自噬。     

不同速率输注瑞芬太尼复合丙泊酚应用于鼻内窥镜手术控制性降压的临床观察

目的 探讨不同剂量瑞芬太尼复合丙泊酚在鼻内窥镜手术中的降压效果. 方法 ASA Ⅰ～ Ⅱ级择期鼻内窥镜手术患者45 例,随机分3组,每组15 例. R1,R2和R3组术中静脉泵入瑞芬太尼速率为0.19,0.28和0.37 μg&#8226;kg^-1&#8226;min^-1,术中以0.11 μg&#8226;kg^-1&#8226;min^-1的基础速率复合输注丙泊酚,根据脑电双频指数（BIS）值调节丙泊酚的输注速度,以维持稳定的麻醉深度. 分别在降压前（t₀） 、降压后20 min（t ₁ ） 、40 min（t₂） 及术毕（t ₃）作血气分析,并记录各时点的平均动脉压（MAP）、心率（HR）. 结果 在t1时点R2组和R3组HR、MAP均有显著下降（P＜0.05）; 而R1组在t ₂时点才出现显著下降（P＜0.05）; R1组和R3组的t ₂时点与t 1时点比较HR 、MAP差异有显著性（P＜0.05）,而R2组两时点HR、MAP差异无显著性（P＞0.05）. R3组术中t ₁、t ₂与术前t ₀比较,氧分压（PaO₂）、二氧化碳分压（PaCO₂）和乳酸（Lac）含量差异有显著性（P＜0.05）,而在其他两组差异无显著性（P＞0.05）. 结论 在鼻内窥镜手术中,以0.28 μg&#8226;kg^-1&#8226;min^-1输注瑞芬太尼复合丙泊酚,降压作用明显、平稳,并且不会造成组织灌注压过低导致组织低氧.     

过氧化氢存在下极谱催化波法测定桑枝中桑色素的含量

目的 考察过氧化氢(H₂O₂)存在下桑色素的极谱催化波,建立高灵敏度测定桑枝中桑色素含量的方法. 方法 采用电分析化学法. 结果 在0.2 mol&#8226;L^-1 醋酸 醋酸钠 (pH值5.3 ± 0.1)和2.0×10-2 mol&#8226;L^-1 H₂O₂最佳介质中,桑色素极谱催化波峰电流与其浓度在6.0×10^-8～3.0×10^-6 mol&#8226;L^-1范围内有线性关系. 结论 该方法灵敏度高、快速、简便,从电化学角度为桑色素清除氧自由基提供实验依据.     

丹参酮化合物对HeLa细胞抑制作用与构效关系探讨

目的 研究丹参酮化合物对HeLa细胞增殖的抑制作用,阐明其结构与细胞毒活性的关联性. 方法 MTT比色法检测不同浓度的丹参酮化合物,分别在24,48,72 h对HeLa细胞增殖的抑制作用. 结果 不同浓度（0.5～16.0 μg&#8226;mL^-1）丹参酮ⅡA、丹参酮I、二氢丹参酮、隐丹参酮对HeLa细胞毒性均有明显剂量和时间依赖性,其作用24 h的IC50值分别为：24.1,20.1,8.4,17.8 μg&#8226;mL^-1;作用48 h的IC₅₀值分别为：8.0,11.1,2.2,8.2 μg&#8226;mL^-1;作用72 h的IC₅₀值分别为：4.6,3.5,1.5,8.1 μg&#8226;mL^-1. 结论 丹参酮化合物对HeLa细胞均具有增殖抑制作用,其中二氢丹参酮对宫颈癌细胞系HeLa的细胞毒性最强. A环为芳香环与D环呋喃环的结构差别使其对HeLa细胞增殖的抑制作用有明显的差异.     

高效液相色谱法测定养血祛风丸中士的宁的含量

目的 建立高效液相色谱法测定养血祛风丸中士的宁的含量. 方法 采用HyperClone BDS C₁₈柱（250 mm×4.6 mm,5 μm）; 流动相：乙腈 0.01 mol&#8226;L^-1庚烷磺酸钠与0.02 mol&#8226;L^-1磷酸二氢钾等量混合溶液（用10%磷酸调节pH值为2.8）（21：79）; 流速1.0 mL&#8226;min^-1; 检测波长：260 nm; 柱温35 ℃. 结果 士的宁浓度的线性范围为4.8～191.8 μg&#8226;mL^-1,r＝0.999 8,回收率为101.30%,RSD＝1.3%. 结论 该方法方便、快速、准确,可用于养血祛风丸的质量控制.     

