BACKGROUND:Doxorubicin (DOX) is a widely used antitumor drug. However, its clinical application is limited for its serious cardiotoxicity. The mechanism of DOX-induced cardiotoxicity is attributed to the increasing of cell stress in cardiomyocytes, then following autophagic and apoptotic responses. Our previous studies have demonstrated the protective effect of Shenmai injection (SMI) on DOX-induced cardiotoxicity via regulation of inflammatory mediators for releasing cell stress.PURPOSE:To further investigate whether SMI attenuates the DOX-induced cell stress in cardiomyocytes, we explored the mechanism underlying cell stress as related to Jun N-terminal kinase (JNK) activity and the regulation of autophagic flux to determine the mechanism by which SMI antagonizes DOX-induced cardiotoxicity.STUDY DESIGN:The DOX-induced cardiotoxicity model of autophagic cell death was established in vitro to disclose the protected effects of SMI on oxidative stress, autophagic flux and JNK signaling pathway. Then the autophagic mechanism of SMI antagonizing DOX cardiotoxicity was validated in vivo.RESULTS:SMI was able to reduce the DOX-induced cardiomyocyte apoptosis associated with inhibition of activation of the JNK pathway and the accumulation of reactive oxygen species (ROS). Besides, SMI antagonized DOX cardiotoxicity, regulated cardiomyocytes homeostasis by restoring DOX-induced cardiomyocytes autophagy. Under specific circumstances, SMI depressed autophagic process by reducing the Beclin 1-Bcl-2 complex dissociation which was activated by DOX via stimulating the JNK signaling pathway. At the same time, SMI regulated lysosomal pH to restore the autophagic flux which was blocked by DOX in cardiomyocytes.CONCLUSION:SMI regulates cardiomyocytes apoptosis and autophagy by controlling JNK signaling pathway, blocking DOX-induced apoptotic pathway and autophagy formation. SMI was also found to play a key role in restoring autophagic flux for counteracting DOX-damaged cardiomyocyte homeostasis.

AIMS:Doxorubicin (DOX) is an effective anthracycline anticancer drug. However, the clinical usage of it is limited due to its severe cardiotoxicity side effects. Metformin (Met) is a kind of first-line antihyperglycemic drug which has a potential protective effect on the heart，it is often used for oral treatment of type 2 diabetes. In this study, we explored whether Met could attenuate cardiotoxicity induced by DOX. MATERIALS AND METHODS:For the sake of exploring the Met protective effect and mechanism, we established the DOX-induced cardiotoxicity models both in H9C2 cells incubated with 5 μM DOX in vitro and Sprague-Dawley rats treated with 20 mg/kg cumulative dose of DOX. KEY FINDINGS:Met is able to inhibit growth inhibition and apoptosis of H9C2 cells induced by DOX. The heart indexes of rats were examined to evaluate the Met cardiotoxicity protection. Met improved the abnormal indexes, serum markers of cardiac heart injury, echocardiography, electrocardiogram, cardiac pathology, cardiomyocyte apoptosis, and oxidative stress markers induced by DOX. Furthermore, in vivo and in vitro studies demonstrated that Met protected against DOX-induced increasing cleaved caspase-3 and Bax. Met also prevented the downregulation of Bcl-2, activated the AMPK pathway, and inhibited the MAPK pathway. SIGNIFICANCE:Met showed protective effects on DOX-induced cardiotoxicity by reducing oxidative stress and apoptosis, as well as regulating AMPK and MAPK signaling pathways.

