铁死亡是一种铁离子依赖性的细胞程序性死亡方式,与铁代谢、脂质代谢及氨基酸抗氧化系统密切相关.铁死亡在肝纤维化发生发展过程中扮演重要角色,可能成为预防和治疗肝纤维化的潜在靶向.研究显示,大量中药成分可通过铁死亡途径抑制肝纤维化.笔者就铁死亡的调控途径、铁死亡在肝纤维化中的作用及治疗策略等方面进行综述,并总结近五年中药单体及活性成分干预铁死亡治疗肝纤维化相关研究,以期为中药干预、抗肝纤维化提供新的思路.

目的:研究氢氧化钙糊剂浸提液对大鼠骨髓间充质干细胞(BMMSCs)成骨向分化的影响,为其临床使用提供理论依据.方法:全骨髓贴壁法分离培养大鼠BMMSCs,流式细胞仪进行细胞鉴定,不同条件诱导细胞后进行茜素红染色、油红O染色及阿利新蓝染色,鉴定细胞成骨、成脂及成软骨分化能力.将氢氧化钙糊剂烘干、研磨成粉后,制作氢氧化钙糊剂浸提液及氢氧化钙糊剂条件培养基(CH),使用不同浓度CH(0、0.02、0.2、1、1.5、2 g/L)培养大鼠BMMSCs,碱性磷酸酶(ALP)活性测定筛选最佳作用浓度用于后续实验.大鼠BMMSCs经CH诱导后,ALP染色、ALP活性测定、茜素红染色检测其ALP活性及矿化能力的变化,Western blot检测细胞成骨相关指标Ⅰ型胶原α1(COL1A1)、ALP、Runt相关转录因子2(RUNX2)、成骨细胞特异性转录因子(OSX)的表达水平变化.结果:成功分离培养了大鼠BMMSCs,具有成骨、成脂及成软骨分化等间充质干细胞特性;0.2 g/L为CH诱导大鼠BMMSCs ALP活性的最佳作用浓度(P＜0.01),使用该浓度CH培养基处理大鼠BMMSCs后,COL1 A1、ALP、RUNX2、OSX表达上调(P＜0.01).结论:氢氧化钙糊剂浸提液可促进大鼠BMMSCs成骨分化.

Bone defects that result from trauma, infection, surgery, or congenital malformation can severely affect the quality of life. To address this clinical problem, a phosphoserine-loaded chitosan membrane that consists of chitosan membranes serving as the scaffold support to accommodate endogenous stem cells and phosphoserine is synthesized. The introduction of phosphoserine greatly improves the osteogenic effect of the chitosan membranes via mutual crosslinking using a crosslinker (EDC, 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide). The morphology of PS-CS membranes was shown by scanning electron microscopy (SEM) to have an interconnected porous structure. The incorporation of phosphoserine into chitosan membranes was confirmed by energy dispersive spectrum (EDS), Fourier Transforms Infrared (FTIR), and X-ray diffraction (XRD) spectrum. The CCK8 assay and Live/Dead staining, Hemolysis analysis, and cell adhesion assay demonstrated that PS-CS membranes had good biocompatibility. The osteogenesis-related gene expression of BMSCs was higher in PS-CS membranes than in CS membranes, which was verified by alkaline phosphatase (ALP) activity, immunofluorescence staining, and real-time quantitative PCR (RT-qPCR). Furthermore, micro-CT and histological analysis of rat cranial bone defect demonstrated that PS-CS membranes dramatically stimulated bone regeneration in vivo. Moreover, H&E staining of the main organs (heart, liver, spleen, lung, or kidney) showed no obvious histological abnormalities, revealing that PS-CS membranes were no additional systemic toxicity in vivo. Collectively, PS-CS membranes may be a promising candidate for bone tissue engineering.

