SD大鼠阳虚便秘模型的建立及评价

目的在中医药理论指导下建立SD大鼠阳虚便秘动物模型并对其进行评价。方法应用山西白醋加活性炭冰水法制作阳虚便秘动物模型。观察模型动物的一般生物学特征变化,进行模型动物的排便和在体结肠肌电等实验,检测模型动物的胃肠激素、肠神经递质等客观指标。结果造模后,模型动物的一般生物学特征表现为:腹胀、披毛蓬松、倦怠蜷缩、体重减轻、大便色黑、干燥;与空白对照组比较,模型动物的首粒排便时间明显延迟,6h内排便粒数明显减少,结肠收缩频率明显降低、结肠收缩幅度减弱,胃肠激素、肠神经递质分泌紊乱;经通便灵药物治疗后,上述症状和指标明显改善。造模后动物恢复6d,与空白对照组比较,其首粒排便时间和6h内排便粒数仍有差异。结论应用山西白醋加活性炭冰水能复制出稳定的SD大鼠阳虚便秘模型。     

总香豆素抗支气管哮喘气道变应性炎症的作用机制

目的:探讨总香豆素抗支气管哮喘气道变应性炎症的作用机制。方法:支气管哮喘气道变应性炎症动物模型采用卵白蛋白致敏哮喘大鼠;激发后d4起,正常组和哮喘模型组灌胃给予等量蒸馏水,补骨脂总香豆素组、补骨脂黑色黏稠物组和蛇床子总香豆素组分别按体重灌胃给药,剂量分别为23．3,25．0和20．0 g·kg~(-1),连续10d。末次给药后2h,各组动物心脏采血测定血清皮质醇(化学发光法)、cAMP/cGMP及IFN-γ/IL-4(ELISA法)。结果:哮喘模型组皮质醇含量明显下降,蛇床子总香豆素皮质醇含量明显提高,其他组变化不明显。哮喘模型组cGMP无明显变化,而cAMP有降低趋势;蛇床子总香豆素组cAMP和cGMP含量及比值明显提高,而补骨脂总香豆素组cGMP无明显变化,但cAMP和cAMP/cGMP比值明显提高。哮喘模型组IL-4明显增加,Th_1/Th_2有降低趋势,而IFN-γ无明显变化;补骨脂黑色黏稠物组IL-4明显降低,IFN-γ增加,且Th_1/Th_2比值增加。结论:总香豆素抗支气管哮喘气道变应性炎症的作用机制与其调节下丘脑-垂体-肾上腺皮质系统有关,并通过调节cAMP/cGMP及Th_1/Th_2比值起作用。     

定痫丸对青霉素致痫大鼠脑组织环核苷酸(cAMP、cGMP)影响

目的观察探讨定痫丸治疗癫痫的作用及作用机制。方法各组大鼠按不同剂量灌胃给药7 d后,采用腹腔注射青霉素制备癫痫急性模型,青霉素注射剂量为600万U.kg-1,分别测定各组大鼠脑组织中环核苷酸(cAMP、cGMP)的含量,并进行各组大鼠脑电波观察。结果定痫丸能明显减轻青霉素的致痫作用,减少模型大鼠癫痫发作的频率,显著升高模型大鼠脑组织中SOD活性、降低MDA含量,显著升高脑组织中cAMP含量、降低cGMP含量。结论定痫丸抗癫痫作用机制与其抑制自由基引起的脂质过氧化反应、增加自由基的清除,调节脑内cAMP、cGMP含量变化有关。     

