Intravenous administration of glutathione protects parenchymal and non-parenchymal liver cells against reperfusion injury following rat liver transplantation

AIM:To investigate the effects of intravenous administration of the antioxidant glutathione (GSH) on reperfusion injury following liver transplantation. METHODS:Livers of male Lewis rats were transplanted after 24 h of hypothermic preservation in University of Wisconsin solution in a syngeneic setting.During a 2-h reperfusion period either saline (controls,n=8) or GSH (50 or 100 μmol/(h·kg),n=5 each) was continuously administered via the jugular vein. RESULTS:Two hours after starting reperfusion plasma ALT increased to 1 457±281 U/L (mean±SE) in controls but to only 908±187 U/L (P<0.05) in animals treated with 100 μmol GSH/(h·kg).No protection was conveyed by 50μmol GSH/(h·kg).Cytoprotection was confirmed by morphological findings on electron microscopy:GSH treatment prevented detachment of sinusoidal endothelial cells (SECs) as well as loss of microvilli and mitochondrial swelling of hepatocytes.Accordingly,postischemic bile flow increased 2-fold.Intravital fluorescence microscopy revealed a nearly complete restoration of sinusoidal blood flow and a significant reduction of leukocyte adherence to sinusoids and postsinusoidal venules.Following infusion of 50μmol and 100 μmol GSH/(h·kg),plasma GSH increased to 65±7 mol/L and 97±18 mol/L,but to only 20±3 mol/L in untreated recipients. Furthermore,plasma glutathione disulfide (GSSG) increased to 7.5±1.0 mol/L in animals treated with 100μmol/(h·kg) GSH but infusion of 50μmol GSH/(h·kg) did not raise levels of untreated controls (1.8±0.5 mol/L vs 2.2±0.2 mol/L). CONCLUSION:Plasma GSH levels above a critical level may act as a “sink” for ROS produced in the hepatic vasculature during reperfusion of liver grafts.Therefore,GSH can be considered a candidate antioxidant for the Drevention of reperfusion injury after liver transplantation,in particular since it has a low toxicity in humans.

High IFN-α expression is associated with the induction of experimental autoimmune uveitis (EAU) in Fischer 344 rat

Th1-response plays a crucial role in determining pathogenesis of organ-specific autoimmune diseases. It is believed that both IL-12 and INF-alpha are initiators to regulate Th1-response. In our experimental autoimmune uveitis (EAU) model, both Lewis and Fischer 344 rats share the same MHC class II molecules, while Lewis rat is EAU susceptible and Fischer 344 rat is EAU resistant. However, under the same condition of immunization, if pertussis toxin (PTX) was injected intraperitoneally as an additional adjuvant, Fischer 344 rat can develop EAU. In this study we investigate which mechanisms are involved in the induction of EAU in CFA+R16+PTX-treated (CRP-treated) Fischer 344 rats. In vivo and in vitro data demonstrated that Th1-cytokine, IFN-gamma mRNA expression was significantly increased in disease target tissue-eyes and in draining lymph node cells of CRP-treated Fischer 344 rat. When IL-12 and IFN-alpha mRNA expression were compared in the experimental groups, only IFN-alpha mRNA expression was associated with EAU development. To distinguish the sources of IFN-alpha producing cells, it was observed that IFN-alpha expression was mainly produced by macrophages. It was further confirmed that normal macrophage from Fischer 344 rat was able to produce significant IFN-alpha in the presence of PTX. The data strongly suggested that IFN-alpha might be involved in initiating Th1-cell differentiation and in turn contribute to the induction of EAU. High IFN-alpha expression induced by PTX may represent a novel pathway to initiate Th1 response in Fischer 344 rat.

