Metabolomics study of Shenfu Injection on mid-stage cardiogenic shock rats by UPLC-Q-TOF/MS

OBJECTIVE To study the effects of Shenfu Injection(SFI) on the potential endogenous markers of experimental mid-stage cardiogenic shock rats by UPLC-Q-TOF/MS and look out the different biomarker groups and the molecular mechanism. METHODS Male SD rats were randomly divided into five groups, which were the normal control group, model group, and 15 m L·kg~(-1) SFI group. Except for the normal control group, others were ligated the root of left anterior descending coronary arteries to establish the mid-stage model of cardiogenic shock rats. One hour after administration of SFI or saline by femoral venous catheter, the blood samples were collected from abdominal aorta, and the plasma were pretreated via protein precipitation and analyzed by UPLC-Q-TOF/MS. Data were processed by PCA and PLS-DA. RESULTS All groups could be distinguished by metabolomics successfully. Compared with the model group, 14 pathological biomarkers have changed significantly(P<0.05). And SFI can significantly increase 2 ingredients, i.e. cysteineglutathione disulfide, oxidized glutathione. It can also significantly decrease 11 ingredients, such as dimethylbenzimidazole, imidazolelactic acid, isovalerylglutamic acid, l-gamma-glutamyl-l-isoleucine, n-acetylglucosamine 6-sulfate, adenosine 3′, 5′-diphosphate, agmatine, diadenosine hexaphosphate, cysteineglutathione disulfide, oxidized glutathione, hexadecanedioic acid, glutamylphenylalanine(P<0.05). According to the metabolic pathways of relevant endogenous markers,it is suggested that SFI may affect the model rats through arginne and proline metabolism, sulfur metabolism, glutathione metabolism and purine metabolism,among them adenosine 3′,5′-diphosphate, agmatine and oxidized glutathione were recognized as the key potential biomarkers of the related metabolic pathway. CONCLUSION This study primarily clarified the therapeutic effect of SFI on mid-stage cardiogenic shock rats, which may be related to improving the levels of endogenous metabolites in serum and then restoring the metabolism to be normal in vivo.     

药理效应法测定参附注射液药动学参数的研究

目的:考察参附注射液在大鼠体内的药动学特征.方法:采用结扎冠状动脉法复制大鼠心源性休克模型,以血压值作为效应指标,选取1.25、2.5、5、10、20、25 mL/kg六个剂量参附注射液做量-效曲线,选择20 mL/kg剂量做时-效曲线,依据时-效曲线和量效曲线求得时间-生物体存量曲线,用DASver 2.0软件分析参附注射液的药动学参数.结果:主要药物动力学参数为t1/2=8.685 min,Ke=0.08 min-1, CL=1.417 L· min -1·kg-1, AUC(0-t) =12.63 mg·L-1·min-1.结论:在大鼠体内静脉注射参附注射液的体存量的表观动力学过程符合一室模型.     

栓必灵对大鼠肠系膜微循环的影响

目的:研究栓必灵对大鼠肠系膜微循环的影响。方法:大鼠分别舌静脉注射栓必灵45、90、180IU/kg ,采用显微录像静画步进法观察给药30min后大鼠肠系膜毛细血管血流速度、血流状态及血管内径的变化。结果:低、中、高剂量组血流速度增加率分别为24.25 %、26. 34 %和25. 88 % ;细动脉内径增加率分别为21. 37 %、27 .13 %和28. 80 % ;细静脉内径增加率分别为6 .70 %、9. 31 %和12 .95 % ;血流状态明显改善。结论:栓必灵有改善大鼠肠系膜微循环的作用。     

Hepatic microvascular effects of terbutaline in experimental cardiogenic shock in rats

1. Experimental myocardial infarction was produced in rats by direct electrical cauterization of the myocardium of left ventricle. This produced cardiogenic shock with the accompanying haemodynamic changes of low cardiac output, low mean arterial pressure, raised central venous pressure and an absence of cardiac arrhythmias. 2. The liver microcirculation was observed using in vivo television microscope method. The diameter and erythrocyte flow velocity in the liver sinusoids were measured quantitatively. 3. During experimental cardiogenic shock 80% of the liver sinusoids were constricted; the remaining 20% showed dilatation. In all these liver sinusoids the erythrocyte flow velocity was only 50% of the pre-shock level. 4. Intravenous injection of the selective beta 2-adrenoceptor agonist terbutaline (0.15 mg/kg) restored the systemic arterial pressure to pre-shcok levels and partially raised the cardiac output. In the liver microcirculation terbutaline restored both constricted and dilated liver sinusoids to pre-shock calibres, but only partially raised erythrocyte flow velocity. 5. It is proposed that during experimental cardiogenic shock, terbutaline produces dilatation in the terminal liver microcirculation by opening sphincters of liver sinusoids and restores sinusoid diameters to pre-shock calibres. Therefore, terbutaline has the capacity to decrease peripheral resistance and unload the circulation during cardiogenic shock.     

