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1.
大鼠实验性脾虚证空肠粘膜吸收细胞ATP酶的细胞化学研究 总被引:4,自引:0,他引:4
用Wistar成年雄性大鼠8只,分为对照组、实验性脾虚组、自然恢复组和中药治疗组。四组动物用水合氯醛麻醉后,经心脏灌流固定,取空肠制成50μm厚的振荡切片,按Robinson和Karnovsky法显示Mg2+-ATPase,然后常规电镜包埋和制片,在H-700型电镜下观察。结果显示,在粘膜吸收细胞微绒毛质膜铈离子(Ce3+)沉淀呈现为电子密度较高的细密颗粒。脾虚组酶反应强度明显低于对照组;自然恢复组酶反应高于脾虚组而低于对照组;中药治疗组酶反应较高,与对照组相似。本文提示,肠粘膜吸收细胞ATP酶活性的降低,使食物的消化吸收减弱 相似文献
2.
实验性氟中毒大鼠中缝背核酶细胞化学研究 总被引:2,自引:0,他引:2
以饮用高氟水(100ppm)的方法制造了雄性大鼠慢性氟中毒模型。对中缝背核神经元Mg2+-腺苷三磷酸酶(Mg2+-ATPase)和硫胺素焦磷酸酶(TPPase)进行了定性、定量分析。结果表明:氟中毒大鼠中缝背核神经元内Mg2+-ATPase、TPPase染色均浅于对照组,定量分析Mg2+-ATPase、TPPase含量均减少。电镜观察氟中毒大鼠神经元内TPPase颗粒减少,同时出现超微结构改变。这些结果提示:高氟可抑制某些酶的活性,并对中枢神经系统有直接损害 相似文献
3.
稀土离子对CaM及Ca2+-Mg2+-ATPase活力及CD研究 总被引:4,自引:0,他引:4
研究了稀土离子(Ln3+)对钙调蛋白(CaM)调控的Ca2+-Mg2+-ATPase的活力影响。结果表明,在CaM和Ca2+-Mg2+-ATPase的体系中,一些Ln3+(La3+、Gd3+)对由CaM调节的Ca2+-Mg2+-ATPase的活力影响呈现双相效应,即Ln3+在低浓度时,能提高激活Ca2+-Mg2+-ATPase的水解活力;在高浓度时,则抑制CaM调节Ca2+-Mg2+-ATPase活力的能力;少数Ln3+(Sm3+)仅表现出抑制效应。在无CaM的Ca2+-Mg2+-ATPase体系中,高浓度的Ln3+抑制Ca2+-Mg2+-ATPase的基础活力。结合圆二色(CD)谱信息对Ln3+和CaM相互作用的分子机制进行了初步的探讨。 相似文献
4.
本文对10例成年Wistar大鼠海马,应用过氧化物酶二氨基联苯胺(DAB)法、碱性磷酸酶(AIP)、镁离子激活的三磷酸腺苷酶(Mg(2+)-ATPase)、钙离子激活的三磷酸腺昔酶(Ca(2+)-ATPase)和5’-核苷酸酶(5’-Nase)等酶组织化学方法显示其微血管,并应用体视学方法测算,比较上述方法显示微血管的效果,结果表明:DAB法显示微血管的效果最好,AIP法次之,Mg(2+)-AT-Pase法再次之。大鼠海马微血管Ca(2+)-ATPase呈弱阳性,5‘-Nase呈阴性。DAB法和Mg(2+)-ATPase法分别适宜作微血管长度密度和血管直径的定量分析。 相似文献
5.
6.
黄卓辉 《植物生理与分子生物学学报》1994,(2)
用2μg/ml玉米素溶液预处理叶绿体或在光活化前于活化液中加入2μg/ml玉米素溶液,观察到玉米素能促进叶绿体膜上耦联因子DTT光活化Mg2+-ATPase及Mg2+GTPase的活力.且对GTPase的促进比例常较ATPase的大些。王米素对OG活化可溶性CF1Mg2+-ATPase活力同样表现出促进作用。用玉米素预处理CF1-β亚基(含微量CF1-α亚基)也观察到它能促进CF1-β亚基催化的Mg2+-ATPase活力。这些结果表明,玉米素在CF1上的作用部位至少有一个在β亚基或α.β亚基交界处调节其催化功能的。 相似文献
7.
