miR-103a对癫痫大鼠海马组织星形胶质细胞活化的影响

为了考察miR-103a对癫痫大鼠海马组织星形胶质细胞活化的影响。本研究通过腹腔注射氯化锂和毛果芸香碱诱导癫痫大鼠模型,对大鼠脑室内注射miR-103a抑制剂来敲低miR-103a的表达;采用免疫组织化学染色检测大鼠海马组织中胶质纤维酸性蛋白(GFAP)的阳性表达;采用RT-qPCR和Western blotting方法检测大鼠海马组织中miR-103a、脑源性神经营养因子(BDNF)、GFAP、TNF-α和IL-6的m RNA和蛋白表达;苏木精-伊红(HE)染色评价海马组织病变程度;Nissl染色检测神经元存活情况;TUNEL染色检测神经元的凋亡。结果显示,癫痫大鼠海马组织中miR-103a被上调。下调miR-103a抑制癫痫大鼠海马组织中GFAP的mRNA和蛋白表达,且抑制癫痫大鼠海马神经元的病理损伤,但能促进癫痫大鼠海马神经元的存活并抑制其凋亡。此外,下调miR-103a还抑制癫痫大鼠海马组织中IL-6和TNF-α的表达,并促进癫痫大鼠海马组织中BDNF的表达。本研究表明,靶向沉默miR-103a可以抑制癫痫大鼠海马组织中星形胶质细胞的活化并改善神经元的病理损伤。     

蝎毒对癫痫敏感性和海马GFAP释放的影响

目的和方法 :本工作用海人酸癫痫模型 ,通过对癫痫大鼠蝎毒治疗后行为变化及脑内胶质原纤维酸性蛋白(GFAP)免疫反应活性的检测 ,对蝎毒抗癫痫反复发作的相关脑区及其机制做以初步探讨。结果 :癫痫大鼠蝎毒治疗三周后 ,能明显减少癫痫发作的例数 ,减轻癫痫发作的程度 ,使发作的潜伏期延长 (P <0 .0 5 )。免疫细胞化学的实验显示 ,蝎毒抗癫痫反复发作的相关脑区是海马。 8例蝎毒治疗的大鼠与实验对照组相比 ,有 6例背侧海马GFAP免疫染色明显减轻 ,未见星形胶质细胞增生 ;CA1区无明显神经元缺失 ;而且与空白对照组相比无显著差异。结论 :癫痫大鼠蝎毒治疗三周后 ,能明显减轻癫痫发作的行为 ,抑制海马星形胶质细胞的增生肥大 ,减轻海马神经元受损的程度。蝎毒抑制海马星形胶质细胞增生很可能是蝎毒抗癫痫反复发作的重要机制之一。     