Regular nicotine intake increased tooth movement velocity,osteoclastogenesis and orthodontically induced dental root resorptions in a rat model

Orthodontic forces have been reported to significantly increase nicotine-induced periodontal bone loss. At present, however, it is unknown, which further （side） effects can be expected during orthodontic treatment at a nicotine exposure corresponding to that of an average European smoker. 63 male Fischer344 rats were randomized in three consecutive experiments of 21 animals each （A/B/C） to 3 experimental groups （7 rats, 112/3）. （A） cone-beam-computed tomography （CBCT）; （B） histology/serology; （C） reverse- transcription quantitative real-time polymerase chain reaction （RT-qPCR）/cotinine serology--（1） control; （2） orthodontic tooth movement （OTM） of the first and second upper left molar （NiTi closed coil spring, 0.25 N）; （3） OTM with 1.89 mg-kg- 1 per day s.c. of L（- ）-nicotine. After 14 days of OTM, serum cotinine and IL-6 concentration as well as orthodontically induced inflammatory root resorption （OIIRR）, osteoclast activity （histology）, orthodontic tooth movement velocity （CBCT, within 14 and 28 days of OTM） and relative gene expression of known inflammatory and osteoclast markers were quantified in the dental-periodontal tissue （RT-qPCR）. Animals exposed to nicotine showed significantly heightened serum cotinine and IL-6 levels corresponding to those of regular European smokers. Both the extent of root resorption, osteoclast activity, orthodontic tooth movement and gene expression of inflammatory and osteoclast markers were significantly increased compared to controls with and without OTM under the influence of nicotine. We conclude that apart from increased periodontal bone loss, a progression of dental root resorption and accelerated orthodontic tooth movement are to be anticipated during orthodontic therapy, if nicotine consumption is present. Thus patients should be informed about these risks and the necessity of nicotine abstinence during treatment.

Endothelin-1, an important mitogen of smooth muscle cells of spontaneously hypertensive rats

OBJECTIVE: To study the features of vascular smooth muscle cell (VSMC) proliferation induced by endothelin-1 (ET-1). METHODS: VSMCs of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats were cultured and treated with ET-1. Basic fibroblast growth factor (bFGF) gene expression was measured using both Northern blot and an enzyme-linked immunoassay. RESULTS: ET-1 resulted in an increase in bFGF transcripts at 8 - 24 h; bFGF levels were significantly higher in VSMCs treated with ET-1 than in those not treated. However, VSMCs growth responses in SHR and WKY were different. Smooth muscle cells of SHR were hyper-responsive to ET-1. Maximal bFGF mRNA levels were elevated 3.5-fold at 4 h of stimulation in WKY and 8-fold at 8h in SHR4. Moreover, the proliferation of VSMCs induced by ET-1 was inhibited by antisense phosphorothioate oligodeoxynucleotides (10 micromol/L AS-bFGF) but not sense bFGF oligomers at the same concentrations, being reduced by 80% in SHR and 40% in WKY vs control, respectively. Furthermore, the effect of AS-bFGF oligomers on SHR SMC proliferation is significantly greater than on WKY SMC proliferation. CONCLUSION: ET-1 may be required for exaggerated vascular growth responses in SHR and bFGF may be involved.

Endothelin receptor antagonist combined with a calcium channel blocker attenuates renal injury in spontaneous hypertensive rats with diabetes

OBJECTIVE: To investigate the effects of the mixed endothelin receptor antagonist, bosentan, combined with the long-acting calcium channel blocker, amlodipine, compared to the angiotensin-converting enzyme inhibitor, cilazapril, on the progressive renal injury in spontaneous hypertensive rats (SHR) with diabetes. METHODS: Diabetic hypertensive rats (SHR-DM) were induced by streptozotozin injected in male SHR (7-week-old),and divided into an untreated and three treated groups: 1) cilazapril treated group; 2) bosentan+amlodipine treated group; and 3) amlodipine treated group. Wistar Kyoto rats (WKY) and SHR rats served as normotensive and hypertensive control, respectively. The mean arterial blood pressure, renal function, endothelin and angiotensin II levels as well as the protein expression of renal extracellular matrix components and transforming growth factor (TGF)-beta1 were determined at the end of the 4th week. RESULTS: Mean arterial blood pressure significantly increased in SHR and SHR-DM rats compared to WKY rats. All the therapies reduced the blood pressure to normal levels. However, the enhanced urinary protein excretion, the decreased creatinine clearance as well as the increased plasma and intrarenal endothelin and angiotens in II levels were found in the untreated SHR-DM and prevented by treatment with bosentan+amlodipine and cilazapril. Similarly, these two kinds of therapies in SHR-DM abolished the overexpression of renal TGF-beta1 by Western blot analysis and reduced the accumulation of collagen type IV, laminin and fibronectin proteins by an immunochemical approach. Amlodipine monotherapy had no detectable effects on the above parameters. CONCLUSION: Bosentan combined with amlodipine can offer similar renoprotective effects on that of cilazapril and may be a potent therapy to attenuate renal injury by reducing renal protein levels of TGF-beta1 in diabetes with a hypertensive state.