四氯化碳致大鼠肝损伤早期血清总胆汁酸、α-谷胱甘肽S转移酶、嘌呤核苷磷酸化酶和鸟氨酸氨基甲酰转移酶的变化

目的 探讨血清总胆汁酸、α-谷胱甘肽S转移酶(α-GST)、嘌呤核苷磷酸化酶(PNP)和鸟氨酸氨基甲酰转移酶(OCT)对肝损伤早期诊断的应用价值。方法 Wistar大鼠分别ig给予CCl₄ 0.02,0.05和0.08 ml·kg^-1,每天1次,连续10 d。于给CCl₄后第1天和第3天采集血清,采用全自动生化仪测定血清谷丙转氨酶(GPT)、谷草转氨酶(GOT)、碱性磷酸酶、总胆红素以及总胆汁酸的水平,采用ELISA测定血清α-GST, PNP和OCT的水平; 于给CCl₄后第10天,测定血清GPT、 GOT、碱性磷酸酶及总胆红素,计算大鼠的肝指数,观察肝组织病理学变化。通过Logistic回归建立回归模型,分别用ROC曲线分析给CCl₄后第1和第3天总胆汁酸、α-GST、PNP和OCT以及GPT和GOT对CCl₄致大鼠肝损伤早期诊断的意义。结果 与正常对照组同时间点相比,给CCl₄第1天,CCl₄ 0.02,0.05和0.08 ml·kg^-1组PNP均升高(P<0.05, P<0.01),CCl₄ 0.08 ml·kg^-1组GPT和总胆汁酸升高(P<0.05, P<0.01); 给CCl₄第3天,CCl₄ 0.05和0.08 ml·kg^-1组GOT, α-GST和OCT升高(P<0.05, P<0.01); 给CCl₄第10天,CCl₄ 0.02,0.05和0.08 ml·kg^-1组大鼠肝指数均显著升高(P<0.01),肝出现严重的肝细胞脂肪变性(P<0.01)。大鼠血清中GPT、GOT、总胆汁酸、α-GST、PNP和OCT的ROC曲线下面积分别为0.786, 0.728, 0.878, 0.629, 0.850和0.571。GPT和GOT联合检测的ROC曲线下面积为0.871;总胆汁酸、α-GST、PNP和OCT以及总胆汁酸、PNP和OCT联合检测的ROC曲线下面积为0.939,且均高于各指标单项检测。结论 总胆汁酸、α-GST、PNP和OCT可作为CCl₄致肝损伤早期的生物标志物,且总胆汁酸、PNP和OCT联合检测在CCl₄致肝损伤早期具有较高的诊断价值。     

复方丹参滴丸对过氧化氢损伤PC12细胞的保护作用

目的 研究复方丹参滴丸对过氧化氢（H₂O₂）损伤PC12细胞的保护作用,探讨复方丹参滴丸治疗缺血性脑血管病的作用机制.方法 采用细胞培养和H₂O₂诱导细胞损伤的方法,观察复方丹参滴丸对PC12细胞的保护作用.结果 复方丹参滴丸 6.25,12.50,25.00 μg&#8226;mL^-1均可明显对抗100和500 μmol&#8226;L^-1 H2O2引起的PC12细胞凋亡（P＜0.01）.结论 复方丹参滴丸可以促进PC12细胞的营养和增殖作用,并有抗H2O2诱导细胞凋亡作用.     