目的:探讨羽毛角蛋白水凝胶与蒙脱石(MMT)复合改性后对骨髓间充质干细胞(BMSCs)的生物学行为及成骨性能的影响.方法:通过改良Shindai法制备羽毛角蛋白,将羽毛角蛋白与MMT混合制备羽毛角蛋白-MMT复合水凝胶.根据MMT/羽毛角蛋白水凝胶质量比例(w/w)分为0％、1％、3％三个组.结合考马斯亮蓝染色和傅里叶红外光谱鉴定所提取的羽毛角蛋白,利用扫描电镜观察羽毛角蛋白-MMT复合水凝胶的表面形态.加入使用含羽毛角蛋白-MMT复合水凝胶浸提液的培养基培养BMSCs,通过CCK-8实验探究其生物安全性,通过碱性磷酸酶(ALP)试剂盒检测其对BMSCs的ALP活性的影响,通过茜素红染色观察其对BMSCs的细胞外基质矿化的影响,通过Western blot检测其对BMSCs的BMP/SMAD信号通路相关蛋白表达水平的影响.结果:结合考马斯亮蓝染色和傅里叶红外光谱证明顺利提取羽毛角蛋白,羽毛角蛋白水凝胶具有天然孔隙,当加入MMT后其形态结构会发生一定程度的改变,当MMT的比例达到3％时,复合水凝胶支架的孔径明显变小.CCK-8实验结果表明,羽毛角蛋白+3％MMT复合水凝胶的浸提液可能对BMSCs的生长有抑制作用.羽毛角蛋白+1％MMT组的ALP染色和茜素红染色明显强于其他两组,说明羽毛角蛋白+1％MMT组的浸提液对BMSCs成骨分化有促进作用.与其他两组比较,羽毛角蛋白+1％MMT组的BMSCs骨形成蛋白-2(BMP-2)、Wnt3a、磷酸化细胞信号转导分子SMAD 1/5/8(p-SAMD 1/5/8)、Runt相关转录因子2(RUNX2)、骨钙素(OCN)和Ⅰ型胶原蛋白(COL-1)蛋白表达较高(P＜0.01).结论:羽毛角蛋白-MMT复合水凝胶可通过BMP/SMAD信号通路促进BMSCs成骨向分化.

目的:研究Vitapex糊剂对根尖牙乳头干细胞(SCAPs)成牙/骨向分化的影响,初步讨论其潜在机制.方法:酶消化法分离并培养SCAPs;制备不同浓度Vitapex条件培养基,通过碱性磷酸酶(ALP)活性测定选取最适浓度;ALP染色、实时荧光定量PCR和Western blot检测成牙/骨向相关基因和蛋白的变化情况;Western blot检测自噬相关蛋白表达水平.结果:0.2 g/L Vitapex条件培养基组ALP活性最高,选取为最适浓度进行后续实验.与对照组比较,该浓度Vitapex条件培养基能显著促进SCAPs细胞ALP、核心结合蛋白因子2(RUNX2)、成骨相关转录因子(OSX)、牙本质涎磷蛋白(DSPP)基因和蛋白表达(P<0.05).同时,0.2 g/L Vitapex组中微管相关蛋白1轻链3(LC3)Ⅱ/LC3Ⅰ及Beclin-1蛋白表达水平升高,P62蛋白表达水平降低(P<0.01).结论:Vitapex可促进SCAPs成牙/骨向分化,并激活自噬.

Let-7 miRNA family has been proved as a key regulator of mesenchymal stem cells’ (MSCs’) biological features. However, whether let-7b could affect the differentiation or proliferation of periodontal ligament stem cells (PDLSCs) is still unknown. Here, we found that the expression of hsa-let-7b was visibly downregulated after mineralization induction of PDLSCs. After transfected with hsa-let-7b mimics or inhibitor reagent, the proliferation ability of PDLSCs was detected by cell counting kit-8 (CCK-8), flow cytometry, and 5-ethynyl-2-deoxyuridine (EdU) assay. On the other hand, the osteogenic differentiation capacity was detected by alkaline phosphatase (ALP) staining and activity, alizarin red staining, Western blot, and quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR). We verified that hsa-let-7b did not significantly impact the proliferation ability of PDLSCs, but it could curb the osteogenic differentiation of PDLSCs. Besides, we predicted CTHRC1 acts as the downstream gene of hsa-let-7b to affect this process. Moreover, the combination of CTHRC1 and hsa-let-7b was verified by dual luciferase reporter assay. Our results demonstrated that the osteogenic differentiation of PDLSCs was enhanced after inhibiting hsa-let-7b, while was weakened after cotransfection with Si-CTHRC1. Collectively, hsa-let-7b can repress the osteogenic differentiation of PDLSCs by regulating CTHRC1.