By regulating IP3/PKC/TRPV4 pathway hyperoside induces endothelium-dependent vasodilatation in rat basilar artery following four vessel occlusion ischemia reperfusion

OBJECTIVE To investigate regulatory effects of hyperoside(Hyp) on IP3/PKC/TRPV4 pathway in rat cerebral basilar artery(CBA) subjected to global cerebral ischemia-reperfusion(I/R). METHODS The model of global cerebral I/R in rats was established by four-vessel occlusions methods. The treated rats were administrated with Hyp(50 mg·kg~(-1)) group, Hyp(50 mg·kg~(-1))+HC-067047(10 mg·kg~(-1)), Hyp(50 mg·kg~(-1))+2 APB(2 mg·kg~(-1)), Hyp(50 mg·kg~(-1))+Bis I(2.5 mg·kg~(-1)), Hyp(50 mg·kg~(-1))+ 2 APB(2 mg·kg~(-1))+Bis I(2.5 mg·kg~(-1)). Hematoxylin-eosin(HE) and Nissl staining were performed and the contents of methane dicarboxylic aldehyde(MDA), neuron-specific enolase(NSE), S100β and the activity of lactic dehydrogenase(LDH) in serum were measured by enzyme-linked immunosorbnent assay(ELISA).The specific blocker N-nitro-L-arginine-methyl-ester(L-NAME) and indomethacin(Indo) were used to delete the prostacyclin(PGI2) and nitric oxide(NO) dependent relaxation. The protein expression level of TRPV4 was detected by Western blotting.Ca~(2+) intensity in vascular smooth muscle cells was measured by confocal laser scanning microscope and flow cytometry was performed to observe the apoptosis of CBA endothelial cells after in vivo administration. RESULTS Hyp induced a dose-dependent relaxation of CBA in IR rats via a PGI2 and NO independent manner, as evidenced by alleviated pathological changes and up-regulated expression of TRPV4 protein in the endothelial cells from cerebral vessels. Hyp significantly reduced the contents of MDA, NSE, S100β and the activity of LDH in serum and decreased the fluorescence intensity of Ca~(2+) in cerebral vascular smooth muscle cells by in vivo administration. The apoptotic rate of endothelial cells in Hyp treated group was significantly less than that in IR group. CONCLUSION Hyp does in fact ameliorate I/R injury by regulating IP3/PKC/TRPV4 pathway.     

药物联合治疗对肾血管性高血压大鼠左室重构影响的对比研究

目的：通过两两药物联合应用治疗高血压,观察并对比药物联合治疗的降压效果,并探讨其对左室重构的作用。方法：用两肾一夹法建立高血压大鼠模型,43只SD大鼠随机分为：假手术组（A组,n=10）,高血压组（B组,n=11）,β-B＋ACEI组（C组,n=11）予以比索洛尔5mg·kg-1.d-1及贝那普利5mg·kg-1·d-1灌胃,ACEI＋CCB组（D组,n=11）予以贝那普利5mg·kg-1·d-1及氨氯地平2.5mg·kg-1·d-1灌胃,饲养20周,期间每两周给大鼠测量鼠尾动脉压和超声心动图,20周后采集标本,用免疫组化法检测胶原Ⅲ、胶原Ⅳ及心钠素（ANF）。结果：用药组血压、胶原Ⅲ、胶原Ⅳ含量较B组明显降低,差异具有统计学意义。与A组比较,B组ANF明显升高（P〈0.05）,各药物治疗组ANF无明显升高;与B组比较,各药物治疗组ANF显著降低（P〈0.01）。结论：在降压和靶器官保护方面,贝那普利和氨氯地平联用疗效优于贝那普利和比索洛尔联用。     

Salvianolic acid A ameliorates AGEs-induced glomerular endothelial dysfunction and protects against diabetic nephropathy

