p53、BCL-2、Caspase-3 等凋亡相关蛋白在大鼠脑损伤中的表达及意义

目的:检测脑外伤大鼠中p53、bcl-2及caspase-3表达,并分析其与脑外伤之间的关系,为脑损伤患者预后提供部分参数依据。方法:建立脑外伤大鼠实验动物模型,用免疫组化方法检测p53、Bcl-2和Caspase-3的表达。结果:脑创伤后在创伤周围区的神经元会发生凋亡,凋亡的发生可能与p53、bcl-2、caspase-3等基因及蛋白的调节有关。结论:脑创伤后在创伤周围区的神经元会发生凋亡,凋亡的发生可能与p53、bcl-2、caspase-3等基因及蛋白的调节有关。     

p53、BCL-2、Caspase－3等凋亡相关蛋白在大鼠脑损伤中的表达及意义

目的：检测脑外伤大鼠中p53、bcl-2及caspase-3表达,并分析其与脑外伤之间的关系,为脑损伤患者预后提供部分参数依据。方法：建立脑外伤大鼠实验动物模型,用免疫组化方法检测p53、Bcl-2和Caspase-3的表达。结果：脑创伤后在创伤周围区的神经元会发生凋亡,凋亡的发生可能与p53、bcl-2、caspase-3等基因及蛋白的调节有关。结论：脑创伤后在创伤周围区的神经元会发生凋亡,凋亡的发生可能与p53、bcl-2、caspase-3等基因及蛋白的调节有关。     

Capsaicin Ameliorates Cisplatin-Induced Renal Injury through Induction of Heme Oxygenase-1

Cisplatin is one of the most potent chemotherapy agents. However, its use is limited due to its toxicity in normal tissues, including the kidney and ear. In particular, nephrotoxicity induced by cisplatin is closely associated with oxidative stress and inflammation. Heme oxygenase-1 (HO-1), the rate-limiting enzyme in the heme metabolism, has been implicated in a various cellular processes, such as inflammatory injury and anti-oxidant/oxidant homeostasis. Capsaicin is reported to have therapeutic potential in cisplatin-induced renal failures. However, the mechanisms underlying its protective effects on cisplatin-induced nephrotoxicity remain largely unknown. Herein, we demonstrated that administration of capsaicin ameliorates cisplatin-induced renal dysfunction by assessing the levels of serum creatinine and blood urea nitrogen (BUN) as well as tissue histology. In addition, capsaicin treatment attenuates the expression of inflammatory mediators and oxidative stress markers for renal damage. We also found that capsaicin induces HO-1 expression in kidney tissues and HK-2 cells. Notably, the protective effects of capsaicin were completely abrogated by treatment with either the HO inhibitor ZnPP IX or HO-1 knockdown in HK-2 cells. These results suggest that capsaicin has protective effects against cisplatin-induced renal dysfunction through induction of HO-1 as well as inhibition oxidative stress and inflammation.     

大鼠脑创伤后caspase-3的过度表达与细胞凋亡的关系

目的：探讨大鼠脑创伤后海马神经组织中casepase-3表达及其在细胞凋亡中的机制。方法：雄性Wistar大鼠72只随机分成对照组和创伤组。用Marmarou方法造成大鼠重型弥漫性颅脑创伤,采用免疫组织化学检测海马CA1区神经细胞casepase-3蛋白表达情况,原位细胞DNA断裂检测末端标记（TUNEL）法观察大鼠海马CA1区神经细胞凋亡动态变化。同时行TUNEL与caspase-3双标染色。结果：对照组海马区神经细胞casepase-3未见明显表达,创伤组海马CA1区神经细胞casepase-3表达在伤后3小时开始升高,伤后3天达高峰（P〈0．01）,伤后7天下降明显。对照组海马区未见TUNEL阳性细胞,创伤组海马区TUNEL阳性细胞伤后3小时开始增多,伤后3天达高峰（P〈0．01）,伤后7天下降。可见创伤组TUNEL染色与caspase-3免疫染色双标阳性的细胞伤后6小时细胞数量逐渐增多,于伤后3天达高峰（P〈0．01）,伤后7天双标阳性细胞数量下降。Casepase-3表达与TUNEL阳性细胞明显相关（P〈0．01）。结论：大鼠脑创伤后casepase-3的过度表达是影响大鼠脑创伤后神经细胞凋亡原因之一,抑制casepase-3活性表达对神经组织起保护作用。     