基于Ca2+-CaMKⅡ信号通路的济川煎治疗阳虚便秘的作用及分子机制

目的 基于Ca²⁺-CaMKⅡ通路观察济川煎对阳虚便秘大鼠的治疗作用及分子机制。方法 采用白醋+活性炭冰水联合复方地芬诺酯片复制阳虚便秘大鼠模型。将模型成功大鼠随机分为5组,即模型组、莫沙必利组(1.88 mg·kg^-1)、济川煎高(8.26 g·kg^-1)、中(4.31 g·kg^-1)、低(2.16 g·kg^-1)剂量组,灌胃给药每日1次,连续7 d。末次给药,记录大鼠状态并评分,进行排便功能测定;分离模型大鼠结肠平滑肌细胞并鉴定,FCM检测结肠平滑肌细胞内Ca²⁺浓度变化;采用ELISA法检测大鼠血清中cAMP、cGMP含量及其比值;采用RT-PCR及Western blot法检测大鼠结肠平滑肌细胞CaM、CaMKⅡ亚型的表达情况。结果 济川煎可明显缓解模型大鼠腹胀,摄食、饮水量、排便减少及体质量降低的症状(P<0.05),评分明显升高(P<0.01);能明显缩短首粒排便时间(P<0.01),增加排便粒数(P<0.05);济川煎...     

大鼠肺纤维化发生发展过程中肺组织环核苷酸的变化及其作用研究

目的 观察纤维化肺组织环核苷酸(cAMP和cGMP)的动态变化,研究环核苷酸的信号作用机制.方法 48只SD大鼠分为对照组、模型组和干预组.模型组和干预组经气管内注入BLM(Smg/kg,0.2～0.3m1)诱导形成肺纤维化,干预组每日经胃管灌注卡托普利(CPT)60mg/kg·d.各组动物于3、7、14、21d各处死4只,观察肺组织病理改变,并同位素放免检测肺组织环核苷酸水平.结果 干预组肺泡炎和肺纤维化程度均轻于模型组;模型组和干预组肺组织cAMP表达呈进行性下降趋势,cGMP表达则正好相反;干预组环核苷酸表达改变小于模型组.结论 纤维化肺组织的cAMP表达降低,cGMP表达升高;肺组织环核苷酸的表达与纤维化的严重程度有关;肺纤维化有效干预提高肺组织cAMP表达,抑制cGMP表达.     

黄柏盐炙前后对肾阴虚大鼠肠道菌群的影响

目的 比较黄柏及其盐炙品对肾阴虚模型大鼠肠道菌群多样性的影响。方法 将70只大鼠随机分成7组,分为空白组、模型组、阳性组、生黄柏高剂量组、生黄柏低剂量组、盐黄柏高剂量组和盐黄柏低剂量组,采用先给药后灌胃给予氢化可的松建立肾阴虚模型。采用Elisa法分析大鼠血液中相应指标,确认模型建立;Illumina Miseq测序分析各组大鼠肠道菌群的差异。结果 除空白组,其余各组cAMP/cGMP比值明显升高,肾阴虚模型建立成功。与空白组相比,模型组大鼠有明显的肾阴虚症状。肾阴虚模型组大鼠肠道菌群结构与空白组差异明显,盐黄柏能改善不同分类水平上肠道菌群的相对丰度,优于生黄柏,并接近空白组。结论 盐黄柏调节肾阴虚大鼠肠道菌群的效果更好。     

育阴软肝颗粒剂对大鼠肝纤维化的治疗作用

目的　研究育阴软肝颗粒剂对大鼠肝纤维化及其阴虚证的治疗作用。方法　采用多因素复制肝纤维化动物造模,通过观测大鼠肝纤维化指标(ALT、AST、肝指数、肝组织病理学、羟脯氨酸及胶原蛋白等)与阴虚指标(进食与饮水量、自主活动、体重、cAMP、cGMP等),研究育阴软肝颗粒剂对实验性肝纤维化大鼠的防治作用。结果　6.2~24.8g·kg-1 剂量范围内的育阴软肝颗粒剂能降低纤维化大鼠血清ALT、AST及肝指数,降低肝组织羟脯氨酸及胶原蛋白的含量,缓解肝组织纤维化病理学变化;同时,育阴软肝颗粒剂能降低肝纤维化大鼠的饮水量,增加体重,减少活动次数与站立次数,降低cAMP及cAMP/cGMP,缓解阴虚症状。结论　多因素复制的肝纤维化大鼠模型有一定的阴虚症状;育阴软肝颗粒剂对肝纤维化及其阴虚证有一定的防治作用。     

