罗汉果甜甙对雄性小鼠的遗传毒性研究

目的探讨罗汉果甜甙对成年雄性小鼠的遗传毒性。方法采用小鼠急毒试验、小鼠精子畸形试验、小鼠骨髓微核试验及生殖、淋巴器官重量指数分析方法。结果罗汉果甜甙对小鼠经口最大耐受剂量为15异／kg。微核试验与精子畸形试验的小鼠均分6组，即罗汉果甜甙0．25g／kg、0．5g,／kg、1g/kg、5g／kg4个剂量组与阴、阳性对照组。其中灌胃剂量为0．25g/kg、0．5g／kg、1g／k时，均来诱发各组的精子畸形率、骨髓微核率增高，与阴性对照组相比均无显著差异（P〉0．05）：当剂量达到5g／kg时诱发的精子畸形率、骨髓微核率均略高于阴性对照组但有显著性差异（P〈0．05）。灌胃罗汉果甜甙各剂量组小鼠的生殖与淋巴器官重量指数与阴性对照组相比无显著变化（P〉0．05）。结论经口最大给药剂量为15g／kg，属于无毒级。罗汉果甜甙以0．25g／kg，0．5g／kg及1g/kg的剂量灌胃小鼠无生殖与遗传毒性。当灌胃剂量达到5g／奴时，则对雄性小鼠有轻微的潜在遗传毒性。     

桑寄生的遗传毒理学研究

目的探讨桑寄生水煎液对成年小鼠及胚胎鼠的遗传毒性。方法采用小鼠急毒试验、小鼠骨髓嗜多染红细胞微核试验、小鼠胚胎肝转移微核试验、小鼠精子畸形试验。结果桑寄生水煎液急毒试验为无毒级,最大用药剂量为40g/kg。桑寄生水煎液灌胃孕鼠和雄鼠40g/kg剂组诱发的胚胎肝微核率、骨髓微核率、精子畸形率均较阴性对照组显著升高（P〈0.05）。桑寄生灌胃剂量为20g/kg时,诱发的胚胎肝微核率较阴性对照组显著升高（P〈0.05）,骨髓微核率与精子畸形率与阴性对照组相比,无显著性差异（P〉0.05）。而10g/kg剂量组诱发的胚胎肝微核率、骨髓微核率、精子畸形率与阴性对照组相比,均无显著性差异（P〉0.05）。结论桑寄生水煎液40g/kg剂量组对成年小鼠和胎鼠均具有潜在的遗传毒性,20g/kg剂量组仅对胎鼠有潜在的遗传毒性,10g/kg剂量组对成年小鼠和胎鼠均无遗传毒性。     

氟西汀对雄性小鼠的遗传毒性研究

目的探讨氟西汀对成年雄性小鼠的遗传毒性。方法采用小鼠急毒试验、小鼠精子畸形试验、小鼠骨髓微核试验及生殖、淋巴器官重量指数分析方法。结果氟西汀对小鼠经1：2最大耐受剂量为15g／kg微核试验与精子畸形试验的小鼠均分6组,即氟西汀1.25mg／kg,2.5mg／kg,5mg／kg和10mg／kg4个剂量组与阴、阳性对照组其中灌胃剂量为1.25mg／kg、2.5mg／kg、5mg／kg时,均未诱发各组的精子畸形率、骨髓微核率增高,与阴性对照组相比均无显著差异（P〉0.05）;当剂量达到10g／kg时诱发的精子畸形率、骨髓微核率均略高于阴性对照组但有显著性差异（P〈O.05）;灌胃氟西汀各剂量组小鼠的生殖与淋巴器官重量指数与阴性对照组相比无显著变化（P＞O.05）。结论经口最大给药剂量为15g／kg,属于无毒级。氟西汀以1.25mg／kg、2.5mg／kg、5mg／kg的剂量灌胃小鼠无生殖与遗传毒性,当灌胃剂量达到30g／kg时,则对雄性小鼠有轻微的潜在遗传毒性。     

柠檬黄对雄性小鼠生殖细胞的影响

探讨柠檬黄对雄性小鼠生殖细胞的影响.选择成年雄性小鼠分别给予柠檬黄0.25 g/kg(低剂量组)、0.5 g/kg(中剂量组)和1 g/kg(高剂量组),连续灌胃染毒5d,通过小鼠精子畸形率、精细胞微核率以及睾丸形态变化等方面来评价柠檬黄对生殖细胞的影响.与对照组相比,高剂量组的柠檬黄能引起小鼠精子的畸形率和精细胞微核...     

