二仙汤对去卵巢大鼠心肌微血管及血液流变学的影响

目的:观察去卵巢大鼠心肌微血管密度、微血管内皮细胞形态及血液流变学的变化并探讨二仙汤的干预作用。方法:选用健康10周龄雌性SPF级SD大鼠32只,随机分为假手术组、模型组、雌激素组(戊酸雌二醇,0.18 mg·kg~(-1)),二仙汤组(9 g·kg~(-1))。去卵巢后2周开始灌胃给药,1次/d,连续16周。二仙汤组及雌激素组分别给予二仙汤或戊酸雌二醇灌胃,假手术组和模型组灌胃等体积纯净水。给药16周末,无创超声心动图(UCG)检测心功能;CD34免疫荧光染色法检测心肌微血管密度;透射电镜观测心肌微血管超微结构;放射免疫分析法检测血浆雌二醇(E_2)含量;酶联免疫吸附测定(ELISA)检测血浆内皮素-1(ET-1),前列环素I_2(PGI_2),血栓素A_2(TXA_2),血管内皮生长因子(VEGF)和血管性血友病因子(vWF)水平;凝固法检测凝血四项;血液流变学检测全血黏度和血浆黏度。结果:与假手术组比较,模型组左室射血分数(EF)显著降低(P0.01),左室短轴缩短率(FS)显著降低(P0.01),左室收缩末期容积(LVVols)显著升高(P0.01);心肌微血管密度显著减少(P0.01);心肌微血管内皮细胞肿胀明显,胞浆空化;血浆E_2含量显著降低(P0.01);ET-1,VEGF,vWF含量显著升高(P0.01),前列环素I_2/血栓素A_2(PGI_2/TXA_2)显著降低(P0.01);血浆活化部分凝血酶原时间(APTT)显著降低(P0.01),纤维蛋白原(FIB)含量显著升高(P0.01);全血黏度、血浆黏度、卡松黏度显著升高(P0.01),全血高、低切指数和红细胞聚集指数明显升高(P0.05)。与模型组比较,二仙汤组大鼠EF,FS明显升高(P0.05),LVVols明显降低(P0.05);微血管密度显著增多(P0.01);心肌微血管内皮细胞水肿改善,运输小泡清晰可见;血浆E_2显著升高(P0.01);ET-1,VEGF显著降低(P0.01),PGI_2/TXA_2显著升高(P0.01);APTT显著升高(P0.01);全血黏度、全血高切相对指数、红细胞聚集指数明显降低(P0.05),卡松黏度、血浆黏度显著降低(P0.01)。结论:二仙汤增加去卵巢大鼠心肌微血管密度、保护微血管内皮细胞结构的完整性,改善其内皮分泌功能和血液流变学,保护心功能。     

基于PKC/P2X3通路探讨正天丸对偏头痛大鼠的保护作用

目的探讨正天丸对偏头痛大鼠的保护作用,并初步探究其可能的作用机制。方法将大鼠分为对照组(0.9%NaCl溶液)、模型组(0.9%NaCl溶液)、正天丸组(1.62 g/kg)、蛋白激酶C(PKC)激动剂组(2 mg/kg TPPB)、正天丸+TPPB组(1.62 g/kg正天丸+2 mg/kg TPPB),每组12只,采用硝酸甘油注射法制备偏头痛大鼠模型,造模后观察大鼠耳红出现、消失的时间以及不同时间段内爬笼、抓头次数;HE染色观察脑组织神经细胞形态学变化;ELISA法检测血清中TNF-ɑ、IL-6、嗜中性粒细胞趋化因子(CINC-1)水平;放射免疫法检测脑组织匀浆中降钙素基因相关肽(CGRP)、5-羟色胺(5-HT)水平;Western blot法检测脑组织PKC、P2X3蛋白表达。结果①与对照组比较,模型组0～45 min、45～75 min、75～120 min内挠头及爬笼次数增加,血清TNF-α、IL-6、CINC-1水平升高,脑组织CGRP水平升高,5-HT水平降低,PKC、P2X3蛋白表达升高(P0.05)。②与模型组比较,正天丸组挠头及爬笼次数均显著减少,耳红出现时间显著延长,耳红消失时间显著缩短,TNF-α、IL-6、CINC-1水平降低,CGRP水平降低,5-HT水平升高,PKC、P2X3蛋白表达降低(P0.05);TPPB组挠头及爬笼次数增加,耳红出现时间显著缩短,耳红消失时间显著延长,血清TNF-α、IL-6、CINC-1水平升高,脑组织CGRP水平升高,5-HT水平降低,PKC、P2X3蛋白表达升高(P0.05);正天丸+TPPB组挠头及爬笼次数、耳红消失时间、血清TNF-α、IL-6、CINC-1水平、脑组织CGRP水平、PKC、P2X3蛋白低于TPPB组,5-HT水平、耳红出现时间高于TPPB组(P0.05)。结论正天丸可缓解偏头痛大鼠神经细胞损伤,缓解头痛,其可能是通过抑制PKC/P2X3通路实现的。     

