��ζ�ӷӶ�H2O2�յ�SH-SY5Y ϸ���������˵ı�������

??OBJECTIVE To investigate the protective effect and possible mechanism of schisanhenol(Sal) in SH-SY5Y cell induced by H₂O₂. METHODS SH-SY5Y cells were treated with Sal (1,10 and 50 ??mol??L^-1) for 4 h and then exposed to H₂O₂ 100 ??mol??L^-1 for 24 h. Cell viability was measured by MTT assay. The expressions of silent information regulator 1(SIRT1), PGC-1?? and p-tau (S396) protein were detected using Western blotting. RESULTS MTT results showed that Sal significantly increased the survival rate of SH-SY5Y cell damaged by H₂O₂. Western blotting analysis showed that H₂O₂ reduced the expressions of SIRT1 and PGC-1?? in SH-SY5Y cells. However, tau protein content was increased by H₂O₂ at p-tau(S396) sites. Sal treatment significantly increased the levels of SIRT1 and PGC-1?? and decreased p-tau(S396) level induced by H₂O₂ in SH-SY5Y cells. CONCLUSION These results suggest that Sal has a protective effect on H₂O₂ damaged SH-SY5Y cells, which is related to up regulating the expressions of SIRT1 and PGC-1?? protein and decreasing the phosphorylation of tau protein.     

miR-204-5p在氧糖剥夺/复糖复氧致SH-SY5Y细胞损伤中的作用研究

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。     

miR-204-5p在氧糖剥夺/复糖复氧致SH-SY5Y细胞损伤中的作用研究

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。     

竹叶提取物抗肿瘤和神经保护作用研究

目的研究竹叶提取物抑制肿瘤细胞增殖的活性及其分子生物学机制。方法选取血管内皮生长因子(VEGF)诱导型肿瘤细胞人肝癌细胞Hep G2和不诱导型肿瘤细胞人神经母细胞瘤细胞SH-SY5Y为模型,用不同质量浓度竹叶提取物处理Hep G2和SH-SY5Y细胞。采用MTT法检测竹叶提取物对2种细胞增殖的抑制作用;采用RT-PCR法检测VEGF和血管内皮生长因子受体2(VEGFR2)基因在Hep G2和SH-SY5Y细胞中的表达差异以及竹叶提取物处理后Hep G2细胞中VEGF基因表达的变化;用荧光倒置显微镜观察竹叶提取物处理后SH-SY5Y细胞神经突触的生长情况;采用RT-PCR法检测竹叶提取物处理后SH-SY5Y细胞中神经生长因子(NGF)表达的变化。结果竹叶提取物对Hep G2细胞增殖的抑制活性高于对SH-SY5Y细胞的增殖抑制作用。进一步研究发现Hep G2细胞中VEGF和VEGFR2基因的表达显著高于SH-SY5Y细胞,且竹叶提取物可显著抑制Hep G2细胞的VEGF表达。适当剂量的竹叶提取物可以刺激SH-SY5Y细胞神经突触的生长,并可上调其NGF的表达。结论竹叶提取物可能通过抑制VEGF的表达影响VEGF诱导型肿瘤细胞的增殖,同时,可能通过上调NGF的表达刺激神经细胞神经突触的生长,竹叶提取物可能具有潜在的抗肿瘤和神经保护的双重功效。     

钾通道在MPP+诱导的α-synuclein转基因细胞凋亡中的作用

 目的帕金森氏病(PD)的主要病理表现为中脑黑质致密部多巴胺能神经元的进行性死亡,为了揭示帕金森氏病中多巴胺能神经元死亡的机制,我们研究了钾离子通道在MPP⁺(1-甲基-4-苯基吡啶)诱导的野生型及α-synuclein转基因SH-SY5Y细胞死亡中的作用。方法MPP⁺孵育野生型及转染A53T突变型α-synuclein基因的SH-SY5Y细胞,或提前孵育钾离子通道抑制剂,全细胞膜片钳技术观察钾电流的改变。结果①MPP⁺可以诱导野生型及转染α-synuclein基因的SH-SY5Y细胞中延迟整流钾电流[I_K(DR)]的增加,并且在α-synuclein转基因细胞中I_K(DR)的增加更显著;②MPP⁺孵育野生型及转染α-synuclein的SH-SY5Y细胞6h即可诱导Ik电流的增加,24～48h Ik电流增加加剧;③四乙铵(TEA)可以抑制MPP⁺诱导的野生型和α-synuclein转基因SH-SY5Y细胞中钾电流的增加。结论钾离子通道的开放及延迟整流钾电流的增加在MPP⁺诱导的帕金森氏病细胞模型中发挥重要作用,为揭示PD中多巴胺能神经元的凋亡提供了新的思路。     