柠檬黄对雄性小鼠生殖细胞的影响

探讨柠檬黄对雄性小鼠生殖细胞的影响。选择成年雄性小鼠分别给予柠檬黄0.25 g/kg（低剂量组）、0.5 g/kg（中剂量组）和1 g/kg（高剂量组）,连续灌胃染毒5 d,通过小鼠精子畸形率、精细胞微核率以及睾丸形态变化等方面来评价柠檬黄对生殖细胞的影响。与对照组相比,高剂量组的柠檬黄能引起小鼠精子的畸形率和精细胞微核率升高（P〈0.05）,而中、低剂量组没有明显的变化（P〉0.05）。睾丸组织切片显示,低、中剂量组与对照组相比没有明显改变,高剂量组小鼠睾丸生精小管管腔内可见精子数量减少,生精上皮细胞层次减少,部分细胞有浓缩、溶解等坏死样变。高浓度的柠檬黄能使雄性小鼠精子畸形率增加,并造成雄性小鼠精细胞微核率上升,有一定的致突变性。     

中药对小鼠骨髓细胞遗传物质影响的实验研究

目的：通过本实验研究探讨孕妇禁忌中药红花、牛膝对小鼠骨髓嗜多染红细胞微核频率的影响。方法：选用体重18－22g的雌性昆明小鼠，随机分为8组，即阴性、阳性对照组分别灌胃生理盐水及腹腔注射环磷酰胺30mg/kg；实验组用红花及牛膝水煎剂分别以10g/kg,2g/kg,1g/kg的剂量灌胃，每天1次，连续5天后断颈取骨髓细胞涂片观察。结果：各用药剂量组诱发小鼠骨髓细胞微核率虽有数目的差异，但与阴性对照组相比无显著性差异（P＞0．05），而与阳性对照组相比均有显著差异，P＜0．01。讨论：本实验结果提示孕妇禁忌中药红花、牛膝虽有影响生育的作用，但是无明显诱发小鼠骨髓微核率增高的作用，表明这两种中药均无损伤遗传物质的作用。     

巴豆对小鼠骨髓及胚胎肝细胞微核率的影响

目的　探讨孕妇禁忌中药巴豆水提液对小鼠骨髓细胞及胚胎鼠肝细胞微核率的影响。方法　采用小鼠骨髓嗜多染红细胞微核 (MN)实验法与小鼠胚胎肝转移微核实验法。结果　当小鼠用药剂量在 1g/kg、 5g/kg时微核率与阴性对照组无明显差异 ,10g/kg时诱发骨髓MN率与阴性对照组比较有显著差异 (P <0 0 1)。而在孕鼠用药 1g/kg、 5g/kg、 10g/kg各剂量组时均可诱发鼠胎肝细胞微核率增高。各组与阴性对照组比较都有显著性差异 (P <0 0 1)。结论　实验表明巴豆水提液在同等剂量作用下诱发胚胎鼠肝细胞微核率明显高于成年鼠骨髓细胞 ,此药可以通过胎盘屏障 ,对胎鼠具有更为显著的致遗传物质损伤作用。     

装潢居室空气中挥发性化学物的遗传毒性研究

目的探讨装潢居室空气中挥发性化合物的遗传毒性.方法采用小鼠骨髓嗜多染性红细胞微核试验和小鼠精子畸形试验检测居室装潢后空气中挥发性化合物的遗传毒性.结果该化合物对小鼠呼吸系统有明显的刺激作用,与阴性对照组比较,高、中、低剂量组的微核率、精子畸形率均有显著差异性.结论居室装潢后室内空气中挥发性化合物具有遗传毒性作用.     

