Pulmonary carcinogenesis induced by ferric nitrilotriacetate in mice and protection from it by Brazilian propolis and artepillin C

In experiments using the renal carcinogen ferric nitrilotriacetate (Fe-NTA) in male ddY mice, primary pulmonary cancers were also induced in bronchiolar and alveolar tissues. 4-Hydroxy-2-nonenal (4-HNE) and 8-hydroxy-2'-deoxyguanosine (8-OHdG), products of oxidative processes, increased in bronchiolar and alveolar cells after administration of Fe-NTA. These substances disappeared after oral administration of propolis or artepillin C, as shown histochemically, and correlated with an anticancer prophylactic effect of propolis and artepillin C. From our investigation, lipid peroxidation seems to play an important role in pulmonary carcinogenesis. Malignant progression from adenoma of bronchiolar or alveolar origin to malignant tumors has been proposed to involve a stepwise transformation. In our study, adenomas developed into adenocarcinomas and large cell carcinomas after treatment with Fe-NTA. In contrast, after oral administration of propolis or artepillin C, adenomas did not progress to carcinomas. Instead of developing into large cell cancers, as induced by Fe-NTA in control mice, adenomas showed remarkable proliferation of macrophages and local anti-oxidant activity after treatment with either propolis or artepillin C. Propolis and artepillin C therefore appear to inhibit lipid peroxidation and the development of pulmonary cancers.     

Renal cysts and associated renal tumours in male ddY mice injected with ferric nitrilotriacetate

In experiments using ferric nitrilotriacetate (Fe-NTA) as a renal carcinogen, multiple renal cysts are often observed in addition to renal tumours. In the present study, we used 3-week-old male ddY mice and examined the relation between renal cysts and cancer development. Four months after the start of Fe-NTA administration, we observed cysts in the renal cortex in all Fe-NTA-treated mice, but not in Fe-free NTA-treated mice. Three types of cysts were observed, but only those which originated from the renal proximal tubules showed multi-layered or papillary growth of cyst epithelial cells. Using histochemical staining, we found a cyst formation-tumour induction sequence, and the supposed cystic-papillary tumour induced by Fe-NTA was of proximal tubular cell origin. We also found that the minimum dose of Fe-NTA capable of inducing renal tumours in ddY mice was 10 mg of iron/kg/day, four times in 2 weeks.     

Intranuclear iron deposition in hepatocytes and renal tubular cells in mice treated with ferric nitrilotriacetate

 Cytoplasmic and intranuclear iron depositions were observed in the livers and kidneys of male and female ddY mice treated for 4–12 weeks with ferric nitrilotriacetate (Fe-NTA), a known renal carcinogen that acts through the production of free radicals. The intranuclear iron deposition consisted of a spherical aggregation of ferritin particles of approximately 10 nm diameter, as revealed by immunohistochemical staining and electron microscopy. Although the incidence of renal tumours was greater in the males than in the females, the incidence of iron depositions did not differ with gender. The most abundant intranuclear iron depositions were observed in the animals treated with Fe-NTA for the longest duration (12 weeks). These findings suggest that the intranuclear production and propagation of free radical reactions are prevented by the trapping of iron in a chemically inert iron form of ferritin. Received: 5 May 1998 / Accepted: 6 August 1998     

Diurnal rhythms of cell proliferation in the early stages of liver carcinogenesis induced in mice by orthoaminoazotoluene

The diurnal rhythm of mitotic activity and of the number of DNA-synthesizing cells was studied by an autoradiographic method, using thymidine-³H, in mouse hepatocytes in the early stages of carcinogenesis of the liver induced by orthoaminoazotoluene. The rhythms of fluctuation of mitotic activity in the control animals in both the first (irregular diffuse hyperplasia) and the second (focal proliferation) stages of carcinogenesis in the liver had a distinct monophasic character with a maximum of the number of mitoses in the early morning and a minimum in the evening or night. Rhythms of DNA-synthesizing cells under normal conditions and in the first stage of carcinogenesis of the liver were monophasic in character with a maximum in the afternoon and evening, respectively. In the second stage of carcinogenesis the rhythm was characterized by the appearance of a second maximum in the early morning. The mean dirunal values of the two indices increased in the second stage of carcinogenesis.Presented by Academician of the Academy of Medical Sciences of the USSR L. M. Shabad.Translated from Byulleten' Éksperimental'noi Biologii i Meditisiny, Vol. 82, No. 7, pp. 856–858, July, 1976.     