当归芍药散通过调控NF-κB炎性通路改善H2O2诱导的SH-SY5Y细胞氧化损伤的作用

目的:研究当归芍药散(Danggui Shaoyaosan,DSS)含药血清对过氧化氢(H2O2)诱导人神经母细胞瘤细胞(SHSY5Y)氧化应激和炎症反应的调控作用。方法:采用噻唑蓝(MTT)比色法测定SH-SY5Y细胞存活率构建最佳时间和最佳剂量的H2O2诱导的细胞损伤模型。实验设空白组,模型组,当归芍药散含药血清高、中、低剂量组(2.5%,5%,10%)。采用MTT比色法检测DSS干预后细胞活率,分光光度法测定抗氧化指标丙二醛(MDA),过氧化氢酶(CAT),超氧化物歧化酶(SOD)和谷胱甘肽(GSH),采用实时荧光定量聚合酶链式反应(Real-time PCR)检测肿瘤坏死因子-α(TNF-α),白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)mRNA表达水平,采用免疫荧光检测核转录因子-κB(NF-κB)p65的核移位情况。结果:H2O2浓度为250μmol·L-1干预24 h后SH-SY5Y细胞存活率约55%为最佳造模条件,DSS高、中、低剂量干预后,与模型组比较,细胞活率呈剂量依赖性上升(P<0.05),MDA明显下调(P<0.05),抗氧化指标CAT,SOD和GSH明显上升(P<0.05);H2O2能显著上调SH-SY5Y细胞炎性因子TNF-α,IL-1β和IL-6 mRNA表达水平,并介导胞质NF-κB的激活和向核移位,当归芍药散含药血清能呈剂量依赖性抑制NF-κB p65的入核并降低IL-1β,IL-6和TNF-α等细胞炎性因子的mRNA表达水平。结论:当归芍药散含药血清可以显著降低H2O2诱导的SH-SY5Y细胞的氧化损伤通过改善其抗氧化状态,同时也能降低炎症反应通过抑制NF-κB信号通路。     

参知健脑方组分对淀粉样β蛋白β25-35诱导的SH-SY5Y细胞凋亡的保护作用

目的:探讨SZJ对淀粉样β蛋白25-35(Aβ25-35)诱导的SH-SY5Y细胞凋亡的保护作用机制。方法:以Aβ25-35诱导SH-SY5Y细胞凋亡,采用MTT比色分析测定细胞存活率,应用Western blot检测Aβ25-35以及SZJ/APP17肽对SH-SY5Y细胞Bax、Bcl-2、Caspase-3表达的影响。结果:25μmol/L的Aβ25-35减低SH-SY5Y细胞存活率,SZJ/APP17肽预处理24h可提高SH-SY5Y细胞存活率,相同浓度Aβ25-35 SZJ预处理组与非处理组比较差异显著(P<0.05);25μmol/L Aβ25-35处理的SH-SY5Y细胞Bax表达增加,Bcl-2表达减少,Caspase-3的表达增高,SZJ/APP17肽预处理能抑制其表达升高,与非预处理组比较差异显著(P<0.05)。结论:SZJ可对抗Aβ25-35对SH-SY5Y细胞的毒性作用,其机制可能与抑制Bax、Caspase-3的表达,促进Bcl-2的表达有关。     

海康灵含药血清在体外对SH-SY5Y神经细胞损伤的保护作用

 目的观察海康灵含药血清(HKL-serum)对SH-SY5Y神经细胞损伤的保护作用。方法采用体外细胞培养的方法,建立SH-SY5Y神经细胞过氧化氢(H₂O₂)损伤模型。通过观察细胞形态,测定细胞存活率(MTT法)、乳酸脱氢酶(LDH)漏出率,检测HKL-serum对SH-SY5Y神经细胞损伤的保护作用。结果同造模组比较,10%,15%HKL-serum能减轻H₂O₂引起的SH-SY5Y神经细胞的损伤,明显提高细胞的存活率,减少LDH的释放量。结论结果显示,HKL-serum对SH-SY5Y神经细胞损伤具有明显的保护作用。     