用2μg/ml玉米素溶液预处理叶绿体或在光活化前于活化液中加入2μg/ml玉米素溶液,观察到玉米素能促进叶绿体膜上耦联因子DTT光活化Mg^2+-ATPase及Mg^2+-GTPase的活力,且对GTPase的促进比例常较ATPase的大些。玉米素对OG活化可溶性CF1Mg^2+-ATPase活力同样表现出促进作用。用玉米素预处理CF1-β亚基(含微量CF1-α亚基)也观察到它促进CF1-β亚基催 相似文献
8.
骤冷与饥饿对小鼠肝脏影响的实验研究 总被引:4,自引:0,他引:4
为探讨饥饿及饥饿与骤冷对动物肝脏的影响,本实验用健康昆明种小鼠25只,随机分成正常组5只,饥饿组10只,饥饿后再予冷刺激组10只(下称骤冷组)。采用组织化学及酶组织化学方法观察糖原(PAS反应)、SDH(琥珀酸脱氢酶),LDH(乳酸脱氢酶),ChE(胆碱酯酶)、Mg2+-ATPase(镁激活三磷酸腺苷酶),ACP(酸性磷酸酶)。结果提示:饥饿时肝脏PAS反应,SDH,Mg2+-ATPase、ChE活性显著下降,而ACP活性明显增强;饥饿后骤冷时PAS(反应)、SDH、ChE更显著下降,而ACP及Mg2+-ATPase活性反而增强。 相似文献
9.
牛磺酸对损伤的心肌肌膜ATPase活性影响的研究 总被引:7,自引:0,他引:7
本研究以异丙肾上腺素(Isoproternol,Iso.)诱导的大鼠心肌损伤为模型,观察牛磺酸(Taurine,Tau)对损伤心肌肌膜上N_a ̄+—K_- ̄+ATPase、C_a ̄(2+)-ATPase、M_g ̄(2+)ATPASE活性的影响。按Dahalla方法分离及鉴定心肌肌膜同时分别测定三种ATPase活性,结果:Iso组,与Tau+Iso组的三种ATPase活性明显低于对照组与Tau组,且同时Tau+Iso组明显高于Iso组但对照组与Tau组之间差别无显著性。结果提示:牛磺酸能提高异丙肾上腺素诱导心肌损伤的心肌膜ATPase活性,可能通过其保护心肌肌膜的结构及功能而实现的。 相似文献
10.
莱氏衣原体膜上Mg~(2+)-ATPase用DOC溶解后,经Sepharose-6B和DEAE-CelluloseDE-52离子交换柱,得到了部分纯化的Mg~(2+)ATPase,并将此ATPase与不同极性头部的磷脂和膜糖脂重组,研究了不同的极性头部的磷脂和膜糖脂对ATPase活性的影响。此酶的活性不依赖酸性磷脂,PG、DPG、大豆磷脂等明显抑制酶活性,中性磷脂DMPC、PE、PC则能增加酶活性,其中尤以非双层脂PE的作用最为明显。从莱氏衣原体膜上提取的糖脂(MGDG,DGDG)单独和ATPase重组时,酶活性增加并不明显,当MGDG和DGDG以等比例混合时,能大大地增加酶活性。这表明Mg~(2+)-ATPase的活性很大程度上与磷脂的表面电荷及磷脂的组成相关。 相似文献
11.
实验性脾虚证大鼠结肠和盲肠组织化学、免疫组织化学研究 总被引:1,自引:0,他引:1
成年雄性Wistar大鼠60只,分正常对照组;脾虚组;自然恢复组和四君子汤治疗组。四组动物取结肠和盲肠分别进行H-E;酶组织化学和5-羟色胺细胞PAP免疫组织化学观察。结果表明,脾虚组结肠和盲肠粘膜上皮的杯状细胞较对照组明显增多,淋巴细胞及淋巴集结也增多。固有层毛细血管充血并有炎症细胞的增多。上皮细胞和腺细胞琥珀酸脱氢酶(SDH)减弱,乳酸脱氢酶(LDH)增强;结肠上皮细胞和腺细胞碱性磷酸酶(AIP)和三磷酸腺苷酶(ATPase)增强,而盲肠上皮细胞和腺细胞AIP和ATPase减弱。结肠和盲肠5-HT细胞免疫组化反应减弱。这些结果表明肠上皮细胞酶的变化和5-HT细胞分泌活性的变化与脾虚证的发生密切相关,可能是导致脾虚证原因之一。经四君子汤治疗后以上各项指标均比自然恢复组更接近于正常对照组,说明此药对消化道粘膜上皮酶活性和5-HT细胞分泌活性恢复正常有明显效果,从而起治疗作用。 相似文献
12.