lncRNA PVT1沉默对高糖诱导的血管内皮细胞增殖、凋亡及氧化应激的影响*

目的: 探讨抑制lncRNA PVT1对高糖诱导的血管内皮细胞的增殖,凋亡和氧化应激的影响。方法: 体外培养人脐静脉内皮细胞(HUVECs),分为四组:对照组(5.5 mmol/L葡萄糖),高糖组(30 mmol/L葡萄糖),高糖+siNC组(30 mmol/L葡萄糖+siNC,细胞转染阴性对照组),高糖+siPVT1组(30 mmol/L葡萄糖+siPVT1,抑制lncRNA PVT1组)。采用荧光定量PCR的方法检测转染后PVT1的表达水平。MTT检测siPVT1(短片段干扰RNA PVT1)对高糖诱导的HUVECs细胞增殖能力的影响。流式细胞术检测siPVT1对高糖诱导的HUVECs细胞ROS和凋亡水平。Western blot检测HUVECs细胞中凋亡相关蛋白如Bax,Bcl-2和cleaved-caspase-3的表达水平。结果: 与对照组比较,转染siPVT1后,PVT1的表达水平显著降低(P＜0.05)。MTT结果显示,与对照组比较,培养24 h和48 h后高糖组中HUVECs细胞增殖活力均显著降低,与高糖+siNC组(阴性对照组)比较,培养24 h和48 h后,高糖+siPVT1组中的HUVECs细胞增殖活力显著增加(P＜0.05)。流式细胞术检测结果表明,与对照组比较,高糖组HUVECs细胞中ROS和凋亡率均显著增加;和高糖+siNC组比较,高糖+siPVT1组中HUVECs细胞中ROS和凋亡率均有减少(P＜0.05)。Western blot结果表明,与对照组比较,高糖组中cleaved-caspase-3和Bax表达水平均显著上调,Bcl-2的表达水平显著下调(P＜0.05,P＜0.01)。与高糖+siNC组比较,高糖+siPVT1组cleaved-caspase-3和Bax表达水平显著下调,Bcl-2的表达显著上调(P＜0.05,P＜0.01)。结论: 抑制lncRNA PVT1可以显著增加高糖诱导的HUVECs细胞增殖活力,减轻氧化应激,抑制细胞凋亡。     

美洛昔康对Aβ诱导Alzheimer’s disease大鼠脑内炎症损伤的影响

目的：研究美洛昔康对β-淀粉样蛋白（Aβ）诱导的阿尔茨海默病（AD）模型大鼠脑内炎症损伤的保护作用,并探讨其抑制炎症作用的机制。方法：Aβ1-40海马注射建立AD大鼠模型。免疫组化法观察大鼠海马核因子κBp65（NF-κBp65）和星形胶质细胞（AS）胶质纤维酸性蛋白（GFAP）表达变化;Western-blot法测定大鼠皮层组织GFAP的表达;ELISA法检测大鼠皮层组织肿瘤坏死因子-α（TNF-α）水平变化;RT-PCR法检测大鼠海马组织白细胞介素-1β（IL-1β）mRNA的表达情况。结果：美洛昔康能抑制AD大鼠海马NF-κBp65和GFAP的表达;降低大鼠皮层TNF-α的含量;抑制AD大鼠海马IL-1βmRNA的表达。结论：美洛昔康通过减少AD模型大鼠海马、皮层组织GFAP表达,抑制AS的增生,降低NF-κBp65的活性,减少炎症因子TNF-α和IL-1β的水平,减轻脑内炎症反应。     

Wogonin preventive impact on hippocampal neurodegeneration,inflammation and cognitive defects in temporal lobe epilepsy

Previous studies demonstrated that the pathophysiological changes after temporal lobe epilepsy (TLE) such as oxidative stress, inflammatory reaction contribute to cognitive defect and neuronal damage. The present study was conducted to evaluate the anticonvulsant effect of wogonin ameliorates kainate-induced TLE, and to investigate the mechanism underlying these effects. Rats were divided into control, wogonin, kainate, and wogonin-pretreated kainate groups. The rat model of TLE was induced by unilateral intrahippocampal injection of 0.4 ug/ul of kainate. The results showed that the cognitive function in TLE rats was significantly impaired, and wogonin treatment improved cognitive function in the Morris water maze (MWM). H & E staining and TUNEL staining showed obvious damage in the hippocampus of TLE rats, and wogonin alleviated the damage. To evaluate the oxidative stress, the expression of MDA and GSH in plasma were detected. Nrf-2 and HO-1 mRNA expression in the hippocampus were detected. The levels of MDA in plasma increased in TLE rats, and the levels of GSH in plasma and Nrf-2, HO-1 in the brain decreased. Treatment with wogonin alleviated these changes. We also detected the mRNA expression of inflammatory mediators like IL-1β, TNF-α, and NF kB in the brain. The inflammatory reaction was significantly activated in the brain of TLE rats, and wogonin alleviated neuroinflammation. We detected the mRNA expression of Bcl-2, Bax, caspase-3, in the hippocampus. The levels of Bcl-2 decreased in TLE rats, Bax and caspase-3 increased, while wogonin alleviated these changes. The present study indicated that wogonin exerted a noticeable neuroprotective effect in kainate-induced TLE rats.     

