常规实验操作对Wistar大鼠的影响

目的常规实验操作对Wistar大鼠的影响。方法采用Wistar大鼠,分别进行腹腔注射、肌内注射、灌胃、尾静脉切割采血、剪尾采血和固定操作,连续处理7d后,测定各组大鼠神经内分泌、免疫、血液系统主要指标及肝脏hsp72mRNA表达量,并与对照组比较,分析这些操作的影响。结果与对照组相比,腹腔注射组、肌内注射组和灌胃组的皮质酮(CORT)、γ-干扰素(IFN-γ)、γ-干扰素/白介素-4(IFN-γ/IL-4)比值极显著升高,白细胞总数(WBC)、中性粒细胞比例(WLCR)、C-反应蛋白(CRP)、β-内啡肽(β-EP)及白介素-2(IL-2)有不同程度的显著降低。与对照组相比,尾静脉切割采血组的CORT、IFN-γ、IFN-γ/IL-4比值极显著升高,WBC、β-EP有不同程度的显著降低;剪尾采血组的CORT、IFN-γ、IFN-γ/IL-4比值有不同程度的显著升高,β-EP、IL-2、CRP极显著降低;固定组的CORT、IFN-γ/IL-4比值、IL-2、CRP极显著升高,WBC、β-EP、IL-4有不同程度的显著降低。与对照组相比,所有实验操作均使热休克蛋白(HSP)hsp72mRNA表达极显著升高。此外,与对照组相比,肌内注射组的红细胞数(RBC)和血红蛋白含量(HGB)显著升高,尾静脉切割和剪尾采血组的HGB和红细胞压积(HCT)极显著降低,固定组的RBC、HGB和HCT均极显著升高。结论本研究中涉及的常规实验操作均引起了大鼠极显著的应激反应,并造成了大鼠神经内分泌系统的紊乱及免疫功能的抑制,从而对动物造成了恶性应激。同时这些操作可能会对涉及的神经内分泌及血液学指标的相关研究产生背景性干扰。     

亚健康状态大鼠的神经-免疫-内分泌机制的研究

目的探讨亚健康状态大鼠的神经-免疫-内分泌的机制。方法取雄性SPF级Wistar大鼠36只,按体重随机分成6组,即多因素组（MF）、多因素干预组（MFT）、热水游泳组（WS）、睡眠不足组（SD）、单纯束缚组（PC）和正常对照组（C）,每组6只。分别采用热水游泳、饮食限制、睡眠剥夺、束缚等方式建立亚健康大鼠模型,多因素干预组每日经口给予300 mg/kg的抗衰老片,在造模5 d后处理动物,并取大鼠血,分别测定血清5-羟色胺（5-HT）、多巴胺（DA）、甲状腺激素T3和T4、睾酮（T）、皮质酮（CORT）、促肾上腺皮质激素（ACTH）、白介素IL-1β、IL-2、IL-6、IL-8和IFN-γ含量以及T淋巴细胞亚群的变化,并取脾脏测定T、B淋巴细胞增殖能力和NK细胞活性变化。结果 （1）与正常对照组比,①造模结束后多因素组大鼠血清5-HT明显降低（P〈0.05）,而热水游泳组和单纯束缚组5-HT含量却升高显著（P〈0.01）,且睡眠不足组和单纯束缚组血清DA含量降低显著（P〈0.05）;同时各亚健康模型大鼠的血清T3、T4、CORT和ACTH含量均显著升高（P〈0.01）,T/C比值降低显著（P〈0.01）,且睡眠不足组血清T降低明显（P〈0.05）;②造模结束后多因素组的大鼠IL-8含量降低显著（P〈0.01）,而热水游泳组和单纯束缚组IL-1β、IL-2、IL-6、IL-8和IFN-γ含量以及睡眠不足组IL-1β、IL-2、IL-6含量均显著升高（P〈0.05,P〈0.01）,同时显著降低各亚健康模型大鼠淋巴细胞增殖转化能力和T淋巴细胞亚群CD3＋、CD4＋和CD4＋/CD8＋比值和NK细胞活性（P〈0.05,P〈0.01）。（2）与多因素组比,抗衰老片干预后多因素干预组大鼠血清DA含量明显升高（P〈0.05）,并能显著提高机体T淋巴细胞亚群CD3＋、CD4＋数目和CD4＋/CD8＋比值（P〈0.01）,降低血清CORT含量。结论亚健康状态能引起大鼠HPA轴反应性失衡,皮质醇释放过度,引起内分泌的紊乱,并能引起淋巴细胞功能降低,出现明显机体免疫抑制现象;而抗衰老片具有抗应激,提高T细胞免疫功能,改善情绪和内分泌紊乱的作用,从而达到改善亚健康体质的目的。     