c-kit在蒽醌类泻剂所致肠动力减退发病机制中的作用

目的 探讨c-kit在蒽醌类泻剂所致肠动力减退发病机制中的作用.方法 健康成年SPF级SD大鼠36只随机均分为模型组、模型恢复组和对照组.模型组和模型恢复组大鼠采用“大黄酸混悬液灌胃法”制作肠动力减退动物模型,对照组用等体积生理盐水灌胃.模型组造模结束后取材,模型恢复组造模结束正常饲养30 d后取材.各组大鼠处死前观察首粒黑便排出时间以检测肠道传输功能,Western blot法检测各组大鼠结肠组织中c-kit的蛋白水平.结果 模型组和模型恢复组大鼠首粒黑便排出时间较对照组明显延长(P＜0.01)；模型组和模型恢复组结肠组织中的c-kit蛋白表达较对照组明显减弱(P＜0.01).结论 蒽醌类泻剂所致肠动力减退可能与结肠组织中c-kit蛋白表达下调有关.     

肠泻停胶囊对实验性结肠炎大鼠的影响

目的:探讨肠泻停胶囊对三硝基苯磺酸(TNBS)诱导大鼠实验性结肠炎的影响.方法:120只SD大鼠随机分为6组,每组20只:正常组、模型组、阳性对照组、肠泻停胶囊高、中、低组.除正常对照组未行造模外,其余五组大鼠均采用TNBS造模.肠泻停胶囊高、中、低组分别灌胃给药(1.80,0.90,0.45 g /kg体质量)、阳性对照组灌胃给予地塞米松剂量(0.2 mg/kg体质量)、其余组灌胃给予0.5%羧甲基纤维素钠溶液连续3 周、4周;观察肠泻停胶囊对大鼠腹泻率、死亡率、血白细胞计数、淋巴细胞百分率、脾脏及胸腺重量、组织形态学评分、组织MPO活性的影响.结果:经肠泻停胶囊干预后各剂量组动物腹泻明显缓解,腹泻率降低,动物死亡率降低,外周血WBC、LYM值及组织MPO值降低,剖检可见结肠组织溃疡面积明显缩小,水肿缓解,坏死减轻,未见肠壁增厚.结论:肠泻停胶囊连续给药,对实验性结肠炎治疗作用显著.     

大鼠和小鼠术后疲劳综合征模型的建立与评价

目的 以SD大鼠和C57BL/6J小鼠为实验对象,建立术后疲劳综合征(POFS)模型.方法 手术切除SD大鼠或C57BL/6J小鼠70％的肝,制备POFS动物模型.于术后不同时间观察动物的一般状态,采用自发活动、抓力、跑台运动和负重游泳实验以及生理信号遥测技术对模型动物的活动性、肌肉力量、运动耐力和睡眠结构进行综合评价...     

Effect of milk on intestinal fluid accumulation and renal injury following mercuric chloride ingestion in rats

To investigate the effect of milk on intestinal fluid accumulation and renal injury following mercuric chloride (HgCl(2)) ingestion, 10 ml kg(-1) of saline or 10 mg kg(-1) of HgCl(2) dissolved in 10 ml kg(-1) of water or raw milk was administered enterally to rats and the mercury content in biological samples was determined by cold vapour atomic absorption spectrometry. Three hours after administration, the intestinal water content in rats that received HgCl(2) in water (group S2) was significantly higher than in rats that received saline (group S1) (P < 0.01) or HgCl(2) in milk (group S3) (P < 0.01). The amount of mercury detected per gram dry weight of small intestine was higher in group S2 than in group S3 (P < 0.05). Seventy-two hours after administration, both the serum urea nitrogen and creatinine concentrations in rats that received HgCl(2 )in milk (group L3) were significantly higher than in rats that received saline (group L1) (P < 0.05) or HgCl(2) in water (group L2) (P < 0.05). Mercury levels in many of the biological samples in group L3 were higher than in group L2 (P < 0.05). Milk may reduce the intestinal cytotoxicity of mercury but it promotes its absorption, which may lessen intestinal fluid accumulation but worsen renal injury.     