Administration of simvastatin after kainic acid-induced status epilepticus restrains chronic temporal lobe epilepsy

In this study, we examined the effect of chronic administration of simvastatin immediately after status epilepticus (SE) on rat brain with temporal lobe epilepsy (TLE). First, we evaluated cytokines expression at 3 days post KA-lesion in hippocampus and found that simvastatin-treatment suppressed lesion-induced expression of interleukin (IL)-1β and tumor necrosis factor-α (TNF-α). Further, we quantified reactive astrocytosis using glial fibrillary acidic protein (GFAP) staining and neuron loss using Nissl staining in hippocampus at 4-6 months after KA-lesion. We found that simvastatin suppressed reactive astrocytosis demonstrated by a significant decrease in GFAP-positive cells, and attenuated loss of pyramidal neurons in CA3 and interneurons in dentate hilar (DH). We next assessed aberrant mossy fiber sprouting (MFS) that is known to contribute to recurrence of spontaneous seizure in epileptic brain. In contrast to the robust MFS observed in saline-treated animals, the extent of MFS was restrained by simvastatin in epileptic rats. Attenuated MFS was related to decreased neuronal loss in CA3 and DH, which is possibly a mechanism underlying decreased hippocampal susceptibility in animal treated with simvastatin. Electronic encephalography (EEG) was recorded during 4 to 6 months after KA-lesion. The frequency of abnormal spikes in rats with simvastatin-treatment decreased significantly compared to the saline group. In summary, simvastatin treatment suppressed cytokines expression and reactive astrocytosis and decreased the frequency of discharges of epileptic brain, which might be due to the inhibition of MFS in DH. Our study suggests that simvastatin administration might be a possible intervention and promising strategy for preventing SE exacerbating to chronic epilepsy.     

