瘦素对糖尿病大鼠糖脂代谢及炎性因子的影响

目的: 探讨瘦素对糖尿病大鼠糖脂代谢及相关炎症因子的影响。方法: 将健康Wistar雄性大鼠60只随机选取10只作为对照组,50只给予高糖高脂饲料喂养加腹腔内注射链脲佐霉素(STZ,25 mg/kg)的方法诱发并建立糖尿病大鼠模型。并随机分为模型组、瘦素低剂量组、瘦素中剂量组和瘦素高剂量组,每组10只。4组大鼠造模成功后均持续给予高糖高脂饲料喂养,瘦素低、中、高剂量组给予20 μg/kg、50 μg/kg和100 μg/kg,连续5 d。GOD-PAP法检测大鼠血糖(FBG),放射免疫法测定胰岛素含量(Ins),全自动生化分析仪测定血清中三酰甘油(TG)、总胆固醇(TC)、低密度脂蛋白(LDL-C)、高密度脂蛋白(HDL-C)。采用酶联免疫方法(ELISA)测定丙二醛(MDA)、白细胞介素-6(IL-6)及肿瘤坏死因子-α(TNF-α)。采用Western blot检测糖尿病大鼠脂肪组织中瘦素表达情况。结果: 与对照组比较,各组大鼠血糖水平均显著升高(P＜0.01);与模型组比较,瘦素中、高剂量大鼠血糖显著降低(P＜0.05,P＜0.01);瘦素高剂量组胰岛素水平显著降低(P＜0.01)。不同剂量瘦素组间比较,给药后三组大鼠FBG及INS无统计学差异(P＞0.05)。与模型组比较,瘦素中、高剂量组TC水平显著下降(P＜0.05,P＜0.01);高剂量组TG、LDL-C水平显著降低(P＜0.05),高剂量组HDL-C水平显著升高(P＜0.01)。不同剂量瘦素组进行组间比较,高剂量组在降低TC、TG、LDL-C水平,升高HDL-C水平优于中、低剂量组(P＜0.05)。Western blot结果显示,与模型组(52.27±10.93)比,瘦素高剂量组100 μg/kg(40.13±9.87)、中剂量组50 μg/kg(44.68±10.23)、低剂量组20 μg/kg(47.35±12.09)脂肪中瘦素表达水平依次降低。结论: 瘦素水平分泌异常是诱发糖尿病因素之一,在给予一定浓度外源性瘦素(100 μg/kg)干预下,能显著降低MDA、TNF-α水平,提高IL-6水平,其机制可能与瘦素在减轻炎症反应、氧化应激,纠正血脂异常紊乱有密切关系。     

贝那普利和氨氯地平对自发性高血压大鼠血压和肠道微生物的影响

目的观察贝那普利和氨氯地平对自发性高血压大鼠(SHR)血压及肠道微生物的影响。方法 18只14周龄雄性SHR随机分为模型组(Mod组,n=6)、贝那普利组[Bph组,盐酸贝那普利灌服0.90 mg/(kg·d),n=6]、氨氯地平组[Aml组,苯磺酸氨氯地平灌服0.45 mg/(kg·d),n=6],另以Wistar-Kyoto(WKY)大鼠作为正常对照组(NC组,n=6), NC组和Mod组每日给予蒸馏水灌胃,干预8周后测定各组大鼠血压,观察结束处死大鼠取空肠粪便,进行16S rDNA V4区扩增子信息采集与分析。结果贝那普利和氨氯地平对SHR具有明显的降压作用,用药后对SHR肠道微生物的门属组成、α多样性、β多样性有一定影响;根据关系热图分析菌群和环境因子血压的关系,拟杆菌属(Bacteroidetes)、厚壁菌属(Firmicutes)和粪球菌属(Coprococcus)与收缩压(SBP)相关,拟杆菌属与平均动脉压(MBP)相关。结论贝那普利和氨氯地平具有明确的降压作用,并对SHR肠道微生物有一定的影响。     

