脂联素对慢性间歇性低氧大鼠颏舌肌收缩功能的影响

目的 探讨慢性间歇性低氧(chronic intermittent hypoxia,CIH)对颏舌肌收缩功能的影响及脂联素(Ad)的干预作用.方法 健康雄性Wistar大鼠39只,随机分为正常氧组(NC组)、CIH组和CIH Ad干预组(CIH+Ad组),每组13只.NC组呼吸正常空气,CIH组与CIH+Ad组接收CIH建模环境(CIH 8 h/d,共35 d),CIH+Ad组织给予颈静脉注射脂联素10 μg/次,2次/周,共5周,NC组与CIH组注射生理盐水0.5 ml/次对照.于实验终止时测定并比较各组大鼠血清Ad浓度及颏舌肌的收缩功能.结果 ①CIH组血清Ad浓度明显低于NC组[(1 210.32±84.20) μg/L,(2 236.43±117.72)μg/L]；②CIH组单刺激收缩最大张力[(0.84±0.072) N/g]、强直收缩最大张力[(3.37±0.29)N/g]、单刺激波峰值张力时间[(93.47±7.4)ms]和1/2松弛时间[(8.79±0.66) ms]较NC组降低(P＜0.05),CIH+Ad组较CIH组改善(P＜0.05)；③三组大鼠强直收缩疲劳指数在第一个20 s下降明显,在此后的100 s,疲劳指数继续下降,但趋势缓慢,NC组、CIH+Ad组抗疲劳性均明显高于CIH组(P＜0.01).结论 CIH可致血清脂联素浓度下降,并影响颏舌肌收缩功能及抗疲劳性能,补充外源性Ad能部分改善CIH对颏舌肌收缩功能及抗疲劳性的影响.     

脂联素缓解慢性间歇低氧所致的颏舌肌线粒体损伤

目的 探讨慢性间歇低氧（CIH）对颏舌肌线粒体的损伤以及脂联素（Ad）的干预作用及机制。方法 45只成年Wistar大鼠随机分为3组：正常对照（NC）组、CIH组及CIH＋Ad组,每组15只。CIH组及CIH＋Ad组的大鼠暴露于同样的间歇低氧环境（8h/d,5周）,而NC组的大鼠则只暴露于正常空气。此外,CIH＋Ad组的大鼠还接受2次/周的Ad静脉注射。结果 与NC组相比,CIH组大鼠的颏舌肌出现以下的损伤性表现：线粒体数量减少、线粒体结构损伤伴Ⅰ型纤维减少（P＜0.05）。但与CIH组相比,CIH＋Ad组的大鼠颏舌肌线粒体结构和功能改善且Ⅰ型纤维的数量有所增加（P＜0.05）。与NC组相比,CIH组大鼠颏舌肌显示LKB1-AMPK-PGC1-α通路蛋白表达下降（P＜0.05）,而CIH＋Ad组较CIH组有明显改善（P＜0.05）。结论 CIH可引起颏舌肌线粒体等损伤,而补充外源性Ad可能通过调节AMPK通路改善上述CIH诱导的颏舌肌病理改变。     

慢性间歇低氧大鼠的血压变化及其颈动脉体中还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶的表达

目的 探讨慢性间歇低氧(chronic intermittent hypoxia,CIH)大鼠的血压变化及其颈动脉体中还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶的表达情况,以明确CIH致血压升高的可能机制.方法 清洁级雄性SD大鼠30只按随机数字表法分为CIH组、慢性持续缺氧组及对照组.尾袖法测量大鼠尾动脉收缩压,采用RT-PCR法检测大鼠颈动脉体中NADPH氧化酶各亚基gp91 phox、p22phox及p47phox mRNA的表达,对颈动脉体行免疫组织化学染色,并对p22phox的表达情况行半定量分析.结果 CIH组大鼠的尾动脉收缩压为[(145±11)mm Hg,1 mm Hg=0.133kPa],较CIH组(129±9)mm Hg及对照组(124±7)mm Hg显著升高(F值为19.895,P＜0.01),CIH组gp91phox、p22phox及p47phox mRNA的表达(分别为2.82±0.51、2.74±0.45和2.88±0.47)较慢性持续缺氧组(分别为2.35±0.42、2.25±0.38和2.41±0.43)及对照组(分别为2.23±0.35、2.16±0.30和2.30±0.36)显著升高(F值分别为5.794、6.854和7.163,P＜0.01)免疫组织化学检查结果显示CIH组p22phox蛋白相对表达量(99±12)较其他两组(分别为38±7和34±8)增多.结论 CIH可刺激大鼠颈动脉体中NADPH氧化酶的表达上调并使血压升高,NADPH氧化酶在颈动脉体中的过度表达可能与OSAHS患者发生高血压病存在相关联系.