Prenatal exposure to di-n-butyl phthalate (DBP) differentially alters androgen cascade in undeformed versus hypospadiac male rat offspring

This study was to compare the alterations of androgen cascades in di-n-butyl phthalate (DBP)-exposed male offspring without hypospadias (undeformed) versus those with hypospadias. To induce hypospadias in male offspring, pregnant rats received DBP via oral gavage at a dose of 750 mg/kg BW/day during gestational days 14–18. The mRNA expression levels of genes downstream of the androgen signaling pathway, such as androgen receptor (AR) and Srd5a2, in testes of undeformed rat pups were similar to those in controls; in hypospadiac rat pups these levels were significantly lower than those of control pups. In contrast, both undeformed and hypospadiac rats had decreased serum testosterone levels, reduced mRNA expression of key enzymes in the androgen synthetic pathway in the testes, and ablated genes of developmental pathways, such as Shh, Bmp4, Fgf8, Fgf10 and Fgfr2, in the genital tubercle (GT) as compared to those in DBP-unexposed controls, albeit hypospadiac rats had a more severe decrement than those of undeformed rats. Although other possibilities cannot be excluded, our findings suggest that the relatively normal levels of testosterone-AR-Srd5a2 may contribute to the resistance to DBP toxicity in undeformed rats. In conclusion, our results showed a potential correlation between decreased testosterone levels, reduced mRNA expression of AR and Srd5a2 and the occurrence of hypospadias in male rat offspring prenatally exposed to DBP.     

Melatonin reduces hepatic mitochondrial dysfunction in diabetic obese rats

Hepatic mitochondrial dysfunction is thought to play a role in the development of liver steatosis and insulin resistance, which are both common characteristics of obesity and type 2 diabetes mellitus (T2DM). It was hypothesized that the antioxidant properties of melatonin could potentially improve the impaired functions of hepatic mitochondria in diabetic obese animals. Male Zucker diabetic fatty (ZDF) rats and lean littermates (ZL) were given either melatonin (10 mg/kg BW/day) orally for 6 wk (M‐ZDF and M‐ZL) or vehicle as control groups (C‐ZDF and C‐ZL). Hepatic function was evaluated by measurement of serum alanine transaminase and aspartate transaminase levels, liver histopathology and electron microscopy, and hepatic mitochondrial functions. Several impaired functions of hepatic mitochondria were observed in C‐ZDF in comparison with C‐ZL rats. Melatonin treatment to ZDF rats decreases serum levels of ALT (P < 0.001), alleviates liver steatosis and vacuolation, and also mitigates diabetic‐induced mitochondrial abnormalities, glycogen, and lipid accumulation. Melatonin improves mitochondrial dysfunction in M‐ZDF rats by increasing activities of mitochondrial citrate synthase (P < 0.001) and complex IV of electron transfer chain (P < 0.05) and enhances state 3 respiration (P < 0.001), respiratory control index (RCR) (P < 0.01), and phosphorylation coefficient (ADP/O ratio) (P < 0.05). Also melatonin augments ATP production (P < 0.05) and diminishes uncoupling protein 2 levels (P < 0.001). These results demonstrate that chronic oral melatonin reduces liver steatosis and mitochondria dysfunction in ZDF rats. Therefore, it may be beneficial in the treatment of diabesity.     

