Effect of Lunei Xiaoyu Decoction on Vascular Endothelial AMPK/eNOS Signal in Balloon Injury Model of Common Carotid Artery in Rats

目的观察颅内消瘀汤对大鼠颈动脉球囊损伤后血管内皮及AMPK、eNOS表达的影响,探讨其可能的作用机制。方法 32只SD大鼠随机分为假手术组、模型组、颅内消瘀汤低剂量(临床等效量)组和颅内消瘀汤高剂量(临床2倍量)组,除假手术组大鼠仅切开皮肤,其余大鼠均行颈总动脉球囊损伤术以成模。成模第4天灌胃给药,模型组及假手术组给予等体积生理盐水,连续给药14 d后取损伤侧颈总动脉,行HE染色观测血管内皮损伤及内膜增生情况,用免疫组化染色检测eNOS表达水平,并用Western blot检测总AMPK、eNOS蛋白水平。结果颈总动脉球囊损伤模型组大鼠血管内膜增生明显、管壁变厚,颅内消瘀汤低、高剂量组大鼠经治疗后损伤血管增生较模型组轻、管壁较薄;e NOS免疫组化染色:模型组阳性细胞较少,经颅内消瘀汤干预后,低、高剂量2组均有不同程度的表达,与模型组比较差异有统计学意义(P<0.01);Western blot检测结果显示模型组总AMPK、eNOS蛋白表达下降,经颅内消瘀汤干预后,均有不同程度增加,与模型组比较差异有统计学意义(P<0.05或P<0.01)。结论颅内消瘀汤可通过上调AMPK/eNOS信号表达,保护大鼠颈动脉球囊损伤血管内皮的作用。OBJECTIVE To observe the effect of Lunei Xiaoyu Decoction on vascular endothelial AMPK, eNOS expression in ballon injury model of common carotid artery in rats, and to explore its possible mechanism. METHODS Thirty two SD rats were randomly divided into sham operation group, model group, low dose of Lunei Xiaoyu Decoction(dose in clinical equivalent) group and high dose of Lunei Xiaoyu Decoction(2 times the amount of clinical) group, except the sham operation group rats only skin incision, the other rats underwent carotid artery balloon injury surgery. On the 4th day after modeling, the rats were administrated orally for 14 d, model group and sham operation group were given equal volume of physiological saline. Then all the rats were killed, HE staining observation of vascular endothelial injury and intimal hyperplasia, immunohistochemistry staining was used to detect the expression level of e NOS, and Western-blot was used to detect the expression level of AMPK and eNOS. RESULTS The endometrial hyperplasia and thickness were obvious after carotid artery balloon injury in rats; and after Lunei Xiaoyu Decoction treatment, the rats' endometrial hyperplasia and thickness were improved. The expression of AMPKand e NOS was significantly higher in Lunei Xiaoyu Decoction groups compared with the model group(P<0.05 or P<0.01). CONCLUSION Lunei Xiaoyu Decoction can upregulate the expression of AMPK/e NOS signal, in order to protecting vascular endothelial after balloon injured carotid artery in rats.福州市科技计划项目(2012-S-155-1

Effect of Activating Blood and Resolving Stasis Therapy on Injuried Vascular wnt / β-catenin Signal Pathway of Rats with Carotid Atherosclerosis

目的:观察活血化瘀法对颈动脉粥样硬化血管内皮损伤模型大鼠wnt/β-catenin信号通路的影响,探讨其可能的作用机制。方法:32只SD大鼠高脂喂养1个月后,随机取8只作为假手术组(仅切开皮肤),其余大鼠均行颈总动脉球囊损伤术,成膜后再随机分为模型组(n=8只)、活血化瘀中药方中剂量(临床等效量)组(n=8只)和高剂量(临床2倍量)组(n=8只)。从术后第4天开始予以相应药物灌胃治疗(模型组、假手术组灌等量生理盐水),2周后取术侧颈总动脉段,采用实时荧光定量PCR法捡测wnt、Fz D1、AXIN1、GSK-3β、β-catenin、VEGF mRNA基因表达水平。结果:与假手术组比较,模型组wnt、Fz D1、AXIN1、β-catenin、VEGF等mRNA水平降低,差异有统计学意义(P<0.01),经活血化瘀中药方干预后,上述各基因mRNA水平均有不同程度升高,尤其是高剂量组与模型组比较差异有统计学意义(P<0.05~0.01);只有GSK-3βmRNA水平各组无明显差异。结论:活血化瘀法可通过上调wnt/β-catenin信号通路表达,使损伤血管VEGFmRNA基因表达水平上升,提示该法可保护颈动脉粥样硬化血管内皮损伤。Objective: To observe the effect of activating blood and resolving stasis on injuried vascular wnt / β-catenin in carotid atherosclerosis mode rats and its possible mechanism. Methods: After one-month high fat-diet,32 SD rats were randomly divided into sham operation group,model group,middle dose( dose in clinical equivalent) group and high dose of( 2 times the amount of clinical) group. Except for the sham operation group,the other rats underwent carotid artery balloon injury surgery. After 3 days of intraperitoneally administered orally,model group and sham operation group were given equal volume of physiological saline,administered continuously for 14 d. Then the expression of wnt,Fz D1,AXIN1,GSK-3β,β-catenin,VEGF mRNA in injured vascular were analyzed by real-time fluorescence quantitative PCR in all groups. Results: Compared with the sham operation group,the expression of Wnt,Fz D1,AXIN1,β-catenin,and VEGF mRNA levels in model group decreased( P < 0. 01); and after treated by activing blood and resolving stasis therapy,the expression of the above genes increased in different degrees,especially in the high dose group( P < 0. 01 ~ 0. 05); but there was no significant difference among groups on the expression of GSK-3β mRNA. Conclusion: The theapy of activating blood and resloving stasis therapy can increase the expression of VEGF mRNA of balloon injured carotid artery vascular through up-regulating wnt / β-catenin signal pathway,which suggested that it can protect the injured vascular of carotid atherosclerosis.福建省教育厅A类科技项目(编号:JA13165);; 福州市科技计划项目(编号:2012-S-155-1);; 福建中医药大学校管科研项目(编号:X2012018