知母皂苷元通过上调PI3K/Akt信号通路减轻淀粉样β蛋白诱导的乳大鼠海马神经元突触损伤

目的 探讨知母皂苷元(Sar)减轻淀粉样β蛋白(Aβ)诱导的海马神经元突触损伤的信号转导机制。方法 取出生0~24 h SD乳大鼠海马神经元,体外培养7 d。海马神经元分别加入磷脂酰肌醇-3-激酶(PI3K)特异性阻断剂LY294002 30 μmol·L^-1或蛋白激酶B(Akt)特异性阻断剂曲西立滨1 μmol·L^-1 1 h后,加Sar 30和100 μmol·L^-1作用1 h,再加Aβ_1-42 50 nmol·L^-1作用24 h。应用突触囊泡蛋白(SYP)免疫荧光染色观察突触的改变。Western蛋白质印迹法检测海马神经元SYP、磷酸化Akt(p-Akt)和磷酸化糖原合成酶3β(p-GSK3β)表达水平的改变。结果 与正常对照组相比,Aβ_1-42组培养的海马神经元SYP, p-Akt和p-GSK3β表达水平明显减低(P<0.01)。与Aβ_1-42组相比,Aβ_1-42+Sar 30和100 μmol·L^-1组海马神经元SYP, p-Akt和p-GSK3β表达水平明显增加(P<0.01)。给予LY294002作用后,SYP和p-Akt表达水平明显降低(P<0.05)。给予曲西立滨作用后,SYP和p-GSK3β表达水平明显降低(P<0.05)。单独给予LY294002,海马神经元SYP表达无变化,p-Akt表达水平明显降低(P<0.01)。单独给予曲西立滨,海马神经元SYP和p-GSK3β蛋白表达水平明显降低(P<0.01)。结论 Sar通过上调PI3K/Akt/GSK3β信号通路对抗Aβ_1-42诱导的海马神经元突触损伤。     

培氟沙星与锌配合物的制备与抗菌活性测定

目的 制备培氟沙星与锌配合物,并观察其抗菌活性. 方法 氯化锌0.136 3 g和培氟沙星0.666 0 g化学反应制备配合物.对配合物进行元素分析和红外光谱分析,并进行体外抗菌活性测定. 结果 配合物各元素的组成比例为：锌(Zn) 13.16%,碳(C) 41.05%, 氢(H)4.02%, 氮(N)8.43%, 氯(Cl) 7.15%, 水(H2O )12.50%. 由元素分析可推知配合物的分子式为Zn₂(C₁₇H₂0FN₃O₃)₂Cl₂(H₂O)₇.培氟沙星在1 713 cm^-1处出现的羧基上C=O伸缩振动吸收峰,在配合物中消失,而在1 572.1,1 343.7 cm^-1则出现了对应于羧基的不对称和对称伸缩振动吸收峰,培氟沙星在1 634 cm^-1出现的萘啶环上羰基(C=O)伸缩振动吸收峰,在配合物中则红移了13个波数至1 621.2 cm^-1.培氟沙星对流感嗜血杆菌和粪链球菌的最低抑菌浓度分别为1.25和2.5 mg&#8226;mL^-1,配合物对流感嗜血杆菌、粪链球菌最低抑菌浓度分别为0.62和1.25 mg&#8226;mL^-1. 结论 该配合物制备方法简单,体外抗菌活性 结果表明, 培氟沙星与锌配合物对流感嗜血杆菌、粪链球菌的抑制作用略高于培氟沙星.     

吲哚美辛致小鼠胚泡着床障碍模型的建立

目的观察不同剂量吲哚美辛对小鼠胚泡着床的影响,建立吲哚美辛致小鼠胚泡着床障碍的动物模型。方法①体重(30±2)g成年♂昆明小鼠,妊娠d3、d4分别于9∶00和16∶00二次颈背部皮下注射0~5mg·kg-1剂量的吲哚美辛溶液,观察其对小鼠着床率和着床点数的影响以确定造模剂量。②选用每次注射433mg·kg-1作为造模剂量,比较对照组和模型组小鼠妊娠d8子宫重量,观察小鼠妊娠d5子宫内膜局部对伊文思蓝反应率和平均反应点数,放免法检测子宫内膜局部6ketoPGF1a的水平。结果①小鼠妊娠d3、d4每天2次皮下注射433mg·kg-1剂量的吲哚美辛开始有明显抗着床效应,着床率和着床点数明显降低(P<001),剂量达5mg·kg-1着床可完全阻断。②与对照组相比,小鼠妊娠d8模型组子宫重量明显减轻(P<001);小鼠妊娠d5模型组子宫内膜局部对伊文思蓝反应率明显降低(P<005),平均反应点数和6ketoPGF1a的水平明显降低(P<001)。结论于小鼠妊娠d3、d4每天2次皮下注射433mg·kg-1剂量吲哚美辛可用来建立胚泡着床障碍的动物模型,其抗着床与其抑制血管渗透性和抑制蜕膜化有关。     