蜂胶醇提物通过抑制C/EBP同源蛋白表达减轻氧化低密度脂蛋白诱导的血管内皮细胞凋亡

目的:研究蜂胶醇提物(ethanol extract of propolis,EEP)对氧化低密度脂蛋白(oxidized low-density lipoprotein,ox-LDL)诱导的血管内皮细胞凋亡的抑制作用,并探讨可能的分子机制。方法:体外培养人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs),给予EEP(7.5、15和30 mg/L)、4-苯丁酸(4-phenylbutyric acid,PBA;4 mmol/L)预处理1 h,再加入ox-LDL(100 mg/L)或衣霉素(tunicamycin,TM;4 mg/L)继续培养24 h。分别采用MTT法和Annexin V-FITC/PI双染法检测细胞活力和凋亡情况;试剂盒测定培养液乳酸脱氢酶(lactic dehydrogenase,LDH)和细胞内caspase-3活性。分别采用Western blot和real-time PCR技术检测内质网应激(endoplasmic reticulum stress,ERS)凋亡途径关键蛋白C/EBP同源蛋白(C/EBP homologous protein,CHOP)和Bcl-2的表达变化。结果:与ERS抑制剂PBA相似,EEP呈剂量依赖性地减轻ox-LDL所诱导的HUVECs损伤,表现为细胞活力增加(P0.01或P0.05),LDH漏出、凋亡率和caspase-3活性降低(P0.05或P0.01),且可抑制ERS诱导剂TM所致的HUVECs活力下降(P0.05),以及LDH漏出、细胞凋亡率和caspase-3活性增加(P0.05或P0.01);与PBA相似,EEP可抑制ox-LDL所诱导的CHOP上调和Bcl-2下调(P0.05或P0.01);另外,与TM组比较,EEP预处理组CHOP蛋白和mRNA表达上调也受到明显抑制(P0.05或P0.01)。结论:EEP可减轻oxLDL所诱导的HUVECs凋亡,其机制可能与抑制CHOP介导的ERS凋亡途径有关。     

Effect of topical application of Brazilian propolis on scratching behavior induced by compound 48/80 in mice

Background and aim: We investigated the effect of topical application of Brazilian propolis on scratching behavior induced by compound 48/80 in mice.

Results: Propolis inhibited compound 48/80-induced scratching behavior when applied immediately after treatment with propolis at a dose of 3?mg/site. Dibucaine 0.3?mg/site also significantly inhibited compound 48/80-induced scratching behavior immediately after application. On the other hand, propolis inhibited compound 48/80-induced scratching behavior even 15, 30 and 60?min after application; however, dibucaine showed no significant inhibition of compound 48/80-induced scratching behavior 15, 30 and 60?min after application. In addition, propolis had no effect on increased vascular permeability just after application, but the drug had a significant effect 15, 30 and 60?min after application. On the contrary, histamine-induced scratching behavior was inhibited significantly by propolis just after application. On the other hand, propolis significantly inhibited histamine release from rat mast cells induced by compound 48/80 at a concentration of more than 10?μg/ml.

Conclusion: From these results, it can be concluded that inhibition of scratching behavior induced by topical application occurred by both its local anesthetic and systemic action through inhibition of histamine release.     

Carcinogenesis induced by polycyclic aromatic hydrocarbons in male and female (CBA×C57/BL)F1 mice

Sex differences were found in the development of skin tumors induced by 3-methylcholanthrene and benz(a)pyrene in (CBA×C57/BL)F₁ mice. Males were more susceptible to the induction of skin tumors than females.Laboratory of Carcinogens, Institute of Nutrition, Academy of Medical Sciences of the USSR. (Presented by Academician of the Academy Sciences of the USSR N. A. Fedorov.) Translated from Byullenten' Éksperimental'noi Biologii i Meditsiny, Vol. 82, No. 10, pp. 1244–1245, October, 1976.     