实验性脾虚证大鼠内分泌腺形态学与细胞化学研究 总被引:3,自引:0,他引:3
成年Wistar大鼠20只,分对照组和实验组。实验组用大黄煎剂灌服42天致实验性脾虚证。对两组睾丸,肾上腺和甲状腺进行常规形态学与细胞化学观察。形态学显示:睾丸,肾上腺和甲状腺的组织和细胞均有不同程度的损伤。细胞化学显示:各内分泌腺均表现酸性磷酸酶(ACP)减弱,个别内分泌腺细胞显示琥珀酸脱氢酶(SDH),三磷酸腺苷酶(ATPase)和PAS反应的变化。结果证明脾虚证不仅消化系统功能低下,而且内分泌功能也受到了影响 相似文献
13.
丹参对鼠缺血后残肝酶组织化学的影响 总被引:2,自引:1,他引:1
研究观察鼠缺血后肝再生时酶组织化学的变化。结果表明缺血后肝再生时肝细胞SDH、G6Pase和Mg2+ATPase活性及PAS反应均在不同程度上低于对照组,而LDH、ACP活性较对照组有不同程度升高。丹参组酶活性及PAS反应基本处于缺血组与对照之间,说明丹参对保护某些酶活性,促进肝再生有一定的作用 相似文献
14.
Properties of (Na+ plus K+)-activated ATPase in rat liver plasma membranes enriched with bile canaliculi. 总被引:4,自引:0,他引:4
Liver plasma membranes enriched in bile canaliculi were isolated from rat liver by a modification of the technique of Song et al. (J. Cell Biol. (1969) 41, 124-132) in order to study the possible role of ATPase in bile secretion. Optimum conditions for assaying (Na+ plus K+)-activated ATPase in this membrane fraction were defined using male rats averaging 220 g in weight. (Na+ plus K+)-activated ATPase activity was documented by demonstrating specific cation requirements for Na+ and K+, while the divalent cation, Ca(2+), and the cardiac glycosides, ouabain and scillaren, were inhibitory. (Na+ plus K+)-activated ATPase activity averaged 10.07 plus or minus 2.80 mumol Pi/mg protein per h compared to 50.03 plus or minus 11.41 for Mg(2+)-activated ATPase and 58.66 plus or minus 10.07 for 5'-nucleotidase. Concentrations of ouabain and scillaren which previously inhibited canalicular bile secretion in the isolated perfused rat liver produced complete inhibition of (Na+ plus K+)-activated ATPase without any effect on Mg(2+)-activated ATPase. Both (Na+ plus K+)-activated ATPase and Mg(2+)-activated ATPase demonstrated temperature dependence but differed in temperature optima. Temperature induced changes in specific activity of (Na+ plus K+)-activated ATPase directly paralleled previously demonstrated temperature optima for bile secretion. These studies indicate that (Na+ plus K+)-activated ATPase is present in fractions of rat liver plasma membranes that are highly enriched in bile canaliculi and provide a model for further study of the effects of various physiological and chemical modifiers of bile secretion and cholestasis. 相似文献
15.
A Columbano G M Ledda-Columbano M G Ennas M Curto M G De Montis M W Roomi P Pani D S Sarma 《Basic and applied histochemistry》1988,32(4):501-510
The effect of a single administration of lead nitrate on the activity of gamma-glutamyltranspeptidase (gamma-GT), adenosine triphosphatase (ATPase), the placental form of glutathione S-transferase (GST-P) and adenylate cyclase (AC), four enzymes widely used as phenotypic markers for preneoplasia, was investigated in the liver of male Wistar rats. The results of the histochemical enzymatic staining indicated that an acute treatment with lead nitrate induces the activity of gamma-GT, mainly in the hepatocytes located around zone I of the liver acinus, with a maximum seen between 72-96 hours. On the other hand, the activity of ATPase was found to be severely inhibited at 2-3 days after treatment, as shown by a strong decrease in the staining of the bile canaliculi of zones II and III. Immunohistochemical analysis revealed that lead nitrate administration also resulted in the appearance in most of the hepatocytes of GST-P, an enzyme whose activity is almost undetectable in normal rat liver, but is elevated in preneoplastic liver lesions. Finally, lead nitrate treatment resulted in an inhibition of AC activity which was maximal after 24 hours. 相似文献
16.