富氢水和乳果糖对环磷酰胺处理小鼠的抗突变作用

目的 探讨饮用富氢水和产氢营养因子乳果糖的抗突变作用.方法 36只昆明小鼠按随机数字表分为6组,每组6只.空白对照组(A组)、富氢水处理组(B组)和乳果糖处理组(C组):这3组小鼠正常喂养3d后,第4～6天分别使用蒸馏水、富氢水、50％乳果糖灌胃(20 ml/kg、1次/d);抗生素对照组(D组)、抗生素+富氢水处理组(E组)和抗生素+乳果糖处理组(F组):这3组小鼠第1～3天随意饮用含抗生素的水,第4～6天分别使用蒸馏水、富氢水、50％乳果糖灌胃(20 ml/kg、1次/d).末次灌胃后小鼠腹腔注射环磷酰胺(0.04 g/kg),24 h后处死小鼠,进行骨髓细胞学检查,计算各组小鼠骨髓嗜多染红细胞微核率.结果 A、B、C、D、E、F组小鼠骨髓嗜多染红细胞微核率分别为(16.00±0.67)‰、(15.11±0.25)‰、(10.17±0.35)‰、(22.39±0.51)‰、(15.44±0.44)‰、(18.67±0.37)‰.与A组比较,C组微核率显著减少,D组微核率显著增加(均P＜0.01).与D组比较,E组和F组微核率显著减少(均P＜o.o1).与C组比较,F组微核率显著增加(P＜0.01).结论 富氢水和乳果糖均可拮抗环磷酰胺的致突变作用,抗生素预处理可以显著降低乳果糖的抗突变作用.     

烟草提取液诱导小鼠骨髓嗜多染红细胞微核的研究

目的 研究烟草提取液对小鼠骨髓嗜多染红细胞的遗传毒性效应.方法 采用30h给受试物法,用不同浓度烟草提取液灌喂实验小鼠,小鼠处死后检测小鼠骨髓嗜多染红细胞微核率.其微核率与环磷酰胺阳性对照组以及生理盐水阴性对照组嗜多染红细胞微核率比较其差异性.结果 发现不同浓度烟草提取液均可诱导小鼠骨髓嗜多染红细胞产生微核,其微核率与阳性对照组比较无显著差异,而远高于阴性对照组.结论 烟草提取液具有较大的遗传毒性.     

Toxicity assessment and analgesic activity investigation of aqueous acetone extracts of Sida acuta Burn f. and

ABSTRACT: BACKGROUND: Sida acuta Burn f. and Sida cordifolia L. (Malvaceae) are traditionally used in Burkina Faso to treat several ailments, mainly pains, including abdominal infections and associated diseases. Despite the extensive use of these plants in traditional health care, literature provides little information regarding their toxicity and the pharmacology. This work was therefore designed to investigate the toxicological effects of aqueous acetone extracts of Sida acuta Burn f. and Sida cordifolia L. Furthermore, their analgesic capacity was assessed, in order to assess the efficiency of the traditional use of these two medicinal plants from Burkina Faso. METHOD: For acute toxicity test, mice were injected different doses of each extract by intraperitoneal route and the LD50 values were determined. For the subchronic toxicity evaluation, Wistar albinos rats were treated by gavage during 28 days at different doses of aqueous acetone extracts and then haematological and biochemical parameters were determined. The analgesic effect was evaluated in mice by the acetic-acid writhing test and by the formalin test. RESULTS: For the acute toxicity test, the LD50 values of 3.2 g/kg and 3.4 g/kg respectively for S. acuta Burn f. and S. cordifolia L. were obtained. Concerning the haematological and biochemical parameters, data varied widely (increase or decrease) according to dose of extracts and weight of rats and did not show clinical correlations. The extracts have produced significant analgesic effects by the acetic acid writhing test and by the hot plate method (p <0.05) and a dose-dependent inhibition was observed. CONCLUSION: The overall results of this study may justify the traditional uses of S. acuta and S. cordifolia .     