脐带间充质干细胞对内毒素血症诱发的大鼠急性肝功能损伤的影响

目的评价脐带间充质干细胞（hUC-MSCs）对内毒素血症诱发的大鼠急性肝功能损伤的影响及其与凋亡机制的关系。 方法6周龄雄性SD大鼠18只,随机分为3组,分别是对照组（C组）、内毒素血症组（M组）和内毒素血症+hUC-MSCs治疗组（M+cells组）,每组6只。大鼠腹腔注射5 mg/kg脂多糖（LPS）诱导内毒素血症模型,并经尾静脉注射含20×10⁶个hUC- MSCs。4 h时检测血清谷草转氨酶（AST）和谷丙转氨酶（ALT）,ELISA方法检测肿瘤坏死因子（TNF-α）、白细胞介素6（IL-6）,HE常规染色鉴定肝脏组织病理,Western Blot法检测肝脏组织抗凋亡蛋白Bcl-2、促凋亡蛋白Bax、凋亡信号调节激酶1（ASK1）、应激活化蛋白激酶即JUN氨基末端激酶（JNK）蛋白的表达。多组间比较采用单因素方差分析,相关分析选用pearson。 结果（1）C组AST、ALT、TNF-α和IL-6浓度分别为（74.66±6.39）U/ L、（40.07±6.07）U/ L、（37.74±3.08）ng/L和（0.42±0.07）ng/L;与M组比较（310.75±9.13）U/L、（107.04±10.04）U/ L、（160.32±4.88）ng/L和（0.90±0.09）ng/L,差异具有统计学意义（P均 < 0.05）,M组AST、ALT、TNF-α、IL-6浓度分别为（310.75±9.13）U/L、（107.04±10.04）U/ L、（160.32±4.88）ng/ L和（0.90±0.09）ng/L,与M+cells组比较（204.49±15.36）U/L、（71.24± 7.34）U/ L、（117.61±9.37）ng/ L和（0.60±0.10）ng/L,差异具有统计学意义（P均 < 0.05）。（2）C组大鼠肝细胞形态正常,可见肝小叶结构清晰,肝汇管区无炎性细胞浸润,M组大鼠肝小叶散在点状坏死肝细胞伴炎性浸润,肝细胞间隙散布增生的Kuffer细胞,M+cells组大鼠肝小叶炎性细胞浸润及肝细胞间隙Kuffer细胞浸润改善。（3）与C组比较,M组大鼠肝脏组织JUN、ASK1和Bax蛋白表达均增高（P均 < 0.05）,Bcl-2蛋白表达降低（P < 0.05）;与M组比较,M+cells组大鼠肝脏组织JUN、ASK1和Bax蛋白表达降低（P均 < 0.05）,Bcl-2蛋白增加（P < 0.05）。（4）单因素相关分析显示大鼠血清ALT、AST与TNF-a指数呈正相关（r值分别为0.9580、0.9865,P均< 0.05）,大鼠血清ALT、AST与IL-6指数呈正相关（r值分别为0.9892、0.9630,P均 < 0.05）,大鼠血清ALT、AST分别与BAX、ASK1、JNK指数均呈正相关（r值分别为0.9993、0.9851、0.7901、0.9864、0.9557、0.7128,P均 < 0.05）,大鼠血清ALT、AST分别与BCL-2指数均呈负相关（r值分别为-0.8824、-0.9338,P均 < 0.05）,大鼠血清TNF-α分别与BAX、ASK1、JNK指数均呈正相关（r值分别为0.9466、0.8958、0.6025,P均< 0.05）,大鼠血清TNF-α与BCL-2指数呈负相关（r = -0.6025,P均 < 0.05）,大鼠血清IL-6分别与BAX、ASK1、JNK指数均呈正相关（r值分别为0.9941、0.9997、0.8679,P均< 0.05）,大鼠血清IL-6与BCL-2指数呈负相关（r = -0.8078,P均 < 0.05）。 结论hUC-MSCs具有减轻内毒素血症大鼠急性肝功能损伤的作用,其机制与抑制肝脏细胞凋亡相关。     

Lentivirus-mediated microRNA-26a-modified neural stem cells improve brain injury in rats with cerebral palsy

This study is launched to investigate the effect of lentivirus-mediated microRNA-26a (miR-26a)-modified neural stem cells (NSCs) in brain injury in rats with cerebral palsy (CP). The successfully constructed miR-26a lentivirus expression vector and empty vector virus were used to modify NSCs. The model of CP with ischemia and anoxia was established in rats. NSCs and miR-26a-NSCs were stereoscopically injected into the cerebral cortex of the modeled rats, respectively. The survival and migration of NSCs infected with recombinant lentivirus expressing green fluorescence in vivo was observed under a light microscope. The neurobehavioral functions, morphology, and ultrastructure of cerebral cortex and hippocampus, apoptosis of brain cells, expression of apoptosis-related protein caspase-3 and Bax, together with the expression of the glial fibrillary acidic protein (GFAP) in cerebral cortex and hippocampus were determined. Expression of miR-26a in NSCs infected with plVTHM-miR-26a increased significantly. After NSCs transplantation, the neurobehavioral status of CP rats was improved, the degree of brain pathological injury was alleviated, the apoptotic index of cells in cerebral cortex and hippocampus and the expression of the apoptotic protein (caspase-3 and Bax) were decreased, the expression of GFAP were significantly decreased. After miR-26a-NSCs transplantation, these aforementioned results further improved or decreased. Our study suggests that miR-26a-modified NSCs mediated by lentivirus can improve brain injury, inhibit apoptosis of brain cells and activation of astrocytes in CP rats.     