Influence of Valsartan on myocardial apoptosis in spontaneously hypertensive rats

OBJECTIVE: To explore the pathogenic changes of myocardial apoptosis in heart hypertrophy during hypertension and evaluate the anti-apoptosis effect of Valsartan. METHODS: Thirty spontaneously hypertensive rats (SHRs) were divided into two groups: 15 treated with Valsartan (20 mg x kg(-1) x d(-1)) (SHR + Valsartan group), the others with placebo (SHR + placebo group), with 15 normal Wistar rats as control. Systolic blood pressure was measured by the tail-cuff method. The observation period was from 8 to 16 weeks of age. Cardiac apoptosis was evaluated by a Terminal Deoxynucleotidyl Transferase-Mediated dUTP-biotin Nick End Labeling (TUNEL) assay. RESULTS: Mean blood pressure values were 127 +/- 2 mm Hg in controls, 163 +/- 6 mm Hg in the SHR + Valsartan group and 193 +/- 7 mm Hg in the SHR + placebo group at 16 weeks of age, whereas the blood pressure in 8-week-old SHR and Wistar rats were 175 +/- 3 mm Hg and 125 +/- 5 mm Hg, respectively. The ratio of the heart weight over body weight declined in Wistar (3.07 +/- 0.03 mg/g) and SHR + Valsartan groups (3.22 +/- 0.19 mg/g) compared with the SHR + placebo group (4.02 +/- 0.31 mg/g) (P

Ventricular remodeling by Scutellarein treatment in spontaneously hypertensive rats

OBJECTIVE: To observe reversal of ventricular remodeling by the protein kinase C inhibitor Scutellarein in spontaneously hypertensive rats (SHRs). METHODS: Twelve SHRs were randomly divided into two groups. Scutellarein and saline (10 mg x kg(-1) x d(-1)) were given by intraperitoneal injection to two groups of rats separately. Systolic blood pressure (SBP) and ventricular weight index (LVW/BW, RVW/BW) were measured. A polarization microscope and an image analyzer system (IAS) were used to observe changes in cardiovascular structure and to count the content of cardiac muscle interstitial collagen. RESULTS: The pathologic changes in the left ventricle in the Scutellarein group rats (SHR(D)) improved to varying degrees, including hypertrophy of the cardiac muscle and collagen volume fraction. CONCLUSION: Scutellarein can reverse ventricular remodeling, improve myocardial stiffness and protect heart cardiac muscle.

Tong-xin-luo capsule inhibits left ventricular remodeling in spontaneously hypertensive rats by enhancing PPAR-γ expression and suppressing NF-κB activity