Regulation of cAMP metabolism in mouse parotid gland by cGMP and calcium.

The interaction of hormones acting via the mobilization of calcium and stimulation of cAMP levels in cells was examined by determining the effects of carbachol and forskolin on cAMP and cGMP accumulation in mouse parotid gland. Treatment of isolated acini with either carbachol (0.01 to 20 microM) or forskolin (1 microM) alone produced little or no increase in cAMP levels; carbachol, however, augmented the effect of forskolin on cAMP accumulation approximately 3- to 4-fold. The effects of carbachol on forskolin-stimulated cAMP levels were further augmented approximately 10-fold in the presence of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (MIX) but not in the presence of "low Km" cGMP-inhibited phosphodiesterase inhibitor milrinone. Augmentation of cAMP levels also occurred in the presence of carbachol plus the beta-adrenergic agonist isoproterenol (0.01 microM). In either the presence or absence of forskolin, carbachol increased cGMP levels independently of the inclusion of MIX and in a fashion parallel to that observed for cAMP accumulation. In the presence of forskolin (1 microM), the concentration of carbachol that produced half-maximal effects on cAMP and cGMP levels was 0.62 and 0.72 microM, respectively. Similar values were obtained in the presence of MIX. Cyclic GMP levels were also enhanced by carbachol plus isoproterenol. Hydroxylamine, as well as dibutyryl-cGMP and 8-bromo-cGMP in combination with forskolin, mimicked the effects of carbachol plus forskolin on cAMP levels. LY83583 (6-anillino-5,8-quinolinedione), an agent that lowers cGMP by inhibiting guanylate cyclase, reduced basal levels of cGMP and also completely prevented the increase in cGMP caused by carbachol plus forskolin. In these experiments, however, the augmentation of forskolin-stimulated cAMP levels by carbachol was reduced by approximately 50%. Additional studies suggest that calcium is also required for carbachol augmentation of forskolin-stimulated cAMP accumulation by effects on the adenylate cyclase complex. Augmentation of cAMP levels by carbachol did not involve effects on cAMP degradation. The results suggest that, when cAMP synthesis is stimulated by forskolin or isoproterenol, the muscarinic agonist carbachol augments cAMP accumulation by mechanisms involving cGMP and calcium in mouse parotid gland.     

cAMP and vascular action of endothelin.

Endothelin, a potent vasoconstricting peptide, induced maximal contraction of the rat tail artery within 3 min of administration. Endothelin also caused a dose-dependent decrease in tissue levels of cAMP. Concentrations of cAMP decreased about 60% 1 min after endothelin (10(-8) M) administration, but returned to basal levels in 5 min. By contrast, tissue levels of cGMP were not changed by endothelin. Modulation of cAMP production may provide one mechanism for the vascular action of endothelin.     

Effects of acute and chronic electroconvulsive stimuli on cAMP and cGMP efflux in the rat striatum and hippocampus

The effects of acute and chronic electroconvulsive stimuli (ECS) on extracellular concentrations of the cyclic nucleotides, cAMP and cGMP, from the striatum and hippocampus of awake rats were studied with in vivo microdialysis in conjunction with radioimmunoassay. Acute ECS, but not acute sham-ECS, significantly increased cAMP and cGMP efflux from the striatum by about 75 and 50%, respectively. Chronic ECS did not influence significantly basal efflux of cAMP or cGMP from the striatum or the hippocampus in comparison to control animals receiving chronically sham-ECS. Administration of a challenge ECS in animals treated chronically with sham-ECS resulted in an increase in cAMP and cGMP concentrations in the striatum by 20%, but it failed to affect significantly efflux of these nucleotides in animals treated chronically with ECS. Similarly, in the hippocampus, administration of a challenge ECS in animals treated chronically with sham-ECS resulted in an increase in cAMP and cGMP concentrations by about 40 and 65%, respectively, whereas it failed to affect significantly efflux of these nucleotides in animals treated chronically with ECS. Thus, acutely administered ECS increases cAMP and cGMP efflux in the striatum and hippocampus of rats, an effect that is greatly diminished in animals chronically receiving ECS. These findings suggest changes in the cAMP and cGMP signal transduction mechanisms in response to acute and chronic ECS that may be related to the therapeutic effects of this antidepressant and antipsychotic treatment.     