�컨��ɫ�ضԸ������յ�������ĸϸ����SH-SY 5Yϸ��Tau�������ữ�ĵ��������о�

??OBJECTIVE To investigate the regulating effects and its possible mechanism of safflower yellow (SY) on tau protein hyperphosphorylation in SH-SY 5Y cell induced by okadaic acid (OA). METHODS Cell viability was measured by MTT colorimetric method. The cell apoptosis was determined with Hoechst 33342 after SH-SY 5Y cell were treated with SY. To determined the phosphorylated tau protein of p-tau(T205), p-tau(S199) sites, and total GSK-3β, PP2A in SH-SY 5Y cell were determined using Western blotting. RESULTS SY significantly increased survival rates of SH-SY5Y cell damaged by OA. Hoechst 33342 showed that SY decreased the apoptosis of SH-SY5Y cells. Western blotting analysis showed that tau protein content was increased compared to control group at p-tau (T205), p-tau (S199) sites. However, SY treatment significantly reduced tau protein content at p-tau (T205) and p-tau (S199) sites. PP2A content was significantly suppressed by OA, significantly increased by SY, GSK-3β content was significantly increased by OA and significantly decreased by SY. CONCLUSION These results suggest that safflower yellow may have neuroprotective effect on OA-induced SH-SY 5Y cells injury, which may be associated with tau hyperphosphorylation at the enzyme level by activation of tau kinases or by a decline in the activities of tau phosphatases.     

注射用血塞通对SH-SY5Y细胞缺糖缺氧损伤模型Lingo-1表达的影响

目的建立SH-SY5Y细胞缺糖缺氧损伤模型,考察注射用血塞通对缺糖缺氧损伤SH-SY5Y细胞存活、凋亡及Lingo-1 mRNA及蛋白表达的影响。方法采用RPMI 1640无糖培养基和三气培养箱缺氧法制备SH-SY5Y细胞缺糖缺氧损伤模型,CCK8法测定细胞存活率,确定最佳缺氧时间及最适注射用血塞通质量浓度,Annexin V-FITC/PI双染法检测SH-SY5Y细胞凋亡率,并通过实时荧光定量PCR(qRT-PCR)方法测定Lingo-1 mRNA表达情况,Western blotting法测定Lingo-1蛋白表达情况。结果 SH-SY5Y细胞缺糖缺氧损伤模型的最佳缺氧时间为16 h,注射用血塞通最适质量浓度为640 mg/L,在该质量浓度条件作用下,血塞通组与模型组相比,SH-SY5Y细胞的凋亡率显著降低,Lingo-1的m RNA表达水平及蛋白表达水平均显著降低。结论当SH-SY5Y细胞受到缺糖缺氧损伤后,细胞凋亡率显著升高,Lingo-1呈高表达状态,血塞通可以明显减少缺糖缺氧损伤SH-SY5Y细胞的凋亡,同时抑制Lingo-1的高表达,具有抑制凋亡和显著的神经保护作用。     

中药复方海康灵含药血清对过氧化氢诱导的神经细胞凋亡的影响

 目的观察海康灵对SH-SY5Y神经细胞凋亡、细胞内线粒体膜电位和钙离子的影响。方法采用血清药理学方法,用流式细胞仪观察海康灵对过氧化氢(H₂O₂)损伤的SH-SY5Y神经细胞凋亡、细胞线粒体膜电位和钙离子的影响。结果与模型组比较,海康灵含药血清能明显抑制H₂O₂引起的SH-SY5Y神经细胞的凋亡,稳定线粒体膜电位,抑制钙离子内流。结论以上结果提示,海康灵可通过抑制胞浆钙离子升高,进而稳定线粒体膜电位,从而发挥其抑制神经细胞凋亡的作用。     

Protection against 1-methyl-4-phenylpyridinium ion (MPP+)-induced apoptosis by water extract of ginseng (Panax ginseng C.A. Meyer) in SH-SY5Y cells

Ethnopharmacological relevance

The present study investigates the protective effects of water extract of ginseng (Panax ginseng C.A. Meyer) against 1-methyl-4-phenylpyridinium ion (MPP⁺)-induced cytotoxicity in SH-SY5Y human neuroblastoma cells and explores the underlying mechanisms. The approach may be used for screening therapeutic agents for degenerative disorders such as Parkinson's disease.