Background Tong-xin-luo capsule （TXL）, used as a traditional Chinese herb, offeres a therapeutic potential for treatment of cardiovascular diseases. It has been shown to exert a variety of pharmacological effects, including antihypertensive effects, and is able to improve ventricular remodeling. However, the mechanisms of its action are not completely understood. The aim of this study was to evaluate the molecular mechanisms of Tong-xin-luo capsule on left ventricular remodeling in spontaneously hypertensive rats （SHR）. Methods Sixteen eight-week-old SHRs were randomized into an SHR group （n=8） and a TXL group （n=8） that were given Tong-xin-luo capsule （1.5 mg·kg^-1·d^-1）. Eight Wistar Kyoto （WKY） rats fed with 0.9% NaCl served as the control group （WKY group）. Systolic blood pressure （BP）, body weight and heart rate were monitored once every two weeks. Ventricular remodeling was detected by histopathological examination. Nuclear factor kappa B P65 （NF-κB P65） and peroxisome proliferators activated receptor y （PPAR-γ） protein and phosphorylated inhibitor kappa a （IκBα） protein were detected by immunohistochemistry and western blot respectively. The physical interaction of the P65-P50 heterodimer with IκBα and NF-κB were measured by co-immunoprecipitation. PPAR-γ mRNA, collagen Ⅰ mRNA and collagen Ⅲ mHNA were measured by real-time PCR.Results TXL inhibited NF-κB P65 expression and ventricular remodeling and suppressed the activation of NF-κB compared with the SHR group （P〈0.01, P〈0.05）. TXL reduced IκBα phosphorylation, increased expression of PPAR-γ protein and enhanced the physical interaction of the P65-P50 heterodimer with IκBα. The mRNA expression of PPAR-γ was enhanced but the mRNA expression of collagen Ⅰ mRNA and collagen Ⅲ mRNA were suppressed by TXL. Conclusions In spontaneously hypertensive rats, TXL could inhibit ventricular remodeling induced by hypertension, and the inhibitory effect might be associated with the process of TXL increasing the expression of PPAR-γ that could result in the inhibition of the activation of NF-κB.

Five novel monoclonal antibodies to thymic epithelial cell surface antigens in rats

OBJECTIVE: To establish monoclonal antibodies (mAbs) against thymic epithelial cells and study the function of epithelial cells during T-cell differentiation in the thymus. METHODS: Hybridomas secreting mAbs against thymic epithelial cells were derived by immunization of Balb/c mice with two thymic epithelial cell lines, TaD3 and FTE. The distribution of antigens recognized by these mAbs was detected by immunochemical staining and cytofluorographic analysis, and the molecular weight of the antigens by immunoblotting. RESULTS: Five specific monoclonal antibodies (mAb) were obtained. On the basis of their distribution in the thymus determined by immunochemical staining, mAb RE-4D8 was regarded as clusters of thymic epithelium staining (CTES) type IIA: mAb RE-12B2, which showed a unique distribution pattern only in the medulla, was CTES type V: mAb RE-5C6 was CTES type IV: mAb RE-6D6 might be CTES type IIB: and mAb RE-1D4 was classified as type V. The molecular weight (MW) of antigen RE-4D8, RE-6D6 and RE-12B2 were 120 kDa, 220 kDa and 35 kDa, respectively. Antigen RE-1D4 is a novel marker of cortical epithelium, several established thymic epithelial cell lines were classified and their original intrathymic locations were determined by these mAbs. Thymic cell lines, TuD3 and FTE were cortical phenotypes whereas TaD3 had a medullar phenotype. CONCLUSIONS: These mAbs clearly demonstrate the heterogeneity of the thymic epithelium; they could detect antigens not only in the cytoplasm but also on the surface of thymic epithelial cells. Our data suggest that these newly established mAbs may help elucidate the interaction between thymocytes and epithelial cells during T cell maturation.

SHR、SHRsp、WKy三品系大鼠血液生化检查参数

目的 检查了解SHR、SHRsp、WKy三品系大鼠血液生化的参数。方法 每个品系分别取年轻大鼠(12周龄 )、年老大鼠 (2 2～ 2 8周龄 )各 14只 ,心脏取血 ,用自动生化分析仪检验血生化及电解质等指标。结果 年老SHR、SHRsp、WKy大鼠血肌酐平均水平较年轻大鼠分别高 8 5 %、9 2 %和 14 2 % (P =0 0 7;P =0 0 7;P =0 0 4 ) ,但血钙、血磷较低。同龄SHR、SHRsp血磷平均水平均较同龄WKy低 2 5 0 % (P =0 12～P <0 0 0 1)。同龄SHR、SHRsp的肌酸磷酸激酶同工酶 (CK MB)及乳酸脱氢酶同工酶 (LDH 1)较WKy升高 1～ 2倍 (P =0 15～P <0 0 0 1)。结论 SHR、SHRsp及Wky三种品系大鼠的血液生化参数多为相似 ,但也有一些差别。