Liver tumors induced in mice by prenatal and postnatal administration of orthoaminoazotoluene

The prenatal and postnatal carcinogenic action of orthoaminoazotoluene (OAAT) on the liver was studied in CBA mice. The carcinogen was administered to the mice by the gastric route in oily solution in three doses each of 4 mg (total 12 mg). Tumors of the liver were found 12 months after administration of OAAT in the postnatal period in females in 89.3% of cases compared with 11.5% in the control and in males in 78.7% of cases compared with 68.5% in the control. In the experimental progenies exposed to OAAT during the last 4–5 days of prenatal development tumors of the liver were found in females at the age of 12 months in 54.8% of cases compared with 5.3% in the control and in males in 81.1% of cases compared with 35.2% in the control. In the experimental progenies, especially in males, malignant neoplasms were found more often than in intact animals or those exposed to OAAT in the postnatal period.Department of Carcinogenic Agents, Oncologic Scientific Center, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR L. M. Shabad.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 85, No. 2, pp. 199–202, February, 1978.     

C1型尼曼-匹克氏症小鼠的肾脏功能及病理变化

目的:探讨Npc1基因突变小鼠肾脏功能及病理生理的变化,为C1型尼曼-匹克氏症(NPC1)患者临床上药物的使用及肾功能的维持提供理论依据。方法:用PCR法确定小鼠基因型;选取出生后第60天(P60)的野生型NCPC1(Npc1~(+/+))和突变型NCPC1(Npc1~(-/-))小鼠,检测血清中丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)和乳酸脱氢酶(LDH)的活性及尿素(Urea)、尿酸(UA)和肌酐(Cr)的含量来评价Npc1-/-小鼠的肝肾功能变化;进一步常规冷冻切片后通过β-半乳糖苷酶染色及Masson染色来分别评价Npc1~(-/-)小鼠肾脏衰老及纤维化情况。结果:与P60的Npc1~(+/+)小鼠相比,Npc1~(-/-)小鼠的体重和肾脏重量显著降低(P0.01);肝肾功能明显减退:ALT、AST及LDH活性显著升高(P0.05),Urea、UA及Cr的含量显著增加(P0.05);冷冻切片染色结果表明肾脏组织衰老明显(P0.01),纤维化程度显著增强(P0.01)。结论:Npc1基因突变导致的脂质异常代谢可加速肾间质纤维化,促使肾脏衰老,最终导致肾功能减退。     

Cys C、SCr、Hb在肾功能改变中的应用

目的:分析胱抑素C(Cys-C)、血肌酐(SCr)、血红蛋白(Hb)在肾功能变化过程的改变情况.方法:回顾性分析201例行肾动态显像患者,记录同时期患者Cys-C、SCr、Hb的水平,并将两者与肾动态显像计算所得GFB<,总>做相关分析.同时以GFB<,总>为肾功能改变的分级标准,将201例患者分成5组,观察Cys-C...     

Serum Cystatin C Is a Determinant of Paraoxonase Activity in Hemodialyzed and Renal Transplanted Patients

Background: Human paraoxonase-1 (PON1) inhibits LDL-oxidation and atherogenesis, and possesses lactonase activity. Decreased PON1 activity was found in hemodialyzed and renal transplanted patients. Cystatin C plays a protective role in atherosclerosis, and is a new, sensitive marker of renal function. The relationship between these two markers in renal failure has not been investigated.Aims: The goal of this study was to clarify the relationship between PON1 activity, cystatin C and homocysteine in chronic renal failure. We also determined the levels of oxidatively modified LDL (oxLDL) and thiobarbituric acid reactive substances (TBARS) to characterize lipid peroxidation.Patients and methods: 74 hemodialized (HD), 171 renal transplanted patients (TRX), and 110 healthy controls (C) were involved in the study. PON1 activity and TBARS levels were measured spectrophotometrically. OxLDL level was determined with sandwich ELISA.Results: There was a negative correlation between PON1 activity and cystatin C level. Homocysteine level correlated negatively with PON1 activity, and positively with cystatin C level. OxLDL and TBARS levels were significantly higher in the HD and TRX groups compared to C.Conclusions: Cystatin C may be a good predictive factor not only for homocysteine levels but for the antioxidant status in patients with renal failure and renal transplantation.     