1Department of Radiology, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, Republic of China; 2Department of Diagnostic Radiology and Nuclear Medicine, University of Maryland School of Medicine, Baltimore, Maryland, USA; 3Department of Oncology, Wuxi Xishan Hospital of Traditional Chinese Medicine, Wuxi, Jiangsu, Republic of China; 4Institute of Oncology, The First Clinical Medical College, Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine Prevention and Treatment of Tumor, Nanjing University of Chinese Medicine, Nanjing, Jiangsu, Republic of China; 5The First Clinical Medical College, Liaoning University of Chinese Medicine, Shenyang, Liaoning, Republic of China; 6Science and Technology Department, Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine Prevention and Treatment of Tumor, Nanjing University of Chinese Medicine, Nanjing, Jiangsu, Republic of China; 7Department of Oncology, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, Republic of China Purpose: Xiaoai Jiedu recipe (XJR), a formula long used by Chinese National Medical Professor Zhou Zhongying, has potent antitumor properties, but the molecular mechanism is still unclear. The aim of the study was to investigate the antitumor mechanism of XJR on hepatocellular carcinoma (HCC) by focusing on miRNA. Methods: Three concentrations of XJR (low, middle, and high) were used to treat tumor xenograft mice models. Microarray technology was used to identify the differential expressed genes after XJR treatment, and bioinformatic tools and luciferase reporter assay to predict the potential pathways. HepG2 cells were transfected with inhibitor of miR-200b3p to detect the effect of miR-200b-3p and Notch1 on tumor growth. Results: XJR effectively exerted anti-HCC effect both in vitro and in vivo. MiRNA chip analysis results showed that the expression of 75 miRNAs was upregulated and 158 miRNAs was downregulated in blood from XJR-treated mice. Further validation by using real-time polymerase chain reaction (RT-PCR) assay showed that the expression of five miRNAs (miR-453, miR-200b-3p, miR-135a-1-3p, miR-1960, miR-378a-5p, and miR-466f) was consistent with the results of miRNA chip analysis. Among them, miR-200b-3p was selected as candidate for further research. Results of the MTT, migration, and wound healing assays showed that down-expression of miR-200b-3p abrogated the effect of XJR on cell growth and metastasis. Luciferase reporter assay confirmed that Notch1 was the direct target of miR200b-3p. XJR significantly decreased Notch1 expression in HepG2 cells, whereas miR200B-3p inhibitor abrogated the XJR-induced decrease in Notch1 expression. Conclusion: This study indicated that XJR could effectively inhibit HCC and might exert its antitumor effect through the miR-200b-3p/Notch1 axis. These findings provided new avenues for the use of XJR for prevention and treatment of HCC.

目的 网络药理学角度解释中药复方“消癌解毒方”中君药、臣药的关系以及该方防治肝癌的有效性.方法 依托中药药理分析平台(TCMSP)、国家健康研究院PubChem平台、Swiss Target Prediction平台寻找到复方“消癌解毒方”中君药白花蛇舌草、臣药太子参、山慈菇所对应的靶点,借助DisGeNET数据库、人类孟德尔遗传研究平台OMIN、治疗靶点数据库TTD、PharmGkb数据库找到与肝癌相关的靶点.通过网络映射分析药物-靶点-疾病的关系.利用CCK-8、划痕实验、流式细胞术、western blot等技术检测相关靶点可行度.结果 通过口服生物利用度(OB)＞30％,类药性(DL)＞0.18的筛选获得有效活性成分13种,药物对应靶点330个,肝癌相关靶点603个.利用生物功能及信号通路分析,消癌解毒方可能参与多种肿瘤发生、细胞凋亡、氧化应激等信号通路.并通过BCL2、CASP8、NCOA2、PTGS2、TGFB1、BAX、CASP3等基因影响肝癌的发生发展.研究显示山慈菇的生物功能富集在淋巴细胞稳态、B细胞稳态、T细胞稳态等相关功能上,太子参富集在了骨髓白细胞分化的正调节、B细胞稳态、T细胞稳态、白花蛇舌草功能则富集在上皮细胞凋亡过程、平滑肌细胞增殖、受体代谢过程、RNA聚合酶Ⅱ转录pri-miRNA、细胞周期等方面.同时通过实验手段验证了网络药理学靶点预测的可行度.结论 从网络药理学、现代生物学角度来评述消癌解毒方中君药、臣药组成是合理、科学的.消癌解毒方防治肿瘤是具有生物学支撑的.

