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1.
该文探讨通窍活血汤含药脑脊液(TQHXD-CSF)对氧糖剥夺/复糖复氧(OGD/R)损伤小鼠海马神经元(HT22)细胞的保护作用及其作用机制是否与调控ASK1/MKK4/JNK信号通路有关.将HT22细胞随机分为5组,分别为空白脑脊液组(control组)、OGD/R组、TQHXD-CSF组、凋亡抑制剂组(Z-VAD-FMK,20μmol·L^(-1))、TQHXD-CSF+凋亡抑制剂组.除control组外,其余各组细胞在氧糖剥夺6 h后迅速复糖复氧12 h进行OGD/R造模并分组给药.CCK8法检测HT22细胞活力,LDH试剂盒检测乳酸脱氢酶释放量,倒置显微镜观察HT22细胞形态.Western blot检测Bax、Bcl-2和caspase-3的蛋白表达情况,实时荧光定量PCR(qRT-PCR)检测Bax、Bcl-2和caspase-3的mRNA水平.再将HT22细胞随机分为6组,分别为control组、OGD/R组、si-NC组、si-ASK1组、TQHXD-CSF组、TQHXD-CSF+si-ASK1组.CCK8法检测细胞活力,细胞动态分析仪动态监测细胞增殖,Hoechst染色和流式细胞术检测HT22细胞凋亡情况,Western blot检测MKK4、p-MKK4、JNK、p-JNK、c-Jun、p-c-Jun、Cyt C、Bax、Bcl-2和caspase-3的蛋白表达情况.结果显示,与OGD/R组相比,TQHXD-CSF可以显著提高细胞存活率,改善细胞形态,降低Bax/Bcl-2、caspase-3蛋白和mRNA的表达量.当沉默ASK1后,与OGD/R组相比,TQHXD-CSF组细胞存活率显著提升,降低细胞荧光强度,降低细胞凋亡率,p-MKK4、p-JNK、p-c-Jun、Cyt C、Bax/Bcl-2和caspase-3的蛋白表达量减少,但其效果不及TQHXD-CSF+si-ASK1组.综上,TQHXD-CSF可抑制OGD/R损伤的HT22细胞ASK1/MKK4/JNK通路介导的细胞凋亡,对缺血再灌注神经元细胞损伤具有保护作用.  相似文献   

2.
目的观察miR-204-5p在氧糖剥夺/复糖复氧(oxygen-glucose deprivation/reoxygenation, OGD/R)致人神经母细胞瘤细胞(SH-SY5Y)损伤中的作用并从炎症/凋亡途径探讨其机制。方法 SH-SY5Y细胞分为control组、OGD/R组、OGD/R+miR-204-5p mimic组、OGD/R+miR-204-5p inhibitor组、OGD/R+miR-204-5p mimic negative control组、OGD/R+miR-204-5p inhibitor negative control组。采用MTT法测定细胞增殖、流式细胞术、TUNEL染色法检测细胞凋亡、酶联免疫法检测炎症因子(IL-10、IL-1β、TNF-α、PGE2)含量、qRT-PCR检测miR-204-5p的表达、western blot检测炎症及凋亡相关蛋白(COX-2、Bcl-2、Bax)的表达。结果与对照组相比,OGD/R组细胞miR-204-5p表达显著降低,IL-1β、TNF-α、PGE2含量增多,IL-10含量减少,COX-2、Bax蛋白水平上升,Bcl-2蛋白水平下降,细胞损伤明显加重;与OGD/R组比较,miR-204-5p mimic下调COX-2、Bax蛋白表达,上调Bcl-2蛋白表达,IL-10含量增加,IL-1β、TNF-α、PGE2含量减少,细胞活力明显增加、凋亡率显著降低,细胞损伤减轻。结论 miR-204-5p对OGD/R致SH-SY5Y细胞损伤具有明显保护作用,其机制可能与减轻细胞炎症和凋亡有关。  相似文献   