Effect of age,castration, and pregnancy on carcinogenesis induced in CBA mice by 1,2-dimethylhydrazine

When injected subcutaneously in a dose of 8 mg/kg weekly into female CBA mice, 1,2-dimethylhydrazine (DMH) induced the development of tumors of the intestine, anal region, uterus, and liver. When DMH was injected into mice aged 12–13 months the appearance of sarcomas of the uterus was observed earlier (at 8 weeks) and the incidence of tumors of the anal region rose more rapidly than in mice aged 3 months. In mice receiving DMH against the background of repeated pregnancies, a statistically significant decrease in the frequency of sarcomas of the uterus was observed (10.3% compared with 48.3% in nonpregnant mice); pregnancy did not affect the frequency of tumors of other organs. Castration had no significant effect on the time of appearance or the frequency of tumors in all situations.Laboratory of Carcinogenic Substances, Department of Carcinogenic Agents, Oncologic Scientific Center, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR L. M. Shabad.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 87, No. 5, pp. 458–460, May, 1979.     

肾癌荷瘤小鼠组织中补体 C5a / C5aR 通路活化与肿瘤高凝状态 的关系及其机制研究

目的 研究肾癌荷瘤小鼠组织中补体 C5a / C5aR 通路活化与肿瘤高凝状态的关系及其机制。 方法 收集 2016 年 7 月至 2019 年 10 月于唐山市人民医院收治的 32 例经病理学活检确诊的肾癌患者的癌组织和 癌旁组织。 采用 Western 印迹法检测组织中 C5a / C5aR 通路活化相关蛋白表达情况。 于 BALB/ c-nu / nu 裸鼠 前肢腋窝皮下接种人 ACHN 肾癌细胞株, 接种成功后随机分为干预组、 模型组, 其中干预组尾静脉注射 C5a 联合 C5aR 阻断剂 (C5aR antagonist, C5aRA), 模型组等体积注射生理盐水, 另取 6 只空白小鼠作为对 照组。 观察小鼠肿瘤生长情况, 检测高凝状态相关指标水平。 脱颈处死小鼠, 收集癌组织, 采用 Western 印迹法检测组织中 C5a / C5aR 通路活化相关蛋白表达情况。 结果 两组小鼠肿瘤体积于荷瘤 6 ~ 21 d 差异具 有统计学意义, 且干预组的肿瘤体积明显低于模型组 ( P< 0. 05)。 3 组小鼠于荷瘤第 1 天的血清 D-D、 vWF、 TF 水平差异无统计学意义, 荷瘤第 14 天、 第 21 天干预组、 模型组血清凝血指标水平明显增加, 其 中干预组均明显低于模型组 (P< 0. 05)。 干预组癌组织中 C5aR、 C5b-9、 FGL2、 P38、 p-P38 的表达水平均 明显低于模型组 (P< 0. 05)。 肾癌患者癌组织中 C5aR、 C5b-9、 FGL2、 P38、 p-P38 的表达水平均明显高于 癌旁组织 (P< 0. 05)。 32 例肾癌患者中高凝状态患者 14 例, 非高凝状态患者 18 例, 高凝状态组癌组织中 C5aR、 C5b-9、 FGL2、 P38、 p-P38 的表达水平均明显高于非高凝状态组 (P< 0. 05)。 结论 补体 C5a / C5aR 通路可通过调控 FGL2 的表达而诱导肾癌的高凝状态, 进而参与肾癌的发病过程。     

