长期服用别嘌醇缓释胶囊对高尿酸血症大鼠肾脏的影响

目的考察长期服用别嘌醇缓释胶囊对高尿酸血症大鼠肾脏改变的影响和机制。方法 90只雄性Wistar大鼠随机分为正常对照组(10只),模型组(16只),别嘌醇缓释胶囊低、高(27、54 mg·kg~(-1)·d~(-1))剂量组(各16只),别嘌醇片低、高(32、64 mg·kg~(-1)·d~(-1))剂量组(各16只),采用腺嘌呤和乙胺丁醇连续灌胃3wk,3 wk后改为隔日1次,共12 wk造成大鼠高尿酸血症。在造模的同时,药物治疗组分别给予低、高剂量的别嘌醇缓释胶囊或别嘌醇片;在wk 12末,处死大鼠,取血和肾脏组织,检测血尿酸、肾脏功能及肾组织的TNF-α表达情况和观察肾脏形态、病理学变化。结果与正常对照组比较,模型组大鼠血尿酸、尿素氮含量和肾脏组织内TNF-α表达增加,肾脏含水量显著增加(P<0.05)。与模型组比较,别嘌醇缓释胶囊高剂量组显著降低大鼠血尿酸、血尿素氮含量和肾脏组织内TNF-α表达(P<0.05);低剂量组能显著降低肾组织含水量(P<0.05);肌酐含量和肾脏指数无明显差异(P>0.05)。与别嘌醇片高剂量组比较,别嘌醇缓释胶囊组降低肾脏指数和肾组织含水量有显著差异(P<0.05,P<0.01)。结论长期应用别嘌醇缓释胶囊治疗高尿酸血症能减缓肾脏的病理改变,其机制可能与其降低血尿酸水平的同时抑制TNF-α介导的炎症反应有关。     

水飞蓟宾对局灶性脑缺血再灌注损伤大鼠保护作用的研究

目的 研究水飞蓟宾(SIL)对局灶性脑缺血再灌注损伤大鼠的保护作用,并探讨其可能的作用机制。方法 取112只清洁级雄性大鼠随机分为6组:假手术组、模型对照组、水飞蓟宾(100、200和400 mg·kg-1)预处理组和尼莫地平(32 mg·kg-1)预处理组,通过中动脉线栓法制备局灶性脑缺血再灌注大鼠模型。再灌注6 h后,进行神经功能评分,分析测定脑组织梗死体积及含水量;通过苏木精-尹红(HE)染色法观察脑组织形态学变化;测定血清中磷酸肌酸激酶(CPK)、乳酸脱氢酶(LDH)含量及总抗氧化能力(T-AOC)水平;测定脑组织中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性和丙二醛(MDA)含量;通过TUNEL染色观察神经细胞凋亡状况并计算凋亡指数(AI),通过Western blot方法测定脑组织中NF-κB蛋白表达并进行半定量分析。结果 与模型组相比,水飞蓟宾(200和400 mg·kg-1)预处理组大鼠神经功能评分显著降低、脑梗死体积和含水量均显著降低,脑组织病理形态学变化及神经细胞凋亡均明显减轻,凋亡指数显著降低;脑组织中SOD、CAT活性显著升高且MDA含量显著降低;血清中CPK,LDH含量显著降低;脑组织中NF-κB蛋白表达量显著降低;水分蓟宾400 mg·kg-1预处理组血清中T-AOC水平显著升高;差异均具有统计学意义(P<0.05,P<0.01)。结论 水分蓟宾对局灶性脑缺血再灌注损伤大鼠具有保护作用,其作用机制可能与水分蓟宾能够改善局灶性脑缺血再灌注损伤大鼠脑组织中抗氧化酶活性、抑制氧化应激损伤有关。     