3.
目的观察miR-204-5p在氧糖剥夺/复糖复氧(oxygen-glucose deprivation/reoxygenation, OGD/R)致人神经母细胞瘤细胞(SH-SY5Y)损伤中的作用并从炎症/凋亡途径探讨其机制。方法 SH-SY5Y细胞分为control组、OGD/R组、OGD/R+miR-204-5p mimic组、OGD/R+miR-204-5p inhibitor组、OGD/R+miR-204-5p mimic negative control组、OGD/R+miR-204-5p inhibitor negative control组。采用MTT法测定细胞增殖、流式细胞术、TUNEL染色法检测细胞凋亡、酶联免疫法检测炎症因子(IL-10、IL-1β、TNF-α、PGE2)含量、qRT-PCR检测miR-204-5p的表达、western blot检测炎症及凋亡相关蛋白(COX-2、Bcl-2、Bax)的表达。结果与对照组相比,OGD/R组细胞miR-204-5p表达显著降低,IL-1β、TNF-α、PGE2含量增多,IL-10含量减少,COX-2、Bax蛋白水平上升,Bcl-2蛋白水平下降,细胞损伤明显加重;与OGD/R组比较,miR-204-5p mimic下调COX-2、Bax蛋白表达,上调Bcl-2蛋白表达,IL-10含量增加,IL-1β、TNF-α、PGE2含量减少,细胞活力明显增加、凋亡率显著降低,细胞损伤减轻。结论 miR-204-5p对OGD/R致SH-SY5Y细胞损伤具有明显保护作用,其机制可能与减轻细胞炎症和凋亡有关。  相似文献   

4.
目的:探讨巴利森苷A(PA)对氧糖剥夺/复糖复氧(OGD/R)损伤的HT22细胞的保护作用。方法:将HT22细胞随机分为6组,分别是空白组(Control组)、OGD/R组、依达拉奉(EDA)组、PA低剂量组(40μmol·L-1)、PA中剂量组(80μmol·L-1)、PA高剂量组(160μmol·L-1)。以连二亚硫酸钠(Na2S2O4)10 mmol·L-1合并无糖DMEM培养基造成氧糖剥夺培养2 h,然后恢复为无血清高糖DMEM培养2 h作复糖复氧处理,以建立体外OGD/R模型。EDA和PA组自氧糖剥夺前24 h即加入相应药物的浓度,一直持续到复糖复氧结束。采用MTT比色法检测细胞存活率;乳酸脱氢酶(LDH)法测定细胞损伤率;采用试剂盒法检测超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-PX)活性、丙二醛(MDA)含量及细胞内的活性氧(ROS)水平;Western blot法检测各组细胞缺氧诱导因子通路相关蛋白HIF-...  相似文献   

5.
线粒体作为细胞能量供应的主要场所,是决定缺血后神经细胞生存和死亡的关键靶区。缺血性脑卒中发生时,及时清除受损线粒体对于改善线粒体功能修复神经损伤至关重要。该文旨在研究中药活性成分藁本内酯(ligustilide, LIG)对缺糖缺氧/复氧(oxygen and glucose deprivation/reperfusion, OGD/R)致HT22细胞损伤时线粒体功能及线粒体自噬的影响。通过体外建立OGD/R模型,经藁本内酯预处理HT22细胞3 h后,采用CCK-8法检测细胞活力;免疫荧光法及流式细胞术检测线粒体功能相关指标如线粒体膜电位、钙离子超载情况、活性氧(reactive oxygen species, ROS)含量变化;Western blot法检测线粒体分裂蛋白Drp1及凋亡执行蛋白cleaved caspase-3表达;免疫荧光观察线粒体标志蛋白TOMM20和自噬发生标志物——溶酶体关联膜蛋白2的共定位情况。结果显示,与模型组相比,藁本内酯可以增加HT22细胞的存活率。进一步实验表明,藁本内酯可以抑制OGD/R损伤后HT22细胞内ROS的释放、钙离子超载及线粒体膜电位的下降,同时促进线粒体分裂蛋白Drp1的表达,增加线粒体与溶酶体关联膜蛋白2的共定位。以上结果表明藁本内酯可以改善OGD/R损伤后的线粒体功能,促进线粒体自噬的发生。当给予线粒体自噬抑制剂mdivi-1后,凋亡蛋白表达升高,表明藁本内酯发挥的神经保护作用可能与促进线粒体自噬有关。  相似文献   