Effects of electroacupuncture at Taichong(LR 3) and Baihui(DU 20)on cardiac hypertrophy in rats with spontaneous hypertension

OBJECTIVE: To investigate the effects of electroacupuncture(EA) at Taichong(LR 3) and Baihui(DU 20)on myocardial hypertrophy in spontaneously hypertensive rats(SHRs).METHODS: Thirty-six SHRs were randomly assigned to model, EA, and Losartan groups, with twelve rats per group. Twelve Wistar Kyoto rats were selected as the normal control group. Systolic blood pressure(SBP) and cardiac function were measured in all rats.Expression levels of factors associated with the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR) pathway were evaluated by Western blotting and real-time PCR.Pathological changes of the heart tissue were observed by hematoxylin-eosin staining.RESULTS: After treatment, enhanced SBP was significantly decreased in the EA and Losartan groups compared with the model group(P < 0.01). Echocardiographic and morphological analyses revealed that enhanced end-diastolic interventricular septal thickness and left ventricular posterior wall thickness, as well as ratio of left ventricular weight to body weight were markedly diminished in the EA and Losartan groups(P < 0.01 or P < 0.05), while reduced left ventricular end-diastolic dimension and left ventricular ejection fraction were significantly ameliorated(P < 0.01). Real-time PCR and western blotting analyses showed that the expression levels of PI3K,Akt, and mT OR in SHRs were significantly up-regulated by EA and Losartan(P < 0.01), while the expression levels of PTEN and ANP were down-regulated(P < 0.01).CONCLUSION: EA at Taichong(LR 3) and Baihui(DU20) inhibited the development of cardiac hypertrophy and improved the cardiac function in SHRs, possibly through regulation of the PI3K/Akt/mTOR signalling pathway.

电针对WKY大鼠抑郁样行为及海马GluR1的影响

目的通过电针"百会""印堂"观察对抑郁大鼠行为及海马Glu R1的影响。方法 21只雄性WKY大鼠被随机分为电针组、假针组和模型组;雄性Wistar大鼠7只分为正常组。电针组取穴为"百会""印堂",假针组给予安慰治疗,模型组和正常组常规饲养,21 d后测定各组大鼠糖水消耗量(SPT)、强迫游泳实验(FST)的改变,用Western blot检测大鼠海马谷氨酸受体(glutamate receptor1,Glu R1)蛋白表达。结果电针组蔗糖水喜好率明显高于假针组(P<0.05),正常组蔗糖水喜好率高于模型组(P<0.05);强迫游泳试验中电针组和正常组不动时间少于假针组和模型组(P<0.05);海马Glu R1的表达量中,电针组高于假针组(P<0.05),正常组高于模型组(P<0.01)。结论电针对WKY抑郁大鼠的行为学核心症状有明显的改善作用,其作用可能与海马Glu R1的表达增加有关。