川芎嗪对顺铂肾损伤大鼠肾细胞凋亡及凋亡相关蛋白表达的影响

目的探讨川芎嗪(TMP)对顺铂(DDP)大鼠肾毒性有无保护作用。方法DDP8mg·kg-1ip诱导大鼠肾损伤,于给予DDP2d前大鼠分别ip50,100mg·kg-1·d-1TMP,连续5d,于给药d5收集各组大鼠尿液,测24h尿蛋白含量,并于末次给药后4h处死大鼠,测血清尿素氮(BUN)和肌酐(Cr)含量,采用原位缺口末端标记法检测肾脏细胞凋亡,免疫组化SP法检测肾脏Bax和Bcl2蛋白表达水平。结果50,100mg·kg-1TMP组可降低DDP所致大鼠24h尿蛋白及血清BUN和Cr含量的升高(P<0.01);TMP两剂量组也可明显减少肾脏细胞凋亡率(P<0.01),显著增强肾脏凋亡相关蛋白Bcl2的表达,减少Bax表达,并明显降低Bax/Bcl2比值(P<0.01)。结论TMP可能通过降低凋亡相关蛋白Bax和增强Bcl2的表达,并降低Bax/Bcl2比值而抑制DDP引起的肾细胞凋亡,使肾脏免受损伤的作用。     

黄芩总黄酮对慢性肾功能衰竭大鼠的肾保护作用及ROS/MAPK/NF-κB信号通路的影响

摘要：目的:探讨黄芩总黄酮对慢性肾功能衰竭大鼠的肾保护作用及活性氧/丝裂原活化蛋白激酶/核因子κB（ROS/MAPK/NF-κB）信号通路的影响。方法:100只SD大鼠随机分为5组:正常对照组、模型组、海昆肾喜组(150 mg·kg-1)、黄芩总黄酮低(100 mg·kg-1)、高(200 mg·kg-1)剂量组,每组20只。除正常对照组外,其余各组建立慢性肾功能衰竭模型。建模成功后灌胃给予相应药物,1次/d,连续4周,正常对照组和模型组给予等体积生理盐水。实验结束后,测定24 h尿蛋白、肾/体比值、肌酐（SCr）、尿素（BUN）、肾小球细胞凋亡水平、肾脏ROS水平、肾脏MAPK、NF-κB基因和蛋白水平。结果:与正常对照组比较,模型组大鼠BUN、SCr、24 h尿蛋白、肾/体比、肾小球凋亡率、肾小球ROS、肾小球MAPK、NF-κB mRNA和蛋白水平显著升高（P<0.05）。与模型组比较,海昆肾喜组、黄芩总黄酮低、高剂量组大鼠BUN、SCr、24 h尿蛋白、肾/体比、肾小球凋亡率、肾小球ROS、肾小球MAPK、NF-κB mRNA和蛋白水平显著降低（P<0.05）,且黄芩总黄酮高剂量组各指标显著低于低剂量组（P<0.05）;黄芩总黄酮低剂量组大鼠各指标显著高于海昆肾喜组（P<0.05）,黄芩总黄酮高剂量组与海昆肾喜组相比差异无统计学意义（P>0.05）。模型组小鼠基底膜、肾小球系膜细胞明显增生肿胀,伴炎症粒细胞浸润;海昆肾喜组及黄芩总黄酮高剂量组肾小球系膜细胞轻度增生,炎症细胞浸润减少;黄芩总黄酮低剂量组仍然可见明显炎症细胞浸润,肾小球肿胀增生。结论:黄芩总黄酮对慢性肾功能衰竭大鼠具有肾保护作用,能明显降低肾小球凋亡率,其机制可能与黄芩总黄酮抑制氧化应激炎症反应通路ROS/MAPK/NF-κB激活有关。     

白芍总苷对糖尿病大鼠肾脏保护作用及部分机制

目的探讨白芍总苷(TGP)对糖尿病大鼠肾脏保护作用及部分机制。方法应用链脲佐菌素诱导大鼠糖尿病模型,同时每日灌胃给予TGP(50,100,200mg·kg-1,连续8wk。结果TGP给药呈剂量依赖性降低糖尿病大鼠AER的增加。TGP给药(50mg·kg-1)大鼠肾小球平均容量(VG)明显低于模型组,肾小管-间质损伤指数(TII)较模型组有所下降,但差异无统计学意义;TGP给药(100,200mg·kg-1VG与TII均明显低于模型组。模型组肾组织MDA含量明显高于对照组,TGP给药(200mg·kg-1)大鼠肾组织MDA含量明显低于模型组。Western blot显示模型组肾组织1α(IV)Ⅳ型胶原蛋白表达较对照组增加2.7倍,TGP给药(50、100、200mg·kg-1)8wk分别可使肾组织1α(IV)Ⅳ型胶原蛋白表达下降47.9%、60.4%与72.9%。模型组肾组织ICAM-1及TGFβ1蛋白表达明显高于对照组,TGP给药(50,100,200mg·kg-1)肾组织ICAM-1与TGFβ1蛋白表达明显低于模型组。结论TGP对糖尿病大鼠肾脏保护作用机制部分与其抑制肾组织中ICAM-1及TGFβ1蛋白表达有关。     