Reactive Transformation and Increased BDNF Signaling by Hippocampal Astrocytes in Response to MK-801

MK-801, also known as dizocilpine, is a noncompetitive N-methyl-D-aspartic acid (NMDA) receptor antagonist that induces schizophrenia-like symptoms. While astrocytes have been implicated in the pathophysiology of psychiatric disorders, including schizophrenia, astrocytic responses to MK-801 and their significance to schizotypic symptoms are unclear. Changes in the expression levels of glial fibrillary acid protein (GFAP), a marker of astrocyte activation in response to a variety of pathogenic stimuli, were examined in the hippocampus of rats treated with the repeated MK-801 injection (0.5 mg/10ml/kg body weight for 6 days) and in primary cultured hippocampal astrocytes incubated with MK-801 (5 or 20 μM for 24 h). Moreover, the expression levels of BDNF and its receptors TrkB and p75 were examined in MK-801-treated astrocyte cultures. MK-801 treatment enhanced GFAP expression in the rat hippocampus and also increased the levels of GFAP protein and mRNA in hippocampal astrocytes in vitro. Treatment of cultured hippocampal astrocytes with MK-801 enhanced protein and mRNA levels of BDNF, TrkB, and p75. Collectively, our results suggest that hippocampal astrocytes may contribute to the pathophysiology of schizophrenia symptoms associated with NMDA receptor hypofunction by reactive transformation and altered BDNF signaling.     

Prolactin induced expression of interleukin-1 alpha,tumor necrosis factor-alpha,and transforming growth factor-alpha in cultured astrocytes

Prolactin (PRL) is a potent mitogen in cultured astrocytes. Because one of the major effects of astrocyte proliferation is the expression of inflammatory cytokines, we examined the effect of PRL-induced mitogenesis on the expression of interleukin-1 (IL-1α), tumor necrosis factor-α (TNF-α), and transforming growth factor-α (TGF-α) in cultured astrocytes. Astrocytes were stimulated with PRL or growth hormone (GH), and the expression of cytokines was determined by immunohistochemistry and Western blot analysis. Following incubation of astrocytes with 1 nM PRL for 6 h, strong positive staining of IL-1α and TNF-α, but not TGF-α, was found. No detectable staining for the above cytokines was found in vehicle, or GH treated astrocytes. When astrocytes were incubated in the presence of 1 nM PRL for 18 h, strong positive staining for IL-1α and TGF-α was found. Immunocytochemical analysis of the expression of TNF-α and IL-1α in PRL stimulated astrocytes suggested that the expression of IL-1α preceded the expression of TNF-α. To confirm this observation, Western blot analyses were performed on extracts from astrocytes incubated with 1 nM PRL. In unstimulated astrocytes, IL-1α levels were not detectable. In astrocytes stimulated with 1 nM PRL, expression of IL-1α was clearly detected after 1 h of incubation, and IL-1α levels continued to increase during the course of the experiment (6 h). In contrast, in astrocytes stimulated with 1 nM PRL, an increase in the expression of TNF-α was first apparent after 2 h of incubation. TNF-α levels peaked 3 to 4 h after the addition of PRL, and returned to near control levels after 6 h. Finally, injection of PRL into a wound site in female rats increased the expression of glial fibrillary acid protein (GFAP), an astrocyte specific protein. These data suggest that PRL can stimulate astrogliosis at the wound site in vivo. These data clearly indicate that PRL can stimulate the expression of TNF-α and IL-1α in cultured astrocytes and suggest that PRL may play a role in the regulation of the neuroimmune response in vivo.     