血清胱抑素C与尿白蛋白排泄率联合检测对原发性高血压患者早期肾损害的诊断价值

目的:探讨血清胱抑素C(CysC)和尿白蛋白排泄率(UAER)联合检测,对高血压患者早期肾损害的诊断价值。方法:测定320例确诊原发性高血压患者内生肌酐清除率(Ccr)并据此分为肾小球功能正常组(A组,Ccr≥80ml/min)和肾小球功能损害组(B组,Ccr<80ml/min)。再测定A组、B组及正常对照组血清CysC浓度UAER和血清肌酐(Scr)水平,分析以上指标单检和联检对早期肾损害阳性检出率差异。结果:A组、B组CysC、UAER和Scr水平均显著高于对照组(P<0.05)。B组高于A组(P<0.01)。A组患者血清CysC、UAER单检阳性率分别为41.84%和36.73%,二者联合检测的阳性率增至58.16%(P<0.01),B组患者血清CysC、UAER单检阳性率分别为90.54%和80.18%,二者联检的阳性率增至97.75%(P<0.01)。结论:血清Cy-sC、UAER联合检测有助于及早发现原发性高血压早期肾损害。     

血清Cyst C用于判断早期肾功能损害的敏感性与特异性研究

目的:评价血清胱蛋白酶抑制剂C(Cyst C)用于判断早期肾功能损害的敏感性与特异性。方法:采用固定时间颗粒增强的免疫透射比浊法(PETIA)测定116例肾科住院病人血清CystC浓度,同时测定血清肌酐(Scr,酶法),以目前国内最常用的内生肌酐清除率(Ccr)推算肾小球滤过率(GFR)的方法为判断肾功能损害与否的指标,计算并比较血清CystC、Scr用于判断肾功能损害的敏感性与特异性。结果:血清CystC诊断肾小球滤过功能受损的敏感性为99%,特异性为40%。而Scr则与之相反,敏感性为80%,特异性为100%。结论:血清CystC诊断肾小球滤过功能受损的敏感性高于Scr,尤其是在早期肾脏损害时更敏感。血清CystC诊断肾小球滤过功能受损的特异性低于Scr,进一步表明在早期肾功能损害时,血清CystC结果已经反映出早期肾功能损害,而此时的Scr值尚不能反映出早期肾功能损害。血清CystC测定在早期预测肾功能损害的发生中有重要的价值。     

巨细胞病毒感染C57BL/6小鼠诱导NK细胞免疫应答的模型研究

目的:研究小鼠巨细胞病毒(murine cytomegalovirus,MCMV)感染C57BL/6小鼠诱导自然杀伤(natural killer,NK)细胞免疫应答的最佳剂量和最佳时间。方法:分别根据剂量和时间效应进行分组,剂量效应：无特定病原体( specific pathogen free,SPF)级8周龄C57...     

美法仑治愈荷瘤小鼠的过程与TNFα的关系

目的探讨单一剂量的美法仑治愈荷瘤野生型C57BL/6小鼠的过程与肿瘤坏死因子α(TNFα)的关系。方法以3种遗传背景相同、肿瘤坏死因子受体1(TNFR1)基因型不同的TNFR1 / 、TNFR1 /-和TNFR1-/-C57BL/6小鼠为实验动物,皮下接种数量相同的小鼠淋巴瘤EL4细胞。接种瘤细胞后12d,给基因型不同的各组荷瘤小鼠腹腔内单次注射7.5mg/kg的美法仑。以荷瘤野生型C57BL/6小鼠(TNFR1 / )为对照,观察美法仑对荷瘤TNFR1 /-C57BL/6小鼠和荷瘤TNFR1-/-C57BL/6小鼠的治疗效应。结果在美法仑(7.5mg/kg)治疗后的1周内,基因型不同的各组荷瘤小鼠肿瘤消退的速度基本相同。在随后的2月内,荷瘤TNFR1 / 和TNFR1 /-C57BL/6小鼠的肿瘤结节逐渐消退、肿瘤治愈;而多数荷瘤TNFR1-/-C57BL/6小鼠的肿瘤结节缩小后又再次出现并逐渐长大、肿瘤复发。结论TNFα与美法仑治愈肿瘤的过程密切相关,其中美法仑的抗肿瘤作用与荷瘤小鼠TNFR1的表达无关,但在美法仑治疗后,机体预防或避免肿瘤复发方面需要TNFR1在机体细胞的表达,而不是在肿瘤细胞的表达。