Context Shenmai Injection (SMI) is usually used to treat atherosclerotic coronary heart disease and viral myocarditis in China. However, the effect of SMI on multidrug resistance has not been reported. Objective To investigate the reversal effect of SMI in adriamycin (ADR) resistant breast cancer cell line (MCF-7/ADR) and explore the related molecular mechanisms. Materials and methods The effect of SMI (0.25, 0.5, 1 mg/mL) to reverse chemoresistance in MCF-7/ADR cells was elucidated by MTT, HPLC-FLD, DAPI staining, flow cytometric analysis, western blotting. At the same time, in vivo test was conducted to probe into the effect of SMI on reversing ADR resistance, and verapamil (10 μM) was used as a positive control. Results The results showed that the toxicity of ADR to MCF-7/ADR cells was strengthened significantly after treated with SMI (0.25, 0.5, 1 mg/mL), the IC50 of ADR was decreased 54.4-fold. The intracellular concentrations of ADR were increased 2.2-fold (p < 0.05) and ADR accumulation was enhanced in the nuclei (p < 0.05). SMI could strongly enhance the ADR-induced apoptosis and increase intracellular rhodamine 123 accumulation in MCF-7/ADR cells. Additionally, a combination of ADR and SMI (5 mg/kg) could dramatically reduce the weight and volume of tumour (p < 0.05). Furthermore, the results revealed that SMI might reverse MDR via inhibiting ADR-induced activation of the mitogen-activated protein kinase/nuclear factor (NF)-κB pathway to down-regulated the expression of P-glycoprotein (P-gp). Discussion and conclusions SMI could potentially be used to treat ADR-resistance. This suggests possibilities for future clinical research.

目的 建立UHPLC法同时测定参麦注射液(红参、麦冬)中人参皂苷Rg1、Re、Rf、Rb1、Rc、Rb2、Rd、Rg3的含有量.方法 该药物的分析采用Agilent Eclipse Plus C18色谱柱(2.1 mm×50 mm,1.8 μm);流动相乙腈-水,梯度洗脱;体积流量0.3 mL/min;柱温30℃;检测波长203 nm.结果 8种人参皂苷在各自范围内线性关系良好(r＞0.997 0),平均加样回收率97.43％～104.24％,RSD 0.37％～5.34％.结论 该方法稳定可靠,重复性好,可用于参麦注射液质量控制.

Background Doxorubicin (DOX) is a chemotherapy drug for malignant tumors. The clinical application of DOX is limited due to its dosage relative cardiotoxicity. Oxidative damage and cardiac inflammation appear to be involved in DOX-related cardiotoxicity. Shenmai injection (SMI), which mainly consists of Panax ginseng C.A.Mey.and Ophiopogon japonicus (Thunb.) Ker Gawl , is widely used for the treatment of atherosclerotic coronary heart disease and viral myocarditis in China. In this study, we investigated the protective effect of Shenmai injection on doxorubicin-induced acute cardiac injury via the regulation of inflammatory mediators. Methods Male ICR mice were randomly divided into seven groups: control, DOX (10 mg/kg), SMI (5 g/kg), DOX with pretreatment with SMI (0.5 g/kg, 1.5 g/kg or 5 g/kg) and DOX with post-treatment with SMI (5 g/kg). Forty-eight hours after the last DOX administration, all mice were anesthetized for ultrasound echocardiography. Then, serum was collected for biochemical and inflammatory cytokine detection, and heart tissue was collected for histological and Western blot detection. Results A cumulative dose of DOX (10 mg/kg) induced acute cardiotoxicity in mice manifested by altered echocardiographic outcome, and increased tumor necrosis factor, interleukin 6 (IL-6), monocyte chemotactic protein 1, interferon-γ, and serum AST and LDH levels, as well as cardiac cytoplasmic vacuolation and myofibrillar disarrangement. DOX also caused the increase in the expression of IKK-α and iNOS and produced a large amount of NO, resulting in the accumulation of nitrotyrosine in the heart tissue. Pretreatment with SMI elicited a dose-dependent cardioprotective effect in DOX-dosed mice as evidenced by the normalization of serum inflammatory mediators, as well as improve dcardiac function and myofibril disarrangement. Conclusions SMI could recover inflammatory cytokine levels and suppress the expression of IKK-α and iNOS in vivo, which was increased by DOX. Overall, there was evidence that SMI could ameliorate DOX-induced cardiotoxicity by inhibiting inflammation and recovering heart dysfunction.