细胞周期调控对大鼠全脑缺血后海马迟发性神经元死亡的影响

目的观察细胞周期调控对大鼠全脑缺血再灌流后海马区迟发性神经元死亡(delayed neuronal death,DND)以及星形胶质细胞的活化、增殖的影响.方法建立大鼠短暂性全脑缺血再灌流模型,利用尼氏染色、TUNEL、免疫组织化学方法观察再灌流后细胞周期素依赖的蛋白激酶(cyclin depedent kinase, CDK)抑制剂Olomoucine对海马DND以及星形胶质细胞活化增殖的影响.结果全脑缺血再灌流后3d、7d、30d海马神经元明显脱失,部分CA1、CA2区神经元凋亡;星形胶质细胞数目增多,GFAP表达上调,应用Olomoucine后TUNEL阳性神经元数目明显减少,幸存神经元数目增加;星形胶质细胞数目无明显增多,GFAP表达明显下调.结论 CDK抑制剂Olomoucine可有效抑制大鼠全脑缺血后海马神经元DND以及星形胶质细胞活化增殖.     

Kinetics of Apoptosis and Expression of Apoptosis-Related Proteins in Rat CA3 Hippocampus Cells After Experimental Diffuse Brain Injury

The present study examined kinetics of apoptosis and expression of apoptosis-related proteins Bcl-2, Bax, and caspase-3 in the CA3 hippocampus cells after diffuse brain injury (DBI) induced experimentally in rats. Percentage of apoptotic cells and expressions of above proteins were examined by flow cytometry and immunohistochemistry. Substantial neuronal apoptosis was documented in the CA3 hippocampus cells after DBI (22.26 ± 2.97 % at 72 h after DBI vs. 2.92 ± 0.88 % in sham-operated animals). Expression of Bc1-2 decreased, while expression of Bax and caspase-3 increased after DBI, with caspase-3 expression peaking after that of Bax (72 vs. 48 h, respectively). Further, the Bc1-2/Bax expression ratio decreased prior to increase of caspase-3 expression. In conclusion, cell apoptosis and altered expressions of Bcl-2, Bax, and caspase-3 are present in the CA3 region of hippocampus after experimental DBI. Changes in the Bc1-2/Bax expression ratio may facilitate activation of caspase-3 and aggravate neuronal apoptosis after brain injury.     

Hepatic oxidative stress,up-regulation of pro-inflammatory cytokines,apoptotic and oncogenic markers following 2-methoxyethanol administrations in rats

2-methoxyethanol (2-ME) is an organic solvent widely used in the manufacture of brake fluids, paints, resins, varnish, nail polish, acetate cellulose, wood coloring, and as a plasticizer in plastics manufacturing. We therefore, investigated its effect on the liver, in a time-course study in male Wistar rats. Animals were orally administered 50 mg/kg body weight of 2-ME for a period of 7, 14, and 21 days. Following 7 days of administration of 2-ME, there was a significant increase in the level of Bax, c-Myc, K-Ras, TNF-α, IL-1β, IL-6, MDA and GPx activity, while the levels of Bcl-2, NO and GSH were significantly reduced compared with control. At the end of 14 days exposure, Bcl-2, and GSH levels, as well as GST activity, were significantly decreased, while levels of Bax, c-Myc, K-Ras, caspase-3, TNF-α, IL-1β, IL-6, MDA and NO were significantly increased compared with control. After 21 days of 2-ME administration, Bcl-2, IL-10, and GSH levels, as well as SOD and GST activities, were significantly decreased, while levels of Bax, c-Myc, K-Ras, caspase-3, p53, TNF-α, IL-1β, IL-6, MDA and NO were significantly increased compared with control. Lastly, liver histopathology confirmed and corroborated the biochemical findings reported above. We therefore, advised that exposures to 2-ME should be strictly avoided as it could trigger hepatic damage through the disorganization of the antioxidant system, up-regulation of inflammatory, apoptotic, and oncogenic markers in rats.     