慢性复合应激对大鼠学习记忆功能及脑ERK表达活化的影响

目的：研究慢性复合应激对大鼠学习记忆的影响,以及大脑细胞外信号调节激酶（ERK）表达活化的变化,探讨慢性应激致学习记忆损害的分子机制。方法：采用低温暴露、足电击、白噪声、束缚、尾部悬吊、睡眠剥夺、水平震荡等刺激方式,建立慢性复合应激大鼠模型。Morris水迷宫实验观察应激对学习记忆的影响;放射免疫法检测血清皮质酮（CORT）含量;Western blot检测ERK的表达。结果：应激组大鼠水迷宫训练潜伏期较对照组延长,应激3周时有所恢复,但4周时大鼠的训练潜伏期再次显著延长（P〈0.05）。同时应激组大鼠血清CORT水平增高,海马、前额皮质P-ERK水平降低（P〈0.05）,两者在应激3周时均出现短时恢复,但4周时再次下调。结论：海马、前额皮质ERK蛋白磷酸化水平的改变可能参与了慢性复合应激损害学习记忆的分子机制。     

大鼠冷束缚应激时胃粘膜损伤与外周β—内啡肽关系的实验研究

目的：探讨冷束缚应激大鼠血浆及胃黏膜中β－内啡肽（β－endorphine)的变化及作用。方法：制作大鼠冷束缚应激模型,以放免法测定应激0,1／4,2,8h血浆及胃黏膜中β－内啡肽的含量,同时测定胃液pH,胃黏膜溃疡指数（UI）,另设两预处理组,分别于应激前15分钟用纳洛酮（1．6mg/kg)和等容量生理盐水颈静脉注射,测定其UI及胃液pH,结果：整个应激过程中血浆及胃黏膜中β－内啡肽的含量无显著变化（P＞0．05）,应激各组胃黏膜UI均显著高于未应激组（P＜0．01）,应激15分钟组胃液pH显著高于未应激组（P＜0．01）,临终前（应激8h组）胃液pH显著高于对照组（P＜0．05）,阻断阴UI及胃液pH较对照组均无显著变化（P＞0．05）,结论：应激过程中血浆及胃黏膜β－内啡肽含量稳定,阻断β－内啡肽不影响胃液pH及UI,外周β－内啡肽变化对应激所致胃黏膜损伤可能不起重要作用。     

维吾尔药神香草调节哮喘大鼠下丘脑-垂体-肾上腺轴功能紊乱的实验研究

目的：观察维吾尔药（维药）神香草对哮喘大鼠模型神经内分泌免疫网络若干组分的影响。方法：取雄性健康Wistar大鼠50只,随机分为正常对照组,哮喘模型组,神香草低、中、高剂量治疗组。采用致敏和雾化的方法制备哮喘模型。采用酶联免疫吸附试验（ELISA）方法检测血清中白介素-4（IL^4）、白介素-6（IL-6）、干扰素-γ（IFN-γ）、皮质酮（CORT）水平;放免法检测血浆中促肾上腺皮质激素（ACTH）的含量;采用实时定量-聚合酶链反应（RT—PCR）法测定下丘脑促肾上腺皮质激素释放激素（CRH）mRNA表达水平。结果：哮喘反复发作时,大鼠下丘脑-垂体-肾上腺（HPA）轴紊乱,哮喘大鼠下丘脑CRHmRNA表达和血浆ACTH无明显变化,实验各组血清CORT水平升高（P〈0．05）,神香草高剂量组血清CORT含量高于低剂量组（P〈0．05）。细胞因子IFN-γ无明显变化,IL-6、IL-4有下降趋势（P〈0．05）。结论：哮喘反复发作的大鼠存在NEI网络的紊乱;神香草可以增强下丘脑．垂体．肾上腺皮质（HPA）轴的功能,改善细胞因子的平衡。这些可能是其治疗哮喘的机制之一。     