Abstract：

Objective Chronic intermittent hypoxia (CIH) occurs in patients with obstructive sleep apnea-hyponea syndrome (OSAHS) and has adverse effects on multiple physiological functions. Previous studies have shown that reflexes arising from carotid bodies mediate CIH evoked circulation-respiratory responses, and reactive oxygen species (ROS) play important roles in eliciting systemic responses to CIH.But very little is known about the molecular mechanisms underlying CIH. NADPH oxidase is the most important sources of ROS. In the present study we examined changes of blood pressure and expression of NADPH oxidase in carotid body in rats exposed to intermittent hypoxia. Methods Thirty healthy male SD rats were randomly divided into 3 groups, a CIH group, a chronic continuous hypoxia group and a control group. The systolic blood pressure (SBP) was measured with tail-cuff method. RT-PCR was used to examine mRNA expressions of NADPH oxidase subunit gp91phox, p22phox, p47phox. Immunohistochemistry and semiquantitative analysis of NADPH oxidase subunits p22phox were done in the carotid body sections of all rats. Results Compared with normal group [ ( 124 ± 7 ) mm Hg, 1 mm Hg = 0. 133 kPa ] and chronic continuous hvpoxia group[ (129 ± 9) mm Hg], the SBP in CIH group [( 145 ± 11 ) mm Hg] was significantly higher( F = 19. 895, P ＜0. 01 ＝, and the expression of NADPH oxidase subunits gp91phox,p22phox,p47phox mRNA in CIH group ( 2. 82 ± 0. 51, 2. 74 ± 0. 45, 2. 88 ± 0. 47, respectively ) were significantly higher than those in chronic continuous hypoxia group ( 2. 35 ± 0. 42, 2. 25 ± 0. 38, 2. 41 ±0. 43, respectively)and normal group(2. 23 ±0. 35, 2. 16 ±0. 30, 2. 30 ±0. 36, respectively) ( F =5.794,6. 854, 7. 163, respectively, P ＜ 0. 01 ). The Immunohistochemistry and semiquantitative analysis showed that the expression of NADPH oxidase subunit p22phox in the carotid body in CIH group ( 99 ± 12 ) were more than those in chronic continuous hypoxia and control groups ( 38 ± 7 and 34 ± 8, P ＜ 0. 05 ).Conclusion CIH upregulates the expression of NADPH oxidase in rat carotid body and elevates the rat SBP. These results indicate that NADPH oxidase up-expression is closely associated with OSAHS patients with hypertension.     

追赶生长大鼠胃泌素及内脏脂肪细胞CCK2R表达变化

24只Wistar大鼠分为正常对照组、限食组、追赶生长组,检测所有大鼠血糖、血脂、血清胃泌素,内脏脂肪体脂比、脂肪细胞CCK2R mRNA和蛋白水平.结果 显示限食组和正常组相比,血清胃泌素水平降低54%(P＜0.05),内脏脂肪体脂比减少55%(P＜0.05),脂肪细胞CCK2R mRNA和蛋白表达下降(2.19±0.18对3.2±0.24,0.11±0.03对0.15±0.04,P＜0.05).追赶生长组血清胃泌素水平分别高于限食组72%和正常组31%(P＜0.05),内脏脂肪体脂比高于限食组114%(P＜0.05),达到正常对照组水平;同时脂肪细胞CCK2R mRNA和蛋白表达高于正常对照组(4.09±0.59对3.2±0.24,0.25±0.05对0.15±0.04,P＜0.05).追赶生长大鼠内脏脂肪优先沉积的机制可能和胃泌素分泌增加及脂肪细胞CCK2R表达增加相关.

Abstract：

Wistar rats(n=24) were divided into normal control group(NC), food restriction group(FR), and catch-up group(RN). Serum glucose,lipids, gastrin, the ratio of visceral fat to body fat, adipocyte CCK2R mRNA and protein levels were determined. Compared with NC group, FR rats had lower serum gastrin and visceral fat formation. The adipocyte CCK2R mRNA and protein levels of FR rats were lower than those of NC rats. Serum gastrin level of RN rats was higher than those of FR and NC rats(P＜0.05). The ratio of visceral fat to body fat in RN rats increased compared with FR rats and was close to that of NC rats. The adipocyte CCK2R mRNA and protein levels of RN rats were higher than those of FR and NC rats. Gastrin and its receptor pathway possibly play a role in the mechanism of visceral fat accumulation in catch-up rats.     

血红素氧合酶-一氧化碳系统对肝硬化大鼠血流动力学的影响

目的 观察血红素氧合酶-一氧化碳系统对肝硬化大鼠全身血流动力学的影响.方法将30只雄性SD大鼠分为对照组(14只)和肝硬化组(16只),肝硬化组大鼠皮下注射50%四氯化碳(以橄榄油稀释,0.003 ml/g),对照组给予相同剂量的橄榄油.12周后,将肝硬化组大鼠分为肝硬化给药组(8只)、肝硬化模型组(8只),对照组大鼠分为正常给药组(7只)、正常对照组(7只).肝硬化给药组和正常给药组大鼠经后颈部皮下注射锌原卟啉(20 μmol/kg),肝硬化模型组和正常对照组予以等渗盐水,用动脉插管生理多导仪记录心率、平均动脉压的变化,门静脉插管测定门静脉压,联二亚硫酸盐还原法测定血浆一氧化碳水平,用比色法测定胆红素生成量.数据间比较用t检验.结果 正常对照组与肝硬化模型组大鼠平均动脉压分别为(18.9±0.9)kPa和(15.6±1.7)kPa,门静脉压分别为(8.8±0.3)cm H2O(1 cm H2O=0.098 kPa)和(16.7±0.8)cm H2O,血浆一氧化碳分别为(10.4±1.3)μmol/L和(18.0±1.9)μmol/L,脾脏血红素氧合酶(HO)活性分别为(6.5±0.9)nmol·mg1·h1和(11.1±0.9)nmol·mg 1·h-1,小肠HO活性分别为(1.3±0.2)nmol·mg1·h-1和(2.5±0.1)nmo1·mg-1·h-1,两组比较,t值分别为4.52、23.10、8.42、9.28、15.10,P值均＜0.01,差异有统计学意义.正常对照组与肝硬化模型组大鼠肝脏HO活性分别为(2.7±0.2)nmol·mg-1·h-1、(2.7±0.1)nmol·mg-1·h-1,差异无统计学意义.肝硬化模型组、肝硬化给药组平均动脉压分别为(15.6±1.7)kPa、(17.3±1.5)kPa,两组比较,t=2.18,P＜0.05,差异有统计学意义,门静脉压分别为(16.7±0.8)cmH2O、(13.2±0.7)cmH2O,两组比较,t=8.53,P＜0.01,差异有统计学意义.结论 HO-CO系统的激活可能是肝硬化血流动力紊乱的重要原因.