Evaluation of Insulin and Ascorbic Acid Effects on Expression of Bcl-2 Family Proteins and Caspase-3 Activity in Hippocampus of STZ-Induced Diabetic Rats

Aims Effects of insulin and ascorbic acid on expression of Bcl-2 family proteins and caspase-3 activity in hippocampus of diabetic rats were evaluated in this study. Methods Diabetes was induced in Wistar male rats by streptozotocin (STZ). Six weeks after verification of diabetes, the animals were treated for 2 weeks with insulin or/and ascorbic acid in separate groups. Hippocampi of rats were removed and evaluation of Bcl-2, Bcl-x_L, and Bax proteins expression in frozen hippocampi tissues were done by SDS-PAGE electrophoresis and blotting. The Bcl-2, Bcl-x_L, and Bax proteins bands were visualized after incubation with specific antibodies using enhanced chemiluminescences method. Caspase-3 activity was determined using the caspase-3/CPP32 Fluorometric Assay Kit. Results Diabetic rats showed increase in Bax protein expression and decrease in Bcl-2 and Bcl-x_L proteins expression. The Bax/Bcl-2 and Bax/Bcl-x_L ratios were found higher compared with non-diabetic control group. Treatments with insulin and/or ascorbic acid were resulted in decrease in Bax protein expression and increase in Bcl-2 and Bcl-x_L proteins expression. The Bcl-2/Bax and Bcl-x_L/Bax ratios were found higher in treated groups than untreated diabetic group. Caspase-3 activity level was found higher in diabetic group compared with non-diabetic group. Treatment with insulin and ascorbic acid did downregulated caspase-3 activity. Conclusions Our data provide supportive evidence to demonstrate the antiapoptotic effects of insulin and ascorbic acid on hippocampus of STZ-induced diabetic rats.     

骨髓基质干细胞移植改善慢性酒精中毒大鼠脑损害的相关机制研究

目的:探讨骨髓基质干细胞(bone marrow stormal cells, BMSCs)静脉移植对慢性酒精中毒大鼠脑保护作用的相关机制。方法:体外分离、培养、扩增SD大鼠BMSCs。成年雄性SD大鼠随机分为慢性酒精中毒组、BMSCs回输组、磷酸缓冲盐溶液(phosphate buffer saline,PBS)回输组和对照组,每组7只。前三组用酒精灌胃8周建立慢性酒精中毒动物模型,对照组不造模(给予蔗糖灌胃),BMSCs回输组和PBS回输组于造模7周时一次性经尾静脉回输BMSCs或PBS。免疫印迹法检测海马Bcl-2、Bax、NGF、BDNF以及信号转导分子p-Akt的表达;反转录PCR检测海马神经生长因子(nerve growth factor, NGF)和脑源性神经营养因子(brain derived neurotrophic factor, BDNF)。结果:BMSCs回输组海马抗凋亡蛋白Bcl-2表达高于其余三组(P0.05);促凋亡蛋白Bax表达低于慢性酒精中毒组(P0.01),与对照组无统计学差异(P=0.989)。BMSCs回输组鼠海马内NGF和BDNF m RNA和蛋白表达、p-Akt蛋白表达均高于其余三组(P0.05)。结论:静脉移植BMSCs能够明显改善慢性酒精中毒大鼠海马的细胞凋亡;其可能与自或旁分泌BDNF和NGF营养因子有关,且可能部分是通过激活PI3K/Akt通路实现。     

Gastrodin Ameliorates Depressive-Like Behaviors and Up-Regulates the Expression of BDNF in the Hippocampus and Hippocampal-Derived Astrocyte of Rats