自噬是细胞在能量缺乏等代谢压力下,通过包裹细胞内的蛋白、细胞器和胞质,实现细胞成分的循环利用,从而维持细胞的正常代谢和生存.近期的研究表明,自噬在肿瘤的发生、发展和治疗中具有重要意义,既可以诱发肿瘤细胞死亡又可以介导死亡逃避.中医药既可以调控肿瘤细胞的自噬性死亡,又可以影响其保护性自噬,为中医药抗肿瘤研究提供了新的思路.

目的 探讨消癌解毒方对H22荷瘤小鼠移植瘤血管生成相关因子及蛋白的影响.方法 建立H22荷瘤小鼠腹水瘤株体内移植肿瘤模型,空白组每日生理盐水灌胃,消癌解毒方低、中、高剂量组每日10,30,90 g·kg-1灌胃,顺铂组每日1 g·kg-1腹腔注射给药,连续给药10 d,记录瘤体体积及移植鼠体重.给药结束,取瘤体组织,称重,计算各给药组的肿瘤抑制率;通过肿瘤组织的全基因组表达谱芯片,Elisa、qRT-PCR、Western-blot观察各给药组对移植瘤的血管生成因子及相关基因STAT1、VEGF、MMP2、TGF-β、CXCL2的影响.结果 与空白组比较,顺铂组及消癌解毒方低、中、高剂量组移植瘤小鼠瘤体重量显著降低(P<0.05),VEGF、MMP2、TGF-β、CXCL2细胞因子水平显著降低(P<0.05);qPCR、western-blot实验结果表明,与空白组比较,消癌解毒方各剂量组STAT1基因及蛋白表达能够显著升高,VEGF、MMP2、TGF-β、CXCL2基因、蛋白的表达显著下降,差异具有统计学意义(P<0.05).结论 消癌解毒方能够有效抑制H22荷瘤小鼠移植瘤生长,其中抑制肿瘤血管新生是其可能的作用机制之一.

目的:通过对Lewis肺癌原位移植小鼠肿瘤组织长链非编码RNA (long noncoding RNA,lncRNA)和mRNA表达谱的检测,探讨麦门冬汤合千金苇茎汤(金方)抗肺癌的作用机制.方法:C57BL/6小鼠随机分为正常组、模型组、金方组(20 g·kg-1 ·d-1),构建小鼠Lewis原位肺癌模型成功后,次日用金方灌胃开始治疗.应用基因芯片技术,检测与金方抗肺癌作用密切相关的差异lncRNA,mRNA,进行聚类分析,并利用t检验和差异表达的倍数变化,筛选出关键lncRNA,mRNA.运用生物信息学方法预测差异lncRNA调控的靶基因,并对其进行功能和通路分析.应用病理组织学技术,检测各组小鼠肿瘤组织光镜下的差异表现.结果:lncRNA,mRNA芯片杂交结果显示,金方调控了887个lncRNA差异表达,其中上调442个,下调445个(P＜0.05).差异表达的mRNA共610个,其中上调376个,下调234个(P＜0.05).使用GO分析方法,发现下调或上调的靶基因主要参与了细胞代谢调控、信号通路调控等生物学过程(biological process,BP).靶基因分子功能(molecularfunction,MF)分析显示,下调或上调的靶基因主要具有DNA结合、蛋白质结合、受体结合和转录因子活性等功能.信号通路预测显示金方可调控Janus激酶/信号转导与转录激活因子(JAK/STAT)等与肺癌的发生发展密切相关的信号通路.病理组织学检查结果证实,与模型组比较,金方能明显改善小鼠肺组织的病理改变.结论:金方可能通过调控多种lncRNA和mRNA的表达及下游相关信号通路来发挥其抗肺癌的作用.