细胞色素C对庆大霉素致大鼠肾小管上皮细胞毒性的影响研究

目的:探讨细胞色素C对庆大霉素致大鼠肾小管上皮细胞毒性的影响及其可能的作用机制。方法:取大鼠随机分为溶媒对照组、庆大霉素组、庆大霉素+细胞色素C组,每组10只,前2组分别腹腔注射生理盐水和庆大霉素(100mg·kg-1·d-1),第3组先静脉注射细胞色素C100mg·kg-1·d-1,30min后腹腔注射庆大霉素100mg·kg-1·d-1。单剂量给药后24h处死大鼠,以荧光偏振免疫发光技术测定后2组大鼠肾脏中庆大霉素的含量,用TUNEL法检测肾小管上皮细胞凋亡情况,用MIAS医学图像分析系统计算凋亡阳性细胞数密度,以苏木精-伊红染色法观察肾脏组织病理学变化。结果:与溶媒对照组比较,庆大霉素组肾小管上皮细胞凋亡的阳性细胞数密度明显增加(P<0.01);与庆大霉素组比较,庆大霉素+细胞色素C组肾脏中庆大霉素的含量降低了33.86%((335.16±99.18)μg·g-1vs.(221.67±71.12)μg·g-1),肾小管上皮细胞凋亡的阳性细胞数密度明显下降((637.78±169.64)n·mm-2vs.(404.75±135.57)n·mm-2,P<0.05),肾小管上皮细胞水肿、空泡变性、肾间质内炎细胞浸润、肾小球充血肿胀等均有所好转。结论:细胞色素C可能通过抑制庆大霉素在肾脏中蓄积来减轻其肾毒性。     

环维黄杨星D对大鼠的肾脏毒性

目的观察环维黄杨星D(CVB-D)对大鼠的肾脏毒性及其可逆性。方法 120只大鼠随机分为正常对照组,CVB-D2.5,5和10mg·kg-1组,每组30只。CVB-D组大鼠分别ip给予CVB-D2个月,并于给药后第1和第2个月末每组各取10只大鼠的眼眶血分离血清,检测尿素氮(BUN)、肌酐(SCr)、TammHorsfall蛋白(THP)、β2微球蛋白(β2-MG);收集24h尿液检测N-乙酰-β-氨基葡萄糖苷酶(NAG)、微量白蛋白(mAlb)、免疫球蛋白G(IgG)、视黄醇结合蛋白(RBP)、β2微球蛋白(β2-MG)和转铁蛋白(TRF);并做肾脏组织病理学检测。作为恢复期观察,停药4周后每组另10只大鼠做同样检查。结果与正常对照组相比,ip给予大鼠CVB-D1个月后,大鼠血清中的β2-MG明显升高(P<0.01),CVB-D5和10mg·kg-1组的THP含量明显降低(P<0.01),CVB-D10mg·kg-1组大鼠血清中BUN含量升高(P<0.01);同时,CVB-D10mg·kg-1组大鼠尿液中NAG,TRF,β2-MG和IgG含量显著升高,CVB-D5和10mg·kg-1组尿液mAlb含量及RBP含量均显著升高(P<0.05,P<0.01)。持续给药2个月后,CVB-D5和10mg·kg-1组大鼠血清中β2-MG含量显著升高(P<0.05),CVB-D5mg·kg-1组BUN含量明显升高(P<0.05),CVB-D10mg·kg-1组THP含量显著降低(P<0.05);同时,CVB-D10mg·kg-1组大鼠尿液中NAG和IgG含量明显升高(P<0.05,P<0.01),CVB-D5和10mg·kg-1组β2-MG和TRF含量明显升高(P<0.01);病理组织切片显示CVB-D2.5mg·kg-1组部分动物肾小球及肾小管出现坏死的现象,CVB-D5和10mg·kg-1组部分动物出现组织自溶现象。在恢复期,血清及尿液中仍有部分指标显著高于正常对照组,病理组织切片显示CVB-D组仍有部分肾单位出现肾间质内少量炎细胞浸润或不同程度淤血的现象。结论长期应用CVB-D可能引起大鼠肾毒性,且病变在停药后不能彻底恢复。     

