Endothelial dysfunction and pathological alterations are pivotal in the pathogenesis of cardiovascular disease. To date, effective interventions for these endothelial changes are lacking. Tumor necrosis factor-alpha (TNF-α) is known to significantly contribute to these alterations. It has been reported the potential of luteolin to mitigate TNF-α-induced inflammation, yet its specific mechanisms and targets still remain to be elucidated. This study aims to investigate the effects and mechanisms of luteolin on TNF-α-induced inflammatory injury in human microvascular endothelial cells, thereby advancing the understanding of luteolin’s medicinal properties. Our findings demonstrate that luteolin notably inhibits TNF-α-induced phosphorylation of Akt, mitogen activated protein kinase (MAPK), and the nuclear factor-kappaB (NF-κB) p65. It significantly reduces the transcriptional activity of NF-κB p65 and AP-1 and decreases the expression of mRNA and proteins related to adhesion molecules and inflammatory mediators. Additionally, luteolin inhibited the reduction in STAT3 phosphorylation. In conclusion, luteolin effectively suppresses TNF-α-induced inflammatory injury in endothelial cells via the Akt/MAPK/NF-κB pathway.

Phytochemical investigation on the bioactive constituents of the tubers of Lepidium meyenii led to two new quinoline derivatives (1 and 2) and one new macamide (5), together with six known compounds (3, 4, and 6–9). The new structures were elucidated by means of the extensive spectroscopic analyses. The NMR data of compound 3 were firstly reported and its structure was affirmed by the X-ray crystallography diffraction. Compound 9 showed moderate AChE inhibitory activity, and 1, 3, and 8 exhibited moderate inhibitory effect on the secretion of TNF-α by RAW264.7 cells.

原矛头蝮蛇(Protobothrops mucrosquamatus)主要分布于我国长江以南和台湾地区,原名烙铁头蛇(Trimeresurus mucrosquamatus),属蝰蛇科原矛头蝮属[1].原矛头蝮蛇毒(Protobothrops mucrosquamatus venom,PMV)主要表现为血液毒性,影响心血管及凝血系统等方面,对组织细胞也有毒性作用[2-3].该蛇伤较为常见,临床表现主要为凝血障碍、头昏恶心、视力模糊和胸闷腹痛等,严重者出现皮下出血、血尿、全身内脏出血甚至失血性休克[4-5].迄今,对PMV的研究主要集中在纤溶酶、类凝血酶、丝氨酸蛋白酶及诱导血小板聚集的蛋白等成分[1-2,6].课题组前期研究表明,PMV具有明显的破坏凝血功能的作用,PMV及其组分能诱导血小板聚集引起全血中血小板计数下降,具有酶切底物S-2251活性,能延长凝血酶时间(TT)和部分活化凝血活酶时间(APTT),破坏人微血管内皮细胞[2,7].此前研究更多采用凝血四项等研究方法,为进一步研究和认识PMV对凝血功能的影响,课题组采用血栓弹力图测定方法来开展PMV对大鼠全血凝血功能影响的研究,以期为进一步认识原矛头蝮蛇伤的机制及临床救治方案和新药研究提供参考.