Aim To determine the effect of cornel iri-doid glycoside ( CIG ) on human hepatocyte cell line (L-02) injured by D-galactosamine (GalN) and tumor necrosis factor-α( TNF-α) .Methods Firstly, CIG was extracted , separated and purified . Cell lesion model injured by D-GalN/TNF-αwas tested by MTT method.T-AOC, SOD, MDA and calcium ion concen-tration were taken as indicators to study the effects of CIG on L-02 cell injured by D-GalN/TNF-α.The ex-pression of p-PERK, p-eIF-2α, caspase-3 protein were detected by Western blot .Results 44 mg · L-1 D-GalN and 100 μg · L-1 TNF-αwere suitable for L-02 cell lesion model.CIG high, middle, low concentra-tion group could significantly increase the L-02 cell ac-tivity by 21％, 13％, 8％, respectively and SOD activity and T-AOC ability as well compared with model group.At the same time, they markedly reduced the MDA activity except the low concentration .Three con-centrations of CIG could reduce the expression of endo-plasmic reticulum stress related protein PERK , eIF-2αand apoptosis-associated protein caspase-3. Conclu-sions CIG could protect L-02 cells injured by D-GalN/TNF-α.Increasing the cellular antioxidant abili-ty, reducing the damage of endoplasmic reticulum stress and the expression of apoptosis-associated protein may be the possible mechanism .

目的:通过消癌解毒方(XAJD)对H22荷瘤小鼠移植瘤肿瘤组织全基因组芯片的检测以及对相关蛋白的表达影响,探讨其抑制肿瘤的作用机制.方法:ICR小鼠随机分组,接种H22荷瘤小鼠腹水,移植瘤模型成功后,将模型小鼠随机分为空白组,XAJD低、中、高剂量组(10,30,90 g·kg-1 ·d-1),顺铂组(1 mg·kg-1·d-1).给药结束,动物处死并取各组动物瘤体组织,无菌生理盐水洗去血渍,称重,计算肿瘤抑制率,取部分新鲜瘤体组织进行全基因组表达谱芯片检测,部分瘤体组织用实时荧光定量PCR(Real-time PCR),蛋白免疫印迹法(Western blot)检测其相关mRNA及蛋白表达.结果:与空白组比较,消癌解毒方低、中、高剂量能够显著降低移植瘤瘤重(P＜0.05),消癌解毒方中剂量组与顺铂组效果优于消癌解毒方低、高剂量组(P＜0.01),抑瘤率分别达42.8％,58.6％;全基因组芯片检测分析发现,基质金属蛋白酶家族(matrix metalloproteinases,MMPs)和组织金属蛋白酶抑制剂(tissueinhibitor of metalloproteinase,TIMPs)相关蛋白在消癌解毒方不同剂量组中均发生了明显的改变.Real-time PCR,Western blot检测显示,与空白组比较,MMP-9 mRNA及蛋白在消癌解毒方低、中、高剂量组及顺铂组中表达均显著降低(P＜0.01),MMP-12 mRNA及蛋白在消癌解毒方中、高剂量组及顺铂组中表达均显著降低(P＜0.01),TIMP2 mRNA及蛋白在消癌解毒方中、高剂量组及顺铂组中表达均明显升高(P＜0.05).结论:消癌解毒方能够抑制H22荷瘤小鼠移植瘤生长,可通过抑制MMPs相关蛋白的表达,降低肿瘤的转移和复发可能是其抗癌机制之一.