Hepatotoxicity induced by radix Sophorae tonkinensis in mice and increased serum cholinesterase as a potential supplemental biomarker for liver injury

Radix Sophorae tonkinensis (S. tonkinensis) is used in Chinese folk medicine to treat sore throats, viral hepatitis, and jaundice. However, little is known about the hepatotoxicity induced by it. This study is to investigate hepatotoxicity induced by radix S. tonkinensis and a potential supplemental biomarker for liver injury through acute toxicity, accumulative toxicity, tolerance test, and sub-chronic toxicity. The contents of cytisine (CYT), matrine (MT), and oxymatrine (OMT) in radix S. tonkinensis extracts were determined simultaneously by the method we developed. In the acute toxicity study, mice were scheduled for single oral gavage at doses of 0, 2.4, 3.2, 4.2, 5.6, 7.5 g/kg of radix S. tonkinensis extracts respectively. Another three groups of mice received radix S. tonkinensis extracts orally in single doses of 0, 4.3, 5.6 g/kg, while the two groups of the hepatic injury model were induced by intraperitoneal injection with 0.1% and 0.2% carbon tetrachloride (CCl₄). Mortality rate, analysis of serum biochemistry, and histopathological examination were used to assess the acute toxicity. In the accumulative toxicity study, mice were treated radix S. tonkinensis extracts orally by the method of dose escalation for 20 days respectively. Accumulative toxicity was assessed by mortality rate. In the tolerance test, half of the mice of test group in the accumulative toxicity were administered the dose of 4.3 g/kg radix S. tonkinensis extracts, and the rest of the mice in the test group were assigned to receive the dose of 5.6 g/kg radix S. tonkinensis extracts. In the sub-chronic toxicity study, mice were treated with daily doses of 0, 0.25, 1.0, 2.5 g/kg radix S. tonkinensis extracts for 90 days. Assessments of body weights, serum biochemical analysis, and histopathological examination were performed. An enzyme-inhibition assay for butyrylcholinesterase (BuChE) and acetylcholinesterase (AChE) of CYT, MT, and OMT was also carried out. The contents of CYT, MT, and OMT in radix S. tonkinensis extracts were 5.63 mg/g, 27.63 mg/g, and 16.20 mg/g respectively. In the acute toxicity study, LD50 of radix S. tonkinensis extracts was 4.3 g/kg. No mice were found dead in the accumulative toxicity study. In the acute toxicity and tolerance test, increased ALT, AST, and CHE levels were observed in a dose-response manner, while the severity of histological changes in liver was shown in a dose-dependent mode. In the sub-chronic toxicity, though there was a decline trend of ALT and AST levels found in 0.25 g/kg, 1.0 g/kg, and 2.5 g/kg radix S. tonkinensis extracts as compared to control, which might be related to weight loss, the severity of histopathological changes in the liver and the increased serum CHE level was shown in a dose-response manner. MT, OMT, and CYT showed inhibitory effects on BuChE and AChE in the enzyme-inhibition assay. The results of this study indicate that radix S. tonkinensis should have hepatotoxicity, and increased serum CHE is a potential supplemental biomarker for liver injury.     

清胰汤对L-精氨酸诱发的重症急性胰腺炎小鼠胰腺p-STAT3表达的影响

目的: 观察L-精氨酸(L-Arg)诱发的重症急性胰腺炎小鼠胰腺组织p-STAT3表达的变化,以及清胰汤对p-STAT3表达的影响,从而探讨STAT3在急性胰腺炎中的作用和清胰汤治疗急性胰腺炎的机制。方法: 健康雄性成年昆明种小鼠30只,随机分为3组(n=10):对照组、模型组和清胰汤组。除对照组外,其余各组给予腹腔注射20 % L-精氨酸(3 g/kg,间隔1 h再注射1次);清胰汤组在第2 次腹腔注射20 %L-Arg 30 min 后给予清胰汤浓缩液灌胃(10 mL/kg),之后每天灌胃2次。在造模后72 h麻醉处死动物检测血清淀粉酶活性;取胰腺组织计算胰腺湿重比,HE染色观察胰腺病理学改变;取肺组织匀浆检测髓过氧化物酶(MPO)的活性,HE染色观察肺病理学改变; Western blotting及real-time PCR分别检测胰腺组织p-STAT3蛋白及单核细胞趋化蛋白-1(MCP－1)mRNA的表达变化。结果: L-Arg诱发急性胰腺炎72 h后,血清淀粉酶活性明显升高、胰腺湿重比增加、肺组织MPO显著增加,与对照组比较差异显著(P<0.05);而清胰汤组血清淀粉酶的活性、胰腺湿重比、MPO水平明显降低,与模型组相比差异显著(P<0.01);模型组72 h胰腺及肺可见明显病理损伤,胰腺组织p-STAT3蛋白及MCP-1 mRNA的表达明显增强;清胰汤治疗组胰腺及肺病理损伤减轻,胰腺组织p-STAT3蛋白及MCP-1 mRNA的表达减少。结论: L-Arg诱发的重症急性胰腺炎小鼠胰腺组织STAT3蛋白表达明显增加,STAT3活化可能参与了L-Arg诱发的急性胰腺炎进展;抑制胰腺STAT3活化是清胰汤治疗急性胰腺炎的作用机制之一。     