Abstract：

Objective To investigate the role of heme oxygenase(HO), a catalyzing enzyme of heme to produce CO, in modulation of systemic circulation in CCl4-induced cirrhotic rats. Methods Saline (vehicle) and ZnPP were s.c. injected into the posterior necks of rats respectively and the rats were then anesthetized by pentobarbital sodium in four hours. Mean arterial pressure (MAP, kPa), heart rate (HR, b/min) and portal pressure (PP, cm/H2O) were measured by indwelling catheter. Plasma CO was determined by Chalmers method. Heme oxygenase acivity was determined by the rate of bilirubin formation. Results The cirrhotic rats showed significant hyperdynamic circulation indicated by decreased mean arterial pressure [MAP,(15.6 ± 1.7) vs (18.9 ± 0.9) kPa, t = 4.52, P ＜ 0.01] and increased portal pressure [PP, (16.7 ± 0.8)vs (8.8 ± 0.3) cm H2O, t = 23.10, P ＜ 0.01] as compared to normal control rats(NS). ZnPP could cause a significant increase in MAP [(17.3 ± 1.5) vs (15.6 ± 1.7) kPa, t = 2.18, P ＜ 0.05] and significant decrease in PP [(13.2 ± 0.7) vs (16.7 ± 0.8) cm H2O, t = 8.53, P ＜ 0.01] in cirrhotic rats. The cirrhotic group presented a significant increase in plasma CO [(18.0 ± 1.9) vs (10.4 ± 1.3) μ mol/L, t = 8.42, P ＜ 0.01] and HO activity in the spleens [(11.1 ± 0.9) vs (6.5 ± 0.9) nmol bilirubin/mg protein/h, t = 9.28, P ＜ 0.01 ] and intestines [(2.5 ±0.1) vs. (1.3 ± 0.2) nmol bilirubin/mg protein/h, t = 15.1, P ＜ 0.01]. ZnPP could cause significant decreases in plasma CO and HO activity in liver, spleen and intestine of both control and cirrhotic rats. Conclusion HO-CO system activation may be an important reason for the hemodynamic disturbance of liver cirrhosis.     

异丙肾上腺素诱导慢性心力衰竭大鼠心肌醛固酮及其核受体的变化

目的 探讨异丙肾上腺素(isoproterenol,ISO)诱导的慢性心力衰竭SD大鼠心肌醛固酮及其核受体的变化.方法 将SD大鼠抽签随机分为心力衰竭组(9只)和对照组(10只),心力衰竭组皮下注射ISO,对照组皮下注射等量生理盐水,心功能采用超声心动图及血流动力学检查,放射免疫法测定血浆及心肌组织醛固酮水平,免疫印迹和免疫组化染色法检测盐皮质激素核受体蛋白表达的变化.结果 心力衰竭组与对照组比较心功能明显下降,左心室射血分数分别为(38.8±4.0)%与(79.4±4.6)%;左心室内压最大上升速率分别为(7164.4±502.6)mm Hg(1 mm Hg=0.133 kPa)/s与(10199.5±462.9)mm Hg/s(均P＜0.01).血浆及心肌组织醛固酮含量明显升高,分别为(0.63±0.06)μg/L与(0.30±0.07)μg/L、(0.41±0.05)μg/kg与(0.08±0.01)μg/kg(均P＜0.01),左心室心肌醛固酮核受体蛋白表达增高(P＜0.01).结论 ISO可诱导SD大鼠出现类似扩张型心肌病的慢性心力衰竭表现,在这种心力衰竭模型中其循环和左心室心肌醛固酮水平明显升高,心肌醛固酮核受体表达上调,可能在心力衰竭的发生、发展中起着重要作用.

Abstract：

Objective To investigate the changes of cardiac aldosterone and mineralocorticoid receptor (MR) in Sprague-dawley (SD) rats with chronic heart failure (CHF) induced by isoproterenol (ISO). Methods The SD rats were randomly divided into CHF group (n=9) and normal control(NC) group (n=10). The experimental CHF group was induced by subcutaneous injection of ISO, and the NC group received same dose injection of sodium chloride. The heart function was evaluated with both echocardiography and hemodynamics. The contents of aldosterone in both plasma and heart were assessed by radioimmunoassay. The expression of MR was measured by Western blot and immunohistochemistry staining. Results Compared with NC group, the heart function was decreased in CHF group, the left ventricular ejection fraction was (38.8%±4.0%) in CHF and(79. 4%±4.6%), in NC group. The maximal rate of increase of ventricular pressure (+dp/dtmax) was (7164.4±502.6) mm Hg(1 mm Hg=0.133 kPa)/s in CHF and (10199.5±462.9) mm Hg/s in NC group (both P＜0. 01 ). The contents of aldosterone both in plasma and heart were higher in CHF group than in NC group [(0.63±0.06)μg/L vs. (0.3±0.07) μg/L, (0.41±0.05) μg/kgvs. (0.08±0.01)μg/kg, both P＜0. 01]. The MR expression was increased in CHF group versus in NC group (P＜0.01). Conclusions The heart function is decreased in rats with CHF induced by ISO, which is similar to dilated cardiomyopathy. The higher levels of aldosterone both in circulation and in heart as and well as MR expression upregulation in heart may play important roles in the pathogenesis of CHF induced by ISO.     