五味子对实验性水上漂浮及高强度运动大鼠垂体-性腺轴的影响

目的研究五味子对实验性水上漂浮及高强度运动大鼠垂体-性腺轴的影响。方法 34只SD大鼠随机分为非应激对照组（A组,n=10）、应激对照组（B组,n=12）和五味子干预组（C组,n=12）。A组不接受任何刺激,在生理盐水灌胃1周后进行取样。B组和C组进行为期10 d的递增负荷跑台训练,分别用生理盐水和五味子灌胃1周后,行3 h水上漂浮及3 h跑台高强度运动,结束即刻取血,测定血清皮质酮（CORT）、睾酮（T）、黄体生成素（LH）;随机取一侧睾丸,HE染色观察组织病理结构;电子显微镜下观察超微结构。结果 （1）B组大鼠与A组相比,睾酮水平显著下降;C组大鼠与B组相比,睾酮水平无明显变化,皮质酮水平显著降低;黄体生成素在3组之间均无显著变化。（2）HE染色示3组大鼠睾丸组织病理无明显变化。（3）电子显微镜观察发现,B组大鼠睾丸Leydig细胞线粒体肿胀,电子密度增高;C组大鼠睾丸Leydig细胞线粒体结构趋向于正常,可见分泌颗粒。结论五味子对实验性水上漂浮及高强度运动大鼠睾酮水平无明显作用,对睾丸超微结构有一定保护作用。     

白头翁汤对炎症性肠病中促炎细胞因子表达的影响

目的：探讨白头翁汤治疗炎症性肠病的分子机制。方法：40只Wistar雄性大鼠随机分为5组（n=8）：正常对照组、模型组、模型＋阳性药物对照组（美沙拉嗪）、模型＋白头翁中、高剂量治疗组。阳性对照组、中药治疗组分别灌胃给药。疗程结束后取大鼠结肠组织提取RNA,利用realtimePCR的方法检测白介素-1β（IL-1β）、白介素-6（IL-6）YL肿瘤坏死因子-α（TNF-α）在各组中的表达变化。结果：相对于模型组,阳性药物及白头翁汤治疗组尤其是高剂量组可有效抑制IL-1β、IL-6及TNF-α的表达。结论：白头翁汤可通过抑制促炎因子的表达从而发挥了在炎症性肠病中的治疗作用。     

大鼠新生期注射谷氨酸单钠成年后应激时的镇痛效应和血浆皮质酮反应

在大鼠新生期注射谷氨酸单钠,观察动物成年后应激时的镇痛效应和血浆皮质酮反应,以新生期注射１０％ＮａＣｌ作为等渗对照。ＭＳＧ组大鼠下丘脑弓状核的β－内啡肽免疫反应细胞减少６０．７％,痛阈和血浆质酮的基础值未受影响,在不可射避的持续电刺激四肢脚底３０ｍｉｎ后,ＭＳＧ组动物应激镇痛效应明显降低,但应激仍可使血浆皮质酮水平明显升高。实验结果提示,ＡＲＣ的β－ＥＮＤ能神经元参与激镇痛,而对垂体－肾上腺皮质系     

低氧暴露对大鼠下丘脑CRF分泌的影响

目的和方法：利用人工模拟低氧及放射免疫测定（RIA）法,观察不同低氧（5km,7km）暴露对大鼠下丘脑分泌促肾上腺皮质激素释放激素（C orticotropin-releasing factor,CRF）的作用和血浆皮质酮水平的变化。比较低氧暴露不同时间（2h,24h,5d,15d）后,大鼠下丘脑CRF分泌和血浆皮质酮的变化。结果：低氧暴露2h,24h后下丘脑的CRF分泌明显增加,血浆皮质酮水平显著升高,这种变化随着低氧程度的加深而增强,随着低氧暴露时间的延长（5d）上述变化减弱。至慢性低氧暴露15d后,下丘脑CRF分泌及血浆皮质酮水平与对照组相比已无显著差异,基本恢复到对照水平。结论：动物暴露于低氧环境后,下丘脑CRF分泌及血浆皮质酮水平变化为：低氧暴露2h,24h后CRF分泌和皮质酮分泌被激活;低氧暴露5d后CRF分泌和皮质酮分泌活动减弱,并开始恢复;至低氧暴露15d后,这种分泌活动基本恢复,进入恢复期。     