OBJECTIVE To investigate the toxic effect and mechnism of kaempferol on human kidney proximal tubular epithelial cells （HK-2） based on high content analysis technology.METHODS HK-2cells were divided into the control group （final concentration of dimethylsulfoxide＜0.01%）,kaempferol 510,20,50 and 100μmol·L -1 and doxorubicin (Dox,final concentration 10μmol·L -1 ) groups.The count of living cells was detected via Hoechst 33342 staining,the cell viability was determined by MTT assay,the levels of reactive oxygen species （ROS）,mitochondrial membrane potential （MMP） and the intracellular calcium ion were detected by fluorescence probes of DCFH-DA,Mito-Tracker Red CMXRos and Fluo-4 AM.The ATP content was measured with the chemiluminescence method,and apoptosis by flow cytometry.The DNA content was detected by Hoechst 33342 staining.The nuclear translocation of NF-κB p65,the expressions of p38 mitogen-activated protein kinases （p38 MAPK）,p-p38 MAPK,extracellular signal regulated kinase （ERK）,p-ERK and c-Jun N-terminalkinases （JNK）,p-JNK protein in associated mitogen activated protein kinases （MAPK） signal pathway,and those of signal transducer and activator of transcription-3 （STAT3）,p-STAT3 protein in janus kinase-2 （JAK2）/STAT3 signal signa pathway were detected by immunofluorescence assay.RESULTS Compared with the control group,the count of living cells in the kaempferol 5,10,20,50 and 100μmol·L -1 groups and the cell viability in the kaempferol 10,20,50 and 100μmol·L -1 groups were significantly deceased （P＜0.05,P＜0.01）.The levels of ROS were significantly increased in the kaempferol 10,20,50 and 100μmol·L -1 groups （P＜0.05,P＜0.01）,while those of MMP in 5,10,20,50 and 100μmol·L -1 groups were obviously decreased（P＜0.01）.The levels of calcium influx in kaempferol 50,100μmol·L -1 groups were increased significantly（P＜0.05,P＜0.01）,while the ATP content in the kaempferol 50 and 100μmol·L -1 groups was obviously decreased （P＜0.05,P＜0.01）.The apoptotic rates in kaempferol 20,50,100μmol·L -1 groups were up-regulated significantly （P＜0.05,P＜0.01）,but the DNA content hardly changed in all the kaempferol groups.The results of immunofluorescence assay showed that kaempferol 50 and 100μmol·L -1 led to the phosphorylation and nuclear translocation of NF-κB p65,the expressions of p-p38 MAPK,p-ERK,p-JNK and JNK of MAPK signaling pathway were up-regulated significantly,and that the expression of p-STAT3protein of JAK2/STAT3 signaling pathway was also increased （P＜0.05,P＜0.01）.CONCLUSION Kaempferol exerts toxic effect on HK-2 cells,and its mechanism is probably related to oxidative stress injury and apoptosis mediated by the up-regulation of ROS levels.

目的 利用高内涵筛选技术研究吴茱萸次碱(rute-acarpine,RUT)的肝毒性及其可能机制.方法 HepG2细胞暴露于不同浓度的RUT后作用不同时间,MTT法检测细胞活力,采用高内涵检测RUT对细胞存活、细胞核面积、线粒体膜电位(MMP)、ROS、钙离子内流、细胞膜完整性(DIR)和MAPK、NF-κB、JAKs-STATs信号通路活化水平的影响,流式细胞检测细胞凋亡.结果 100 μmol·L-1 RUT明显抑制了HepG2细胞的活力(P＜0.01),表现为RUT作用24 h后,细胞核面积减少,核形态不均一,作用48 h后存活细胞数量明显减少(P＜0.01),并伴随着细胞的早期凋亡(P＜0.01);从6 h开始,100 μmol·L-1 RUT组细胞内活性氧和钙离子水平明显升高(P＜0.01),细胞膜完整性明显降低(P＜0.01);100 μmol·L-1 RUT 作用 24 h 后,ERK1/2、JNK、STAT3 和p38的磷酸化水平明显升高(P＜0.01,P＜0.05),总蛋白未见明显变化;在3 h可检测到c-Jun、c-Fos的表达上调,与对照组相比,差异具有显著性(P＜0.01);在3 h时间点,可明显检测到p-NF-κB p65的表达上调(P＜0.01),但NF-κB p65入核不明显.结论 吴茱萸次碱在100 μmol·L-1的条件下对HepG2细胞表现出细胞毒性,其毒性机制主要与氧化应激和炎症反应导致的细胞损伤有关.