Materials and methods

SH-SY5Y human neuroblastoma cells were used to analyze the protective effects of water extract of ginseng (WEG) against multiple parameters such as MPP⁺-induced viability, oxidative injury, expression of Bax, Bcl-2, cytochrome c and cleaved caspase-3.

Results

WEG exerted inhibitory effect on cell death, overproduction of ROS, elevated Bax/Bcl-2 ratio, release of cytochrome c and activation of caspase-3 expression in MPP⁺-treated SH-SY5Y cells.

Conclusions

WEG exhibited significant protective effects against MPP⁺-induced cytotoxicity in SH-SY5Y cells possibly through the suppression of ROS generation and the inhibition of mitochondria-dependent apoptotic pathway.     

The Novel p53‐Dependent Metastatic and Apoptotic Pathway Induced by Vitexin in Human Oral Cancer OC2 Cells

Vitexin, identified as apigenin‐8‐C‐D‐glucopyranoside, a natural flavonoid compound found in certain herbs such as hawthorn herb, has been reported to exhibit anti‐oxidative, anti‐inflammatory, anti‐metastatic and antitumor properties. The aim of this study was to investigate the possible existence of p53‐dependent pathway underlying vitexin‐induced metastasis and apoptosis in human oral cancer cells, OC2 cells. Vitexin decreased cell viability significantly. Meanwhile, the expression of tumor suppressor p53 and a small group of its downstream genes, p21 ^WAF1 and Bax, were upregulated. The p53 inhibitor pifithrin‐α (PFT‐α) knockdown of the signaling of p53 led vitexin to lose its antitumor effect and inhibited the expression of p53 downstream genes, p21^WAF1 and Bax. Vitexin had anti‐metastatic potential accompanied with increasing plasminogen activator inhibitor 1 (PAI‐1) accumulation and decreasing matrix metalloproteinase‐2 expression. Our present study evidenced, by using p53 inhibitor PFT‐α, PAI‐1 and peroxisome proliferator‐activated receptor γ are downstream genes of p53 in vitexin‐induced signaling. MAPK inhibitor PD98059 decreased the OC2 cells viability significantly. The expression of p53 and its downstream genes p21 ^WAF1 and Bax were enhanced by blocking the activation of p42/p44 MAPK in response to treatment with vitexin. Moreover, p42/p44 MAPK played a negative role in p53‐dependent metastasis and apoptosis. We give evidence for the first time that the novel p53‐dependent metastatic and apoptotic pathway induced by vitexin in human oral cancer OC2 cells. Copyright © 2012 John Wiley & Sons, Ltd.     

葛根素对MPP+诱导的SH-SY5Y细胞线粒体 途径凋亡的保护作用

目的:探讨葛根素对MPP+诱导SH-SY5Y细胞损伤的保护机制.方法:台盼蓝染色检测细胞存活率;PI单染流式细胞术分析细胞凋亡率、细胞周期分布;ELISA和Western blotting法检测胞浆Cyt C的变化;ELISA方法检测Caspase-3,Caspase-8和Caspase-9酶活性;DNA ladder分析药物对细胞凋亡的干预作用.结果:在12.5～50 μmol·L-1,葛根素对MPP+诱导的SH-SY5Y细胞损伤具有一定的保护作用,并呈一定的量效关系;PI单染流式细胞术检测结果显示MPP+诱导SH-SY5Y细胞发生凋亡,并阻滞细胞于G2/M期,预先加入不同剂量的葛根素可以逆转MPP+对SH-SY5Y细胞损伤作用.Western blotting和ELISA分析结果均显示葛根素能够抑制MPP+诱导SH-SY5Y细胞的Cyt C的释放.不同剂量的葛根素也能够抑制MPP+诱导SH-SY5Y细胞的Caspase-3和Caspase-9的活性,但对Caspase-8的活性影响不明显;DNA ladder分析也显示葛根素可以抑制MPP+诱导SH-SY5Y细胞凋亡.结论:葛根素可以提高MPP+诱导的SH-SY5Y损伤的细胞存活率,抑制细胞凋亡的发生,其机制可能与调节线粒体Caspase通路和干预细胞周期有关.     