大鼠脑创伤后caspase-3的过度表达与细胞凋亡的关系

目的:探讨大鼠脑创伤后海马神经组织中casepase-3表达及其在细胞凋亡中的机制。方法:雄性Wistar大鼠72只随机分成对照组和创伤组,用Marmarou方法造成大鼠重型弥漫性颅脑创伤,采用免疫组织化学检测海马CA1区神经细胞casepase-3蛋白表达情况,原位细胞DNA断裂检测末端标记(TUNEL)法观察大鼠海马CA1区神经细胞凋亡动态变化。同时行TUNEL与caspase-3双标染色。结果:对照组海马区神经细胞casepase-3未见明显表达,创伤组海马CA1区神经细胞casepase-3表达在伤后3小时开始升高,伤后3天达高峰(P0.01),伤后7天下降明显。对照组海马区未见TUNEL阳性细胞,创伤组海马区TUNEL阳性细胞伤后3小时开始增多,伤后3天达高峰(P0.01),伤后7天下降。可见创伤组TUNEL染色与caspase-3免疫染色双标阳性的细胞伤后6小时细胞数量逐渐增多,于伤后3天达高峰(P0.01),伤后7天双标阳性细胞数量下降。Casepase-3表达与TUNEL阳性细胞明显相关(P0.01)。结论:大鼠脑创伤后casepase-3的过度表达是影响大鼠脑创伤后神经细胞凋亡原因之一,抑制casepase-3活性表达对神经组织起保护作用。     

高浓度藏红花溶液导致大鼠肝损伤中caspase-3、bcl-2、NF-kB表达及意义

目的：通过研究高浓度藏红花溶液引起肝损伤大鼠肝功能、肝组织病理及肝组织中半胱氨酸蛋白酶-3（caspase-3）,bcl-2,NF-kB表达,探讨半胱氨酸蛋白酶-3（caspase-3）,bcl-2,NF-kB在高浓度藏红花溶液导致的肝损伤中的作用及机制。方法：清洁级雄性大鼠45只,随机分为3组,每组15只,分别标记为A组（藏红花组）;B组（酒精组,阳性对照）;C组（生理盐水组,阴性对照）。分别给予藏红花高浓度水煎浓缩溶液、60%酒精、生理盐水灌胃共6周。第6周末,心腔取血测定ALT、AST。肝组织HE染色,免疫组化方法检测细胞凋亡相关蛋白Caspase-3及调控基因Bcl-2、NF-kB的表达。结果：藏红花组大鼠肝功能ALT、AST升高,肝组织正常结构消失,肝细胞肿胀坏死,部分碎裂,凋亡蛋白caspase-3,bcl-2,NF-kB表达增加（69.6%±16.7%vs 5.3%±1.6%;55.4%±14.5%vs4.5%±2.8%;44.1%±12.6%vs2.5%±1.9%;P〈0.05）。结论：高浓度藏红花溶液可以引起大鼠肝损伤,肝组织caspase-3,bcl-2,NF-kB表达增加,细胞凋亡机制参与了藏红花肝损伤过程。     

Curcumin Ameliorates Streptozotocin‐Induced Heart Injury in Rats

Heart failure (HF) is one of diabetic complications. This work was designed to investigate the possible modulatory effect of curcumin against streptozotocin‐induced diabetes and consequently HF in rats. Rats were divided into control, vehicle‐treated, curcumin‐treated, diabetic‐untreated, diabetic curcumin–treated, and diabetic glibenclamide–treated groups. Animal treatment was started 5 days after induction of diabetes and extended for 6 weeks. Diabetic rats showed significant increase in serum glucose, triglycerides, total cholesterol, low‐density lipoprotein‐cholesterol, very low density lipoprotein‐cholesterol, nitric oxide, lactate dehydrogenase, cardiac malondialdehyde, plasma levels of interleukin‐6, and tumor necrosis factor‐alpha, and also showed marked decrease in serum high‐density lipoprotein‐cholesterol, cardiac reduced glutathione, and cardiac antioxidant enzymes (catalase, superoxide dismutase, and glutathione‐S‐transferase). However, curcumin or glibenclamide treatment significantly mitigated such changes. In conclusion, curcumin has a beneficial therapeutic effect in diabetes‐induced HF, an effect that might be attributable to its antioxidant and suppressive activity on cytokines.     