Effect of a bradykinin potentiating fraction, from venom of the Egyptian scorpion, Buthus occitanus, on the ovaries and endometrium of mice

An isolated fraction with bradykinin potentiating activity, from the venom of the Egyptian scorpion Buthus occitanus, was injected i.p. in female mice (35 days old). Five days after the injection of a single sublethal dose (1 micrograms/g), the number of primary multilaminar and secondary follicles of the ovaries was increased. Graafian follicles were observed in 50% of the treated mice but in none of the control mice. The number and size of the uterine glands and endometrium were increased in treated mice. Concomitantly, estradiol was increased in the circulating blood, while progesterone was within the normal limits. The enhancement of cellular growth by the isolated venom fraction with bradykinin potentiating activity, could be attributed to an enhancement of some growth factor(s) responsible for stimulating the ovarian follicles and uterus.     

Effects of diethylstilbestrol on ovarian follicle development in neonatal mice

Previous results show that diethylstilbestrol (DES) causes polyovular follicles through estrogen receptor (ER) β and increases the number of follicles, suggesting that DES might affect follicular growth and development. Effects of neonatal DES exposure on follicle development were precisely examined in the ovaries of C57BL/6J and ERβ knockout (βERKO) mice. In the DES-exposed C57BL/6J mice, both primary follicle (PmF) progression from primordial follicles at 5 days of age and secondary follicle (SF) progression from PmFs at 10 days of age were delayed as compared with those in the oil-exposed controls. These results indicate that DES may suppress follicle development in neonatal mouse ovaries. DES exposure also decreased the number of follicles in 5-day-old C57BL/6J, WT and βERKO mice, suggesting that DES inhibits follicle formation and development through ERα in the neonatal mouse ovaries.     

Nicotine induced ovarian and uterine changes in albino mice

Nicotine at the dose level of 0.3 mg/100 g body weight was administered to normal cycling mice for 15 days through oral and intraperitoneal routes. At autopsy on 16th day significant reduction in the ovarian and uterine weight was observed. Histological observations showed decrease in the number and size of Graafian follicles, corpora lutea and increase in the atretic follicles in the ovary. The uterus showed absence of endometrial glands, decrease in the height of myometrium, endometrium and its epithelial cells. The total cholesterol content of the ovary and uterus is increased whereas the protein content is decreased. This antagonistic action of nicotine to gonadotrophins is discussed.     

Cigarette smoke exposure leads to follicle loss via an alternative ovarian cell death pathway in a mouse model

Cigarette smoking among reproductive-aged women is increasing worldwide. Cigarette smoking is a lifestyle behavior associated with important adverse health effects including subfertility and premature ovarian failure. We previously demonstrated that cigarette smoke (CS) exposure in mice decreases the primordial follicle pool; however, the mechanism of action is largely unknown. Therefore, the present study was designed to elucidate the mechanisms underlying CS exposure-induced ovarian follicle loss. CS exposure induced a significant decrease in the relative ovarian weight and the number of primordial and growing follicles. Oxidative stress, as shown by increased Hsp25 and decreased superoxide dismutase 2 protein expression, was found in mice exposed to CS for 8 weeks. Exposure decreased Bcl-2 but failed to induce apoptosis. An increased number of autophagosomes in granulosa cells of ovarian follicles together with increased expression of Beclin-1 and microtubule-associated protein light chain 3, key regulatory proteins in the autophagy (Atg) pathway, was found in CS-exposed mice compared with the control group. Taken together, our results suggest that CS exposure does not induce apoptosis but rather activates the Atg pathway ultimately leading to ovarian follicle loss. We further postulate that Atg is a novel mechanism of toxicant-induced ovarian follicle loss.     