褪黑素对吗啡戒断大鼠脑内cAMP和cGMP含量的影响

目的 :观察褪黑素 (MT)对吗啡戒断大鼠不同脑区cAMP和cGMP含量的影响。方法 :以剂量递增法连续皮下注射吗啡建立吗啡依赖模型 ,采用放射免疫学方法测定脑内cAMP和cGMP的含量。结果 :(1)MT对大鼠吗啡戒断症状具有明显的抑制作用 ;(2 )与对照组比较 ,吗啡依赖大鼠的纹状体、间脑、中脑、脑桥和海马内cAMP含量显著增高 (P <0 0 5 ,P <0 0 1) ,cGMP含量显著下降 (P <0 0 5 ,P <0 0 1)。与吗啡依赖组比较 ,催促戒断大鼠海马和纹状体内cAMP的含量显著升高 (P <0 0 5 ) ,而cGMP含量显著下降 (P <0 0 5 ,P <0 0 1) ,其他部位则无明显变化 ;(3)褪黑素急性治疗可使吗啡戒断大鼠纹状体、间脑、中脑、脑桥和海马内cAMP含量明显下降 (P <0 0 5 ,P <0 0 1) ,cGMP含量明显增高 (P <0 0 5 ,P <0 0 1)。结论 :MT可显著抑制大鼠吗啡戒断反应 ,并与调节中枢cAMP和cGMP含量有关。     

Regional rat brain content of adenosine 3',5'-cyclic monophosphate and guanosine 3',5'-cyclic monophosphate after acute and subacute treatment with ethanol

Acute administration of ethanol (1.0, 2.0, and 5.0 g/kg, ip) to naive male rats (Sprague-Dawley) caused a dose-dependent depression of cerebellar guanosine 3′,5′-cyclic monophosphate (cGMP), and a reduction in cortical cGMP at the highest dose of ethanol. The cGMP content was not altered in the anterior hypothalamus, posterior hypothalamus, or striatum. On examining adenosine 3′,5′-cyclic monophosphate (cAMP) levels, only the area of the striatum was reduced (5.0 g/kg, ethanol). In these acute experiments there was a negative correlation between blood ethanol concentrations and body temperatures. An elevated environmental temperature (31 ± 1°C) to prevent hypothermia from ethanol administration indicated that hypothermia was not a contributory factor. In the subacute experiments animals showed at the end of 24 hr of ethanol inhalation less hypothermia than naive animals with similar blood ethanol concentrations, a reduction in cGMP in the cerebellum and cortex, but no alteration in regional brain cAMP. When the animals were injected with ethanol (2.0 g/kg, ip) 48 hr after removal from the chamber (ethanol vapor, 24 hr), ethanol produced no significant reduction in body temperature (tolerance), but a decrease in cerebellar cGMP. The cAMP content of the tissues was similar to control animals. Ethanol administration (2.0 g/kg, ip) 48 hr later to animals which were previously exposed to air only in the chamber (24 hr) demonstrated a reduction in body temperature as compared with tolerant animals, a decrease in cerebellar cGMP, and no depletion of cAMP in regional sections of the brain. From this in vivo study the data seem to suggest that brain cyclic nucleotides, particularly cAMP, may have a limited role in ethanol-induced intoxication and tolerance to the hypothermic effect of ethanol.     