推拿对慢性应激大鼠抑郁行为的影响及其机制

目的: 探讨推拿对慢性应激大鼠抑郁行为的影响及其作用机制。方法: 制备慢性轻度不可预见性的应激大鼠模型^[1-2],造模21 d后,进行推拿治疗14 d。分组:空白对照组、模型组、推拿组、氟西汀组,每组10只。每日推拿膀胱经重要穴位10 min(间隔2 min,共2次)。通过体质量检测、旷场、糖水消耗实验和水迷宫实验评价抑郁模型大鼠行为学改变情况;蛋白质免疫印迹法(Western blot)法检测大鼠海马及前额叶皮质组织中ERK/P-ERK、BDNF蛋白表达情况。结果: 模型组与空白组比较,大鼠的体质量、旷场、糖水消耗实验和水迷宫数据均显著下降(P＜0.01),P-ERK、BDNF蛋白含量均显著降低(P＜0.01);推拿组和氟西汀组与模型组比较,大鼠的体质量、旷场实验、糖水消耗实验和水迷宫实验数据均显著上升(P＜0.01),推拿组和氟西汀组大鼠海马及前额叶皮质组织中P-ERK、BDNF蛋白含量均显著升高(P＜0.05,P＜0.01),氟西汀组升高更为显著(P＜0.01)。结论: 推拿可上调大鼠海马及前额叶皮质组织中ERK蛋白的磷酸化水平,激活ERK信号通路,促进效应蛋白BDNF的表达,改善慢性应激大鼠的抑郁行为。     

牛磺酸上调基因1(TUG1)对肝纤维化的作用及其机制*

目的:探讨牛磺酸上调基因1(TUG1)在肝纤维化中的作用机制。方法:按照文献建立TGF-β1(5 ng/ml)刺激的活化肝星状细胞模型和经典的1%DMN(1 ml/kg/d)致大鼠肝纤维化模型,将肝纤维化大鼠和活化肝星状细胞(HSC)均分为模型对照组、阴性对照组(沉默TUG1阴性对照)、siRNA干扰组(TUG1基因沉默组)。实验结束后利用苏木精-伊红(HE)染色检测大鼠肝脏组织病理变化;采用逆转录-聚合酶链反应(RT-PCR)法、蛋白免疫印记(Western blot)分别测定大鼠肝组织及活化肝星状细胞中α-平滑肌肌动蛋白(α-SMA)、TUG1、I型胶原蛋白(collagenI)、基质金属蛋白酶-2(MMP-2)、金属蛋白酶组织抑制因子(TIMP-1)、Smad2、Smad3表达水平。结果:肝组织病理学检查显示,沉默TUG1能够明显缓解肝脏纤维化病理改变,Western blot结果显示,沉默TUG1能够显著降低大鼠肝组织和活化肝星状细胞中TUG1、α-SMA、collagenI、MMP-2、TIMP-1、Smad2、Smad3基因与蛋白表达水平(P＜0.05)。与模型对照组相比,阴性对照组的TUG1、α-SMA的蛋白与基因水平明显升高(P＜0.05)。与模型对照组和阴性对照组相比,siRNA干扰组中TUG1, α-SMA, collagenI, MMP-2, TIMP-1, Smad2 and Smad3的蛋白和基因水平显著降低(P＜0.05),而在模型对照组和阴性对照组中TUG1, α-SMA, collagenI, MMP-2, TIMP-1, Smad2 and Smad3的蛋白和基因表达水平之间差异无显著性。结论:TUG1在肝纤维化组织和活化的肝星状细胞中显著上调,沉默TUG1可能通过抑制转化生长因子-β1(TGF-β1)/Smad信号通路改善1%DMN致大鼠肝纤维化病理损伤,降低活化肝星状细胞中纤维化相关蛋白水平,发挥抗肝纤维化的作用。     