6.
目的研究丹灯通脑胶囊含药脑脊液对缺糖缺氧再复糖复氧(OGD/R)损伤大鼠脑微血管内皮细胞(BMECs)的保护作用。方法 SD大鼠每日灌胃给予丹灯通脑胶囊(490 mg/kg)2次,连续4 d,以制备含药脑脊液,原代培养大鼠BMECs,并对其进行Ⅷ因子相关抗原免疫荧光鉴定。然后,利用倒置显微镜观察BMECs形态,CCK-8法观察细胞活力,通过测定跨内皮细胞电阻(TEER)来评价BMECs通透性变化,取细胞上清检测BMECs总一氧化氮(NO)和内皮素1(ET-1)水平,以及BMECs组织型纤溶酶原激物(t PA)、纤溶酶原激活剂抑制物1(PAI-1)水平。结果含药脑脊液可显著对抗OGD/R所造成的损伤,明显改善BMECs形态。与模型组比较,含药脑脊液组细胞活力显著升高,TEER值显著增加,细胞通透性降低,细胞培养上清液中总NO、t PA水平明显增加,ET-1、PAI-1是水平显著降低。结论丹灯通脑胶囊含药脑脊液对OGD/R损伤大鼠BMECs具有保护作用。  相似文献   

7.
目的研究苦碟子注射液对氧糖剥夺/复氧(OGD/R)损伤大鼠C6胶质细胞的保护作用及对IκBα/NF-κB蛋白活化的影响。方法将C6胶质细胞分为空白对照组、OGD/R模型组、苦碟子注射液组,采用OGD/R方法建立C6胶质细胞损伤模型。MTT法确定氧糖剥夺/复氧时间和苦碟子注射液的最佳有效浓度;LDH漏出率检测细胞死亡率;Western Blot法检测p-IκBα/IκBα、p-NF-κB/NF-κB蛋白表达。结果 C6胶质细胞OGD/R时间为OGD24h/R6h,苦碟子注射液最佳浓度为1.25%。OGD/R损伤后,细胞IκBα和NF-κB的表达上调及磷酸化蛋白增加;苦碟子注射液能够显著降低细胞死亡率,降低细胞p-IκBα和p-NF-κB蛋白高表达。结论 OGD/R损伤C6胶质细胞,促进了IκBα/NF-κB磷酸化,苦碟子注射液可抑制C6胶质细胞中IκBα/NF-κB磷酸化活化,降低细胞死亡率,对胶质细胞起到保护作用。  相似文献   

8.
目的:研究PC12细胞OGD/R后自噬与损伤的经时性变化以及黄芪甲苷的干预效应。方法:探讨PC12细胞OGD/R后不同时间自噬的变化及与细胞损伤的关系,初步确定细胞发生自噬性损伤的时间点,再制作PC12细胞OGD/R自噬性损伤模型,研究黄芪甲苷抗自噬性损伤的作用。结果:复糖复氧6~36 h后,细胞存活率降低、LDH漏出率增加;用3-MA预处理后,可使复糖复氧6~12 h细胞存活率降低、LDH漏出率增加,复糖复氧后24~36 h相反。激光共聚焦和western-blot检测显示,复糖复氧后6 h LC3、LC3-Ⅱ/LC3-Ⅰ比值增加,至24 h达高峰;p62蛋白表达随再复糖复氧时间的延长逐渐降低。黄芪甲苷对OGD 2 h复糖复氧24 h的PC12细胞自噬具有显著抑制作用,且呈剂量依赖性。结论:PC12在在OGD复糖复氧6~12 h,自噬减轻神经细胞的损伤,而在24~36 h后加重细胞损伤;黄芪甲苷可通过抑制细胞过度自噬诱导的细胞损伤,从而发挥对受损细胞的神经保护作用。  相似文献   