Two new quinolizidine-based alkaloids, ochrocephalamines E (1) and F (2) were isolated from Oxytropis ochrocephala Bunge (Fabaceae). Their structures were elucidated by extensive spectroscopic analysis and electronic circular dichroism (ECD) calculations. Compound 1 represented the first 14-nor matrine with 6/6/6/5 ring system, while compound 2 shared the aloperine-type scaffold including the carbonyl group at C-10 postition. The inhibitory activities of 2 against the secretion of HBsAg and HBeAg were respectively 22.34 +/- 4.38% and 18.00 +/- 5.01% at the noncytotoxic concentration of 1 x 10-5 mol/L, which were more active than the positive control hyperoside (16.09% and 12.23%). The binding mode between compound 2 and HBV core protein was illustrated by molecular docking. (c) 2022 Elsevier Ltd. All rights reserved.

目的 研究黄芩苷对脂多糖(LPS)致人微血管内皮细胞(HMEC)炎症反应的保护作用及作用机制.方法 采用LPS作用HMEC,建立炎症反应模型;以不同浓度的黄芩苷预处理细胞,然后将细胞暴露于LPS,ELISA检测炎症因子ICAM-1、IL-6和MCP-1的含量;荧光观察Ca2+内流并采用流式细胞仪检测细胞内Ca2+水平;免疫荧光法检测NF-κB p65入核情况;双荧光素酶报告基因方法检测NF-κB核内转录活性;Western blot检测NF-κB p65、p-NF-κB p65及TLR4的表达.结果 HMEC暴露于LPS后,出现了明显的Ca2+内流,NF-κB p65发生磷酸化,核内转录活性上调,ICAM-1、IL-6及MCP-1表达上调.不同浓度的黄芩苷均能抑制LPS刺激HMEC后产生的Ca2+内流、NF-κB p65入核及核内转录活性上调,减少ICAM-1、IL-6、MCP-1的表达,下调NF-κB p65、p-NF-κB p65及TLR4的表达,且抑制作用呈现一定的量效关系.结论 黄芩苷能抑制LPS诱导的HMEC炎症反应,其机制与抑制Ca2+内流和NF-κB信号通路的活化,从而降低炎症反应的水平有关.

<span id="ChDivSummary" name="ChDivSummary" class="abstract-text">目的研究银杏双黄酮对高糖致人微血管内皮细胞（HMEC）炎症反应的干预作用及其可能的分子机制。方法 MTT比色法检测细胞活力,采用ELISA检测细胞培养上清中可溶性黏附分子ICAM-1和炎症介质（IL-6、TNF-α）的含量变化;荧光免疫法和流式细胞仪测定HMEC细胞表面ICAM-1的表达;双荧光素酶报告基因检测NF-κB p65核内转录活性;Western blot检测NF-κB p65及p-NF-κB p65、Bax、Bcl-2的表达水平。结果 HMEC暴露于高糖后,NF-κB信号通路活化,NF-κB p65磷酸化及核内转录活性明显上调,HMEC表达ICAM-1和TNF-α、IL-6明显升高,凋亡相关蛋白Bax/Bcl-2的比值出现上调。1×10^-5、1×10^-6、1×10^-7 mol·L^-1的银杏双黄酮对上述炎症反应相关指标的上调均有明显的干预作用,且呈现一定的量效关系。结论银杏双黄酮对高糖刺激HMEC炎症反应有明显的干预作用,其机制可能与抑制NF-κB信号通路活化,从而降低炎症反应水平有关。</span>