基于代谢组学探讨电针对WKY抑郁大鼠的抗抑郁作用

目的采用液质联用非靶向代谢组学技术探究电针抗抑郁的作用机制方法将24只雄性WKY大鼠随机分为模型组、电针组、假针组,每组8只。Wistar雄性大鼠8只设为对照组。电针组于“百会”“足三里”进行电针治疗,每次20 min,1周5次,连续治疗3周。假针组取穴同电针组,毫针不刺破皮肤,毫针与电针仪导线相连,但无电流通过。最后通过旷场实验、强迫游泳和新奇抑制摄食实验评价行为学改变;对照组、模型组和电针组大鼠粪便采用高效液相-色谱质谱系统分析代谢物及组成的变化,探究抑郁症核心生物标记物的代谢途径差异,以评价电针的抗抑郁作用机制。结果与对照组相比,模型组在旷场试验中的直立次数减少(P<0.001),中央运动距离减少(P<0.05);强迫游泳的不动时间延长(P<0.001);新奇抑制摄食潜伏摄食时间增加(P<0.001)。与假针组相比,电针组在旷场试验中的直立次数增加(P<0.001),中央运动距离增加(P<0.05);强迫游泳的不动时间减少(P<0.001);新奇抑制摄食潜在摄食时间减少(P<0.001)。通过模式识别分析发现,WKY大鼠代谢产物发生显著差异,电针后的代谢产物也出现明显差异。对显著性差异代谢产物分析发现,甜菜苷在模型组中升高,在电针组中降低;环己基氨基磺酸酯、10-氢过氧-H4-神经前列腺素、15(S)-羟基二十碳三烯酸、地奥司明在模型组中降低,在电针组中升高。KEGG通路结果发现显著差异代谢产物主要参与了脂质代谢、氨基酸代谢、生物素降解与代谢等代谢途径。结论电针能改变抑郁大鼠的抑郁样行为,可能是通过调节脂质代谢、氨基酸代谢、生物素降解与代谢等代谢途径发挥抗抑郁的治疗作用。

SHR和WKY大鼠主动脉hsp 70mRNA水平的比较研究

本工作应用热激蛋白70KD(hsp 70)核酸分子杂交方法,检测了:1.自发性高血压(SHR)主动脉和离体培养的主动脉平滑肌细胞受热激后hsp 70mRNA水平的变化;2.不同细胞培养时间(3个月与6周)的SHR、WKY主动脉hsp 70mRNA水平。结果提示SHR主动脉hsp mRNA水平增加,SHR细胞培养受热激(42℃,15min)后2h,hsp 70mRNA水平明显高于WKY鼠者,6周较3个月的SHR细胞hsp 70mRNA水平高;6周的SHR细胞较同期和3个月的WKY细胞hsp 70mRNA高。推论SHR血管平滑肌细胞对热敏感,原癌基因c-myc和抗癌基因p^(53)可能参与hsp 70表达调控,并共同参与SHR细胞增殖的调节。

Proliferation of aortic smooth muscle cells and renin-angiotensin system in SHR rats

目的：探讨ＳＨＲ大鼠主动脉平滑肌细胞（ＡＳＭＣ）异常增殖和肾素血管紧张素系统（ＲＡＳ）的关系．方法：测定血管紧张素Ｉ（Ａｎｇ）、卡托普利（Ｃａｐ）、沙拉新（Ｓａｒ）对培养的ＳＨＲ、ＷＫＹＡＳＭＣ增殖和Ａｎｇ、血管紧张素转化酶（ＡＣＥ）的影响．结果：Ａｎｇ在２％血清培养基中可刺激ＳＨＲＡＳＭＣ增生．ＳＨＲＡＳＭＣ分裂增殖能力比ＷＫＹ强，ＳＨＲＡＳＭＣＲＡＳ处于高功能状态．Ｃａｐ长期（４周）干预显著抑制ＳＨＲＡＳＭＣ异常增殖和Ａｎｇ、ＡＣＥ活性，Ｓａｒ长期干预同样抑制ＳＨＲＡＳＭＣ的增殖和ＡＣＥ活性，但Ａｎｇ水平反而升高．Ｃａｐ短期（２４小时）干预不影响两种大鼠ＡＳＭＣＲＡＳ．结论：Ｃａｐ和Ｓａｒ长期干预通过减少ＳＨＲＡＳＭＣＡｎｇ生成或阻断Ａｎｇ和特异受体结合，抑制其异常增殖．