OBJECTIVE Diabetic nephropathy(DN)has been one of the most common complications of diabetes and the leading cause of end-stage renal disease.Glomerular hyperfiltrationis central in earlystage of DN and leads to the progression of renal architectonic and functional abnormalities.Salvianolic acid A(SalA)has been proved to protect diabetic complications such as hepatic fibrosis and neuropathy.The present study was designed to investigate the effects of SalA on glomerular endothelial dysfunctionand diabetic nephropathy.METHODS Primary glomerular endothelial cells were subjected to assess permeability under injury of advanced glycation end-products(AGEs).AGEs-induced changes of Rho A/ROCK pathway and cytoskeleton rearrangement were assessed bywestern blotandimmunofluorescence.The beneficial effects of SalA on diabetic nephropathy were investigated in a rat model induced by high-fat and high-glucose diet combined with low dose of streptozocin(35 mg·kg~(-1),ip).Renal function and architectonic changes were evaluated by biochemical assay and PAS staining.RESULTS SalA 3μMameliorated AGEs-induced glomerular endothelial permeability(P<0.05)and suppressed rearrangement of cytoskeleton through inhibiting AGE-RAGE-Rho A/ROCK pathway.SalA1 mg·kg~(-1)markedly reduced endothelium loss(P<0.01)and glomerular hyperfiltration(P<0.05)in diabetic kidney.Subsequently,SalA 1 mg·kg~(-1) suppressed glomerular hypertrophy and mesangial matrix expansion,eventually reduced 24 h-urinary albumin and ameliorated renal function by decreasing blood urine nitrogen(BUN),serum creatinine(Scr)and serum n-acetyl-β-d-glucosaminidase(NAG).AGEs-RAGE-Nox4-induced oxidative stress was suppressed by the treatment of SalA 1 mg·kg~(-1).CONCLUSION SalA ameliorated AGEs-induced glomerular endothelial hyperpermeability,and effectively protected against early-stage diabetic nephropathy by reducing hyperfiltration and alleviating renal structural deterioration through inhibiting AGEs and its downstream pathway.Thus,SalA might be a promising therapeutic agent for the treatment of diabetic nephropathy.     

活血益智片对血管性痴呆大鼠记忆障碍及海马Bcl-2蛋白表达的影响

目的研究活血益智片对血管性痴呆大鼠(VaD)认知功能及海马神经元凋亡的影响。方法Wistar大鼠腹腔注射硝普钠2.5mg·kg-1后,立即反复夹闭大鼠双侧颈总动脉,制备血管性痴呆大鼠模型,观察活血益智片组(1.55,3.1,6.2g.kg-1)及氢麦角碱组(5.4mg·kg-1)连续给药28天后,对大鼠学习记忆能力及海马中SOD、MDA活性及Bcl-2、Bax蛋白表达的影响。结果活血益智片组与模型组比较,可提高大鼠的学习记忆能力及SOD活性,降低MDA含量;增加Bcl-2及减少Bax蛋白表达。结论活血益智片可通过调节Bcl-2及Bax表达而发挥抗凋亡作用。     

Long-term observation of pial microcirculatory parameters using an implanted cranial window method in the rat

The aim of this study was to confirm whether our improved closed cranial window (CCW) method could be used for long-term microscopical observation of pial microcirculation intravitally in the rat. We investigated chronological changes in three microcirculatory parameters: permeability of blood-brain barrier, leukocyte behavior, and plasma velocities in the pial venules, immediately after implantation (control group) and at one and four weeks after implantation in different age-matched rats (implanted group). No extravasation of sodium fluorescein from pial venules was confirmed in any observation period. The number of endothelial-adhering leukocytes in the implanted group kept within the physiological range, being similar to those of the control group. The velocities of fluorescent microspheres flowing in pial venules showed no noticeable changes between the two groups. These findings suggest that our CCW method allows long-term observation of the pial microcirculation without any pathophysiological changes in the evaluated parameters up to four weeks after the implantation.     