右美托咪定联合罗哌卡因连续股神经阻滞对全膝关节置换术后患者镇痛效果、炎性因子和神经递质的影响

摘要 目的：探讨右美托咪定联合罗哌卡因连续股神经阻滞对全膝关节置换术（TKA）后患者镇痛效果、炎性因子和神经递质的影响。方法：选取2017年2月~2019年12月期间我院收治的行TKA的患者98例,采用随机数字表法分为对照组和研究组,各49例。对照组给予罗哌卡因连续股神经阻滞,研究组在对照组的基础上联合右美托咪定,比较两组患者镇痛效果、炎性因子、神经递质和不良反应。结果：两组术后1h、6h、12h、24h、48h静息及运动状态的视觉疼痛模拟评分法（VAS）评分均呈先升高后降低趋势（P<0.05）;研究组术后1h、6h、12h、24h、48h静息及运动状态的VAS评分均低于对照组（P<0.05）。两组术前、术后24h、术后48h白介素-1β（IL-1β）、白介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）呈先升高后下降趋势,但研究组术后24h、术后48hIL-1β、IL-6、TNF-α低于对照组（P<0.05）。两组术前、术后24h、术后48h P 物质、β-内啡肽及孤啡肽呈先升高后下降趋势,但研究组术后24h、术后48h P 物质、β-内啡肽及孤啡肽低于对照组（P<0.05）。两组不良反应发生率对比未见统计学差异（P>0.05）。结论：行TKA患者采用罗哌卡因联合右美托咪定连续股神经阻滞,术后镇痛效果确切,可减少炎性因子和神经递质水平,且不增加不良反应发生率,安全性佳。     

Adaptation of corticosterone-but not beta-endorphin-secretion to repeated blood sampling in rats

Effects of short-term repeated blood sampling on the secretion of corticosterone (CORT) and beta-endorphin (beta-END) were evaluated in male Wistar rats. Blood was drawn from the tail vein of conscious rats four times within 2 h both at the peak and trough period of the diurnal corticosterone secretion cycle. All rats were well accustomed to the procedure. The main findings were: (1) At both sampling intervals, CORT increased significantly in response to the first sampling and declined to baseline values in successive samples. (2) beta-END also increased significantly in response to the first sampling but remained elevated in successive samples. (3) Intensities of initial CORT and beta-END responses correlated positively with each other and with the baseline beta-END values. Feedback inhibition of CORT secretion with sustained elevation of beta-END titres suggests a moderate stress intensity of the repeated blood sampling procedures. In general, due to lack of short-term feedback inhibition, beta-END seems to reflect the effects of repeated administration of moderate intense stressors more closely than CORT.     

Effects of liquid carbohydrate ingestion on markers of anabolism following high-intensity resistance exercise

We examined the effects of liquid carbohydrate (CHO) supplementation on markers of anabolism following high-intensity resistance exercise. Nine resistance-trained men consumed either CHO or placebo (PLC) 10 minutes before and immediately following 2 resistance exercise sessions. Cortisol (CORT), insulin (INS), ammonia (AMM), and glucose (GLU) were measured before, immediately after, and 1.5 and 4 hours after exercise. Urinary nitrogen (NH(+3)) was measured 24 hours before and after exercise. There was a significant difference in INS levels immediately after exercise and 1.5 hours after exercise. No significant differences were observed for CORT, AMM, GLU, or NH(+3)between treatments. Significant within-group differences were found for the PLC group: CORT before compared with immediately after exercise; INS before compared with immediately after exercise and before compared with 1.5 hours after exercise; and AMM before compared with immediately after exercise and before compared with 1.5 hours after exercise. Significant within-group differences were found for the CHO group: CORT immediately after compared with 1.5 hours after exercise and immediately after compared with 4 hours after exercise; INS before compared with 1.5 hours after exercise; and AMM before compared with immediately after exercise. Liquid CHO ingestion leads to a more favorable anabolic environment immediately following a resistance exercise bout; however, our indirect measures of protein degradation were not altered by CHO ingestion.     