Bertone V Tarantola E Ferrigno A Gringeri E Barni S Vairetti M Freitas I 《European journal of histochemistry : EJH》2011,55(1):e5
Biliary complications often lead to acute and chronic liver injury after orthotopic liver transplantation (OLT). Bile composition and secretion depend on the integrated action of all the components of the biliary tree, starting from hepatocytes. Fatty livers are often discarded as grafts for OLT, since they are extremely vulnerable to conventional cold storage (CS). However, the insufficiency of donors has stimulated research to improve the usage of such marginal organs as well as grafts. Our group has recently developed a machine perfusion system at subnormothermic temperature (20°C; MP20) that allows a marked improvement in preservation of fatty and even of normal rat livers as compared with CS. We sought to evaluate the response of the biliary tree of fatty liver to MP20, and a suitable marker was essential to this purpose. Alkaline phosphatase (AlkP, EC 3.1.3.1), frequently used as marker of membrane transport in hepatocytes and bile ducts, was our first choice. Since no histochemical data were available on AlkP distribution and activity in fatty liver, we have first settled to investigate AlkP activity in the steatotic liver of fatty Zucker rats (fa/fa), using as controls lean Zucker (fa/+) and normal Wistar rats. The AlkP reaction in Wistar rats was in accordance with the existing data and, in particular, was present in bile canaliculi of hepatocytes in the periportal region and midzone, in the canals of Hering and in small bile ducts but not in large bile ducts. In lean ZR liver the AlkP reaction in Hering canals and small bile ducts was similar to Wistar rat liver but hepatocytes had lower canalicular activity and besides presented moderate basolateral reaction. The difference between lean Zucker and Wistar rats, both phenotypically normal animals, could be related to the fact that lean Zucker rats are genotypically heterozygous for a recessive mutated allele. In fatty liver, the activity in ductules and small bile ducts was unchanged, but most hepatocytes were devoid of AlkP activity with the exception of clusters of macrosteatotic hepatocytes in the mid-zone, where the reaction was intense in basolateral domains and in distorted canaliculi, a typical pattern of cholestasis. The interpretation of these data was hindered by the fact that the physiological role of AlkP is still under debate. In the present study, the various functions proposed for the role of the enzyme in bile canaliculi and in cholangiocytes are reviewed. Independently of the AlkP role, our data suggest that AlkP does not seem to be a reliable marker to study the initial step of bile production during OLT of fatty livers, but may still be used to investigate the behaviour of bile ductules and small bile ducts. 相似文献
17.
Nobukazu Araki Yoichiro Takashima Takashi Makita 《Histochemistry and cell biology》1995,104(4):257-265
The redistribution and fate of colchicine-induced alkaline phosphatase (ALPase) in rat hepatocytes were investigated by electron microscopic enzyme cytochemistry and biochemistry. ALPase activity markedly increased in rat hepatocytes after colchicine treatment (2.0 mg/kg body weight, intraperitoneal injection). At 20–24 h after colchicine treatment, the liver showed the highest activity of ALPase. Thereafter, ALPase activity decreased and returned to normal levels at 48 h. In normal hepatocytes from control rats, ALPase activity was seen only on the bile canalicular membrane. However, at 20–24 h after colchicine treatment, colchicine-induced ALPase was redistributed in the sinusoidal and lateral (basolateral) membranes as well as in the bile canalicular membrane. At 30–36 h after colchicine treatment, ALPase activity on the basolateral membrane gradually decreased. In contrast, ALPase in the bile canalicular membrane increased along with the enlargement of bile canaliculi, suggesting that ALPase in the basolateral membrane had been transported to the bile canalicular membrane. Furthermore, ALPase-positive vesicles, cisternae and autophagosome-like structures were frequently seen in the cytoplasm. ALPase was also positive in some lysosomal membranes. ALPase in hepatocytes at 48 h after colchicine treatment returned to almost the same location as in control hepatocytes. Altogether, it is suggested that excessively induced ALPase is at least partially retrieved by invagination of the bile canalicular membrane and then transported to lysosomes for degradation. In addition, this study indicates that excess plasma membrane might be a possible origin of autophagosomal membrane. 相似文献
18.