Metabolic profiles of corydaline in rats by ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry

1. Corydaline, an isoquinoline alkaloid obtained from the rhizomes of Corydalis yanhusuo, exhibits anti-acetylcholinesterase, anti-angiogenic, anti-allergic and gastric-emptying activities. In this study, a rapid and reliable ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS) method was developed and employed for the comprehensive study of the metabolites of corydaline in rats.

2. Altogether, 43 metabolites were identified in the plasma (11), bile (9), urine (34) and feces (21) of rats after oral administration of corydaline at a dose of 4.5mg/kg.

3. It was demonstrated that demethylation, hydroxylation, sulfation and glucuronidation were the major metabolic transformation pathways. Among these, two metabolites were identified as tetrahydropalmatine and isocorybulbine, and 33 phase I and phase II products were inferred to be new metabolites arising from the in vivo metabolism of corydaline.

4. Importantly, this research provides scientific and reliable support for full understanding of the metabolic profiles of corydaline and the results could help to elucidate its safety and efficacy.     

角膜电刺激对糖尿病大鼠前部缺血性视神经病变模型的影响

观察并评估角膜电刺激对糖尿病大鼠前部缺血性视神经病变（AION）模型的影响。方法：实验 研究。健康雄性Sparague-Dawley大鼠40只，随机分组后抽出8只作为正常大鼠组。余下32只先予 以链脲佐菌素腹腔注射建立糖尿病大鼠模型，将造模成功的大鼠随机抽出8只作为糖尿病组，余下 24只糖尿病大鼠采用孟加拉玫瑰红联合532 nm激光方法建立AION大鼠模型。将24只造模成功的 AION大鼠随机分成3组，每组8只，分别为AION模型组，不予任何处理；电刺激组，予以角膜电刺 激（刺激参数为：电流1 mA，频率20 Hz，波宽1 ms/phase，刺激时间1 h，隔日1次，刺激2周）；假电 刺激组，电极安放位置与电刺激组相同，仅不接通电源。2周后5组大鼠进行眼底照相、光学相干断 层扫描和视觉诱发电位，然后处死，行视网膜及视神经冰冻切片，苏木精伊红染色观察。数据采用 单因素方差分析和LSD-t检验进行分析。结果：正常大鼠组视盘上半部视网膜厚度为（211±13）μm， 糖尿病大鼠组为（206±16）μm，AION模型组为（240±54）μm，假电刺激组为（216±11）μm，电刺 激组为（198±4）μm，5组视盘上半部视网膜厚度差异有统计学意义（F=2.854，P=0.038）。其中AION 模型组视盘上半部视网膜厚度高于正常组、糖尿病组、电刺激组，差异均有统计学意义（P<0.05）； 正常组与糖尿病组差异无统计学意义，AION模型组与假电刺激组未见明显差异。视觉诱发电位示 AION模型组N1潜伏期较电刺激组延长，差异有统计学意义（t=4.1，P<0.001）；AION模型组P1潜伏 期较正常组、糖尿病组、假电刺激组、电刺激组延长，差异均有统计学意义（t=4.1、2.5、2.6、3.2， P<0.05）；电刺激组N1-P1波幅大于假电刺激组，差异有统计学意义（t=4.0，P<0.001）。结论：角膜电 刺激能促进糖尿病大鼠前部缺血性视神经病变模型肿胀的视盘变薄，加速视盘水肿的消退，同时在 一定程度上改善视功能。     