Multiple dexamethasone treatment affects morphometric parameters of gonadotrophic cells in adult female rats

Exposure to glucocorticoids leads to numerous changes in various biological systems including the reproductive system. The aim of the present work was to find out whether dexamethasone (Dx) treatment of adult female rats would influence the histological and morphometric characteristics of the pituitary gonadotrophic cells (luteinizing--LH cells and follicle stimulating--FSH cells). One group of female Wistar rats received Dx injections on three consecutive days in doses 1.0, 0.5 and 0.5 mg/kg b.w. respectively, while the control rats were treated with equivalent volumes of saline. Experimental and control animals were sacrificed 24 h and 72 h after the last injection. The peroxidase-antiperoxidase (PAP) immunocytochemical procedure was used to study the LH and FSH cells. The stereological and morphometric analyses showed that multiple Dx treatments of female rats significantly decreased the volume of LH cells and the volume of their nuclei 24 h and 72 h after the last Dx injection in comparison with control values. At 24 h after Dx treatment, the volume density of LH cells was significantly increased, but at 72 h differences between the experimental and control groups were insignificant. The increase in number of LH cells per unit area (mm2) was significant at both timepoints (24 h and 72 h). Stereologic and morphometric characteristics of FSH cells was changed after Dx treatment in the same manner as that of LH cells, except for the volume density, where a significant increase was established 24 h and 72 h after the last Dx application. These results clearly demonstrate that 24 h and 72 h after the last of three Dx injections there were changes in the immunocytochemical and morphometric features of gonadotrophic cells.     

大鼠一次性力竭跑台运动模型的建立及动态评价

目的跑台急性运动疲劳动物模型的建立及评价。方法选取清洁级雄性Wistar大鼠24只(8周龄)作为实验对象。采用多级递增负荷跑台运动方案(跑台坡度为0°,负荷分为三级)建立一次性力竭跑台运动动物模型。尾静脉取血,分别测定大鼠在安静、运动30 min、运动90 min、力竭、恢复30 min、恢复90 min各时间点外周血葡萄糖(GLU)、乳酸(LD)、尿素(BU)、丙二醛(MDA)浓度和肌酸激酶(CK)、超氧化物歧化酶(SOD)活性。结果一次性力竭运动过程中大鼠行为能力和运动能力、血液代谢产物及能量物质呈现阶段性的动态变化。外周血LD、BU、MDA浓度及CK活性均较安静时显著性增高(P<0.01,P<0.05);GLU浓度、SOD活性较安静时显著降低(P<0.01,P<0.05)。各指标的变化特征说明大鼠已达到运动疲劳状态。结论建立了大鼠一次性力竭跑台运动模型,并客观动态评价了大鼠在运动疲劳产生、发展、恢复等不同阶段各指标的变化特点及规律。该模型可用于后续运动疲劳机制的相关研究。     

p53 expression in brain after middle cerebral artery occlusion in the rat.

The purpose of this study is to determine whether the p53 protein, a product of the p53 tumor suppressor gene, that has been associated with the 72 kDa heat shock protein (hsp72), is expressed in ischemic brain. Adult Wistar rats (n = 5) were subjected to 120 minutes of middle cerebral artery occlusion. Twelve hours after reopening the artery, brain tissue was analyzed to determine the extent of neuronal damage (hematoxylin and eosin), and the distribution of p53 and hsp72 (monoclonal antibodies). Our data demonstrate that p53 is expressed in regions of neuronal necrosis; in contrast, morphologically intact neurons express hsp72. The data suggest that the presence of p53 is associated with cell death and that hsp72 may regulate p53 function.     

Short-term treadmill running in the rat: what kind of stressor is it?