碘过量对实验性自身免疫性甲状腺炎大鼠骨代谢的影响

目的 探讨碘过量对实验性自身免疫性甲状腺炎(EAT)大鼠骨代谢的影响.方法 雌性Lewis大鼠36只,体质量为(131±15)g,按体质量随机分为3组:对照组、EAT组和EAT+高碘组,每组12只.以不同含碘量(0.9、0.9、18.0 mg/kg)的饲料喂养各组大鼠,并用猪甲状腺球蛋白(pTG)和完全弗氏佐剂(CFA)对EAT组和EAT+高碘组大鼠进行免疫以建立EAT模型.2周后观察大鼠甲状腺病理改变,测定血清甲状腺自身抗体[甲状腺球蛋白抗体(TGAb)、甲状腺微粒体抗体(TMAb)]和甲状腺激素[三碘甲腺原氨酸(T3)、甲状腺素(T4)]以及骨代谢相关指标[骨钙素(BGP)、抗酒石酸盐酸性磷酸酶(TRAP)、Ⅰ型前胶原羧基末端前肽(PICP)、Ⅰ型胶原羧基吡啶并啉交联肽(ICTP)、胰岛素样生长因子-1(IGF-1)、护骨素(OPG)、核因子κB受体活化因子配体(RANKL)]水平.结果 EAT组和EAT+高碘组大鼠甲状腺组织均呈现炎细胞浸润,局部滤泡结构破坏,其中EAT+高碘组以甲状腺滤泡扩张、融合为主.EAT组和EAT+高碘组大鼠血清TGAb、TMAb、T3和T4水平[(63.01±12.36)%、(60.62±11.24)%,(3.78±1.43)、(125.12±16.00)pmol/L和(75.00±15.44)%、(72.15±15.00)%,(3.69±0.91)、(149.40±20.67)pmol/L]高于对照组[(4.47±1.04)%、(5.73±1.01)%,(0.75±0.12)、(76.91±9.30)pmol/L,P均＜0.05],EAT+高碘组大鼠血清TGAb、TMAb和T4水平较EAT组升高(P均＜0.05).EAT组大鼠血清BGP、PICP和IGF-1水平[(1.70±0.31)、(11.31±1.52)μg/L,(0.31±0.06)mg/L]较对照组[(8.60±0.33)、(14.28±3.10)μg/L,(1.16±0.02)mg/L]降低(P均＜0.05),血清TRAP、ICTP、OPG和RANKL水平[(19.88±3.60)ng/L,(2.43±0.82)、(22.36±2.80)、(1.35±0.23)μg/L]较对照组[(14.57±3.56)ng/L,(0.50±0.20)、(1.61±0.34)、(0.10±0.02)μg/L]升高(P均＜0.05);与EAT组比较,EAT+高碘组大鼠血清PICP和OPG水平[(8.03±1.84)、(16.80±3.79)μg/L]明显降低(P均＜0.05).结论 EAT时大鼠的破骨细胞分化与成熟增强,导致骨吸收增强.碘过量可抑制EAT大鼠成骨细胞和破骨细胞活性,呈现低转化型骨质疏松.

Abstract：

Objective To explore the effect of iodine excess on bone metabolism in experimental autoimmune thyroiditis (EAT) rats. Methods We selected 36 female Lewis rats with body weight of (131 ± 15)g,and divided them into 3 groups randomly: control group, EAT group and EAT + high iodine group, assuring 12 rats in every group. These rats were fed fodder with different concentration of iodine(0.9,0.9, 18.0 mg/kg), and rats in EAT group and EAT + high iodine group were immunized with pig thyroglobulin(pTG) and complete Freund's adjuvant(CFA) to create EAT model. After two weeks, the pathological changes of the thyroid tissues were observed,and the serum thyroid autoantibody[thyroglobulin antibody(TGAb) and thyroid microsomal antibody(TMAb)], the thyroid hormone levels[triiodo thyronine(T3) and thyrine(T4)] and some relevant data of bone metabolism[bone gla protein (BGP), tartrate-resistant acid phosphatase (TRAP), C-terminal propeptide of type Ⅰ procollagen (PICP),C-terminal telopeptide of type Ⅰ collagen (ICTP), insulin-like growth factor- 1 ( IGF- 1 ), osteoprotegerin (OPG) and receptor activator of NF-κB ligand (RANKL)] were measured. Results Inflammatory cell infiltration and local follicular structural damage were observed in the thyroid tissues of EAT rats in EAT group and EAT + high iodine group, and the pathological changes of EAT + high iodine group were mainly thyroid follicular expansion and integration. The level of serum TGAb, TMAb, T3 and T4 of EAT rats in EAT group and EAT + high iodine group[ (63.01 ± 12.36)%, (60.62 ± 11.24)%, (3.78 ± 1.43), (125.12 ± 16.00)pmol/L and (75.00 ± 15.44)%,(72.15 ± 15.00)%, (3.69 ± 0.91 ), (149.40 ± 20.67)pmol/L] were higher than those of the control group[ (4.47 ±1.04)%, (5.73 ± 1.01 )%, (0.75 ± 0.12), (76.91 ± 9.30)pmol/L, all P ＜ 0.05], and the level of serum TGAb,TMAb and T4 of EAT rats in EAT + high iodine group were higher than those of the EAT group(all P ＜ 0.05).The level of serum BGP, PICP and IGF- 1 in EAT group[ ( 1.70 ± 0.31 ), ( 11.31 ± 1.52) μg/L, (0.31 ± 0.06 ) mg/L]were lower than those of the control group[ (8.60 ± 0.33), (14.28 ± 3.10)μg/L, (1.16 ± 0.02)mg/L, all P ＜0.05], and the level of serum TRAP, ICTP, OPG and RANKL[ ( 19.88 ± 3.60)ng/L, (2.43 ± 0.82), (22.36 ± 2.80),( 1.35 ± 0.23 )μg/L] were higher than those of the control group[ ( 14.57 ± 3.56)ng/L, (0.50 ± 0.20), (1.61 ± 0.34),(0.10 ± 0.02)μg/L, all P ＜ 0.05]; compared with EAT group, the level of PCIP and OPG in EAT + high iodine group [ (8.03 ± 1.84), ( 16.80 ± 3.79)μg/L] were obviously decreased(all P ＜ 0.05). Conclusions The reinforcement of differentiation and maturation of osteoblast in the EAT rats results in the increasing of bone resorption. The activity of osteoblast and osteoclast of the EAT rats are inhibited by excessive iodine, showing a low conversion-type osteoporosis.     

