复方肾华片对急性脑缺血再灌注损伤大鼠肾小管Na+-K+-AT酶的调节作用

Objective:To observe the effect of Compound Shenhua Tablet(复方肾华片,SHT) on the sodiumpotassium-exchanging adenosinetriphosphatase(Na~+-K~+-ATPase) in the renal tubular epithelial cells of rats with acute ischemic reperfusion and to investigate the mechanisms underlying the effects of SHT on renal ischemic reperfusion injury(RIRI).Methods:Fifty male Wistar rats were randomly divided into the sham surgery group,model group,astragaloside group[150 mg/(kg-d)],SHT low-dose group[1.5 g/(kg·d)]and SHT high-dose group[3.0 g/(kg·d)],with 10 rats in each group.After 1 week of continuous intragastric drug administration,surgery was performed to establish the model.At either 24 or 72 h after the surgery,5 rats in each group were sacrificed,blood biochemistry,renal pathology,immunoblot and immunohistochemical examinations were performed,and double immunofluorescence staining was observed under a laser confocal microscope.Results:Compared with the sham surgery group,the serum creatinine(SCr) and blood urea nitrogen(BUN) levels were significantly increased,Na~+-K~+-ATPase protein level was decreased,and kidney injury molecule-1(KIM-1) protein level was increased in the model group after the surgery(P0.01 or P0.05).Compared with the model group,the SCr,BUN,pathological scores,Na~+-K~+-ATPase,and the KIM-1 protein level of the three treatment groups were significantly improved at 72 h after the surgery(P0.05 or P0.01).And the SCr,BUN of the SHT low- and high-dose groups,and the pathological scores of the SHT high-dose group were significantly lower than those of the astragaloside group(P0.05).The localizations of Na~+-K~+-ATPase and megalin of the model group were disrupted,with the distribution areas overlapping with each other and alternately arranged.The severity of the disruption was slightly milder in three treatment groups compared with that of the model group.The results of immunofluorescence staining showed that the SHT high-dose group had a superior effect as compared with the astragaloside group and the SHT low-dose group.Conclusions:The SHT effectively alleviated RIRI caused by ischemic reperfusion,promoted the recovery of the polarity of renal tubular epithelial cells,and protected the renal tubules.The therapeutic effects of SHT were superior to those of astragaloside as a single agent.     

Solubilization and separation of divalent cation dependent adenosinetriphosphatase in native gradient polyacrylamide slab gel

A new method for the simultaneous solubilization and separation of different cation dependent adenosinetriphosphatases after non-denaturing polyacrylamide gel electrophoresis is described. Using a gradient system, 3 distinct divalent cation dependent adenosinetriphosphatase bands (Mg2+-, Ca2+-, and Mg2+ + Ca2+-dependent) could be separated on the same gel from NP-40 solubilized brain microsomal preparations. This indicates that these 3 adenosinetriphosphatases represent distinct molecular species.     

Lymphocyte ATP-ase Activity in Treated Cases of Hodgkin's Disease

The ATP-ase activity was determined in lymphocytes isolated from peripheral blood in 41 patients with Hodgkin's disease and 50 healthy controls. All patients were previously treated with irradiation or cytostatic drugs and 17 patients were under maintenance therapy at the time of investigation. A significantly increased ATP-ase activity was found in lymphocytes from patients with Hodgkin's disease. The individual activities were unrelated to the clinical stage of the disease, but correlated to the histological classification of the lymphatic tissue. Significantly lower lymphocyte ATP-ase activity was found in patients under maintenance treatment with immunosuppressive drugs, especially if the patients had previously been irradiated. It is suggested that the ATP-ase activity of circulating lymphocytes is related to the immunological activity against the presence of the malignant cells in Hodgkin's disease.     

