Aluminum-mediated metabolic changes in rat serum and urine: a proton nuclear magnetic resonance study

The toxic effects of Al(3+) have been studied in 90-days AlCl(3) orally treated male albino rats (n = 7) using (1)H NMR spectroscopy-based metabolic profile of rat serum and urine, serum enzyme tests, behavioral impairment, and histopathology of kidney and liver. Metabolic profile of 90-days Al(3+)-treated rat sera showed significantly elevated levels of alanine, glutamine, beta-hydroxy-butyrate, and acetoacetate and significantly decreased level of acetone when compared with that of control rats. However, metabolic profile of 90-days Al(3+)-treated rat urine showed significantly decreased levels of citrate, creatinine, allantoin, trans-aconitate, and succinate and significantly increased level of acetate when compared to control rats. The overall perturbations observed in the metabolic profile of serum and urine demonstrate the impairment in the tricarboxylic acid cycle, liver and kidney metabolism, which was further reinstated by clinical chemistry and histopathological observations. Moreover, "in vivo" behavioral impairment has also been observed as the indication of aluminum neurotoxicity.     

Evaluation of the micronucleus assay in bone marrow and peripheral blood of rats for the determination of cigarette mainstream-smoke activity

The mammalian in vivo micronucleus assay is widely used as part of the genotoxicity testing battery required during the development of new drugs. As such, the in vivo micronucleus assay has been used in a battery of assays for the assessment of cigarette ingredients or design modifications to help ensure that there is no increase in risk or any new risk introduced by these additions or modifications. The present series of studies was conducted to optimize and evaluate this assay for the assessment of the effects of mainstream smoke on the micronucleus frequency in the bone marrow and peripheral blood of rats. In a first experiment, the optimal conditions for performing the micronucleus assay in these tissues were determined. This was done by use of two compounds known for their micronucleus-inducing activity, i.e., the clastogen cyclophosphamide and the aneugen colchicine. In a second experiment, the effects of tube restraint on untreated control rats were investigated. In a third experiment, the optimal conditions were used to assess the clastogenic/aneugenic activity of cigarette smoke in Sprague-Dawley rats. The rat micronucleus assay in both bone marrow and peripheral blood is able to detect clastogenic and aneugenic activity. The flow cytometric determination of micronucleated cells in rat blood is at least as sensitive as determinations in bone marrow. No statistically significant differences were observed in micronucleus frequencies between rats with and without the additional stress of tube restraint; however, the cautious approach would be to use a fresh-air-exposed group (with tube restraint) as the negative control in inhalation experiments. Using the conditions identified as optimal in the above-mentioned experiments, the micronucleus assay was not able to detect effects induced by smoke from conventional cigarettes. Nevertheless, the micronucleus assay will remain a valuable tool as part of a testing battery used to investigate possible adverse effects related to product modifications.     

<Emphasis Type="Italic">Hemidesmus indicus</Emphasis> Protects against ethanol-induced liver toxicity

Alcoholic liver disease (ALD) is one of the most common diseases in modern society. A large number of studies are in progress aiming to identify natural substances that would be effective in reducing the severity of ALD. Although there are currently a number of drugs on the market, their long-term use can have numerous side effects. Hemidesmus indicus is an indigenous Ayurvedic medicinal plant used in soft drinks in India. In this study, we examined the effects of its ethanolic root extract on experimental liver damage in order to evaluate its hepatoprotective effects against hepatotoxicity induced in rats by ethanol at a dosage of 5 g/kg body weight for 60 days. The H. indicus root extract was given at a dose of 500 mg/kg body weight for the last 30 days of the experiment. The animals were monitored for food intake and weight gain. The liver was analysed for the degree of lipid peroxidation using thiobarbituric acid reactive substances (TBARS) and antioxidant status using the activities of glutathione-depedendant enzymes. The degree of liver damage was analysed using serum marker enzyme activities, the total protein, albumin, globulin, ceruloplasmin and liver glycogen contents, and the A/G ratio. The Fourier transform infrared spectra (FT-IR) of the liver tissues were recorded in the region of 4000–400 cm⁻¹. The ethanol-fed rats showed significantly elevated liver marker enzyme activities, lipid peroxidation levels and reduced antioxidant levels as compared to the control rats. Oral administration of H. indicus for the latter 30 days resulted in an increased food intake and weight gain, decreased TBARS levels, near normal levels of glutathione-dependent enzymes, increased total protein, albumin, globulin and liver glycogen contents, an increased A/G ratio, and decreased liver marker enzyme activities and ceruloplasmin levels. The relative intensity of the liver FT-IR bands for the experimental groups were found to be altered significantly (p < 0.05) compared to the control samples. For the group that had H. indicus co-administered with ethanol, the intensity of the bands was near normal. Moreover, the results of the FT-IR study correlated with our biochemical results.     