心脑肾康对肾缺血再灌注损伤模型大鼠的保护作用研究

目的:探讨心脑肾康对肾缺血再灌注损伤模型大鼠的保护作用。方法:将60只大鼠随机分为假手术组(不夹闭双肾动脉、0·1%二甲基亚砜生理盐水)、对照组(0·1%二甲基亚砜生理盐水)、阳性对照组(肾复康胶囊)以及心脑肾康低、中、高剂量组,分别测定血清肌酐(Scr)、尿素氮(BUN)及肾组织内丙二醛(MDA)、超氧化物歧化酶(SOD)、活性氧(ROS)、一氧化氮(NO)的含量,并观察组织病理形态学改变。结果:与对照组比较,3个剂量的心脑肾康组于术前给药7d、术后再灌注1次,均能使肾缺血再灌注损伤模型大鼠中Scr、BUN、MDA、ROS、NO含量降低,SOD含量升高;可减轻肾组织病理形态学改变(P<0·05),且呈剂量依赖性。其中,高剂量组效果最好,优于阳性对照组(P<0·05)。结论:心脑肾康对肾缺血再灌注损伤模型大鼠具有保护作用,其机制可能与改善肾功能、抗自由基损伤有关。     

曲美他嗪对庆大霉素致大鼠肾毒性的预防作用研究

目的:探讨曲美他嗪对庆大霉素致大鼠肾毒性的预防作用。方法:将大鼠分为溶媒对照组,曲美他嗪低、高剂量组(5、10mg.kg-1.d-1),庆大霉素组(100 mg.kg-1.d-1)和低、高剂量联用组(曲美他嗪5或10 mg.kg-1.d-1+庆大霉素100 mg.kg-1.d-1),每组6只,曲美他嗪灌胃给予2 d,第3天开始曲美他嗪灌胃30 min后腹腔注射庆大霉素,无对应药物的注射生理盐水,连续给药7 d,末次给药后24 h处死大鼠。采用荧光偏振免疫分析法测定各组大鼠肾组织中庆大霉素的浓度;缺口末端标记(TUNEL)法观察各组肾脏细胞凋亡情况;苏木精-伊红(HE)染色观察肾脏组织病理学改变。结果:与庆大霉素组比较,高剂量联用组大鼠肾组织中庆大霉素浓度明显降低(P<0.05),低、高剂量联用组大鼠肾脏细胞凋亡数目均明显降低(P<0.05或P<0.01);庆大霉素组大鼠肾脏可见炎性细胞浸润等病变形态;低、高剂量联用组肾脏组织病理变化均有好转。结论:曲美他嗪可能通过降低庆大霉素在肾脏中的蓄积来减轻庆大霉素引起的肾毒性。     

葛根素增强胰岛素抵抗-高血压大鼠胰岛素敏感性作用机制研究

目的观察葛根素对胰岛素抵抗—高血压大鼠胰岛素敏感性的作用及其机制。方法用喂饲高脂高糖高盐的方法建立胰岛素抵抗—高血压(IRH)病理模型,随机分为模型卡托普利组(Cap)、葛根素注射液(Pue)高、中、低三个剂量组、模型阴性对照组及正常空白对照组,分别给药后,测定大鼠血压、血清的甘油三酯(TG)、胆固醇(TC)、肿瘤坏死因子-α(TNF-α)、空腹血浆血糖(FPG)及胰岛素(F ins)含量的变化,并计算胰岛素敏感指数(ISI)。结果模型组的血压、血糖、Ins均有显著升高,ISI降低(P<0.05～0.01);Cap和Pue 80mg·kg^-1组能降低TNF-α含量,纠正IRH大鼠高胰岛素血症(P<0.05～0.01),提高低下的ISI,差异有高度显著性(P<0.01);而Pue其余两个剂量组,对F ins、TNF-α含量和ISI作用,差异均未见显著性(P>0.05)。Cap和Pue 80mg和40mg·kg^-1组,均能降低IRH大鼠TG,TC(P<0.05～0.01);Pue 20mg·kg^-1组对上述各指标,则差异均未见显著性(P>0.05)。结论Pue增强胰岛素抵抗-高血压大鼠胰岛素敏感性和降血压,改善胰岛素抵抗可能是通过减少TNF-α分泌,改善脂质代谢,葡萄糖分解等途径实现的。     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.