9.
目的探讨毛蕊异黄酮苷(CG)通过调控沉默信息调节因子1(SIRT1)信号通路对氧糖剥夺再灌注(OGD/R) 海马神经元细胞损伤的保护作用。方法将海马神经元细胞在无糖培养基中缺氧8 h 后,在完全培养基中复氧 6 h 建立OGD/R 模型。将细胞分为4 组:正常对照组、氧糖剥夺再灌注组(OGD/R 组,模型组)、SIRT1 特异性 抑制剂组(EX527 组)、EX527+毛蕊异黄酮苷组(EX527+CG 组)。细胞处理结束后,采用CCK-8 法测定细胞存 活率;分光光度法检测细胞上清液中乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)和丙二醛(MDA)的含量;荧光 法检测细胞中活性氧(ROS)含量、细胞凋亡率;Western Blot 及ELISA 法检测SIRT1 信号通路中相关蛋白的表 达水平。结果与OGD/R 组比较,EX527 组的细胞活力、SOD 含量及SIRT1、叉头转录因子1(FOXO1)、B 淋 巴细胞瘤-2(Bcl-2)、过氧化物酶体增殖物激活受体γ 共激活因子-1α(PGC-1α)蛋白表达水平均明显降低(P< 0.05),LDH 及MDA 含量、细胞凋亡率、Bax 蛋白表达水平明显升高(P<0.05)。与EX527 组比较,EX527+CG 组的细胞活力及FOXO1、Bcl-2 和PGC-1α 蛋白表达水平均有升高,SOD 含量及SIRT1 蛋白表达水平明显升高 (P<0.05);MDA 含量有降低,LDH 含量、细胞凋亡率及Bax 蛋白表达水平明显降低(P<0.05)。结论毛蕊 异黄酮苷缓解海马神经元OGD/R 损伤的机制与调控SIRT1 信号通路有关。  相似文献   

10.
[目的]探讨心脑舒通胶囊对氧糖剥夺/复氧(OGD/R)损伤的原代神经元细胞活力及微管相关蛋白2(MAP-2)表达的影响。[方法]取新生24 h内SD大鼠的大脑皮质进行神经元原代培养,并通过免疫荧光法进行MAP-2目的蛋白鉴定;建立神经元OGD/R损伤的体外模型,CCK-8检测不同浓度的心脑舒通胶囊对体外正常培养和OGD/R损伤后神经元细胞活力的影响;选取最佳浓度的心脑舒通胶囊进行MAP-2的荧光表达强度实验,通过倒置荧光显微镜进行拍照处理,并通过软件对神经元突触长度进行测量及统计处理,Western blot法检测MAP-2的蛋白表达情况。[结果]神经元原代培养成功,且MAP-2染色阳性。CCK-8结果显示心脑舒通胶囊在10、25、50μg/mL浓度对正常培养神经元细胞有细胞毒作用(P<0.05),因此选取低于10μg/mL浓度的心脑舒通胶囊进行后续实验。而在OGD/R损伤条件下,与模型组比较,心脑舒通胶囊在3.125、6.25μg/mL浓度能显著增加OGD/R损伤神经元的细胞活力(P<0.05);免疫荧光实验结果显示,与对照组比较,模型组MAP-2荧光表达强度及神经元突触长度明显降低(P<0.05);与模型组比较,心脑舒通胶囊在3.125μg/mL浓度能显著增加MAP-2的荧光表达强度及神经元突触长度(P<0.05)。Western blot实验结果显示,与对照组比较,模型组MAP-2蛋白表达显著降低(P<0.05),与模型组比较,心脑舒通胶囊能显著增加MAP-2的蛋白表达(P<0.05)。[结论]心脑舒通胶囊能促进OGD/R损伤神经元活力,其作用机制与促进细胞骨架蛋白MAP-2表达及突触稳定性有关。  相似文献   

11.
汤卫国  王奇志  印敏  王鸣  陈雨  张建华  冯煦 《中草药》2014,45(15):2172-2177
目的建立微波消解-电感耦合等离子体质谱(ICP-MS)直接稀释测定脉络宁注射液中25种矿物质元素(Mg、Ca、Fe、Cu、Zn、Mn、Al、B、Ba、Co、Cr、K、Li、Mo、Na、Ni、P、Pb、Sr、Th、Ti、V、As、Cd和Hg)的方法。方法分别对微波消解条件和测试条件进行考察;样品经微波消解后,采用电感耦合质谱仪测定25种矿物质元素,并对测定方法学进行考察。结果确定最佳消解条件为3步缓慢升温:400 W 80℃升温10 min,保留5 min;600 W 120℃升温10 min,保留5 min;900 W 200℃升温20 min,保留20 min;25种矿物质元素在各自的线性范围内线性关系良好,r≥0.999 6,精密度、稳定性和重复性试验的RSD均符合定量分析要求;加标回收率为94.7%~106.1%,RSD在0.34%~2.79%。脉络宁注射液中检测出Mg、Ca、Fe、Cu、Zn、Mn、Al、B、Ba、Co、Cr、K、Li、Mo、Na、Ni、P、Pb、Sr、Th、Ti、V,未检出As、Cd和Hg。结论该方法简便、迅速、准确,适用于脉络宁注射液中25种矿物质元素的同时测定。  相似文献   