免疫性疾病、急性肺损伤、缺血再灌注损伤等病征的发生与补体异常激活密切相关.抗补体药物研究是新药开发的热点之一.本研究旨在从中华眼镜蛇毒中发现并分离纯化获得新的抗补体活性蛋白质,并对其理化性质和生物学活性加以研究.在抗补体活性追踪的指导下,采用蛋白质层析技术对眼镜蛇毒进行分离纯化;利用MALDI-TOF-MS、SDS-PAGE和葡聚糖凝胶过滤法测定目标蛋白质的纯度及分子量;等电聚焦凝胶电泳法测定其等电点;采用Edman降解法测定目标蛋白质的N-端氨基酸序列;测定目标蛋白质对补体经典途径和旁路途径的抑制活性以及可能的机制;采用MTT法和SRB法检测目标蛋白质对肿瘤细胞的杀伤作用;测定目标蛋白质对多种来源红细胞的溶血活性;采用KB平板扩散法检测抗菌活性.结果 表明,通过SP Sephadex C-25阳离子交换层析和RP-HPLC C18反相层析,从中华眼镜蛇毒中分离纯化获得一个均一的抗补体蛋白质,将其命名为CTX-CI.还原性SDS-PAGE测得CTX-CI的表观分子量为12.7 kD,凝胶过滤法测得分子量为9.7 kD,MALDI-TOF-MS测得精确分子质量为7.0 kD;变性条件下测得CTX-CI的等电点为9.81;N-端氨基酸序列为LKCH.相关活性测定结果表明,CTX-CI能有效抑制人血清补体经典途径,其IC50为0.046 g/L,但对补体旁路途径无明显抑制作用;机制研究表明,CTX-CI能抑制补体经典途径C3转化酶的形成.同时,CTX-CI对肿瘤细胞株A549、K562和MCF-7细胞表现出抑制作用,其IC50分别为0.32 g/L、0.58 g/L、0.63 g/L;对豚鼠红细胞有轻微的溶血作用;能抑制枯草芽孢杆菌和藤黄微球菌的生长.综上所述,本研究从中华眼镜蛇毒中分离纯化出一个新的抗补体蛋白质-CTX-CI,其理化性质和生物学活性表明其属于细胞毒素,CTX-CI能明显抑制补体经典途径,其机制与抑制经典途径C3转化酶的形成有关.

Ochrocephalamines B-D (1-3), composed of fused quinolizidine and octahydroquinoline rings, were isolated from Oxytropis ochrocephala Bunge. Ochrocephalamine B (1) has a unique bridged tetracyclic ring skeleton fused with a lactam ring. The structures of 1-3 were elucidated using spectroscopic and computational approaches. Ochrocephalamine C (2) and D (3) demonstrated potent anti-HBV activities and are more potent against the secretion of HBeAg than that of HBsAg.

Ellagic acid (EA), a naturally occurring polyphenolic compound, is commonly known for its anti-inflammatory properties. The low bioavailability greatly limits the clinical applications of EA. In this study, a biodegradable hollow zein nanoparticle with an average diameter of about 70 nm was developed to mediate oral delivery of EA. The inner core of the nanoparticle consists of EA/sodium carbonate (EA/Na2CO3) prepared by coprecipitation, which was further encapsulated in hollow zein nanoparticles with triethyl citrate as a natural plasticizer. The optimized ellagic acid-hollow plasticized zein nanoparticles (EA-HTZN) exhibited a small dimension of 72 nm with a PDI of 0.131, a drug loading capacity as high as 326 mgg(-1) at an equilibrium concentration of 5.0 mgmL(-1). EA-HTZN had high drug loading and prevented their precipitation at simulated physiological environment. The EA-HTZN significantly improved permeation ability in vitro. Oral administration of EA-HTZN showed effective against inflammation related to suppression of pro-inflammatory cytokines (TNF alpha, IL1 beta) overproduction in carrageenan-induced mouse paw edema model. Pharmacokinetic parameters of optimized formulation revealed 3.6- and 2.1-fold increase in bioavailability as compared to EA suspension and EA solid nanoparticle, respectively. Together, these results demonstrated the successful formulation of EA-HTZN and their potential to improve oral delivery through high drug loadings and good stability.

Objective To investigate the effect of compound GY3 (GY3) on proliferation and apoptosis of breast cancer Michigan cancer foundation-7 (MCF-7) cells,and regulating downstream factors of adenosine 5'-monophosphate-activated protein kinase/cyclooxygenase-2 (AMPK/COX-2) signal pathway.Methods MTT assay was used to measure the inhibition rate of different concentration of GY3,5-minoimidazole-4-carboxyamide ribonucleoside (AICAR),celecoxib and AICAR combined with celecoxib on MCF-7 cells.Cell apoptosis rate was detected by FCM assay.The expressions of acetyl-coa carboxylase product of phosphorylation,phosphorylated acetyl-Co A carboxylase(P-ACC) and COX-2 were assayed in MCF-7 cells cultured with different concentration of GY3 and GY3 + Compound C by Western blot.Results GY3 inhibited MCF-7 cell growth in a concentration dependent manner (P ＜0.01,P ＜0.05).GY3 induced apoptosis of MCF-7 cells(P ＜0.05).GY3 could increase P-ACC and down regulate COX-2 expression(P ＜0.01,P ＜0.05).AMPK inhibitor Compound C could restore activation of AMPK and inhibition of COX-2 by GY3.Conclusion GY3 could inhibit COX-2 expression by activating AMPK,and subsequently induce apoptosis of MCF-7 cells.