妊娠期母体甲状腺功能减退对胎鼠骨骼肌胰岛素抵抗和线粒体功能的影响

摘要 目的：研究妊娠期母体甲状腺功能减退对胎鼠骨骼肌胰岛素抵抗和线粒体功能的影响。方法：构建妊娠期母体甲状腺功能减退小鼠模型,制备胎鼠骨骼肌线粒体,同时选取健康正常胎鼠做本次实验的对照组。采用酶联免疫吸附法和放射免疫分析法测定两组小鼠甲状腺功能;通过全自动生化分析仪检测两组胎鼠胰岛素抵抗结果;利用Clark氧电极测定密闭反应体系评价两组胎鼠线粒体功能结果,并分析母体甲状腺功能减退与胎鼠骨骼肌胰岛素抵抗和线粒体功能的相关性。结果：两组小鼠甲状腺功能结果、两组胎鼠骨骼肌胰岛素抵抗和线粒体结果对比分析之间均有显著差异（P<0.05）。甲减组小鼠血清中促甲状腺激素（Thyroid Stimulating Hormone,TSH）、丙氨酸转氨酶（alanine aminotransferase,ALT）和门冬氨酸氨基转移酶（aspartate aminotransferase,AST）均较对照组低,胎鼠的空腹血糖（fasting blood-glucose,FBG）、动态3（state3,ST3）、动态4（state4,ST4）呼吸速率和呼吸控制比（respiratory control,RCR）也均较对照组低;而甲减组小鼠游离甲状腺素（free thyroxine,FT4）却较对照组高,胎鼠的胰岛素（insulin,INS）和胰岛素抵抗结果（homeostasis model assessment of insulin resistance,HOMA-IR）也较对照组高（P<0.05）。且母体甲状腺功能减退指标中FT4和RCR、ALT和FBG以及RCR之间有负相关关系,母体甲状腺功能减退的其他指标则与胎鼠骨骼肌胰岛素抵抗和线粒体功能的其他相关指标之间呈正相关关系（P<0.05）。结论：妊娠期母体甲状腺功能减退会降低胎鼠骨骼肌胰岛素抵抗和线粒体功能,影响胎鼠的正常发育。     

去卵巢小鼠肠道菌群和血脂的变化

目的探讨去卵巢对小鼠肠道菌群和血脂的影响。方法 12只10周龄C57BL/6小鼠随机分为2组:假手术组(SHAM组)和去卵巢组(OVX组),每组6只,进行12周的喂养。每2周测定小鼠体质量,12周后测肝脏指数、血清三酰甘油水平和游离脂肪酸水平,小肠进行病理学检查,收集小鼠粪便并在Illumina MiSeq测序平台进行16S rRNA基因测序检测。结果与SHAM组相比,OVX组小鼠体质量、肝脏指数、血清三酰甘油水平和游离脂肪酸水平明显增加(t=4.745,t=15.090,t=11.140,t=4.038,均P0.01)。与SHAM组相比,OVX组小鼠肠道Shannon Wiener指数明显降低(t=4.520,P0.01);在SMB53属(P=0.001)、拟杆菌属(P=0.017)、优杆菌属(P=0.018)、粪球菌属(P=0.019)、赖氨酸芽胞杆菌属(P=0.020)的丰度上差异有统计学意义。结论去卵巢小鼠血脂升高和肠道菌群失衡,提示肠道菌群可能是预防和治疗雌激素缺乏后脂质代谢异常的潜在靶点。     

Protective effect of quercetin on some hematological parameters in rats exposed to cadmium

ABSTRACT

We investigated the effects of quercetin (Q) on some hematological parameters and determined the percentage of alpha-naphthyl acetate esterase (ANAE) positive lymphocytes in rats that had been exposed to cadmium (Cd). Thirty male Wistar albino rats were divided into four groups: control (C), quercetin (Q), cadmium (Cd) and Q + Cd (CdQ). Blood samples were taken to assess erythrocytes (RBC), leukocytes (WBC), hemoglobin levels (Hb), hematocrit values (Hct), platelets (PLT), alpha-naphthyl acetate esterase (ANAE) positive lymphocytes. RBC, Hb, Hct; the number of PLT significantly decreased in the Cd group. To the contrary, these parameters were increased significantly in the CdQ group compared to the Cd group. Although we found a significant increase in total WBC count and neutrophil percentage, the number of lymphocytes decreased in the Cd group compared to the other three groups. Also, the percentage of peripheral blood ANAE positive lymphocytes decreased significantly in the Cd group (p < 0.05). Q exhibits positive effects on some hematological characteristics and the percentage of ANAE positive lymphocyte in cases of acute CD toxicity.     