脂联素抑制内质网应激干预慢性间歇性缺氧所致肾损伤

目的探讨脂联素(Ad)对慢性间歇性缺氧(CIH)所致肾损伤的干预作用及相关机制。方法 60只成年Wistar大鼠随机分成4组:正常对照(NC)组、NC+Ad组、CIH组和CIH+Ad组。其中CIH组和CIH+Ad组大鼠接受CIH处理4个月。其余2组接受正常空气处理,同时NC+Ad组和CIH+Ad组大鼠接受Ad(10l人開)治疗,每周2次,持续4个月。荧光显微镜下观察活性氧(ROS)的水平。TUNEL染色检测肾脏细胞凋亡情况。Western blotting检测各组大鼠肾脏组织中GRP78、CHOP、IRE1、PERK、pro-ATF6蛋白的表达,以反映各组大鼠内质网应激情况。采用SPSS 17.0软件进行统计学分析。计量资料用均数±标准差(±s)表示,组间比较用t检验。结果实验满4个月时,组间比较示各参数在NC与NC+Ad组间差异均无统计学意义(P均0.05)。与NC组和NC+Ad组比较:(1)ROS水平在CIH组显著增高,而CIH+Ad组低于CIH组,但仍高于NC组和NC+Ad组(尸均0.05);(2)肾细胞凋亡率和反映凋亡的caspase-12和caspase-3蛋白水平在CIH组明显增加,CIH+Ad组较CIH组明显减少,但仍然高于NC组和NC+Ad组(P均0.05);(3)CIH组肾脏组织的GRP78、CHOP、IRE1、PERK蛋白水平明显增加,在CIH+Ad组明显减少,但仍高于NC组和NC+Ad组(P均0.05);pro-ATF6蛋白水平在CIH组明显降低,在CIH+Ad组有所增加,但仍然低于NC组和NC+Ad组(P均0.05)。结论 CIH可以通过激活ROS和ERS相关的细胞凋亡途径导致肾脏损伤,而补充外源性Ad后,可能通过抑制ROS,进而抑制ERS,保护肾脏细胞。     

慢性阻塞性肺疾病大鼠肺树突细胞数量及成熟度变化与慢性炎症的关系

目的 观察香烟烟雾暴露对大鼠肺组织树突细胞数量、成熟度及肺组织慢性炎症变化的影响.方法 将30只雄性F344大鼠按随机数字表法分为香烟烟雾暴露组(暴露组)、断烟组和健康对照组(对照组),每组10只.采用香烟烟雾暴露法建立大鼠COPD模型,HE染色法检测大鼠气道炎症病理评分及肺泡平均内衬间隔,免疫组织化学ABC法观察大鼠肺组织中CD11c+、CD86+和CD8+ T细胞的分布及数量变化,流式细胞术检测CD11c+/CD86+和CD11c+/主要组织相容性复合体(MHC)Ⅱ+与CD11c+树突细胞比值.结果 暴露组和断烟组大鼠肺组织中出现COPD特征性病理改变,气道炎症病理评分[(390±33)分和(324±28)分]及肺泡平均内衬间隔[(68±11)μm和(58±9)μm]明显高于对照组[(56±13)分和(36±6)μm],差异均有统计学意义(F值分别为459.85和34.03,均P＜0.05).暴露组和断烟组大鼠CD11c+树突细胞阳性率[(1.47±0.12)%和(1.30±0.17)%]及CD86+树突细胞阳性率[(1.26±0.18)%和(1.02±0.08)%]均明显高于对照组[(0.96±0.08)%和(0.65±0.03)%],差异均有统计学意义(F值分别为6.55和30.26,均P＜0.05);暴露组和断烟组大鼠CD8+T细胞阳性率[(2.72±0.15)%和(2.35±0.23)%]均明显高于对照组[(1.39±0.11)%],差异有统计学意义(F=16.07,P＜0.05);暴露组和断烟组大鼠CD11c+/CD86+树突细胞及CD11c+/MHCⅡ+树突细胞占CD11c+树突细胞比例[(5.5±0.4)%和(4.8±0.4)%]及[(4.2±0.3)%和(3.3±0.3)%]明显低于对照组[(8.0±0.5)%和(6.1±0.5)%],差异均有统计学意义(F值分别为14.34和12.82,均P＜0.05).暴露组与断烟组上述各项指标比较,差异均无统计学意义(t值为1.10～2.11,均P＞0.05).结论 香烟烟雾暴露诱导COPD大鼠肺组织中树突细胞数量明显增加,成熟度明显下降,断烟后此趋势无明显变化,且以CD8+ T细胞浸润为主的慢性炎症反应持续存在,提示树突细胞数量变化及成熟异常可能参与了COPD慢性炎症迁延进展.