Activation of p53 signaling and inhibition of androgen receptor mediate tanshinone IIA induced G1 arrest in LNCaP prostate cancer cells

Our group previously reported that tanshinone IIA induced apoptosis via a mitochondria dependent pathway in LNCaP prostate cancer cells. In the present study, the roles of androgen receptor (AR) and p53 signaling pathways were investigated in tanshinone IIA-induced G1 arrest in LNCaP cells. Tanshinone IIA significantly inhibited the growth and proliferation of LNCaP cells by colony formation and BrdU incorporation assays, respectively. Tanshinone IIA induced cell cycle arrest at G1 phase and down-regulated cyclin D1, CDK2 and CDK4. Furthermore, tanshinone IIA activated the phosphorylation of p53 at Ser 15 residue and its downstream p21 and p27. Additionally, tanshinone IIA suppressed the expression of AR and prostate specific antigen (PSA). Conversely, silencing p53 using its specific siRNA reversed cyclin D1 expression inhibited by tanshinone IIA. However, knockdown of AR had no effect on the p53/p21/p27 signaling pathway activated by tanshinone IIA in LNCaP cells. In AR siRNA-transfected cells, tanshinone IIA did not cause cell cycle arrest and reduce cyclin D1, implying that AR is essential to induce G1 arrest by tanshinone IIA in LNCaP cells. Taken together, the findings suggest that tanshinone IIA induces G1 arrest via activation of p53 signaling and inhibition of AR in LNCaP cells.     

半边旗提取物5F诱导HepG2细胞凋亡与p53活化及血管内皮生长因子抑制有关

目的观察半边旗提取物Ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic-acid(5F)对人肝癌细胞HepG2增殖的影响,并探讨p53、血管内皮生长因子(VEGF)及Caspase-3在5F诱导的HepG2细胞凋亡中的作用。方法采用MTT分析检测5F对HepG2细胞增殖的影响,并通过细胞凋亡检测ELISA试剂盒分析经5F处理的HepG2细胞胞浆核小体片段,以确定5F能否诱导细胞凋亡,采用Hoechst/PI分析鉴定凋亡细胞核形态。通过免疫印迹(Western blotting)分析测定p53及VEGF蛋白表达水平,并通过Caspases-3分光光度法检测试剂盒检测Caspase-3活性。结果通过细胞活性分析证实,5F对HepG2的细胞毒作用随着5F质量浓度的升高而增强。5F诱导HepG2产生胞浆核小体片段,并且该诱导作用具有剂量依赖性。5F处理后,凋亡变化,如染色质浓缩,被Hoechst/PI染色所确定。5F处理后HepG2细胞核内p53表达水平显著提高,而胞浆VEGF表达水平却下降,同时,Caspase-3活性通过浓度依赖方式增强。结论5F所诱导的HepG2细胞凋亡与p53及Caspase-3活化、VEGF负调控有关。5F可能具有抗癌尤其是抗肝细胞癌价值。     

SuHeXiang Wan essential oil alleviates amyloid beta induced memory impairment through inhibition of tau protein phosphorylation in mice

SuHeXiang Wan (SHXW), a traditional Chinese medicine, has been used orally for the treatment of seizures, infantile convulsions and stroke. Previously, we reported the effects of a modified SHXW essential oil in terms of sedative effect, anticonvulsant activity and antioxidative activity. The purpose of this study was to evaluate the potential beneficial effects of SHXW essential oil in neurodegenerative diseases such as Alzheimer's disease (AD). SHXW essential oil was extracted from nine herbs. The mouse AD model was induced by a single injection of amyloid β protein (Aβ(1-42)) into the hippocampus. The animals were divided into four groups, the negative control group injected with Aβ(42-1), the Aβ group injected with Aβ(1-42), the SHXW group inhaled SHXW essential oil and received Aβ(1-42) injection, and the positive control group administered with docosahexaenoic acid (DHA, 10 mg/kg) and with subsequent Aβ(1-42) injection. Mice were analyzed by behavioral tests and immunological examination in the hippocampus. An additional in vitro investigation was performed to examine whether SHXW essential oil inhibits Aβ(1-42) induced neurotoxicity in a human neuroblastoma cell line, SH-SY5Y cells. Pre-inhalation of SHXW essential oil improved the Aβ(1-42) induced memory impairment and suppressed Aβ(1-42) induced JNK, p38 and Tau phosphorylation in the hippocampus. SHXW essential oil suppressed Aβ-induced apoptosis and ROS production via an up-regulation of HO-1 and Nrf2 expression in SH-SY5Y cells. The present study suggests that SHXW essential oil may have potential as a therapeutic inhalation drug for the prevention and treatment of AD.     