Melatonin Ameliorates Injury and Specific Responses of Ischemic Striatal Neurons in Rats

Studies have confirmed that middle cerebral artery occlusion (MCAO) causes striatal injury in which oxidative stress is involved in the pathological mechanism. Increasing evidence suggests that melatonin may have a neuroprotective effect on cerebral ischemic damage. This study aimed to examine the morphological changes of different striatal neuron types and the effect of melatonin on striatal injury by MCAO. The results showed that MCAO induced striatum-related dysfunctions of locomotion, coordination, and cognition, which were remarkably relieved with melatonin treatment. MCAO induced severe striatal neuronal apoptosis and loss, which was significantly decreased with melatonin treatment. Within the outer zone of the infarct, the number of Darpp-32+ projection neurons and the densities of dopamine-receptor-1 (D1)+ and dopamine-receptor-2 (D2)+ fibers were reduced; however, both parvalbumin (Parv)+ and choline acetyltransferase (ChAT)+ interneurons were not significantly decreased in number, and neuropeptide Y (NPY)+ and calretinin (Cr)+ interneurons were even increased. With melatonin treatment, the loss of projection neurons and characteristic responses of interneurons were notably attenuated. The present study demonstrates that the projection neurons are rather vulnerable to ischemic damage, whereas the interneurons display resistance and even hyperplasia against injury. In addition, melatonin alleviates striatal dysfunction, neuronal loss, and morphological transformation of interneurons resulting from cerebral ischemia.     

Sodium Thiosulfate Ameliorates Oxidative Stress and Preserves Renal Function in Hyperoxaluric Rats

Background

Hyperoxaluria causes crystal deposition in the kidney, which leads to oxidative stress and to injury and damage of the renal epithelium. Sodium thiosulfate (STS, Na₂S₂O₃) is an anti-oxidant, which has been used in human medicine for decades. The effect of STS on hyperoxaluria-induced renal damage is not known.

Methods

Hyperoxaluria and renal injury were induced in healthy male Wistar rats by chronic exposure to ethylene glycol (EG, 0.75%) in the drinking water for 4 weeks. The treatment effects of STS, NaCl or Na₂SO₄ were compared. Furthermore, the effects of STS on oxalate-induced oxidative stress were investigated in vitro in renal LLC-PK1 cells.

Results

Chronic EG exposure led to hyperoxaluria, oxidative stress, calcium oxalate crystalluria and crystal deposition in the kidneys. Whereas all tested compounds significantly reduced crystal load, only STS-treatment maintained tissue superoxide dismutase activity and urine 8-isoprostaglandin levels in vivo and preserved renal function. In in vitro studies, STS showed the ability to scavenge oxalate-induced ROS accumulation dose dependently, reduced cell-released hydrogen peroxide and preserved superoxide dismutase activity. As a mechanism explaining this finding, STS was able to directly inactivate hydrogen peroxide in cell-free experiments.

Conclusions

STS is an antioxidant, which preserves renal function in a chronic EG rat model. Its therapeutic use in oxidative-stress induced renal-failure should be considered.     

4-Hydroxybenzyl Alcohol Ameliorates Cerebral Injury in Rats by Antioxidant Action

4-hydroxybenzyl alcohol (4-HBA), one of the phenolic constituents found in many herbal medicinal plants, exhibits beneficial effects in neurological disorders. In the present study, we evaluated 4-HBA’s role in transient cerebral ischemia and its potential mechanism. Pre-treatment with 4-HBA (50,100 mg/kg) significantly reduced the cerebral infarct size and improved the neurological symptoms. Morphological examinations showed 4-HBA reduced the number of degenerated neurons. Oxidative stress was evaluated superoxide dismutase (SOD) activity and malondialdehyde (MDA) level. Anti-oxidative mechanisms were studied by Immunofluorescence staining and western immunoblot analysis. 4-HBA increased the expression of NAD(P)H: quinone oxidoreductase1 (NQO1) and ultimately inhibited oxidative stress. In addition, we evaluated the time course expression of NQO1, which was upregulated in the ischemic brain beginning at 1 h. Taken together, these results suggested that 4-HBA ameliorated cerebral injury in rats, This neuroprotective effect is likely related to its antioxidant activities.     