12.
The disturbance and instability of laterality are obstacles to diagnostics and therapy. Correction prior to starting therapy is required. They also predispose toward defined health problems and unspecific diseases. Numerous research activities provide evidence of the relevance of undisturbed laterality in diagnostics and therapy. Techniques of testing and therapeutic corrections will allow for optimized therapy success.  相似文献   

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14.
The Journal of Acupuncture and Tuina Science, a journal with an international scope (IS SN 1672-3597, CN 31-1908/R, Bimonthly), is embodied by 'Springer Verlag' Database, Index Copernicus (IC) and Chinese Scientific and Technical Paper and Citations Data (CSTPCD). You can search full text on http://www, springerlink, com/content/1672 -3597.  相似文献   

15.
正Beverages based on Ilex paraguariensis A.St.-Hil.are used in the south Brazil and other Latin American countries located at the so-called southern cone.It is known as chimarro or mate in south and southeast Brazil,tererêin  相似文献   

16.
This paper deals with an interdisciplinary study covering historic, botanical, phytochemical, pharmacological and clinical aspects of rhubarb and related species, to lay stress on the correlation between plant phylogeny, chemical constituents and purgative activity.

It was found that the official rhubarbs were exclusively restricted in the Sect. Palmata e.g. Rheum palmatum R. palmatum var. tanguticum R. officinale; the following criteria may serve as their standard, viz., the presence of sennoside derivatives and rhein, the occurrence of the reduced form of rhein and aloe-emodin, the leaves with any kind of palmate incision. Comprehensive multivariate analyses showed that there is a very close relationship between the leaf incision, existence of sennosides or rhein and purgative activity.  相似文献   


17.

Background

At present, the state-of-the-art medication in treating idiopathic taste disorder (gustatory dysfunction) is zinc. If zinc medication was unsuccessful, not tolerated or if it is contraindicated the persons affected can hardly be helped.

Objectives

In a randomized placebo-controlled clinical trial an attempt was made to determine the efficacy of acupuncture.

Methods

The treatment group was treated with acupuncture needles, the placebo group with an inactive acupuncture-laser. Endpoints of the study were changes in the taste test, the quality of life and the severity of symptoms of depression (BDI) as well as mood changes (ZMS). Satisfaction with the success of the respective therapy was also evaluated. Data were collected before and after therapy and again for the treatment group 10 weeks after completion of the treatment period.

Findings

Treatment with real acupuncture showed a significant improvement in gustatory function as well as a significant improvement in psychological symptoms. Greater satisfaction with the treatment results was evident in the treatment group. The treatment results of acupuncture remained stable over a period of ten weeks after completion of the treatment.

Conclusion

Acupuncture was effective in the therapy of idiopathic taste disorders and therefore it is an effective alternative when zinc medication is not tolerated or unsuccessful. But it is also suitable as a therapy of first choice. Acupuncture can be considered as an extension of the therapeutic spectrum in the difficult treatment of taste disorders.  相似文献   

18.

Background

Experimental and clinical studies postulate a neurobiological mechanism of acupuncture by modulating central pain systems, neurohormones and -transmitters. It seems to be quite likely that the autonomic nervous system (ANS) plays an important mediating role in this neuromodulation.

Aim

In measuring heart rate variability (HRV), as a non-invasive approach to the ANS-function, the object of this study was to monitor probable acupuncture-induced changes of autonomic balance. The innovative technology of laser-needle acupuncture offers for the first time a stimulation method which fulfils the demands on a placebo-controlled acupuncture trial not requiring controversial sham acupuncture.