To investigate the molecular mechanism for the induction of apoptosis by luteolin in human lung cancer H460 cells,and to provide a new scientific basis for its treatment of lung cancer.H460 cells in logarithm growth phase were selected and randomly divided into various groups:a blank control group and different concentrations of luteolin (LTL)groups.The cell viability was detected by CCK-8 assay,while the cell apoptosis was measured by flow cytometry analysis.The mRNA expression of lung cancer related genes in H460 cells were determined by PCR array.The protein levels of p53,p21,Bax and Bcl-2 were detected by Western blot.The expression levels of microRNA-34a (miR-34a) in H460 cells were assessed by qRT-PCR.Then,the miR-34a-5p was transfected into H460 cells,to observe its effects on cell apoptosis and related protein expression.The results showed that luteolin exhibited the anti-cancer effect in H460 cells through the induction of cell apoptosis and inhibition of cell proliferation,which may be mediated through the activation of the p53 signal pathway,up-regulation of miR-34a-5p,and subsequent regulation of apoptosis-related protein expression such as Bax/Bcl-2.

研究异甘草素对人肺癌NCI-H460细胞增殖、周期及凋亡的影响,并探讨其相关分子机制为其治疗肺癌提供新的科学依据。取对数生长期人肺癌NCI-H460细胞,用不同浓度异甘草素处理,采用CCK-8法检测细胞增殖能力;流式细胞术检测细胞周期、凋亡的变化;PCR array检测肺癌相关基因mRNA表达;Western-blot检测p-p65,IκBα、IκBβ、Bax、Bcl-2表达量变化;细胞免疫荧光实验观察异甘草素对NF-κB p65分布的影响;结果显示,异甘草素能有效抑制人肺癌H460细胞的增殖和诱导细胞周期阻滞,并促进其凋亡,其机制可能是通过抑制NF-κB信号通路的活化,抑制NF-κB p65入核使其不能发挥转录活性,最终影响其下游凋亡相关蛋白Bax/Bcl-2的表达来实现的。

To explore the distribution characteristics and correlations among the natures,flavors and channel entries of pregnancy contraindicated Chinese herbs.Methods Chinese Materia Medica (《中华本草》) and CNKI were searched,and 1136 pregnancy contraindicated Chinese herbs were found.Data on the herbs was analyzed by association rules and correspondence analysis.Results The four natures of pregnancy contraindicated Chinese herbs are most warm,cold and neutral;as for the five flavors,bitter is most,then acrid and sweet;for the channel entries,the liver channel is most,then lung,spleen,stomach,kidney,large intestine and heart.The high-frequency nature and flavor combinations are acrid-warm,bitter-cold,bitter-warm,and bitter-neutral.The high-frequency nature and channel entry combinations are warm-liver,cold-liver,neutral-liver and cold-lung.The high-frequency flavor and channel entry combinations are bitter-liver,acrid-liver,bitter-lung and sweet-liver.Conclusion During pregnancy,more attention should be paid on acrid-warm and bitter-cold Chinese herbs,so as the warm-liver and bitter-liver ones.

目的 从线粒体途径探讨山茱萸活性成分5-羟甲基糠醛对肿瘤坏死因子α(TNF-α)和D-氨基半乳糖(D-GalN)所致肝细胞凋亡的保护作用 方法 体外培养人正常肝L02细胞,用TNF-α(100μg/L)和D-GalN(44 mg/L)诱导肝细胞损伤模型,采用CCK-8细胞增殖检测试剂盒检测细胞活力;流式细胞仪检测5-羟甲基糠醛对细胞凋亡的影响;Western-blot检测线粒体途径凋亡标志蛋白Apaf-l、AIF、cytochrome C、Bcl-2、Bcl-XL、Bax表达水平的变化;分光光度法检测Caspase-3、Caspase-9的活性.结果 与TNF-α和D-GalN模型组相比,不同浓度的5-羟甲基糠醛作用后,可显著减轻L02肝细胞的增殖抑制、降低细胞的凋亡率,并使促凋亡蛋白Apaf-l、AIF、cytochrome C、Bax的表达量明显下降,而使抗凋亡蛋白Bcl-2、Bcl-XL的表达量显著上升,同时使细胞凋亡的关键蛋白酶Caspase-3、Caspase-9的活性显著降低.结论 山茱萸羟甲基糠醛对TNF-α和D-GalN所诱导的肝细胞凋亡具有抑制作用,其机制可能与线粒体介导的凋亡通路以及BCL-2家族成员的表达调控有关.