达格列净抑制2型糖尿病小鼠心血管及肾脏并发症的研究

摘要 目的：探讨达格列净对2型糖尿病小鼠心、肾的保护作用。方法：选取24只6周龄的雄性2型糖尿病模型(C57BLKS/J-leprdb/leprdb, db/db)小鼠,随机等分成达格列净投药组和对照组,另选取同周龄雄性非糖尿病的(C57BLKS/J-leprdb/+, db/m)小鼠12只作为正常组。检测小鼠血糖后,从第7周开始对投药组小鼠进行为期10周的达格列净用药,其余小鼠给予同等计量生理盐水,期间定期监测血糖、血压、尿糖以及各项代谢相关指标。投药结束后分离心脏及肾脏组织,组织切片进行染色观察。结果：与对照组相比,投药组达格列净用药后第1周血糖值显著降低(P < 0.01),用药9周后糖耐量测试结果显示血糖值几乎接近正常小鼠组水平,但各组间血压值无明显差异,心肌间质纤维化、炎性细胞浸润、氧化应激水平明显下降,同时肾小球硬化、炎性细胞浸润和氧化应激程度明显得到改善。结论：达格列净用药不仅能显著降低2型糖尿病模型小鼠血糖,还能有效抑制糖尿病引起的心血管及肾损害。     

低氧性肺动脉高压大鼠肺小动脉Siah1的表达变化及意义

目的:探讨低氧性肺动脉高压(HPH)大鼠肺小动脉Siah 1的动态表达变化及作用.方法:40只成年雄性Wistar大鼠随机分为对照组和低氧3 d、7d、14d、21 d组,每组8只,常压间断低氧复制HPH大鼠模型.测各组大鼠平均肺动脉压(mPAP)、右心室肥大指数(RVHI)、血管形态学指标;原位杂交、RT-PCR检测肺内Siah1 mRNA表达,免疫组化、Western blot检测其蛋白质水平.结果:①低氧7d后,肺小动脉出现血管重塑,且mPAP明显上升;低氧14d后,肺小动脉重塑更明显,mPAP达高峰.RVHI在低氧14d后明显增加.②Siah1 mRNA在对照组肺小动脉壁呈弱阳性表达,低氧7d后显著增高,低氧14d后达高峰.低氧21d后下降但仍高于对照组.免疫组化显示,Siahl蛋白在对照组肺小动脉壁呈弱阳性表达,低氧7d后显著增高,低氧14d后达高峰,低氧21 d后下降但仍高于对照组.③大鼠肺小动脉壁Siah1蛋白与Siah1 mRNA呈正相关;Siah1 mRNA、Siah1蛋白与mPAP、重塑指数、RVHI均呈正相关.结论:慢性低氧诱导肺小动脉壁Siah1表达增加,进而在HPH发病过程中发挥一定的作用.     

Amantadine Ameliorates Dopamine-Releasing Deficits and Behavioral Deficits in Rats after Fluid Percussion Injury

Aims

To investigate the role of dopamine in cognitive and motor learning skill deficits after a traumatic brain injury (TBI), we investigated dopamine release and behavioral changes at a series of time points after fluid percussion injury, and explored the potential of amantadine hydrochloride as a chronic treatment to provide behavioral recovery.

Materials and Methods

In this study, we sequentially investigated dopamine release at the striatum and behavioral changes at 1, 2, 4, 6, and 8 weeks after fluid percussion injury. Rats subjected to 6-Pa cerebral cortical fluid percussion injury were treated by using subcutaneous infusion pumps filled with either saline (sham group) or amantadine hydrochloride, with a releasing rate of 3.6mg/kg/hour for 8 weeks. The dopamine-releasing conditions and metabolism were analyzed sequentially by fast scan cyclic voltammetry (FSCV) and high-pressure liquid chromatography (HPLC). Novel object recognition (NOR) and fixed-speed rotarod (FSRR) behavioral tests were used to determine treatment effects on cognitive and motor deficits after injury.