Method

19 healthy voluntary subjects underwent 3 treatments in randomized sequence, consisting of placebo-, 1-point- (PC 6) and simultaneous 3-point- (PC 6, LR 3, SI 3) laser-needle-stimulation over 20 minutes. Before, during and after placebo-/verum laser-needle acupuncture, HRV-data for normalized HF(high frequency)- and LF(low frequency)-Power (nHF, nLF) and their quotient nLF/nHF obtained by power spectrum analysis (FFT), were measured and analyzed statistically at 5 different time points (ANOVA repeated measures, p < 0,05).

Results

During both verum applications, the 1-point- (PC 6) as well as the simultaneous 3-point- (PC 6, LR 3, SI 3) laser-needle stimulation, significant changes of the ANS-activity were found in comparison to the placebo application. The most significant modification of HRV was an increase of nHF (p < 0,05 (PC 6), p < 0,0001 (PC 6, LR 3, SI 3)) as an expression of growing vagal activity during acupuncture.

Conclusion

Laser-needle acupuncture causes in 1-point-stimulation (PC 6) as well as in simultaneous 3-point-stimulation (PC 6, LR 3, SI 3) significant changes in HRV in comparison to placebo application. HRV-monitoring seems to be a suitable approach in exploration of acupuncture-induced changes of ANS and could possibly be established in combination with laser needle acupuncture as a standard for further scientific and clinical acupuncture investigations which are greatly needed.  相似文献   

19.
瑞香科植物广泛分布于世界热带和温带地区,其中多种植物可供药用。在对该科植物的研究中发现,黄酮类、木脂素和香豆素等酚性化合物的低聚体为其重要的特征性成分,药理活性主要表现在抗肿瘤、抗炎、抗氧化、抗菌和抗病毒等方面。对已发现的酚性聚合体的化学结构、植物来源及药理活性进行综述,为瑞香科植物今后的研究和开发利用提供参考。  相似文献   

20.
目的采用HPLC梯度洗脱法同时测定沉香化滞丸中沉香四醇、柚皮苷、橙皮苷、新橙皮苷、和厚朴酚、大黄素、厚朴酚、木香烃内酯、去氢木香内酯、大黄酚、大黄素甲醚11种成分。方法采用Thermo Syncronis C18色谱柱(250 mm×4.6 mm,5μm),流动相为水-乙腈,梯度洗脱:0~10 min,20%乙腈;10~20 min,20%~40%乙腈;20~24 min,40%乙腈;24~26 min,40%~52%乙腈;26~30 min,52%乙腈;30~31 min,52%~90%乙腈;31~35 min,90%乙腈;35~40 min,90%~100%乙腈;40~43min,100%乙腈;43~45min,100%~20%乙腈;检测波长215nm,体积流量1.0m L/min,柱温30℃,进样量20μL。结果各成分在43 min内分离良好,沉香四醇、柚皮苷、橙皮苷、新橙皮苷、和厚朴酚、大黄素、厚朴酚、木香烃内酯、去氢木香内酯、大黄酚、大黄素甲醚的线性范围分别为1.4~13.6、10.0~200.0、31.5~315.0、1.0~120.1、1.8~50.6、0.93~10.1、1.8~30.0、0.2~40.3、1.8~18.1、1.7~25.0、0.45~10.70μg/mL;样品中各成分的平均回收率均在98.90%~100.87%;11种成分精密度RSD在0.55%~1.54%;供试品溶液在30 h内稳定性良好,RSD在0.75%~1.94%;重复性RSD在0.39%~1.73%。6批次样品中沉香四醇、柚皮苷、橙皮苷、新橙皮苷、和厚朴酚、大黄素、厚朴酚、木香烃内酯、去氢木香内酯、大黄酚、大黄素甲醚质量分数分别为92.0~201.0、511.5~9 033.0、5 475.0~12 635.5、54.5~5 095.5、192.0~2 137.5、117.0~391.5、106.5~1 281.5、13.0~136.5、93.5~199.0、177.0~1 207.0、33.5~251.5μg/g。结论本方法准确、快速、简便,重复性好,精密度高,适用于沉香化滞丸中多种活性成分的定量分析。  相似文献   

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