胸痹通胶囊对大鼠缺血心肌的保护作用研究

目的探讨胸痹通胶囊对大鼠急性缺血心肌的保护作用机制。方法将动物随机分为3组,即正常对照组(A组)、模型对照组(B 组)、模型治疗组(C)组,制作异丙基肾上腺素性心肌缺血模型,测定各组大鼠血浆中超氧化物歧化酶的总活性(T-SOD)、血栓素 A_2、前列腺素 PGI_2的中间代谢产物(TXB_2、6-Keto-PGF_(la))、血浆内皮素(ET)、血浆降钙素基因相关肽(CGRP)、心肌心钠素(ANF)及心肌 ATP 酶活性等的活性,并进行组间比较。结果心肌缺血(模型对照组)大鼠血浆 T-SOD 活性降低、TXB_2、ET 含量增加、6-Keto-PGF_(la)、CGRP 减少,心肌 ANF 减少,ATP 酶活性降低。而模型治疗组(胸痹通 XBTC 组)上述变化则明显改善。结论 XBTC 可通过提高 SOD、ATP 酶的活性以及纠正缺血时血管活性物质的释放而发挥心肌保护作用。     

24例老年肺心病患者红细胞钙－ATP酶和钙镁含量的变化

目的观察老年肺心病患者红细胞膜钙－ＡＴＰ酶（钙泵）活性和红细胞内Ｃａ２、Ｍｇ２＋含量的变化，初步探讨其发生机制及意义。方法采用生化方法和原子吸收法测定上述指标，并与非老年肺心病患者及老年健康者比较。结果老年肺心病组（２４例）红细胞钙泵活性（μｍｏｌｐｉ·ｍｇ－１／ｈ）、Ｃａ２＋含量（μｍｏｌ／ｇ）、Ｍｇ２＋含量（μｍｏｌ／ｇ）分别为０．２３±０．１０、７．４６±１．８２、８．２１±０．８４；非老年肺心病组（２０例）分别为０．３２±０．１１、６．０１±１．５２、８．７４±０．７２；老年健康组（１２名）分别为０．４２±０．１５、４．５１±１．４４、９．０２±１．０８。老年肺心病组与后两组比较，红细胞钙泵活性及Ｍｇ２＋含量降低，Ｃａ２＋含量增高（Ｐ值＜０．０５、０．０１或０．００１）；在肺心病伴呼吸衰竭和心力衰竭时上述异常改变更为明显。结论红细胞膜钙泵活性降低及其细胞内Ｃａ２＋、Ｍｇ２＋含量异常变化，在老年人肺心病的发病中可能具有重要作用     

蒿甲醚对小鼠体内日本血吸虫磷酸化酶、乳酸脱氢酶、三磷酸腺苷酶和磷酸葡萄糖脱氢酶的影响

目的:观察蒿甲醚(Art)对日本血吸虫磷酸化酶(PP)、乳酸脱氢酶(LDH)、6-磷酸葡萄糖脱氢酶(G-6-PDH)和三磷酸腺苷酶(ATPase)的影响。方法:感染32-38天的小鼠于灌服Art 100-300mg·kg~(-1)后24-72h剖杀,收集雌()、雄虫(),按NADH和NADPH的形成和无机磷的释放量测定虫的上述4种酶。结果:感染小鼠用Art 300mg·kg~(-1)治疗后24-48h,、虫的总PP和PPa(激活型)活力明显增加,而PPb则无或仅有轻度增加。上述小鼠用Art 100-300mg·kg~(-1)治疗后24-72h,LDH和G-6-PDH的抑制率分别为9％-59％()和41％-75％()及22％-42％()和74%-80%()。用300mg·kg~(-1)治疗后24h,仅虫的Mg~2-ATPase明显受抑制,48h后,Ca~(2 )-ATPase、Mg~(2 )-ATPase和Na~ -K~ -ATPase分别抑制17％()和19％(),32％()和48％(),及29％()和44％()。结论:Art引起血吸虫PPa活力的增加,使虫的糖原分解,并抑制LDH使虫糖酵解的终产物乳酸明显减少。此外,对血吸虫虫的G-6-PDH有明显的抑制作用。     