去铁酮在大鼠体内的药代动力学与组织分布

目的研究去铁酮(DFP)在大鼠体内的药代动力学和组织分布。方法雄性Wistar大鼠ig给予DFP35,70和140mg.kg-1后,于不同时间点收集血液和组织样本。采用高效液相色谱法测定大鼠血浆及组织中的DFP的含量,运用DAS2.0药代动力学智能分析软件拟合房室模型,并进行药代动力学参数计算。结果大鼠ig给予DFP35,70和140mg.kg-1后,体内药代动力学过程符合二室模型,t1/2α分别为23.3,22.2和20.9min,t1/2β分别为53.3,50.9和46.3min,Cl分别为0.017,0.021和0.016L.min-1.kg-1。大鼠ig给予DFP70mg.kg-1后,DFP在胃和肝中浓度较高,60min时肝中DFP含量可达(359.22±31.16)μg.g-1,其他组织含量较低。结论DFP在大鼠体内吸收和消除较迅速,在体内组织分布广。     

UPLC-MS法测定大鼠血浆中积雪草苷的浓度及其药代动力学研究

目的建立测定积雪草苷血浆药物浓度的超高效液相色谱-电喷雾离子化-质谱(UPLC-ESI-MS)联用的分析方法,探讨其在大鼠体内的药动学。方法SD大鼠8只,单剂量静注(40mg·kg-1)积雪草苷,用UPLC-MS法测定给药后的血浆中药物浓度,并用DAS软件求算其药代动力学参数。结果积雪草苷的血药浓度在0.038～7.6mg·L-1范围内线性关系良好,最低检测限为38μg·L-1,提取回收率大于95%,日间、日内RSD均小于10%。大鼠单剂量静注积雪草苷40mg·kg-1后,血药浓度-时间曲线呈二室模型。主要药动学参数AUC(0-t)、T12β、CL、Vd分别为:(81443.67±57156.81)μg·L-1·min-1、(23.44±9.60)min、(0.19±0.07)L·min-1·kg-1、(8.92±6.68)L·kg-1。结论该方法操作简便、快速、灵敏、专属性强,可用于积雪草苷的体内大批量样品定量分析及药代动力学研究。     

黄芪注射液对内毒素诱导大鼠肠系膜微循环障碍改善作用的实验研究

目的探讨静脉给予黄芪注射液对内毒素血症肠系膜微循环障碍的改善作用。方法 Wistar大鼠30只为对照组、模型组、治疗组,每组10只。采用静脉注入脂多糖（LPS）（5mg·kg-1·h-1）复制内毒素血症模型,治疗组给予静脉注射黄芪注射液（5ml·kg-1·h-1）,用微循环观察系统每20分钟动态观察细静脉粘附白细胞,细静脉血管壁过氧化物的动态变化。在100min观察结束后,计数肠系膜间质内肥大细胞脱颗粒率。取外周血,用流式细胞仪测定粒细胞粘附分子CD11b和CD18的表达。结果模型组在LPS滴注20min后,黏附于大鼠肠系膜细静脉壁上的白细胞数和管壁过氧化物依存的DHR的荧光强度显著增加,100min时计数肠系膜间质内肥大细胞脱颗粒率显著地增加（P〈0.05）。流式细胞仪测定外周血粒细胞黏附分子CD11b和CD18的表达明显增加（P〈0.05）。治疗组白细胞与肠系膜细静脉的血管壁黏附;细静脉壁过氧化物依存的DHR荧光强度的增加,肠系膜间质内肥大细胞脱颗粒率,外周血粒细胞黏附分子CD11b和CD18的表达明显受到抑制（P〈0.01）。结论黄芪注射液对内毒素血症肠系膜微循环障碍有改善作用。可能其抑制粒细胞黏附分子CD11b和CD18表达及肥大细胞脱颗粒相关。     

磷脂对葛根素体外吸收和药效学的影响

探讨磷脂对葛根素吸收和药理作用的影响。方法：用ＨＰＬＣ法测定葛根素含量,观察磷脂对大鼠离休肠管吸收葛根素的影响,比较ｉｖ乳化葛根素与葛根素对家兔血液流变学和微循环指标作用的不同。结果：大鼠离体肠管对乳化葛根素和葛根素的扩散常数 Ｄ分别为 ３ ×１０－７和 １×１０－７ｍ２／ｓ;乳化葛根素改善家兔血流变和微循环指标作用均强于葛根素（Ｐ＜０．０１）。结论：磷脂可提高大鼠离休肠管对葛根素的吸收量,有增强改善家兔血液流变学和微循环指标的作用。     