电针对WKY大鼠抑郁样行为及缰核GluR1的影响

目的:通过电针"百会"、"印堂"观察对抑郁大鼠行为及缰核GluR1的影响。方法:21只雄性WKY大鼠被随机分至电针组、假针组和模型组;雄性Wistar大鼠7只分为正常组。电针组取穴为"百会"、"印堂",假针组给予安慰治疗,模型组和正常组常规饲养,21日后测定各组大鼠体重、糖水消耗量(SPT)、强迫游泳实验(FST)及旷场实验(OFT)的改变,用Western blot检测大鼠缰核谷氨酸受体(glutamate receptor1,GluR1)蛋白表达。结果:1电针组体重明显增加,高于假针组(P<0.05),正常组体重明显高于模型组(P<0.01);2电针组蔗糖水喜好率明显高于假针组(P<0.05),正常组蔗糖水喜好率高于模型组(P<0.05);3强迫游泳试验中电针组和正常组不动时间少于假针组和模型组(P<0.05);4旷场实验中电针组和正常组水平运动和垂直攀爬得分高于假针组和模型组(P<0.05);5缰核GluR1的表达量中,电针组低于假针组(P<0.05),正常组低于模型组(P<0.01)。结论:电针对WKY抑郁大鼠的行为学核心症状有明显的改善作用,其作用可能与缰核GluR1的表达减少有关。

Effect of Taichong(LR 3) acupuncture in spontaneously hypertensive rats

OBJECTIVE: To evaluate the effects of Taichong(LR3) acupuncture points(acupoints) on the expression of glucose transporter protein 1(GLUT1) in the hypothalamus of spontaneously hypertensive rats(SHRs) as measured by combined positron emission tomography and computed tomography(PETCT).METHODS: Spontaneously hypertensive rats(SHR)were divided into model, Taichong(LR 3) acupuncture, and sham groups. Additionally, Tokyo Wistar rats were used as the control group. Changes in blood pressure were recorded in different groups of rats before and after the corresponding treatment. Hematoxylin and eosin(HE) staining was used to study basic morphological changes, and immunohistochemistry was used to determine GLUT1 expression in the hypothalamus. Further,PET-CT was utilized to elucidate the antihypertensive mechanism after acupuncture at the Taichong(LR 3) acupoints.RESULTS: PET-CT indicated activation of the hypothalamus. Measurement of blood pressure showed that acupuncture at the Taichong(LR 3) acupoints lowered blood pressure. HE staining did not show any significant pathological changes, although differences in cell number were observed. Immunohistochemical analysis indicated a GLUT1 downregulation in the SHRs of the Taichong(LR 3) acupuncture group after the treatment.CONCLUSION: Acupuncture at Taichong(LR 3) acupoints lowered blood pressure in SHRs, with possible mechanisms being changes in cell number and GLUT1 expression in the hypothalamus.

Mechanism and effect of Shijueming (Concha Haliotidis) on serum calcium in spontaneously hypertensive rats

OBJECTIVE： To observe the impact of Shijueming （Concha Haliotidis） on spontaneously hypertensive rats via blood pressure, serum calcium, vascular smooth muscle membrane L-type calcium channel α1 C subunit （CaL-α1C）, plasma membrane calci- um-ATPase （PMCA） mRNA expression, and the L-type calcium channel in vascular smooth muscle cells. METHODS： Twelve-week-old male rats with sponta- neous hypertension were divided into three groups： a Shijueming （Concha Haliotidis） group （group 1）, a nifedipine group （group 2）, and a dis- tilled water group （group 3）. All were given a four-week treatment. Blood pressure and dissocia- tive serum calcium were examined before treat- ment. Blood pressure was taken every week during treatment. Atomic absorption spectrometry was used to examine dissociative serum calcium. Re-verse transcription-polymerase chain reaction was used to examine the expression of CaL-α1C and PM- CA1 mRNA. The patch clamp technique was used to examine the electrophysiological characteristics of the vascular smooth muscle cell calcium chan- nels. RESULTS： After treatment, blood pressure of the Shijueming （Concha Halioticlis） group lowered but not significantly （P〉0.05）. Blood pressure of the nifedipine group lowered significantly （P〈0.05）. Blood pressure of the distilled water group re- mained high. The concentration of serum calcium in the Shijueming （Concha Haliotidis） and the dis- tilled water groups lowered （P〈0.05）. Expression of CaL-α1C mRNA in the nifedipine group decreased compared with the distilled water group （P〈0.01）. There was the decreasing trend in the Shijueming （Concha Haliotidis） group, but it was not statistically significant. Shijueming （Concha Haliotidis） also had effects on the expression of PMCA1 mRNA but with- out statistical significance. However, there was a significant decreasing effect on vascular smooth muscle cell Ica-L flow. CONCLUSION： This study indicated that Shijuem- ing （Concha Haliotidis） could increase serum calci- um and decrease blood pressure. It may work by in- fluencing calcium channels, expression of PMCA1 mRNA, and regulating ion calcium channels and calcium-ATPase.