The role of RIP1 and RIP3 in the development of aplastic anemia induced by cyclophosphamide and busulphan in mice

This study aimed to investigate the role of RIP1 and RIP3 in the pathogenesis of aplastic anemia (AA) induced by cyclophosphamide and busulphan in mice. Animals were randomly divided into three groups: the control group, the AA group, and the Nec-1 group. Mouse AA model was established by intraperitoneal injection of cyclophosphamide (40 mg/kg/d) and busulfan (20 mg/kg/d) for 12 days. The Nec-1 group mice received intraperitoneal injection of Nec-1 (2 mg/kg/d) for 12 days prior to intraperitoneal injection of cyclophosphamide (40 mg/kg/d) and busulfan (20 mg/kg/d) for 12 days. The control mice received intraperitoneal injection of equal volume of saline. At 12 h after the last intraperitoneal injection, blood and bone marrow tissues were collected from mice. Peripheral blood cells were analyzed using hematology analyzer and the histological changes of bone marrow tissues were examined using scanning electron microscopy (SEM). The levels of RIP3 and RIP3 in bone marrow were measured using Western blot analysis and the interaction of RIP1 and RIP3 proteins was investigated on the basis of immunoprecipitation analysis. ELISA was used to measure the levels of IL-6, TNF-α, and FLT-3L in bone marrow tissue supernatant. Apoptosis and necrosis of bone marrow cells were analyzed using flow cytometry. Western blot showed that the expression of RIP1 and RIP3 was significantly increases in AA mice compared to the normal controls. Immunoprecipitation detected the pro-necrotic RIP1-RIP3 complex, suggesting that RIP1 and RIP3 mediated necroptosis may involved in the damage of bone marrow cells. Compared to the AA mice, Nec-1 group mice exhibited significantly increase of peripheral blood cells and mononuclear cells in bone marrow tissues and decrease of the apoptosis/necrosis of bone marrow cells. In addition, we observed significant decrease of IL-6, TNF-α, and FLT-3L in bone marrow tissue supernatant in the Nec-1 group mice compared to AA mice. Our results suggest that Nec-1 can prevent the development of AA by inhibiting bone marrow cells necrosis and the production of inflammatory mediators. RIP1 and RIP3-mediated necroptosis may involve in the pathogenesis of AA induced by cyclophosphamide and busulfan in mice.     

地榆鞣质的制备及其对环磷酰胺致小鼠白细胞减少症影响的初步研究

目的 制备地榆鞣质提取物,观察其对白细胞减少症的影响.方法 采用明胶沉淀丙酮解析法制备地榆鞣质提取物.选用KM小鼠,按体重和性别随机分成空白组、模型组、阳性组(rhG-CSF组)和地榆鞣质高、中、低剂量(20、10、5 mg/kg)组,用环磷酰胺复制小鼠白细胞减少症模型,并以白细胞数(WBC)、脾脏系数和骨髓DNA含量作为检测指标,研究地榆鞣质的升白作用.结果 所制地榆鞣质纯度达90％;药理研究显示,地榆鞣质高、中、低剂量组均能显著升高白细胞数量(P＜ 0.05),且地榆鞣质高、中剂量组升高骨髓DNA的作用优于阳性药(rhG-CSF)组.结论 采用明胶沉淀丙酮解析制备法可获得纯度较高的鞣质部位,鞣质具有显著的升高白细胞及保护骨髓DNA的作用.     