脾虚证小鼠胸腺细胞周期和免疫细胞化学的实验研究 总被引:7,自引:0,他引:7
本文用FCM和免疫细胞化学技术检测了小鼠胸腺细胞的活动周期,Brdu+细胞和表达Thyl.2抗原的T细胞。小鼠包括对照组,利血平诱发的脾虚组,自然恢复组和健脾汤治疗后复健组。FCM和免疫细胞化学结果显示,脾虚组小鼠G1期细胞堆积,S期和G2+M期细胞减少。同时胸腺内Brdu+细胞和T细胞也相应减少。经过健脾汤治疗后,除G1期细胞外,S期和G2+M期细胞,Brdu+细胞和T细胞均显著增加,并且几乎达正常水平。但是自然恢复组在恢复细胞活动周期正常化,以及恢复Brdu+细胞和T细胞方面比复健组缓慢。说明健脾汤对脾虚小鼠的胸腺细胞有促进DNA合成和细胞增殖功能。 相似文献
19.
Electron microscopic cytochemical characterization of bile canaliculi and bile ducts in vitro. 总被引:2,自引:0,他引:2
M Oda M J Phillips 《Virchows Archiv. B, Cell pathology including molecular pathology》1975,18(2):109-118
Electron microscopic cytochemical localization of Mg++-activated adenosine triphosphatase (Mg++-ATPase) and 5-nucleotidase (AMPase) was investigated in bile canaliculus-rich and bile duct-containing fractions isolated from rat liver. Comparative cyochemical studies between prefixed and non-prefixed fractions revealed that the activity of both enzymes could be detected in the fractions under appropriate experimental conditions. However, the cytochemical activity of AMPase was much more sensitive to glutaraldehyde than that of Mg++-ATPase. Mg++-ATPase and AMPase reaction products were localized primarily on bile canalicular microvilli, that is, along the outer (luminal) surface of canalicular plasma membranes, but they were never observed on bile ductal microvilli. AMPase was also detectable on lateral hepatic plasma membranes. Mg++-ATPase demonstrated by the cytochemical technique described is a reliable enzyme marker for isolated bile canalicular membranes. At high magnification, Mg++-ATPase reaction product was also observed on the microfilaments surrounding isolated bile canaliculi. The possibility that the reaction product on the pericanalicular microfilaments may result from the hydrolysis of ATP byan actomyosin ATPase-like enzyme associated with these filaments is briefly discussed. 相似文献
20.
目的观察脾虚型肠易激综合征(IBS)模型WHBE兔环核苷酸水平的变化。方法取WHBE兔和日本大耳白兔各18只,分别随机分成两组,即正常对照组6只,模型对照组12只,采用湿热应激加灌服番泻叶煎剂的方法造成脾虚证肠易激综合征(tBs)免模型,其中模型对照组造模后处理6只,剩余兔自然恢复10d,观察实验兔的血浆环核苷酸水平的变化。结果与正常对照组比较,WHBE兔模型对照组血浆cAMP和cGMP水平均显著升高(P〈0.05,P〈0.01),且自然恢复10d后血浆cAMP和cGMP水平仍高于正常对照组(P〈O.05,P〈O.01);日本大耳白兔模型对照组造模后及自然恢复10d后血浆cAMP和cGMP水平略有所升高,但差异均不显著(P〉O.05);且模型对照组和自然恢复组cAMP/cGMP比值均略低于正常对照组,但差异不显著(P〉O.05)。结论本实验脾虚证IBS模型兔的血浆环核苷酸水平随阴阳盛衰而变化,具有双重特点:既有阴虚特点,又有阳虚特点,且WHBE兔更适合于脾虚证IBS模型研究理想的实验动物。 相似文献