Prenatal exposure to an environmentally relevant phthalate mixture accelerates biomarkers of reproductive aging in a multiple and transgenerational manner in female mice

Phthalates are known endocrine-disrupting chemicals that are found in many consumer products. Our laboratory previously developed a relevant phthalate mixture consisting of six phthalates and found that it disrupted female fertility in mice. However, it is unknown if prenatal exposure to phthalate mixtures can accelerate reproductive aging and if this occurs in multiple generations. Thus, we tested the hypothesis that prenatal exposure to a mixture of phthalates accelerates biomarkers of reproductive aging in multiple generations of female mice. Pregnant CD-1 mice were orally dosed with vehicle control or a phthalate mixture (20 μg/kg/day-500 mg/kg/day) daily from gestational day 10 to birth. Adult F1 females born to these dams were used to create the F2 and F3 generations by mating them with unexposed males. At 13 months, estrous cyclicity was monitored and ovaries and sera were collected for analysis. In the F1 generation, the mixture decreased testosterone and inhibin B levels, but increased follicle-stimulating hormone and luteinizing hormone levels compared to control. In the F2 generation, the phthalate mixture decreased the percent of antral follicles and testosterone hormone levels compared to control. In the F3 generation, prenatal exposure to the phthalate mixture increased ovarian weight, increased the time in metestrus/diestrus, altered follicle numbers, and decreased the levels of luteinizing hormone compared to control. Collectively, these data suggest that prenatal exposure to a phthalate mixture may accelerate several biomarkers of reproductive aging in a multi- and transgenerational manner in female mice.     

萘哌地尔衍生物BWYJ抗前列腺增生的作用

目的观察萘哌地尔衍生物BWYJ对前列腺增生模型的作用。方法采用激素法建立去势大鼠和未去势小鼠前列腺增生模型,通过小鼠前列腺湿重,计算前列腺指数。光镜及透射电镜下,分别观察小鼠前列腺组织形态学及超微结构变化;TUNEL法检测BWYJ对大鼠前列腺细胞凋亡的影响。结果BWYJ5、10、20mg·kg-1组均可降低BPH小鼠前列腺湿重指数(P<0.05),光镜及电镜结果表明,BWYJ5、10、20mg·kg-1组均可抑制小鼠组织结构增生性变化,且BWYJ10、20mg·kg-1组使腺腔直径、腺体表面积变小(P<0.05)。TUNEL检测发现,大鼠前列腺凋亡细胞检出率较低,与模型组比较,BWYJ各剂量组差异无统计学意义(P>0.05)。结论BWYJ具有抗小鼠及大鼠良性前列腺增生作用。     

Early effects of ovotoxicity induced by 4-vinylcyclohexene diepoxide in rats and mice

The industrial chemical 4-vinylcyclohexene diepoxide (VCD) causes specific destruction of oocyte-containing small preantral follicles (primordial and primary) in ovaries of rats and mice. The mouse seems more susceptible to ovotoxic effects of VCD than the rat. The purpose of this study was to better understand these species differences in susceptibility to VCD by comparing the initial rates of VCD-induced follicle damage and loss in response to dosing in both species. Female Fischer 344 rats and B6C3F1 mice (age, Day 28) were dosed daily (vehicle or 80 mg/kg, i.p.) for 6, 8, 10, or 12 d. Ovaries collected after the final dose were prepared for histologic evaluation. Primordial and primary follicles in ovarian slices were counted and classified as healthy or atretic. A VCD-dependent increase (P < 0.05) in percent atretic primordial follicles was first observed 4 h after the final dose in mice on Day 8 (VCD-treated, 44.4 +/- 3.1% vs. control, 26.9 +/- 5.4%). Conversely, in rats, this significant increase was not seen until Day 10 (VCD-treated, 44.3 +/- 1.3% vs. control, 23.1 +/- 4.0%). A VCD-dependent increase in percent atretic primary follicles was not observed in either species before Day 12. There was no significant effect on growing or preantral follicles on any day in either species. Significant loss of primordial and primary follicles (P < 0.05) was first measured on day 12 in both rats and mice. However, when compared with controls, follicle loss on that day was greater (P < 0.05) in mice (64.2 +/- 4.5%) than in rats (34.7 +/- 4.9%). Once VCD-dependent follicle loss was observed, the rate of follicle damage was similar in rats and mice, and was fairly constant in response to each dose. VCD-induced follicle damage in mice, as with rats, also displayed morphologic characteristics of atresia (apoptosis). In summary, follicle destruction seems to be similar in rats and mice; however, follicle damage is initiated earlier and to a greater extent in mice than in rats. Additionally, ovotoxic effects of VCD seem to initially directly target primordial follicles. These results provide temporal evidence that mice are more susceptible to VCD-induced ovotoxicity than rats.     