煅烧骨的安全性

背景：经高温处理的煅烧骨具有类似自然骨的连续微孔结构,良好的生物相容性和降解性。 目的：观察牛煅烧骨的生物相容性、细胞相容性及毒性。 方法：①细胞相容性实验：将牛煅烧骨与第3 代已诱导的Wistar大鼠骨髓间充质干细胞复合培养。②溶血实验：将煅烧骨浸提液、生理盐水与双蒸水加入兔血中。③凝血实验：将煅烧骨加入兔血浆中。④急性毒性实验：在昆明种小鼠尾静脉分别注射煅烧骨浸提液、生理盐水。⑤微核实验：在小鼠腹腔分别注射煅烧骨浸提液、生理盐水与环磷酰胺。⑥局部刺激性实验：将煅烧骨浸提液、生理盐水分别注射于兔两侧脊柱皮下。⑦热源检测实验：在兔耳静脉注射煅烧骨浸提液。⑧皮下植入实验：将煅烧骨材料植入Wistar大鼠背部皮下。 结果与结论：煅烧骨材料无细胞毒性,具有良好的细胞及血液相容性;对皮肤、肌肉无刺激作用;对心、肝、肾重要器官无毒性作用;皮下植入后对周围组织无刺激作用,能够部分降解吸收并被机体组织替代;无致热作用,对凝血功能无影响,对小鼠骨髓细胞无抑制及毒性作用。     

Influence of pretreatment with immunosuppressants on O-demethylation of dextromethorphan in isolated perfused rat liver.

Changes in the immune system induced by exogenous or endogenous factors may be accompanied with modifications of the activity of the drug metabolising enzymes in the liver. Some immunostimulatory agents are known to suppress the oxidative metabolism mediated by cytochromes P450. Possible effects of substances which suppress the immune responses of the organism have not been fully understood yet. The present study was undertaken to investigate the influence of immunosuppressants cyclophosphamide and dexamethasone on the CYP2D 1-dependent metabolism of dextromethorphan (DEM) in the isolated perfused liver from male rat donors (Wistar albino, 250-310 g). Recirculatory perfusion system was used with Williams' medium E (Sigma Chemicals Co.) as a perfusion medium (120 mL). DEM was administered as a 1 mg bolus into the perfusion solution at the start of each experiment after 20 min preperfusion. Samples of perfusate for HPLC determination of DEM and its O-demethylated metabolite dextrorphan (DOR) were taken at 15 min intervals for 120 min. The results have shown a rapid conversion of DEM to DOR in the isolated rat liver preparations. Pharmacokinetic parameters in the livers from intact rats were as follows: t1/2 DEM = 19.1+/-4.10 min, k(m) = 0.035+/-0.008 min(-1), Cl(m) = 4.21+/-0.97 mL x min(-1), AUC(DOR) = 2160+/-201 microg x min x L(-1). Pretreatment of rats with dexamethasone (4 mg/kg/day, iv, x 3 days) led to a significant increase in the concentration of dextrorphan in the recirculating solution, but it did not substantially change the kinetic constants of DOR formation (km = 0.036+/-0.004 min(-1), Cl(m) = 4.27+/-0.43 mL x min(-1)). Cyclophosphamide (100 mg/kg, ip, 1 dose on day 5 before perfusion) induced nearly twofold increase in the DOR concentrations in perfusate and thus highly significant (p < 0.01) changes of the kinetic parameters characterizing the increased rate of conversion of DEM to DOR (t1/2 DEM = 12.1+/-0.90 min, km = 0.055+/-0.004 min(-1), Cl(m) = 7.09+/-1.37 mL x min(-1), AUC(DOR) = 3602+/-154 microg x min x L(-1)). Considering that both cyclophosphamide and dexamethasone belong to the most widely used immunosuppressive drugs, their potential to promote the CYP2D-mediated metabolism might have a clinical impact in combined therapy of autoimmune or other diseases.