电针对WKY模型大鼠抑郁样行为及缰核β-CaMK Ⅱ蛋白的影响

目的:观察电针对WKY抑郁模型大鼠行为学及缰核β-CaMK Ⅱ蛋白的影响。方法:将24雄性WKY大鼠随机分为电针组(EA)、假针组(sham EA)、模型组(model)。Wistar雄性大鼠8只设为正常组(normal)。21 d后通过旷场实验、糖水偏好实验、强迫游泳实验对大鼠进行行为学评价,用免疫荧光法检测大鼠缰核中β-CaMK Ⅱ蛋白表达。结果:与正常组比较,模型组旷场实验的水平运动、垂直运动和蔗糖水摄入比明显下降(P<0.05),强迫游泳的不动时间和缰核β-CaMK Ⅱ表达均增加(P<0.05)。与假针组比较,电针组旷场实验的水平运动和垂直运动、蔗糖水摄入比均明显增加(P<0.05),强迫游泳的不动时间和缰核β-CaMK Ⅱ蛋白表达减少(P<0.05)。结论:电针对WKY抑郁模型大鼠的行为学核心症状有明显的改善作用,能明显降低缰核中β-CaMK Ⅱ蛋白表达。这可能是电针治疗抑郁症的机制之一。

Upregulation of the angiotensin-converting enzyme 2-angiotensin-(1-7)-Mas receptor axis by a combination of Yinyanghuo(Herba Epimedii Brevicornus) and Cheqianzi(Semen Plantaginis) improves erectile function in spontaneously hypertensive rats

OBJECTIVE:To evaluate the effects of a combination of Yinyanghuo(Herba Epimedii Brevicornus)(HEB)and Cheqianzi(Semen Plantaginis)(SP)on erectile dysfunction caused by essential hypertension in spontaneously hypertensive rats(SHRs),and to elucidate the role of the angiotensin-converting enzyme 2-angiotensin-(1-7)-Mas receptor(ACE2/Ang[1-7]/Mas receptor)axis in this process.METHODS:A total of 24 SHRs were randomly assigned to three groups:SHR-control,low-dose(12.5 g/kg)and high-dose(25 g/kg)HEB+SP(HEBSP).Eight Wistar-Kyoto rats were used as normal controls.HEBSP was administered by oral gavage for 28 d.Erectile function was measured once a week using the Heaton test.After 4 weeks of treatment,the corpus cavernosum was harvested from each rat to measure nitric oxide(NO),nitric oxide synthase(e NOS)and Ang(1-7)levels,as well as ACE2,Mas receptor and neuronal nitric oxide synthase(n NOS)protein expression.RESULTS:After 4 weeks of treatment,HEBSP significantly increased erectile function in the treated group compared with SHR-control group(P<0.01).Additionally,HEBSP treatment significantly increased cavernosal levels of Ang(1-7),e NOS and NO.Moreover,HEBSP significantly elevated the expression levels of ACE2,Mas receptor and n NOS.These beneficial effects were elevated in the high-dose HEBSP group.CONCLUSION:HEBSP improved erectile function in SHRs by upregulating the ACE2/Ang(1-7)/Mas receptor axis,e NOS and n NOS pathways.