阿加曲班对大鼠肢体缺血再灌注损伤肠系膜微循环影响的实验研究

目的探讨阿加曲班对大鼠肢体缺血再灌注损伤肠系膜微循环的影响,为缺血再灌注损伤的防治提供新的思路。方法采用大鼠肢体缺血再灌注模型,将14只Wistar大鼠随机分为每组7只：对照组和阿加曲班组,每组7只。2组大鼠均于再灌注2h后,酶联免疫吸附（ELISA）法测定血浆P-选择素、细胞间粘附分子-1（ICAM-1）浓度;应用BI-2000医学图像分析系统观察和记录肠系膜微循环改变,指标包括肠系膜微动脉管径（AD）和微静脉管径（VD）、肠系膜微动脉血流速度（AFV）和微静脉血流速度（VFV）以及白细胞粘附情况。结果大鼠肢体缺血再灌注2h后,阿加曲班组血浆P-选择素浓度、血浆ICAM—1浓度显著低于对照组（P〉0．05）。与对照组相比,阿加曲班组肠系膜微循环障碍较轻,AD和VD与对照组之间差异无统计学意义（P〉0．05）,AFV和VFV明显高于对照组（P〈0．05）,白细胞粘附较对照组轻（P〈0．05）。结论阿加曲班能够降低血浆P-选择素和ICAM-1浓度,进而改善大鼠肢体缺血再灌注损伤时的肠系膜微循环障碍．     

Identification of bioactive anti-angiogenic constitutes targeting tumor endothelial cells in Shenmai Injection using multidimensional pharmacokinetics

OBJECTIVE To identify the bioactive anti-angiogenic constitutes targeting tumor endothelial cells(TECs) in Shenmai Injection(SMI). METHEODS For pharmacokinetic(PK) studies, Balb/c mice harboring human colorectal cancer(Lo Vo) xenografts were treated with SMI 10 m L·kg~(-1) daily for 1 or 8 d. Multidimensional PK profiles of ginsenosides in plasma, subcutaneous tumors, and TECs were investigated. For PD studies, the tumor-bearing micewere treated with SMI 10 mL ·kg~(-1) daily, Rd 0.5 and 5 mg·kg~(-1) daily, Rb1 5 mg·kg~(-1) daily and Rg1 5 mg·kg~(-1) daily for 14 d.Intravital multi-photon imaging and CD31 immunofluorescence staining were used to evaluate the number of microvessels and braches. Double staining of CD31 and α-SMA was performed to evaluate pericytes coverage ratios around vessels.ELISA was performed to determine the concentrations of VEGF and FGF in tumor tissues. For synergistic anti-tumor study, the tumor-bearing mice were treated with SMI 10 mL ·kg~(-1) daily, Rd 5 mg·kg~(-1) daily with or without 5-FU 15 mg·kg~(-1) every 3 d for 20 d. HPLC-MS/MS was used to determine the concentrations of 5-FU in plasma and tumor tissues.RESULTS SMI decreased the number of microvessels(P<0.05) and vessel branches(P<0.05) and improved vascular pericytes coverage(P<0.05). PK studies showed that the concentrations of protopanaxadiol-type(PPD) ginsenosides(Rb1, Rb2/Rb3, Rc, and Rd) in both, plasma and tumors, were higher than those of protopanaxatriol-type(Rg1 and Re)and oleanane-type(Ro) ginsenosides. Among PPD ginsenosides, Rd exhibited the greatest concentrations in tumors and TECs after repeated injection. In fact, the proportion of Rd in the detectable components of SMI gradually increased in the following order: SMI formula(2.8%), plasma(16.0%), tumor tissues(34.3%), and TECs(40.3%). In vivo bioactivity results showed that Rd 5 mg·kg~(-1) daily significantly decreased the number of microvessels(P<0.05) and vessel branches(P<0.05) and increased pericytes coverage(P<0.05) while Rd 0.5 mg·kg~(-1) daily, Rb1 and Rg1 had no significant effect on them. Rd 5 mg·kg~(-1) suppressed the expression of VEGF and FGF simultaneously. Rd 5 mg·kg~(-1) enhanced the antitumor effect of 5-FU via increasing the distribution of 5-FU in tumor tissues(P<0.05) in xenograft mice. CONCLUSION Ginsenoside Rd may be the major bioactive anti-angiogenic constituent targeting TECs after SMI treatment.     