Aim To study the development of acute lung inflammation in mice induced by activation of the complement alternative pathway and the changes of the related indicators, and to provide an ideal pathological&nbsp;model of acute lung inflammation in mice for drug screening and intervention. Methods Cobra venom factor( CVF) was used to activate complement alterna-tive pathway of SPF Kunming mice by intravenous&nbsp;injection. According to different sampling time, the mice were divided into 15 min, 30 min, 1 h, 2 h, 6 h group, and the parallel PBS control groups were set at the same time. Lung coefficient, lung water content, myeloperoxidase ( MPO ) activity, BALF cell number and protein content were tested. The pathological changes of lung tissue were observed by HE staining. The concentration of IL-6 , TNF-α, P-selectin and ICAM-1 in bronchoalveolar lavage fluid ( BALF ) and serum were determined by ELISA. Results CVF caused pulmonary inflammatory cell infiltration in mice obviously. Compared with PBS groups, MPO activity of lung tissue, BALF cell and the protein concentration were significantly increased. The contents of IL-6, TNF-α, P-selectin in BALF and serum were in-creased, and the content of ICAM-1 in serum was also increased. The content of P-selectin in BALF reached the first peak at 30 min point, the content of IL-6 and&nbsp;TNF-α in BALF reached the first peak at 1 h point, but the indicators had no further changes at 2 h point, and all the indicators rose again at 6 h point. The lev-els of IL-6 and TNF-α in serum reached peak at 1 h point,then the content showed lower levels at the sub-sequent time points. The levels of P-selectin and ICAM-1 in serum increased along the time. Lung coef-ficient, lung water content and ICAM-1 of the BALF showed no significant alteration. Conclusion The ac-tivation of the complement alternative pathway can lead to acute lung inflammation in mice and the inflammato-ry response is the most obvious at 30 min to 1 h. The study could provide an ideal pathological model of a-cute lung inflammation in mice for drug screening and intervention.

A new lathyrane diterpenoid ester, euphordracunculin C (1), together with a phenolic amide, 4-hydroxy-N-[3-(4-hydroxy-3-methoxyphenyl)-1-oxo-2-propen-1-yl]-3-methoxybenzamide (2), was isolated from the aerial parts of Euphorbia dracunculoides. The structure of the new compound was elucidated on the basis of spectroscopic analysis including 1D and 2D NMR techniques.

目的:分析北五味子总木脂素中的挥发性成分,并对其抗烟草花叶病毒(TMV)活性和抗肿瘤活性进行研究.方法:运用GC-MS技术分析北五味子总木脂素中挥发性化学成分;采用活体半叶枯斑法和MTT法分别研究了其抗TMV活性和抗肿瘤活性.结果:从北五味子总木脂素中检测出58个化合物,鉴定出其中43个,含量较多的有五味子素(13.974％),香芹蒎酮(8.064％),去氢香树烯(6.063％),缬草萜烯醇(5.416％),戈米辛A(4.732％)等,其中非木脂素成分含量为67.06％.抗TMV活性和抗肿瘤活性数据显示北五味子总木脂素对A549细胞增殖有一定的抑制作用,对TMV有较好的抑制作用.结论:运用GC-MS方法对北五味子总木脂素中的低极性成分进行分析检测,该分析方法简便、快捷、准确、可靠,这为分析中药五味子总木脂素中非木脂素成分提供了一种更加高效的检测方法,同时为五味子中总木脂素品质的化学和生物功能研究提供了科学依据.