Gastrodin (GAS), a main constituent of a Chinese herbal medicine Tian ma, has been shown to be effective in treating various mood disorders. The purpose of the present study was to assess the effects of GAS on alleviating depressive-like behaviors in a rat model of chronic unpredictable stress (CUS) and regulating the expression of BDNF in the hippocampus and hippocampal-derived astrocyte from Sprague–Dawley (SD) rats. Following CUS, rats were intraperitoneally administered gastrodin (50, 100, or 200 mg/kg daily) or vehicle for 2 weeks. Rats were then experienced sucrose preference test and forced swim test. The expressions of GFAP and BDNF in the hippocampus were evaluated. In addition, hippocampal astrocytes were isolated from neonatal SD rats and exposed to different concentrations of GAS (sham, 5, 10, 20, 50 and 100 μg/mL) for 48 and 72 h before the cell viability and the levels of pERK1/2 and BDNF were analyzed. Furthermore, the cell viability was also tested after exposure to serum-free condition that contain different concentrations of GAS for 48 and 72 h. GAS administration (100 and 200 mg/kg daily) reversed depressive-like behaviors in rats exposed to CUS paradigm and restored the expression of GFAP and BDNF in the hippocampus. Moreover, in vitro experiments revealed that GAS did not increase the cell viability of astrocytes but protected it from 72 h’s serum-free damage at the dosage 20 μg/mL. Increased levels of ERK1/2 phosphorylation and BDNF protein were also observed after GAS (20 μg/mL) treatment for 72 h. These results indicate that gastrodin possesses antidepressant effect. The changes of the astrocyte activation and the level of BDNF may play a critical role in the pharmacological action of GAS.     

Contribution of inducible nitric oxide synthase and cyclooxygenase-2 to apoptosis induction in smooth chorion trophoblast cells of human fetal membrane tissues

We examined the contribution of apoptosis- and oxidative stress-associated genes to apoptosis induction in trophoblast cells of human fetal membrane tissues undergoing apoptosis during in vitro incubation. RT-PCR analyses demonstrated an increased level of HO-1, Mn-SOD, Cox-2, iNOS, TNFalpha, TNFR1, IL-1beta, IL-6, Bax, Bak, and Bad gene expression, while Bcl-2 mRNA expression level decreased. Western blot analyses demonstrated an increase in iNOS, Cox-2, and HO-1 protein levels; a decrease in pro-caspase-3 and 9, proform-PARP, and Apaf-1 protein levels; a leakage of cytochrome c from the mitochondria. An antioxidative reagent, general and selective Cox-2 inhibitors, and an iNOS inhibitor suppressed in vitro progression of the apoptosis. Furthermore, an NO donor reagent induced apoptosis in primary cultured trophoblast cells. Therefore, we concluded that the induction of apoptosis in the smooth chorion trophoblasts is mediated through oxidative stress induction followed by mitochondria damage, suggesting that iNOS and Cox-2 play an important role in the apoptosis induction in trophoblasts of human fetal membrane tissues.     

川陈皮素对糖尿病肾病大鼠的治疗作用研究

为观察川陈皮素对糖尿病肾病的治疗作用,本研究选用120只SD大鼠适应性喂养2周后分为正常组(20只)和糖尿病肾病造模组(100只),成功构建糖尿病肾病模型后选取50只,分为模型组,川陈皮素低剂量、中剂量和高剂量组以及阳性药物贝那普利组,每组10只,治疗6周后处死,收集尿液检测24h-尿量和24h-尿蛋白,收集血液检测血糖、胰岛素、血脂、肾功能指标和炎性因子的变化特点,收集肾脏检测肾脏病理学以及肾脏组织中凋亡相关蛋白Bcl-2、Bax、Caspase-3表达水平。结果显示,模型组有明显肾小球增大、部分系膜增生和间质纤维化,相较于模型组,贝那普利组和川陈皮素三个剂量组肾小球病变减轻;与模型组相比,贝那普利组和川陈皮素低、中、高剂量组UCr、24h蛋白尿、BUN、Scr、血糖、TG、TC、IL-1、IL-6和TNF-α明显降低(P<0.05),胰岛素含量明显升高(P<0.05);与模型组相比,贝那普利组和川陈皮素低、中和高剂量组Bax和Caspase-3明显降低(P<0.05),Bcl-2明显升高(P<0.05)。上述研究表明,川陈皮素对糖尿病肾脏损害大鼠肾功能具有明显的保护作用。