The use of short-term (1-5 days) treadmill running is becoming increasingly common as a model to study physiological adaptations following the exercise. Although the beneficial effects of acute exercise seem clear, a paucity of data exist describing potential markers of stress in response to forced running. We subjected male and female Sprague-Dawley rats to 0, 1, 2, 5, or 10 days of treadmill running. Twenty-four to 32 h after the last bout of exercise animals were killed and examined for training-induced changes in several physiological variables. No effect of skeletal citrate synthase activity was observed in the male animals after any duration, and only at 10 days of running did females show a significant increase in citrate synthase. Myocardial heat shock protein 72 (HSP72) content was higher in male rats than female rats, and exercise led to increased HSP72 in both sexes, although the time course was different between males and females. Animals displayed several markers of systemic stress in response to the treadmill running, and this was done in a sex-dependent manner. Serum corticosterone was significantly elevated in both sexes 24 h after exercise in three of four exercise groups. Corticosterone-binding globulin was higher in females, and decreased after running in female rats. Body and spleen weights decreased in males (but not females) in response to the exercise training, and running did not alter adrenal gland weights in either sex. These data indicate that in response to short-term treadmill running both male and female rats show signs of systemic stress, but that the pattern of changes occurs in a sex-specific manner.     

烯啶虫胺、 毒死蜱和高效氯氰菊酯对烟粉虱地中海隐种hsp70 mRNA的诱导表达

为深入理解杀虫剂胁迫对烟粉虱Bemisia tabaci适应逆境能力的影响, 本研究运用实时荧光定量PCR技术测定了LC₂₅, LC₅₀和LC₇₅ 3种浓度的烯啶虫胺、 毒死蜱和高效氯氰菊酯分别处理对烟粉虱地中海隐种B. tabaci Mediterranean成虫体内热激蛋白hsp70表达水平的影响。结果表明： 在低温（15℃）条件下, LC₅₀和LC₇₅的烯啶虫胺、 毒死蜱、 高效氯氰菊酯可显著诱导烟粉虱地中海隐种hsp70的表达量增加, 此后随时间延长hsp70表达量逐渐下降, 到72 h时恢复到对照水平, 但LC₂₅的3种杀虫剂对hsp70的表达无明显影响; 在常温（25℃）下, 较高浓度（LC₅₀和LC₇₅）的烯啶虫胺、 毒死蜱和高效氯氰菊酯处理24 h同样可促进hsp70表达量的增加, 然后hsp70表达量逐渐下降, 到72 h时恢复至正常水平, 但低浓度(LC₂₅)的3种药剂处理后hsp70表达量随时间延长而增加, 到72 h时达到最高; 在高温（32℃）条件下, LC₂₅和LC₅₀的烯啶虫胺、 毒死蜱和高效氯氰菊酯处理24 h可显著增强hsp70的表达水平, 此后随时间的延长hsp70表达恢复至正常水平。杀虫药剂诱导的hsp70表达量增加增强了烟粉虱地中海隐种对杀虫药剂和高温的耐受能力, 这可能是导致其在我国快速扩张的原因之一。     

Stress-induced alternations in CuZnSOD and MnSOD activity in cellular compartments of rat liver

Exposure to different stressors initiates generation of reactive oxygen species (ROS), which create harmful environment for cellular macromolecules. Superoxide dismutases (SODs) represent the first line of antioxidant defense. Hence, any alternation in their function might be potentially damaging. To better define the role of SODs, we investigated the CuZnSOD activity in cytosolic and the nuclear fraction as well as mitochondrial MnSOD activity in the liver of Wistar male rats after exposure to 2 h of acute immobilization (IM) or cold (4°C) stress, 21 days of chronic social isolation (IS) or their combination (chronic stress followed by acute stress). Serum corticosterone (CORT) was monitored as an indicator of the stress response. Acute IM stress, with elevated CORT level, led to increased hepatic CuZnSOD activity in the nuclear fraction. Chronic isolation stress, where CORT was close to control value, did not change the CuZnSOD activity either in nuclei or in cytosolic fraction, while combined stress IS+Cold led to increased cytosolic CuZnSOD activity. MnSOD activity in mitochondrial fraction was decreased in all treated groups. Data have shown that different stressors have diverse effect on hepatic CuZnSOD and MnSOD activity as well as on serum CORT level. Increased nuclear CuZnSOD activity after acute stress represents physiological response since the named activity protects cells against oxidative stress, while chronic IS stress compromises CuZnSOD function, suggesting an inefficient defense against ROS. Observed decrease of MnSOD activities indicate inadequate elimination of ROS after acute or chronic stress, which is characteristic of the oxidative stress.