萆苓祛痛方对糖尿病痛风大鼠骨骼肌组织SIRT3蛋白表达及URAT1 mRNA的影响

目的:探讨萆苓祛痛方对糖尿病痛风大鼠骨骼肌组织去乙酰化酶3(SIRT3)蛋白表达及尿酸盐转运体1(URAT1) mRNA的影响。方法:选择健康雄性大鼠40只,除正常组外,其余组予高脂饲料喂养并联合小剂量链脲佐菌素(STZ)溶液40 mg·kg-1腹腔注射1次,以血糖≥16. 7 mmol·L-1,为糖尿病模型。4 d后关节腔注射5%尿酸钠溶液1次,诱导痛风模型,模型成功后,分为萆苓祛痛方组(萆苓组,10 g·kg-1),吲哚美辛组(5 mg·kg-1),吡格列酮组(10 mg·kg-1),均连续给药21 d,正常组、模型组予等量生理盐水;采用蛋白免疫印迹法(Western blot)测定骨骼肌组织SIRT3蛋白表达;实时荧光定量聚合酶链式反应(Real-time PCR)检测骨骼肌组织URAT1 mRNA表达,并进行病理检查,取血测定血糖(GLU),血尿酸(UA)及C反应蛋白(CRP)含量。结果:与正常组比较,模型组GLU,UA及CRP明显升高(P 0. 01);与模型组比较,萆苓组、吡格列酮组血糖下降(P 0. 05);各药物组UA及CRP明显下降(P 0. 01)。与正常组比较,模型组骨骼肌SIRT3蛋白表达量显著降低(P 0. 01);与模型组比较,萆苓组骨骼肌SIRT3蛋白表达量显著提高(P 0. 01),与西药组比较无明显差异;条带图的结果同样显示,与正常组比较,模型组表达亮度明显减弱,药物组表达亮度明显增强;与正常组比较,模型组关节组织URAT1 mRNA相对表达量明显升高(P 0. 01);与模型组比较,各药物组URAT1 mRNA相对表达量显著下调(P 0. 01)。电泳图同样提示,正常组表达亮度减弱,模型组表达亮度显著增强,萆苓组、西药组表达亮度明显减弱。关节病理提示,与正常组比较,模型组大鼠关节病理损伤严重,可见大量炎细胞浸润及纤维增生,滑膜细胞变性、坏死。与模型组比较,萆苓祛痛方关节病变程度明显减低,见少量炎细胞浸润,滑膜上皮轻度增生。结论:具有泻浊解毒通络作用的萆苓祛痛方可显著提高糖尿病痛风大鼠骨骼肌组织SIRT3的蛋白表达量,下调URAT1 mRNA的表达量,减轻骨骼肌组织病理损伤,减低血清炎症因子CRP的含量,降低模型大鼠的血糖、血尿酸水平,有保护关节功能的作用。     

Pyridostigmine bromide and its relation to Gulf War illness

Abstract

Pyridostigmine bromide acts as a reversible cholinesterase inhibitor that is used at relatively high doses in treatment of Myasthenia gravis and in low dose regimens as prophylaxis against nerve agents poisoning during the Gulf War. The manifestation of late nonspecific symptoms commonly called Gulf War illness has led to the discussion about the role of pyridostigmine bromide in the pathogenesis of this illness. In our study, we described plasma absorption profile of pyridostigmine bromide after p.o. administration in rats; subsequently, changes in blood biochemical and oxidative stress markers were measured. Pyridostigmine bromide was applied p.o. at the dose of 5.82?mg/kg b.w. according to the previously published recommendations. The absorption of pyridostigmine was relatively fast; the C_max in plasma was 110.20?±?15.12?ng/ml at T_max of 197.12?±?17.14?min. The bioavailability expressed as AUC_total was 44,348?±?7608?min ng/ml. The prolongation of pyridostigmine in circulation is in agreement with relatively long half-life that was 179.00?±?28.54?min. Several blood biochemical markers were altered, including glucose, creatinine, creatine kinase, alanine aminotransferase, aspartate aminotransferase, interleukin-6, triglycerides, and cholesterol. However, the changes could be considered as mild. Thiobarbituric acid reactive substances and ferric reducing ability of plasma indicate suppression of basal metabolism. The results of blood biochemical and oxidative stress markers imply that long-term use might possibly change the basal metabolism and cause cellular damage with inflammatory changes.