AMP依赖的蛋白激酶及PI3K/Akt信号途径在颈动脉狭窄形成过程中的变化

目的 观察AMP依赖的蛋白激酶（AMPK）及PI3K/Akt信号途径在颈动脉狭窄形成过程中的变化。方法 取SD大鼠经高脂喂养1个月后行颈总动脉球囊损伤术作为模型组（n=24）,继续喂养14 d后取损伤侧颈总动脉,行HE染色观测血管内皮损伤及内膜增生情况,应用荧光定量PCR法检测AMPK及PI3K/Akt mRNA水平,用Western印迹检测p-AMPK、PI3K、p-Akt蛋白水平,另取8只正常SD大鼠作为假手术组。结果 模型组大鼠血管内膜增生明显、管壁变厚;AMPK/PI3K/Akt mRNA及蛋白水平较假手术组显著下降（P〈0.01）。结论 AMPK/PI3K/Akt在颈动脉狭窄的病理生理进程中起到一定的作用。福州市科技计划项目（No.2012-S-155-1

r DNA sequence analysis and morphological redescription of Empoasca onukii from the tea growing area of Fujian

重新描述了小贯小绿叶蝉的形态结构,尤其是头部色斑、翅脉、腹部内突、下生殖板及其刚毛着生位置,并增加了足部(刺毛列)特征描述。通过PCr技术克隆得到该物种的r dnA序列,包括部分18S(1 843bP)和28S序列(667 bP)以及完整的5.8S(155 bP)、ITS1(3 114 bP)和ITS2序列(1 008 bP)。序列分析表明,18S,5.8S,28S与其他物种间具有90%~95%的序列一致性;而ITS1和ITS2序列变异性非常大。碱基组成比率分析显示ITS1以及ITS2具有AT偏好性,前者A+T占66.0%,后者占65.1%。与亲缘种的比较分析显示ITS1和ITS2具有丰富的多态位点,并且ITS2更适用于近缘种的分子鉴定。The morphological structures of Empoasca onukii,the cephalic splashes,vein,abdominal apodemes,subgenital plate and the arrangement of its bristles,and the characteristics of legs were redescribed,especially the arrangement of spines were added.Ribosomal DNA( r DNA) including partial of 18S( 1 843 bp),28S( 667 bp) and complete 5.8S( 155 bp),ITS1( 3 114 bp),ITS2( 1 008 bp) were isolated by PCR technology.Sequence analysis indicated that 18 S,5.8S and 28 S had high sequence identity( 90%- 95%) with other insects; however,ITS1 and ITS2 showed much variability.The analysis of base composition showed that ITS1 and ITS2 were more preferences for AT( contained 66.0% and 65.1% A + T,respectively).Comparison with the phylogenetic species revealed that ITS1 and ITS2 from E.onukii had rich polymorphic loci and ITS2 was more suitable for molecular identification of closely related species.国家自然科学基金(81171595

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本文提出以混合稀土氯化物作为Ｄｉｅｌｓ－Ａｌｄｅｒ反应催化剂，催化三种有工业应用价值的环加成反应。实验结果表明，混合稀土氯化物对具有含氧基团底物的Ｄｉｅｌｓ－Ａｌｄｅｒ反应催化效果显著，其中混合重稀土氯化物比混合轻稀土氯化物表现出更高的催化活性

混合稀土氯化物催化的Diels-Alder反应

本文提出以混合稀土氯化物作为Ｄｉｅｌｓ－Ａｌｄｅｒ反应催化剂，催化三种有工业应用价值的环加成反应。实验结果表明，混合稀土氯化物对具有含氧基团底物的Ｄｉｅｌｓ－Ａｌｄｅｒ反应催化效果显著，其中混合重稀土氯化物比混合轻稀土氯化物表现出更高的催化活性