Abstract：

Objective To evaluate the changes in the number and maturation of lung tissue dendritic cells (DCs) and to assess the chronic inflammation in a cigarette smoke-induced COPD model in rats.Methods Thirty male F344 rats were randomly divided into 3 groups (n = 10):a control group, a smoke-exposure group and a smoking cessation group.Rat lung pathomorphological changes were observed by hematoxylin-eosin (HE) stain.Lung tissue CD11c+ DCs, CD85+ DCs and CD8+ T cell numbers were observed by immunohistochemisty method.Flow cytometry was used for detection of CD11c+/CD86+ DCs and CD11c+/MHCⅡ + DCs proportions.Results The airway inflammatory pathological score and the mean linear intercept (MLI) obtained from he smoke-exposure group and the smoking cessation group (390 ± 33,324 + 28 ) and[(68 ± 11 ) μm, (58 ± 9) μm]were higher than those in the control group ( 56 ± 13 ) and ( 36 ± 6 ) μm( F =459.85 and 34.03, all P ＜0.05 ).In the smoke-exposure group and the smoking cessation group, the positive rate of CD11c+ DCs[(1.47 ±0.12)%, (1.30 ±0.17)%], and the positive rate of CD86+ DCs [( 1.26 ± 0.18 ) %, ( 1.02 ± 0.08 ) %]were higher than those in the control group[( 0.96 ± 0.08 ) %,(0.65 ± 0.03 ) %]( F = 6.55 and 30.26, all P ＜ 0.05 ), but there was no significant difference between the smoke-exposure group and the smoking cessation group ( t = 1.10 and 1.47, all P ＞ 0.05 ).In the smoke-exposure group and the smoking cessation group, CD8+ T positive rate[(2.72 ±0.15)%, (2.35 ±0.23)%]was higher than that in the control group[(1.39 ±0.11)%](F = 16.07, P ＜0.05).CD11c+/CD86+ DCs and CD11c+/MHC Ⅱ+DCs percentages[(5.5 ±0.4)%, (4.8 ±0.4)%],[(4.2 ±0.4)%, (3.3±0.3 )%]decreased in the smoke-exposure group and the smoking cessation group as compared to the control group[(8.0±0.5 ) %, (6.1 ± 0.5 ) %]( F = 14.34 and 12.82, all P ＜ 0.05 ).There was no significant difference between all the above index from the smoke-exposure group and the smoking cessation group ( t = 1.10 and 2.11, all P ＞ 0.05 ).Conclusions Cigarette smoke exposure induced increased DCs transmigrated and influenced the maturation of DCs in COPD rats.Even after smoking cessation, non-specific chronic inflammation was still present, suggesting that DCs number and maturation abnormality may be involved in the chronic inflammation of COPD.     

Effects of long-term tea polyphenols consumption on hepatic microsomal drug-metabolizing enzymes and liver function in Wistar rats

AIM: To investigate the effects of long-term tea polyphenols (TPs) consumption on hepatic microsomal drug-metabolizing enzymes and liver function in rats.METHODS: TPs were administered intragastrically to rats at the doses of 833 mg.kg-1.d-1 (n=20) and 83.3 mg.kg-1@d-1 (n=20) respectively for six months. Controlled group (n=20)was given same volume of saline solution. Then the contents of cytochrome P450, bS, enzyme activities of aminopyrine N-demethylase (ADM), glutathione S-trasferase (GST) and the biochemical liver function of serum were determined.RESULTS: The contents of cytochrome P450 and b5 in the livers of male rats in high dose groups (respectively 2.66±0.55,10.43±2.78 nmol.mg MS pro-1) were significantly increased compared with the control group (1.08±1.04, 5.51±2.98nmol.mg MS pro-1; P＜0.01, respectively). The enzymatic activities of ADM in the livers of female rats in high dose groups (0.91±0.08 mmol@mg MS pro-1min-1) were increased compared with the control group (0.82±0.08 mmol.mg MS pro-1.min-1; P＜0.05). The GST activity was unchanged in all treated groups, and the function of liver was not obviously changed.CONCLUSION: The antidotal capability of rats' livers can be significantly improved after long-term consumption of TPs.There are differences in changes of drug-metabolizing enzymes between the sexes induced by TPs and normal condition.     

慢性间歇低氧对大鼠颏舌肌超微结构和线粒体功能的影响

目的 探讨慢性间歇低氧(CIH)对大鼠颏舌肌细胞超微结构及线粒体功能的影响及脂联素(Ad)的干预作用.方法 健康雄性Wistar大鼠39只,随机分成健康对照组(A)、CIH组(B)、CIH加Ad干预组(C),每组13只.A组大鼠保持呼吸空气,B组与C组均接受CIH环境(CIH 8 h/d,共5周),但C组同时经静脉注射...     

Adiponectin protects the kidney against chronic intermittent hypoxia-induced injury through inhibiting endoplasmic reticulum stress

Purpose

The current study was carried out to assess the effects of chronic intermittent hypoxia (CIH) on the kidney, the intervention roles of adiponectin (Ad), and the associated mechanisms.

Methods

Sixty Wistar rats were randomly divided into four groups: the normal control (NC), normal control plus Ad supplement (NC + Ad), CIH, and CIH plus Ad supplement (CIH + Ad) groups. The rats in both CIH and CIH + Ad groups were submitted to a CIH environment for 4 months, while the rats in NC and NC + Ad groups were housed with the normal air for 4 months. In addition, the rats in NC + Ad and CIH + Ad groups were treated with an intravenous injection of Ad at a dosage of 10 μg per injection, twice a week, for four successive months.

Results

The production level of reactive oxygen species (ROS) and the protein levels of endoplasmic reticulum (ER) stress, as well as the cell apoptosis level in kidney, were all higher in the CIH group than in the NC and NC + Ad groups (all p < 0.05). However, the ROS production, the protein of ER stress, and cell apoptosis levels in kidney were all lower in the CIH + Ad group than those in the CIH group (all p < 0.05).

Conclusion

Ad could protect against CIH-induced renal cell apoptosis through inhibiting ROS-related ER stress.

     

核因子-κB在慢性间歇性低氧对大鼠左心功能影响中的作用

目的了解慢性间歇性低氧对大鼠左心功能的影响,探讨NF-κB在其发生机制中的作用。方法将24只SD大鼠随机分为3组：正常对照组（NC）、慢性间歇性低氧组（CIH）,CIH＋PDTC（吡咯烷二硫代氨基甲酸盐,NF-κB抑制剂）组。CIH组每天白天置于间歇性低氧箱（最低氧浓度5%～7%）8h,共5周;CIH＋PDTC组每天腹腔注射PDTC100mg/kg,饲养环境与CIH组相同;正常对照组给予相似的处理,但是维持空气氧浓度不变。测量大鼠体重、血压、心率,超声心动图评价心功能;最后提取大鼠心肌组织核蛋白,Western blot法测量NF-κB蛋白水平的表达。结果第5周时CIH组和CIH＋PDTC组体重低于NC组。CIH组和CIH＋PDTC组血压明显高于NC组[分别为（136.3±6.8）、（134.3±6.7）和（122.3±4.1）mmHg,P〈0.01],CIH组和CIH＋PDTC组两组间差异无统计学意义。CIH组LVEF低于NC组[分别为（73±6）%和（86±4）%,P〈0.001],CIH＋PDTC组[（84±4）%]较CIH组明显升高（P〈0.001）,与NC组比较没有明显差异（P=0.1 17）。各组心率无明显统计学差异。NF-κB蛋白水平的表达CIH组高于NC组和CIH＋PDTC组,NC组与CIH＋PDTC组间没有统计学差异。结论慢性间歇性低氧大鼠左心功能减低,NF-κB可能参与其作用机制。     