甘草酸二铵对大鼠心肌缺血再灌注损伤脂质过氧化及心肌酶活性的影响

目的:观察甘草酸二铵(DG)对大鼠心肌缺血再灌注损伤脂质过氧化及心肌酶活性的影响。方法:雄性wistar大鼠30只,随机分为假手术组、缺血再灌注组和DG20mg·kg-1组。每组10只。采用在体大鼠心肌缺血30min再灌注60min损伤模型,再灌注60min后分别用比色法测定心肌丙二醛(MDA)含量、超氧化物歧化酶(SOD)、三磷酸腺苷酶(ATP酶)、血清磷酸肌酸激酶(CPK)和乳酸脱氢酶(LDH)水平,并用酶组织化学方法检测心肌组织琥珀酸脱氢酶(SDH)的活性。结果:DG能显著降低心肌组织中MDA含量和SDH的活性(P<0.05,P<0.01),提高SOD和ATP酶活性(P<0.05,P<0.01),并减少心肌CPK和LDH的释放(P<0.05,P<0.01)。结论:DG具有保护大鼠心肌缺血再灌注损伤的作用,其作用机理可能与其降低心肌脂质过氧化,增强心肌细胞SOD、SDH和ATP酶活性有关。     

亚低温对颅脑创伤大鼠脑线粒体活性氧和ATP酶合成能力的影响

目的:研究亚低温治疗对颅脑创伤(TBI)大鼠线粒体活性氧(ROS)生成和ATP酶合成能力的影响。方法:72只动物随机分为假手术组、常温TBI组(肛温36℃～37℃)、亚低温TBI组(肛温31℃～33℃,亚低温持续2h),后2组利用液压打击(FPI)制作中度TBI模型。各组于脑外伤后2h,24h,3d和7d断头取伤侧大脑组织,差速离心法提取伤侧脑线粒体,酶荧光法检测线粒体ATP合成能力,荧光法测定线粒体ROS的生成。结果:常温TBI组脑创伤后线粒体ATP合成能力显著下降,24h降至最低,至7d时仍显著低于假手术组,ROS在各时间点均显著升高,3d时为最高。亚低温TBI组线粒体ATP合成能力在各时间点均高于常温TBI组,除2h外,其他时间点ROS低于常温TBI组。结论:颅脑创伤后线粒体ATP合成能力显著下降,ROS显著升高,亚低温可以改善线粒体ATP酶合成能力,抑制活性氧大量生成,从而保护脑组织。     

何首乌的抗衰老作用研究

目的：探讨何首乌对抗D－半乳糖所致亚急性衰老的机制。方法：将小鼠30只随机分为3组：对照组、衰老模型组和何首乌治疗组。衰老模型组、何首乌治疗组用D－半乳糖皮下注射，1次／d，连续6周造成小鼠衰老。第3周时，何首乌治疗组给小鼠灌胃何首乌。第6周末处死。测定不同组织脂褐质（lipofuscin,LPF)、Na^ /K^ -ATP酶（Na^ /K^ -ATPase）、超氧化物歧化酶（superoxide dismutase,SOD）和丙二醛含量。结果：何首乌可降低小鼠脑组织和肾组织的LPF含量，升高心肌Na^ /K^ -ATPase活性和肝脏SOD活性。结论：何首乌具有抗衰老作用，其机制是通过多种途径而实现的。     

(Na+-K+)-ATPase inhibitors: Implications for new drug discovery

The (Na⁺-K⁺)-ATPase is a ubiquitous membrane-bound enzyme that actively transports Na⁺ out of the cell in exchange for a smaller ratio of extracellular K⁺. The current report focuses on the role of modifiers of (Na⁺-K⁺)-ATPase activity in the development of new pharmacological agents. More versatile biological test systems are proposed. Possible use of Na⁺-K⁺-ATPase activity modulators in treatment of hypertension and other diseases are discussed. It is concluded that the ubiquitous distribution and crucial role of the enzyme in normal and diseased cell function merits a critical biological and chemical reappraisal of the enzyme beyond the current narrow viewpoint dealing with the application of glycosides in the treatment of congestive heart failure.