表没食子儿茶素没食子酸脂联合阿米福汀对小鼠卵巢的辐射保护作用

目的 探讨表没食子儿茶素没食子酸脂（EGCG）联合阿米福汀（WR-2721）对小鼠卵巢的辐射保护作用。方法 150只ICR雌性小鼠采用随机数字表法分成Sham组、Control组、WR组、EGCG组与WR+EGCG组，每组30只。除Sham组外，其余小鼠均接受6 Gy X线照射，在治疗后第8小时、7天和14天，分别测定每组小鼠的体质量、血细胞计数、卵巢指数及细胞凋亡，观察卵巢形态变化。结果 Control组小鼠辐射后体质量下降，Sham组小鼠体质量持续增长，WR、EGCG、WR+EGCG组小鼠体质量缓慢上升，差异有统计学意义（P<0.05）。Control组小鼠辐射后白细胞、红细胞、血小板水平较其余各组均下降，差异有统计学意义（P<0.05），各组小鼠血红蛋白水平辐射后变化不大（P>0.05）。Control组卵泡数量明显减少，出现大量闭锁卵泡或空卵泡，WR、EGCG组存在少量的闭锁卵泡或空卵泡，WR+EGCG组未见闭锁卵泡或空卵泡。Control组小鼠卵巢指数显著下降，EGCG组及WR+EGCG组小鼠卵巢指数高于Control组，差异有统计学意义（P<0.05）。Control组TUNEL染色阳性的闭锁卵泡及凋亡颗粒细胞最多。与Sham组比较，接受辐射的卵巢颗粒细胞凋亡指数均升高，Control组最高，EGCG组和WR组凋亡指数升高介于Control组及WR+EGCG组之间，差异有统计学意义（P<0.05）。结论 EGCG联合阿米福汀可以增强小鼠卵巢的辐射保护作用。     

Differences between rats and mice in the involvement of the aryl hydrocarbon receptor in 4-vinylcyclohexene diepoxide-induced ovarian follicle loss

Repeated dosing with the occupational chemical 4-vinylcyclohexene diepoxide (VCD) selectively depletes small pre-antral follicles in the ovaries of rats and mice via apoptosis. The aryl hydrocarbon receptor (AhR) plays a role in mediating the effects of several xenobiotics. Therefore, this study was designed to investigate a potential role of the AhR in VCD-induced ovotoxicity. Female F344 rats, C57BL/6 mice, or AhR-deficient (-/-, AhRKO) mice were dosed daily (15 days) with vehicle, VCD (80 mg/kg, i.p.) and/or the AhR antagonist, alpha-naphthoflavone (ANF; 80 mg/kg, i.p.). Compared with controls, VCD caused a 60% reduction (P < 0.05) in primordial and primary follicles in mice and rats. Concurrent dosing with ANF protected against the VCD-induced follicle loss in rats, but not in mice. As with AhR-intact mice and rats, VCD induced a 70% loss (P < 0.05) of small pre-antral follicles in AhRKO mice. AhR mRNA expression was increased (P < 0.05) by VCD dosing in small pre-antral follicles isolated from ovaries of rats but not mice. AhR protein in rats was increased by VCD dosing in oocyte nuclei in primordial and primary follicles when measured by immunofluorescence and confocal microscopy. In rat small pre-antral follicles, apoptosis-associated caspase-3-like activity was increased (P < 0.05) by VCD treatment, decreased (P < 0.05) by ANF treatment, and unaffected by VCD plus ANF treatment. VCD had no effect on expression of GST Ya1 or GST Ya2 mRNA or CYP 1A1 protein in rats. Taken together, these findings demonstrate a difference between rats and mice in the potential involvement of AhR as related to VCD-induced ovotoxicity. Whereas, AhR appears to be involved in rats, no evidence for a similar role in mice was obtained. Overall, these findings point out that there can be mechanistic species differences in ovarian responses to xenobiotic chemicals.