Ginsenoside Rb1 attenuates adjuvant-induced arthritis in rats through inactivation of NF-κB signaling pathway

OBJECTIVE To investigate the anti-arthritic effect and mechanism of action of ginsenoside Rb1 on adjuvant-induced arthritis(AIA) in rats. METHODS Male SD rats were received 0.1 m L injections of FCA(10 g·L~(-1)) emulsion into the right hind metatarsal foot pad for arthritis induction. After that, rats were randomly divided into six groups, namely control group, untreated group, dexamethasone(DEX, 2.5 mg·kg~(-1)) group, low(5 mg·kg~(-1)), medium(10 mg·kg~(-1)) and high(20 mg·kg~(-1))doses of ginsenoside Rb1 groups, and treated intraperitoneally at the above dosage once a day for2 weeks. After treatment, paw swelling and arthritis indexes were evaluated, the thymus and spleen index were calculated as well. HE staining were used to observe the joint histopathology in rats. Rat ELISA kits were used to determinate the TNF-α, IL~(-1)β and IL-6 levels. Western blotting were used to detect the related protein expression of NF-κB signaling pathway in the tissues of inflamed joints. RESULTS Rb1 significantly decreased the paw swelling and arthritis index,Compared with AIA group. HE staining results revealed that medium and high doses of Rb1 significantly reduced synovial inflammatory cell infiltration, synovial lining hyperplasia and bone destruction, compared with AIA group. Elisa results showed that Rb1 significantly decreased the TNF-α, IL~(-1)β and IL-6 levels(P<0.05, P<0.01). Western blotting results revealed that the expression of p-IκB and p-P65 were significantly reduced in 20 mg·kg~(-1) of Rb1 group, compared with AIA group(P<0.05, P<0.01). CONCIUSION Rb1 manifests therapeutic anti-inflammatory effects on rats with AIA, potential y through a mechanism of inhibiting activation of the NF-κB.     

芝麻素与维生素E联用对代谢综合征

目的:观察芝麻素与维生素E联用对代谢综合征大鼠肾脏的保护作用并探讨两药联用的协同关系.方法:采用高脂高糖饮食24周诱导大鼠代谢综合征,第9周(57 d)口服含芝麻素(30 mg·kg~(-1)·d~(-1))、芝麻素+维生素E[(30+20)、(15+20)mg·kg~(-1)·d~(-1)]和维生素E(20mg·kg~(-1)·d~(-1))饲料16周.24周末称体重和左肾湿重;测血糖、血脂、血压、肾功能、肾皮质氧化和抗氧化指标;HE和Masson染色观察肾脏形态及胶原沉积;免疫组化法表达诱导型一氧化氮合酶和硝基酪氨酸.结果:(1)模型组肾功能明显损害,肾小球发生硬化和肾间质纤维化,并出现大量炎症细胞浸润,肾小球和肾间质胶原沉积,脂质过氧化物损伤因子MDA、NO_2~-/NO_3~-和OH~-含量升高,iNOS蛋白和硝基酪氨酸表达明显上调,抗氧化酶保护因子T-SOD、CAT、GSH-Px活性显著降低;(2)芝麻素+维生素E[(30+20mg/kg)]组能明显降低血糖、血脂和血压,提高肾皮质总超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化酶活性,减少丙二醛、NO_2~-/NO_3~-和羟自由基含量,下调诱导型一氧化氮合酶和硝基酪氨酸,减轻肾小球与肾间质胶原沉积,逆转肾小球硬化和肾间质纤维化,改善肾功能,并且优于单用芝麻素组和维生素E组(P＜0.01或P＜0.05).结论:芝麻素(30 mg/kg)与维生素E(20 mg/kg)联用具有协同抗氧化和抗代谢综合征大鼠肾脏损伤作用.     

线粒体通透性转换孔在冠脉微循环障碍中的研究进展

由于冠脉微循环障碍的发生机制比较复杂,临床上治疗冠脉微循环的药物疗效均未得到充分肯定,线粒体通透性转换孔(mitochondrial permeability transition pore,mPTP)的开放因会引起冠脉微循环细胞功能异常而成为当前治疗冠脉微循环障碍的热点研究靶标。本文就近几年国内外关于mPTP在冠脉微循环障碍中的作用研究进展进行简要讨论。     