Results

Sequential dopamine-release deficits were revealed in 6-Pa-fluid-percussion cerebral cortical injured animals. The reuptake rate (tau value) of dopamine in injured animals was prolonged, but the tau value became close to the value for the control group after amantadine therapy. Cognitive and motor learning impairments were shown evidenced by the NOR and FSRR behavioral tests after injury. Chronic amantadine therapy reversed dopamine-release deficits, and behavioral impairment after fluid percussion injuries were ameliorated in the rats treated by using amantadine-pumping infusion.

Conclusion

Chronic treatment with amantadine hydrochloride can ameliorate dopamine-release deficits as well as cognitive and motor deficits caused by cerebral fluid-percussion injury.     

The Expression of CUGBP1 After Spinal Cord Injury in Rats

Neurochemical Research - CUG-binding protein 1, a member of the CELF (CUGBP and embryonic lethal abnormal vision-like factor) family of RNA-binding proteins, is shown to be multifunctional,...     

Mixed Organic Solvents Induce Renal Injury in Rats

To investigate the injury effects of organic solvents on kidney, an animal model of Sprague-Dawley (SD) rats treated with mixed organic solvents via inhalation was generated and characterized. The mixed organic solvents consisted of gasoline, dimethylbenzene and formaldehyde (GDF) in the ratio of 2∶2:1, and were used at 12,000 PPM to treat the rats twice a day, each for 3 hours. Proteinuria appeared in the rats after exposure for 5–6 weeks. The incidences of proteinuria in male and female rats after exposure for 12 weeks were 43.8% (7/16) and 25% (4/16), respectively. Urinary N-Acetyl-β-(D)-Glucosaminidase (NAG) activity was increased significantly after exposure for 4 weeks. Histological examination revealed remarkable injuries in the proximal renal tubules, including tubular epithelial cell detachment, cloud swelling and vacuole formation in the proximal tubular cells, as well as proliferation of parietal epithelium and tubular reflux in glomeruli. Ultrastructural examination found that brush border and cytoplasm of tubular epithelial cell were dropped, that tubular epithelial cells were partially disintegrated, and that the mitochondria of tubular epithelial cells were degenerated and lost. In addition to tubular lesions, glomerular damages were also observed, including segmental foot process fusion and loss of foot process covering on glomerular basement membrane (GBM). Immunofluorescence staining indicated that the expression of nephrin and podocin were both decreased after exposure of GDF. In contrast, increased expression of desmin, a marker of podocyte injury, was found in some areas of a glomerulus. TUNEL staining showed that GDF induced apoptosis in tubular cells and glomerular cells. These studies demonstrate that GDF can induce both severe proximal tubular damage and podocyte injury in rats, and the tubular lesions appear earlier than that of glomeruli.     

Taurine Ameliorates Renal Oxidative Damage and Thyroid Dysfunction in Rats Chronically Exposed to Fluoride

Excessive exposure to fluoride poses several detrimental effects to human health particularly the kidney which is a major organ involved in its elimination from the body. The influence of taurine on fluoride-induced renal toxicity was investigated in a co-exposure paradigm for 45 days using five groups of eight rats each. Group I rats received normal drinking water alone, group II rats were exposed to sodium fluoride (NaF) in drinking water at 15 mg/L alone, group III received taurine alone at a dose of 200 mg/kg group IV rats were co-administered with NaF and taurine (100 mg/kg), while group V rats were co-administered with NaF and taurine (200 mg/kg). Administration of taurine significantly reversed the fluoride-mediated decrease in absolute weight and organo-somatic index of the kidney in the exposed rats. Taurine significantly prevented fluoride-induced elevation in plasma urea and creatinine levels in the exposed rats. Moreover, taurine restored fluoride-mediated decrease in the circulatory concentrations of triiodothyronine, thyroxine, and the ratio of triiodothyronine to thyroxine. Taurine ameliorated fluoride-mediated decrease in renal antioxidant status by significantly enhancing the antioxidant enzyme activities as well as glutathione level in the exposed rats. Additionally, taurine inhibited fluoride-induced renal oxidative damage by markedly decreasing the hydrogen peroxide and malondialdehyde levels as well as improved the kidney architecture in the treated rats. Collectively, taurine protected against fluoride-induced renal toxicity via enhancement of thyroid gland function, renal antioxidant status, and histology in rats.