[Obj ective]Comparing the difference in volatile components in fresh leaves at different growth stages and products of Duyun Maoj ian tea among different grades,and researching its antibacterial activity.[Method]The volatile components of fresh leaves of 3 kinds of Duyun Maoj ian tea and 4 kinds of processed tea were extracted by SDE,and analyzed by GC-MS and various components were identified by spectral library Nist2005 and Wiley275,the relative content of each component was determined by peak are-a normalization method.The AGAR plate method of drilling hole was used to determine the antibacterial activities of the volatile oil of 7 samples against 6 bacterial species.[Result]124 kinds of volatile chemical components were detected from the 7 samples in which 18 kinds were the 14same.73 kinds of volatile compo-nents were detected from 3 kinds of fresh tea leaves and 40 kinds were the same ingredients.93 kinds of volatile components were detected from 4 kinds of processed tea and 3 5 kinds were the same.7 samples had inhibitory effect&nbsp;on Bacillus subtilis.Except one bud and one leaf early development,the other 6 samples had inhibiting effect on Staphylococcus aureus.[Conclusion]The processing of Duyun Maoj ian tea had certain influence on volatile compo-nents,volatile components differed at different growth stages and in different grades of processed tea.The volatile components of Duyun Maoj ian tea had certain antibacterial activity.

Extensive phytochemical investigation of the aerial parts of Euphorbia dracunculoides led to the isolation of 19 structurally diversified diterpenoids with five different carbon skeletons, including six new lathyrane glycosides (1–6), three new cyclomyrsinanes (7–9), three new premyrsinanes (10–12), and one new 10,18-dihydromyrsinane (13). Their structures were elucidated by means of extensive spectroscopic analysis, and the structure of 7 was further confirmed by X-ray crystallography. Compounds 1–6 are the first examples of C-17-glycosylated lathyrane diterpenoids. Selected isolates were evaluated for their cytotoxicity against five cancer cell lines. Only the tiglianes exhibited moderate activity.

目的设计、合成具有抗H2O2损伤PC12细胞活性的熊果酸(Ursolic Acid,UA)衍生物.方法通过苄基化、磺酰化、烯化等反应对熊果酸C1-3、C28位点进行修饰,合成系列1α,2α,3α-三羟基熊果酸及其衍生物;采用MTT法对目标产物的细胞毒性、H2O2损伤PC12细胞保护作用进行测试.结果合成1,2,3-三羟基熊果酸及其衍生物9个(7-15),MTT活性测试结果显示化合物10细胞毒性比UA小,浓度为1.0×10-5mol/L时,对H2O2损伤PC12细胞抑制率为13.10％;13、15无细胞毒性,化合物15对H2 O2损伤PC12细胞的保护作用强于阳性对照维生素E,浓度为1.0 × 10-4mol/L时,抑制率为40.18％.结论1α,2α,3α-三羟基熊果酸对H2O2损伤的PC12细胞具有一定的保护作用,尤其是C28位被二乙胺、二乙醇胺取代后活性明显增强.

Two diarylheptanoids, musaitinerins A and B, one heterodimeric phenylphenalenone musaitinerone and four known phenylphenalenones, identified as 4-hydroxy-2-methoxy-9-phenyl-1H-phenalen-1-one, musanolone E, hydroxyanigorufone and irenolone were isolated from the fruits of Musa itinerans Cheesm. Their structures were elucidated using spectroscopic analyses. The antimicrobial activity of these compounds was evaluated against Escherichia coli, Staphylococcus aureus and Candida albicans; the cytotoxic activity of these compounds was also evaluated against human erythromyeloblastoid leukemia (K562) and human alveolar carcinoma epithelial (A549) cell lines, respectively. Musaitinerone and musanolone E exhibited weak effects against the A549 cell line, as compared with adriamycin. However, these two compounds did not exhibit any growth inhibition against K562 cells, S. aureus, E. coli or C. albicans. The other compounds were inactive against all of the tested cell lines and microorganisms, even at concentrations as high as 50 μM.