从钙离子调控改变探讨兔肾素血管紧张素系统阻断剂逆转甲状腺素促心肌肥厚的机制

目的 研究甲状腺素诱导的肥厚心肌细胞内钙调控变化及其机制,探讨肾素血管紧张素系统(RAS)阻断剂逆转甲状腺素促心肌肥厚的机制.方法 腹腔注射左旋甲状腺素(L-Thy)建立兔甲状腺素性心肌肥厚模型,经胸二维超声心动图测定室间隔厚度(IVS)、左心室内径(LV)及左心室后壁厚度(LVPW)后收集心肌标本,光镜及透射电镜下观察心肌细胞结构改变,Fluo3/AM荷载共聚焦显微镜检测细胞内钙离子浓度,逆转录聚合酶链半定量检测L型Ca2+通道(LCC)、兰尼碱受体(RyR)、肌浆网钙泵(SERCA)mRNA表达,对硝基苯磷酸酯(P-nitrophenyl phosphate,P-NPP)法检测SERCA活性,免疫组织化学法检测IP3R蛋白表达.结果 甲状腺素可诱导心肌肥厚、心肌细胞结构改变及纤维组织增生,甲状腺素组细胞内钙离子浓度明显高于对照组,分别为(576.2±41.7)nmol/L比(314.6±35.6)nmol/L(P<0.01);甲状腺素组RyR mRNA、SERCA mRNA及IP3R蛋白较对照组表达上调,分别为1.19±0.21比0.73±0.15(P<0.01)、1.01±0.08比0.76±0.09(P<0.01)、65.3±13.7比47.9±10.2(P<0.01);LCC mRNA表达和SERCA活力降低,分别为0.48±0.11比0.75±0.16(P<0.01)、(0.062±0.013)μmol·min-1·g-1比(0.133±0.022)μmol·min-1·g-1(P<0.01).咪达普利和缬沙坦干预可显著抑制甲状腺素诱导的心肌细胞肥厚,降低细胞内钙离子浓度,咪达普利组和缬沙坦组细胞内钙离子浓度分别为(376.4±32.5)nmol/L和(392.6±41.2)nmol/L均显著低于甲状腺素组(576.2±41.7)nmol/L(P<0.01);咪达普利和缬沙坦干预后LCC mRNA表达高于甲状腺素组,分别为0.68±0.14比0.48±0.11(P<0.01)、0.64±0.13比0.48±0.11(P<0.01);咪达普利和缬沙坦干预增加SERCA的活力,分别为(0.115±0.019)μmol·min-1·g-1比甲状腺素组(0.062±0.013)μmol·min-1·g-1(P<0.01)、(0.109±0.015)μmol·min-1·g-1比甲状腺素组(0.052±0.013)μmol·min-1·g-1(P<0.01),但对RyR mRNA、SERCA mRNA及IP3R蛋白表达无影响.结论 RAS和细胞内钙超载可能参与甲状腺索性心肌肥厚的发病机制,SERCA活力降低可能是导致甲状腺素性肥厚心肌细胞内钙超载的重要因素.咪达普利和缬沙坦可以改善L-Thy诱导的心肌重构和细胞内钙超载并对SERCA活力具有保护作用.     

Adiponectin protects rat heart from left ventricular remodeling induced by chronic intermittent hypoxia via inhibition of TGF-β/smad2/3 pathway

Objective

Obstructive sleep apnea syndrome (OSAS) is associated with many cardiovascular disorders. Chronic intermittent hypoxia (CIH) is the primary player in OSAS of the many associated factors. This study was in order to investigate the effects of the Adiponectin (Ad) on left ventricular remodeling induced by CIH.

Methods

Forty-five rats were randomly divided into three groups: normal control (NC) group, CIH group and CIH plus Ad supplemented (CIH + Ad) group. After 35 days’ CIH exposure, masson analysis was used to detect the left ventricular fibrosis and western blot was used to measure the protein expression of collagen I, collagen III and TGF-β/smad2/3 pathway. Gene analysis by RT-PCR was used to study the MMP2 and TIMP2.

Results

After CIH exposure, the fibrosis of left ventricular in CIH group was significantly remarkable than that in both NC and CIH + Ad groups (P<0.05), although statistical difference existed between NC and CIH + Ad groups (P<0.05). In addition, the protein expression of collagen I as well as collagen III and the ratio of mRNA levels of MMP2/TIMP2 were the highest in CIH group but the lowest in NC group, with CIH + Ad group in between. There was a significant difference among three groups (all P<0.05). The TGF-β/smad2/3 pathway was activated obviously in CIH group, but less noticeably in CIH + Ad group (P<0.05) with a significant difference in the two groups.

Conclusions

The present study showed that Ad could ameliorate the left ventricular remodeling induced by CIH via inhibition of the expression of TGF-β/smad2/3 pathway.     