The relation of oxidative stress and apoptosis to histopathologic alterations in the lungs as a result of global cerebral ischemia

ABSTRACT

Heart attack and oxygen deficiency may cause necrosis in the brain and other tissues. We investigated the histopathological effects of nitric oxide (NO) on ischemia/reperfusion in lung and hippocampus using a rat brain bilateral occlusion ischemia model. Male rats were assigned to sham (SH), ischemic preconditioning (PC), global ischemia (GI) and ischemic reperfusion (IR) groups. Before ischemia was induced, blood was drawn to induce hypovolemic hypotension and for blood gas testing. After sacrifice, samples of hippocampus were harvested. Sections were examined using hematoxylin and eosin (H & E) staining and immunostaining using primary antibodies for GFAP, S100β, iNOS, eNOS and the TUNEL method. Following ischemia, we found evidence of gliosis induced oxidative stress and apoptosis in the hippocampus. No significant differences were detected between the SH and PC groups. In the GI and IR groups, apoptosis and necrosis were observed in the hippocampus. Lung sections were stained with H & E and Masson’s trichrome (MT) and immunostained for iNOS and eNOS. The TUNEL method was used to detect apoptosis. Interstitial edema, vascular congestion, intra-alveolar hemorrhage, perivascular edema, neutrophil infiltration and disruption of alveoli were observed after global ischemia and ischemic reperfusion. Inflammatory cells were detected in the connective tissue. The IR and GI groups exhibited significantly more apoptotic cells than the SH or PC groups. Free radicals, such as nitric oxide (NO), that appear following ischemia and reperfusion in the brain may also injure the lungs. Increased NO in both lung and brain tissue suggests that apoptosis in these organs can be induced by reactive nitrogen species.     

Inhibitory effects of acetylsalicylic acid on exocrine pancreatic carcinogenesis

Abstract

We investigated short (6 months) and long (12 months) term inhibitory effects of low (200 ppm) and high (400 ppm) dosages of acetylsalicylic acid (aspirin) on exocrine pancreatic carcinogenesis. It is known that exocrine pancreatic carcinogenesis can be detected by the presence of atypical acinar cell foci (AACF) in pancreas. We investigated possible inhibitory effects of acetylsalicylic acid in an azaserine-treated rat model. AACF were produced in rats by injection with azaserine according to previous studies. Our findings showed that the number, volume and diameter of pancreatic AACF were reduced after acetylsalicylic acid application. These observations suggest that acetylsalicylic acid may exert a protective effect against neoplastic development of pancreatic acinar cells in azaserine injected rats. Our findings corroborate reports in the literature concerning the effects of aspirin in reducing neoplastic development.     

A histological,immunohistochemical and biochemical study of the effects of pomegranate peel extracts on gibberellic acid induced oxidative stress in adult rat testes

ABSTRACT

Gibberellins are commonly used plant growth regulators that exhibit deleterious effects on various animal tissues. We investigated the histological, immunohistochemical and biochemical effects of gibberellic acid (GA3) on rat testes as well as the possible protective role of pomegranate peel extract (PPE). We used 28 adult male rats divided into control, PPE treated, GS3 treated and GA3 + PPE treated groups. Testis specimens were analyzed for superoxide dismutase (SOD) and catalase (CAT) activity, and examined histologically. We also investigated the androgen receptor using immunohistochemistry. The GA3 treated group exhibited significantly decreased SOD and CAT levels and area percent of androgen receptor. Seminiferous tubules (ST) were widely separated and the germinal epithelium was separated from the basement membrane in some tubules. Areas of vacuolation, degenerated germ cells with pyknotic nuclei and large multinucleated cells were observed. Ultrastructurally, primary spermatocytes exhibited vacuolated cytoplasm, degenerated mitochondria and hyperchromatic nuclei. Degenerated early spermatids with a ruptured or hyperchromatic nucleus were found. Spermatozoa exhibited abnormalities of the head and tail. The cytoplasm of Sertoli and Leydig cells exhibited dilated smooth endoplasmic reticulum. A significant improvement of the biochemical, histological and immunohistochemical alterations was observed in the GA3 + PPE treated group compared to the GA3 treated group.