5-Hydroxymethylfurfural(5-HMF), a water-soluble compound extracted from wine-processed Fructus corni, is a novel hepatic protectant for treating acute liver injury. The present study was designed to investigate the protective effect of 5-HMF in human L02 hepatocytes injured by D-galactosamine(Gal N) and tumor necrosis factor-α(TNF-α) in vitro and to explore the underlying mechanisms of action. Our results showed that 5-HMF caused significant increase in the viability of L02 cells injured by Gal N/TNF-α, in accordance with a dose-dependent decrease in apoptotic cell death confirmed by morphological and flow cytometric analyses. Based on immunofluorescence and Western blot assays, we found that Gal N/TNF-α induced ER stress in the cells, as indicated by the disturbance of intracellular Ca2+ concentration, the activation of protein kinase RNA(PKR)-like ER kinase(PERK), phosphorylation of eukaryotic initiation factor 2 alpha(e IF2α), and expression of ATF4 and CHOP proteins, which was reversed by 5-HMF pre-treatment in a dose-dependent manner. The anti-apoptotic effect of 5-HMF was further evidenced by balancing the expression of Bcl-2 family members. In addition, the knockdown of PERK suppressed the expression of phospho-PERK, phospho-e IF2α, ATF4, and CHOP, resulting in a significant decrease in cell apoptosis after the treatment with Gal N/TNF-α. 5-HMF could enhance the effects of PERK knockdown, protecting the cells against the Gal N/TNF-α insult. In conclusion, these findings demonstrate that 5-HMF can effectively protect Gal N/TNF-α-injured L02 hepatocytes against ER stress-induced apoptosis through the regulation of the PERKe IF2α signaling pathway, suggesting that it is a possible candidate for liver disease therapy.

Aim To study effects of loganin and morro-niside on aging astrocytes induced by D-galactose. Methods Cortex astrocytes of newly born rats were cultured in vivo and indentified by immunofluorescence method.Firstly,appropriate D-galactose concentration was selected and effects of loganin and morroniside on proliferation activity of aging astrocytes induced by D-galactose were determined by MTT test.Then SOD, MDA were taken as indicators to study the effects of loganin and morroniside on aging astrocytes induced by D-galactose.Thirdly,growth factors like gliar cell line-derived neurotrophic factor (GDNF),basic fibro-blast growth factor (bFGF)tested by ELISA method and expression of Bax,caspase-3,phospho-extracellu-lar signal-regulated kinese (p-ERK1 /2),phospho-mi-togen-actived protein kinase /extracellular signal-regu-lated kinase kinase (p-MEK1 /2)were taken as indi-cators to discuss the potential protection mechanism. Results Loganin and morroniside exerted certain effects on proliferation of aging astrocytes induced by D-galactose,improving SOD,GDNF,bFGF release and lowering MDA release significantly (P <0.05 ). The expressions of Bax and caspase-3 proteins had no difference between model group and loganin group, morroniside group.While the expressions of p-ERK1 /2,p-MEK1 /2 of loganin group,morroniside group were improved significantly compared with model group.Conclusion Loganin and morroniside have protective effects on aging astrocytes induced by D-ga-lactose,and increase the proliferation ability.Protec-ting their antioxidant systems and improving the expres-sion of p-ERK1 /2,p-MEK1 /2 proteins are possible mechanisms.

Objective To investigate the antidepressant-like effects of extracts isolated from rhizomes of Cyperus rotundus L.(CR) for the first time. Methods Three models,the mouse forced swim test(FST),tail suspension test(TST) and open field test(OFT),were used to observe antidepressant-like effects of the 95% ethanol extract(XF-E) and the water extract(XF-W) isolated from CR. Results Firstly,in bioactive extract screening,compared with the model group(CMC-Na),XF-E(2 g/kg) significantly reduced the immobility time(IT) during FST and TST,while XF-W(2 g/kg) did not.Meanwhile,there was no significant difference among groups in crossed squares during OFT.Secondly,in dose-effect relationship assay,compared with the model group(CMC-Na),XF-E(1,2 g/kg) significantly reduced IT during FST and TST. Conclusion The present study indicates that XF-E can be bioactive extract of CR in antidepressant-like effect and appears in dose-dependent manners,for the first time.