五味子醇甲对Aβ1-42诱导SH-SY5Y细胞的RAGE-ROS-凋亡通路的影响

目的探讨五味子醇甲(Schisandrin,SCH)对Aβ1-42诱导SH-SY5Y的RAGE/ROS/凋亡通路的影响。方法MTT法检测control组、SCH高中低剂量组的细胞活力,探讨SCH高中低剂量组对SH-SY5Y细胞的保护作用。MTT法检测control组、Aβ组、Vitamin E组及SCH高中低剂量组细胞活力、双染法观察细胞凋亡和DHE染色法检测ROS含量,探讨SCH对Aβ1-42诱导的SH-SY5Y的细胞损伤的保护作用。Western-blot法检测control组、Aβ组、SCH组RAGE蛋白,探讨SCH对Aβ1-42诱导SH-SY5Y的RAGE影响。结果与control组比较,SCH各剂量组细胞活力均显著增强(P<0.01)。与Aβ组比较,各给药组细胞活力均明显增强(P<0.01)和细胞凋亡率均明显下降(P<0.01);各给药组检测ROS的荧光强度均减弱(P<0.01),SCH(5.0μg/mL)组RAGE蛋白表达降低,有显著性差异(P<0.05)。结论SCH防治阿尔茨海默病的抗氧化作用机制可能与降低ROS含量、细胞凋亡率和下调RAGE蛋白表达有关。     

青蒿酯钠诱导人肿瘤细胞凋亡及其分子机制的探讨

目的 研究青蒿酯钠诱导人肝癌细胞凋亡作用及探讨青蒿酯钠诱导人肝癌细胞凋亡的分子机制.方法 肝癌细胞(BEL-7402)经药物处理后,用荧光显微镜,透射电镜和流式细胞仪分析诱导凋亡作用,采用Western blot检测p53,p21和bcl-2.结果 与对照组相比,经青蒿酯钠处理后,肝癌细胞中p53,p21蛋白表达水平无明显变化,而bcl-2蛋白表达水平降低.结论 青蒿酯钠可诱导人肝癌细胞(BEL-7402)凋亡,其凋亡的分子机制是p53非依赖性的,即与p53,p21无关,而与凋亡调节基因bcl-2下调有关.     

川芎嗪烟酸酯对神经细胞氧化损伤的保护作用

 目的观察川芎嗪烟酸酯(TMPN)对H₂O₂引起的体外培养大鼠脑皮层神经细胞及人神经母细胞瘤细胞(SH-SY5Y)氧化性损伤的保护作用,并初步探讨其作用机制。方法H₂O₂诱导体外培养大鼠脑皮层神经元及SH-SY5Y细胞进入氧化应激状态,观察TMPN对神经细胞氧化损伤的保护作用。结果TMPN能显著升高氧化损伤神经细胞的存活率,减少乳酸脱氢酶(LDH)的漏出,对H₂O₂诱导的神经细胞内游离钙浓度([Ca²⁺]_I)的升高有明显的抑制作用,并能减少胞内脂质过氧化物丙二醛(MDA)的生成,提高谷胱甘肽过氧物酶(GSH-Px)含量及超氧化物歧化酶(SOD)活性。结论TMPN可以有效保护活性氧对培养大鼠脑皮层神经元及SH-SY5Y细胞的氧化损伤,维持细胞的正常生理功能,其机制可能与TMPN抑制H₂O₂诱导的[Ca²⁺]_I异常升高,减少脂质过氧化物生成,增加抗氧化物质的含量有关。