Effect of contingent auditory stimuli on concurrent schedule performance: an alternative punisher to electric shock

This study explored whether load auditory stimuli could be used as functional punishing stimuli in place of electric shock. Three experiments examined the effect of a loud auditory stimulus on rats’ responding maintained by a concurrent reinforcement schedule. In Experiment 1, overall response rate decreased when a concurrent 1.5 s tone presentation schedule was superimposed on the concurrent variable interval (VI) 180-s, VI 180-s reinforcement schedule. On the contrary, response rate increased when a click presentation schedule was added. In Experiment 2, the extent of the response suppression with a 1.5 s tone presentation varied as a function of the frequency of the reinforcement schedule maintaining responses; the leaner the schedule employed, the greater the response suppression. In Experiment 3, response suppression was observed to be inversely related to the duration of the tone; response facilitation was observed when a 3.0-s tone was used. In Experiments 1 and 2, a preference shift towards the alternative with richer reinforcement was observed when the tone schedule was added. In contrast, the preference shifted towards the leaner alternative when the click or longer duration stimulus was used. These results imply that both the type and duration of a loud auditory stimulus, as well as the reinforcement schedule maintaining responses, have a critical role in determining the effect of the stimuli on responding. They also suggest that a loud auditory stimulus can be used as a positive punisher in a choice situation for rats, when the duration of the tone is brief, and the reinforcement schedule maintaining responses is lean.     

体外定向诱导大鼠骨髓基质干细胞向成骨细胞分化的实验研究

目的研究大鼠骨髓基质干细胞的生长特点和诱导条件下的成骨能力。方法通过密度梯度离心和贴壁培养法分离成年大鼠骨髓基质干细胞,应用含地塞米松、p甘油磷酸纳和维生素c的诱导分化培养液定向诱导传代细胞向成骨细胞分化并检测碱性磷酸酶活性和细胞矿化作用。结果原代培养基质干细胞首先形成细胞集落,14d时集落间接近融合;传代细胞体积变大,约5～7d传代一次。诱导条件下,细胞碱性磷酸酶活性明显增高,并出现了矿化结节。结论骨髓基质干细胞易于分离培养及体外扩增,成骨能力肯定,可作为骨组织工程的种子细胞。     

光刺激移植视网膜诱导大鼠上丘FOS的表达

目的 用光反复刺激移植视网膜,观察脑内上丘c-fos的表达情况,检测移植视网膜能否对光刺激作出反应。方法 设实验组与对照组,用免疫组化方法显示脑内上丘c—fos的表达。结果 实验组移植视网膜与非移植视网膜大鼠在光反复刺激后1小时,脑内上丘出现明显的c-fos免疫反应阳性神经元,停止光刺激后2～3小时阳性神经元数量达到高峰。结论 光刺激移植视网膜能诱导脑内上丘c—fos的表达,提示移植视网膜具有对光刺激作出反应的能力。     

6-羟多巴胺纹状体内注射制作大鼠帕金森病模型的研究

目的 为拓宽6-OHDA损毁多巴胺能神经元所制备大鼠帕金森病模型的应用范围,采用多位点纹状体内注入6-OHDA的途径来制备模型。方法 研究用SD大鼠,两个针道内四点定位注射,每点注射3μg／μ16-OHDA3μl。结果 术后两周出现缓慢旋转,4周旋转行为达到7转／分并保持稳定;形态学染色可见损毁1周后注射侧黑质酪氨酸羟化酶免疫组化阳性细胞减少20％,2周后减少38％,3～4周减少70％以上,6周后损伤趋缓。高效液相-电化学法活体检测纹状体内多巴胺的代谢产物3、4-二羟基苯乙酸(DOPAC)和高香草酸(HVA),发现注射侧和非注射侧相比含量分别下降98．33％和96．05％;组织匀浆检测损毁侧黑质多巴胺含量下降了73％以上,3、4-二羟基苯乙酸(DOPAC)含量下降60％。结论 纹状体内注射6-OHDA能够制备帕金森病大鼠模型。     

大鼠隔区接受海马一氧化氮合酶(NOS)阳性神经元的投射

目的逆行追踪大鼠海马NOS阳性神经元向隔区的投射。方法用HRP逆行追踪与NADPH-d组化方法相结合进行研究。结果背、腹、后海马均有NOS阳性神经元投射至隔区各亚细胞群,后海马NOS阳性神经元向隔外侧核(sl)、隔三角核和隔伞核(ts,sf)的投射量,占后海马至隔外侧核、隔三角核和隔伞核投射量的80％左右。结论大鼠隔区接受海马NOS神经元的投射。     

灵光注射液对失血性休克大鼠心脏和肝脏的影响

目的　观察灵光注射液 (复方樟柳碱 )对失血性休克再灌注大鼠心脏和肝脏损伤的影响。方法　将5 6只雄性大鼠随机分 4组 ,分别设为假休克组 (8只 )、模型组 (16只 )、灵光注射液低剂量组 (16只 )和高剂量组 (16只 ) ,除假休克组外 ,大鼠均经历 4kPa ,70min的失血性休克 ,在休克复苏后 6h和 12h各组分别处死半数动物 ,检测血清CK、CK MB、LDH、ALT、AST ,心脏和肝脏做组织学和超微结构检查。结果与结论　灵光注射液对大鼠失血性休克再灌注引起的心脏和肝脏功能和形态损伤均有明显的治疗作用 ,其机制可能与改善微循环、清除氧自由基和保护生物膜作用有关。