脂联素对大鼠慢性间歇性低氧所致心肌重塑的影响

目的：探讨慢性间歇性低氧（chronic intermittent hypoxia,CIH）对心肌重塑的影响及脂联素（adiponectin, Ad）的干预作用。方法将45只Wistar大鼠随机分为3组：正常对照组（NC）,CIH组和CIH＋Ad组。CIH 35d后,使用马松染色分析方法检测左室纤维化程度及使用Western blot方法来衡量Ⅰ型胶原蛋白、Ⅲ型胶原蛋白和TGF-β/smad2/3通路蛋白的表达。通过RT-PCR方法来研究基质金属蛋白酶-2（MMP2）/基质金属蛋白酶的组织抑制剂-2（TIMP-2）的mRNA表达比值情况。结果慢性间歇性低氧处理后,CIH组左心室的纤维化程度显著高于NC组和CIH＋Ad组（P＜0.05）,但NC组和CIH＋Ad组之间差异具有统计学意义（P＜0.05）。CIH组Ⅰ型胶原蛋白和Ⅲ型胶原蛋白和MMP2/TIMP-2的mRNA比值表达最高,NC组表达最低,CIH＋Ad组居中,3组之间均差异具有统计学意义（P＜0.05）。TGF-β/smad通路蛋白在CIH组中表达显著高于NC组和CIH组（P＜0.05）,且NC组和CIH＋Ad组差异具有统计学意义（P＜0.05）。结论慢性间歇性低氧可引起左室重构,而Ad可能通过抑制TGF-β/smad2/3通路改善此损害。     

改良心导管测定大鼠肺血管阻力的方法

目的 建立一种简单、可重复性强的测定正常及肺动脉高压大鼠肺血管阻力的方法.方法 雄性Sprague-Dawley( 180 ～ 200 g)大鼠45只,随机分为3组:正常对照组、低剂量野百合碱组(50 mg/kg)和高剂量野百合碱组(60 mg/kg).给予大鼠颈背部皮下一次性注射野百合碱建立肺动脉高压大鼠模型.大鼠肺动脉压力采用自制改良的末端呈圆弧形的PE- 50导管行右心导管法测定.心输出量利用热稀释法原理检测.肺血管阻力由平均肺动脉压力除心输出量得出.结果 三组大鼠肺动脉压力、心输出量及肺血管阻力的检测总成功率均分别为98％、100％和96％,组间差异无统计学意义.注射野百合碱21 d后,低、高剂量野百合碱组大鼠的平均肺动脉压均显著高于对照组[(43.1±0.8)mm Hg(1 mm Hg=0.133 kPa)、(54.8±2.2) mm Hg比(17.4±1.0) mm Hg,P均＜0.001],且高剂量组明显高于低剂量组(P＜0.001).低、高剂量野百合碱组大鼠心输出量均明显低于对照组[(77.5±6.9) ml/min、(71.0+6.7)ml/min比(126.8±3.9) ml/min,P均＜0.001],低、高剂量组间差异无统计学意义.低、高剂量野百合碱组大鼠肺血管阻力均显著高于对照组[(0.56±0.06) mm Hg·min-1·ml-1、(0.76±0.08)mm Hg·min-1·ml-1比(0.13±0.01)mm Hg- min-1·ml-1,P均＜0.001],且高剂量组明显高于低剂量组(P=0.01).结论 采用此方法检测大鼠肺血管阻力准确、可靠、操作性强,具有推广价值.     

不同频率间歇低氧下大鼠肝脏氧化应激损伤及Tempol的干预作用

目的 探讨不同频率间歇低氧(IH)对大鼠肝脏氧化应激损伤差异和Tempol的干预作用及可能机制.方法 应用慢性间歇低氧(CIH)大鼠模型,模拟OSAS周期性间歇低氧/再氧和病理生理过程.56只雄性Wistar大鼠随机分为不同频率IH组(IH1,IH2,IH3,IH4,频率依次为10、20、30、40次/h),30T组(...     

链脲佐菌素诱导的1型糖尿病大鼠肾脏脂联素受体的表达

目的 观察链脲佐菌素(STZ)诱导的糖尿病大鼠在不同时期其血清脂联素和脂联素受体(AdipoR)在肾脏组织的表达水平.方法 64只雌性SD大鼠按随机数字表法分为对照组和实验组(分别为2、6、10、12周,共8组):实验组大鼠32只,一次性空腹腹腔注射STZ 60 mg/kg,诱导糖尿病大鼠模型;对照组大鼠32只,腹腔注射等体积的枸橼酸缓冲液.分别于糖尿病大鼠成模后第2、6、10、12周两组各取8只,测体重、肾重、空腹血糖、24 h尿白蛋白定量;心内采血,离心取血清,检测血肌酐,空腹血清胰岛素;ELISA方法检测血、尿脂联素浓度.取左肾常规病理组织行糖原染色,在光镜下观察肾脏的病理组织学改变,免疫组化SP法检测肾脏2种受体AdipoR1和AdipoR2表达.结果 (1)实验组6、10、12周大鼠血清和尿脂联素高于对照组,差异有统计学意义(P＜0.01);且血清脂联素与24 h尿白蛋白排泄率、尿脂联素呈正相关(r值分别为0.806、0.696,均P＜0.01);尿脂联素与24 h尿白蛋白定量呈显著正相关(r=0.728,P＜0.01);逐步回归分析提示血清脂联素受24 h尿白蛋白定量影响最大(β=0.806,P＜0.01);(2)免疫组化半定量分析显示,AdipoR1和AdipoR2在正常大鼠肾组织中均有表达,主要分布于肾小管上皮细胞和肾小球内皮细胞.实验组造模成功6周时肾脏组织AdipoR1和AdipoR2表达,与对照组比较表达有所增强(F值分别为11.68、23.20,均P＜0.01),且与血清脂联素呈显著正相关(r值分别为0.666、0.684,均P＜0.01).结论 血清和尿脂联素水平与糖尿病肾病病程和AdipoR呈正相关,推测脂联素通过AdipoR直接作用于肾脏,尤其是作用于肾小管,在1型糖尿病肾脏病变中发挥保护作用.