棱曼中毒后家兔脑内环核苷酸和钙调素水平的变化

用放射免疫测定法测定家兔梭曼中毒后小脑cAMP,cGMP含量,发现随梭曼剂量的增加,cAMP、cGMP含量明显升高,而cAMP/cCMP值明显降低。其中当梭曼剂量为25μg/kg时,小脑,桥-延脑cAMP,cGMP含量在中毒30min时最高,cAMP/cGMP值最低,随后开始恢复,在中毒120min时接近对照水平。用PDE法测定家兔大脑皮层的钙调素(CaM)含量(有活性的CaM含量),发现中毒15min时,CaM含量已明显降低,中毒30min时未见恢复。值得提出的是,动物中毒症状的轻重变化过程与cAMP,cGMP含量及cAMP/cGMP值的升降变化过程相吻合。结果提示,梭曼可能通过影响中枢cAMP/cGMP值发挥毒性作用。     

Effects of protein kinase inhibitor (1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H7) on protein kinase C activity and adrenergic stimulation of cAMP and cGMP in rat pinealocytes

Protein kinase C is thought to be involved in the adrenergic regulation of pineal function. In this tissue, norepinephrine increases cAMP and cGMP accumulation through a synergistic dual receptor mechanism involving alpha 1- and beta-adrenergic receptors; the available evidence indicates that the alpha 1-adrenergic stimulation activates protein kinase C, and that this potentiates beta-adrenergic stimulation of pineal cAMP. The role of protein kinase C in the regulation of cGMP is unclear. In the present report, we determined whether an inhibitor of protein kinase C, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H7), inhibits pineal protein kinase C and the adrenergic stimulation of pineal cAMP and cGMP. H7 (10(-4) M) reduced soluble protein kinase C activity by 40%. Treatment of intact pinealocytes with H7 for 0-240 min reduced the effects of subsequent norepinephrine (NE) stimulation of cAMP and cGMP accumulation by at least 25%. H7 also inhibited 25-30% the maximum stimulation of both cAMP and cGMP produced by concurrent treatment with isoproterenol and two agents which elevate intracellular Ca2+, ouabain and A23187. However, H7 did not reduce the effects of selective beta-adrenergic activation, indicating that H7 was probably inhibiting the effects of NE by blocking alpha 1-adrenergic potentiation of beta-adrenergic stimulation, not beta-adrenergically activated mechanisms. H7 also reduced the stimulation of cAMP accumulation produced by the combined treatment of isoproterenol and an activator of protein kinase C, 4-beta-phorbol 12-myristate, 13-acetate, which is consistent with the view that H7 is acting by inhibiting protein kinase C activity. These observations are in agreement with the conclusion that potentiation of beta-adrenergic stimulation of cAMP by alpha 1-adrenergic agonists, protein kinase C activators, or [Ca2+]i elevating agents involves protein kinase C. In addition, these results are of special interest because they point to the possibility that protein kinase C is involved in the regulation of cGMP accumulation.     

慢性吗啡处理对大鼠交感神经节cAMP、cGMP水平的影响

目的观察慢性吗啡(Mor)处理对大鼠交感神经节——颈上神经节(SCG)环腺苷酸(cAMP)和环鸟苷酸(cGMP)含量的影响。方法剂量递增法连续皮下注射(sc)吗啡5 d建立慢性吗啡依赖大鼠模型,纳洛酮(Nal)催促戒断法和55℃热水甩尾试验判断模型依赖性和耐受性,125I放射性免疫测定法测定SCG中cAMP和cGMP的含量。结果①吗啡急性给药组大鼠SCG中cAMP含量较正常对照组降低(P<0.05);②吗啡依赖组大鼠SCG中cAMP含量回到正常对照组水平,并有增高趋势(与正常对照组比较,P>0.05;与吗啡急性给药组比较,P<0.05);③吗啡戒断组大鼠SCG中cAMP含量较正常对照明显增高(P<0.05);④各组大鼠SCG的cGMP含量差异无显著性(P>0.05)。结论慢性吗啡处理大鼠交感神经节cAMP存在适应性上调现象。