BN大鼠与Wistar大鼠Ⅰ型超敏反应敏感性的比较

目的比较BN大鼠和Wistar大鼠在Ⅰ型超敏反应中的敏感性,建立一种灵敏可靠的Ⅰ型超敏反应检测体系。方法BN大鼠和Wistar大鼠分别隔天sc不同剂量的卵白蛋白(10,20和40μg.kg-1),共5次,正常对照组sc给予生理盐水。首次注射后第21天取血清,用ELISA法测定血清总免疫球蛋白E(IgE)水平,通过被动皮肤过敏反应实验检测其特异性IgE水平;第22天检测激发后大鼠血压、血清中组胺和类胰蛋白酶浓度的变化。结果与正常对照组比较,BN大鼠在卵白蛋白10,20和40μg.kg-1下血清总IgE和特异性IgE显著增加,血压下降,血清组胺和类胰蛋白酶浓度增加;Wistar大鼠仅在卵白蛋白40μg.kg-1组出现上述变化。结论与Wistar大鼠相比,BN大鼠用于Ⅰ型超敏反应的检测更为灵敏。血压和血清总IgE、特异性IgE、组胺及类胰蛋白酶浓度等可作为Ⅰ型超敏反应重要的检测指标。     

Ginkgo Leaf Extract and Armillariella Mellea Powders Oral(银杏蜜环口服溶液) attenuates inflammation of microglia in habenular nucleus throughCX3CL1-CX3CR1 axis in post stroke depression

OBJECTIVE The Ginkgo Leaf Extract and Armillariella Mellea Powders Oral(Yinxingmihuan Koufu Rongye, YXMH), a representative drug for "Treating both Brain and Heart", showed considerable clinical effects in ischemic cardiovascular and cerebral vascular diseases. Recently, it is reported that YXMH has the potential for treating myocardial and cerebral ischemia related mental disorders, such as post stroke depression(PSD) and chronic heart disease(CHD) associated anxiety disorder. However, its mechanism has not been clearly elucidated. Meanwhile, increasing evidence revealed that there are close functional links between depression and habenular nucleus. The present study investigates the underlying mechanism of YXMH on attenuating the inflammation of microglia in habenular nucleus through CX3 CL1-CX3 CR1 axis in in a rat model of PSD. METHODS Rats were randomly devided into sham group,model group, Ginaton group(18 mg·kg~(-1)), Armillariella Mellea group(600 mg·kg~(-1)), Fluoxetine group(10 mg·kg~(-1)), YXMH high-dose group(618 mg·kg~(-1)) and YXMH low-dose group(309 mg·kg~(-1)). The PSD model was induced by transarterial microembolization combined with sleep deprivation(2-Chloro-D-phenylalanine, PCPA, IH, 200 mg·kg~(-1), for 3 times,before the behavior test) in SD male rats. Then rats were treated with corresponding medicaments through gavage once a day until 3 weeks later, followed by body mass measurement, neurological deficit score evaluation, gripping strength and thermal withdrawl latency measurement, as well as depression related behavioral indicators, the open field test(OFT) and sucrose preference test. The pathological morphological changes of habenular nucleus was observed by HE staining, the expression of IBA-1 was measured and analyzed by immunohistochemistry staining, and alterations of proteins and genes related to the CX3 CL1-CX3 CR1 axis were analyzed using Western blotting(CX3 CL1, CX3 CR1) and real-time polymerase chain reaction(PCR)(CX3 CL1, CX3 CR1). RESULTS Compared with the sham group, rats in the model group manifested as decreased body mass, deficient neurological behavior and gripping strength, reduced locomotor activity and sugar water consumption, as well as elevated thermal withdrawl latency(P<0.05, P<0.01). Meanwhile, the pathological morphology of the habenular nucleus on the ischemic hemisphere showed significant neuronal degeneration, microglial proliferation, inflammatory cells and glia cells infiltration, together with up-regualted expression of IBA-1, CX3 CL1, CX3 CR1 protein and CX3 CL1, CX3 CR1 mRNA. YXMH attenuated inflammation of microglia in habenular nucleus through improving pathological morphology, inhibiting IBA-1 activation, down-regulating the expression of CX3 CL1 and CX3 CR1 proteins and genes, and thus improved the behavior performance of ischemic injury and depression. CONCLUSION YXMH ameliorates neurological deficit and depressive behavior in rat model of PSD induced by transarterial microembolization combined with sleep deprivation, and the mechanism is probably related to attenu-ating inflammation of microglia in habenular nucleus through CX3 CL1-CX3 CR1 axis.