目的:建立了同时测定参麦注射液中11个人参皂苷含量的超高效液相色谱方法,并对参麦注射液样品进行含量测定.方法:采用Aglient Eclipse plus C18(2.1 mm×50 mm,1.8 μm)色谱柱,乙腈-水为流动相,梯度洗脱,流速0.3 mL· min-1,柱温30℃,检测波长203 nm.结果:人参皂苷Rg1、Re、Rf、Rb1、Rc、Rb2、Rb3、Rd、20(S)-人参皂苷Rh1、20(S)-人参皂苷Rg2、20(S)-人参皂苷Rg3的线性范围分别为0.095～0.380 mg·mL-1(r=0.992 3)、0.055～0.220 mg· mL-1(r=0.992 2)、0.025～0.099 mg· mL-1(r=0.999 9)、0.004～0.019 mg· mL-1(r=0.999 3)、0.005～0.022 mg· mL-1(r=0.999 4)、0.156～0.626 mg· mL-1(r=1.000)、0.067～0.268 mg· mL-1(r=-1.000)、0.058～0.234 mg· mL-1(r=1.000)、0.007～0.031 mg· mL-1(r=0.999 8)、0.032～0.130 mg· mL-1(r=1.000)、0.007～0.028 mg· mL-1(r=0.9992),平均加样回收率(n=6)为91.4％～100.8％,RSD为0.34％～3.4％.样品中上述11个人参皂苷的含量范围分别为0.138 7～0.173 0、0.086 8～0.108 7、0.033 0～0.043 5、0.009 9～0.013 4、0.011 0～0.016 0、0.175 8～0.295 4、0.085 0～0.133 9、0.066 6～0.118 4、0.008 4～0.015 0、0.033 3～0.065 5、0.009 5～0.016 8 mg· mL-1.结论:该方法重复性好,稳定可控,可用于参麦注射液中人参皂苷类成分的含量测定.

目的 建立快速检测Beagle犬血浆中多西他赛浓度的液相色谱-质谱联用方法.方法 取100μL犬血浆,以200μL含有50 μg·L-1内标(紫杉醇)的乙腈溶液沉淀蛋白,10 000 r·min-15 min离心2次,取上清液进样10 μL.色谱柱:Agilent Eclipse Plus C18(150 mm× 2.1 mm,5μm),柱温:30℃,流动相:甲醇-水相梯度洗脱,流速0.2 mL· min-1.ESI源正离子SRM模式,定量的子离子:多西他赛[M+H]+ m/z:830.3→549.3(26 eV),紫杉醇[M+H]+ m/z:876.3→308.1(27 eV).结果 多西他赛的线性范围为5～1000 μg· L-1,标准曲线的相关系数r≥0.996 5,提取回收率为89.9％～114.8％,方法回收率为94.0％～97.5％,批内变异系数＜9.0％,批间变异系数＜10.8％.结论 本方法简便、迅速、灵敏,方法学各项指标均符合要求,适用于犬血浆中多西他赛浓度的检测.

To investigate the possibility of tedizolid phosphate's application in the treatment of intracranial infection, a preclinical comparative pharmacokinetic study was designed. Based on the assumption that the classic efflux transporters P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP) may participate in the transportation of TDZ, two groups of rats were intravenously administered 6 mg/kg tedizolid phosphate alone or 6 mg/kg tedizolid phosphate combined with 1 mg/kg elacridar which was an inhibitor of P-gp and BCRP. Plasma and cerebrospinal fluid samples were collected according to a pharmacokinetic schedule. All the plasma and cerebrospinal fluid samples were assessed with a validated LC-MS/MS method. The penetration ratio of tedizolid from the blood to cerebrospinal fluid was calculated, and a comparison of the penetration ratios between the two groups was made. The mean Cmax of tedizolid in the CSF in the tedizolid phosphate group and the tedizolid phosphate combined with elacridar group was 154 ng/mL and 300 ng/mL, respectively, and the mean penetration ratio of tedizolid in the tedizolid phosphate group and the tedizolid phosphate combined with elacridar group was 2.16% and 3.53%, respectively. The relatively high Cmax in the CSF proved the possibility of tedizolid phosphate's application in the treatment of intracranial infection, and the higher penetration ratios, Cmax, csf and AUCcsf of the rats in co-administered elacridar group than those in the single-administration group indicated that the transporters P-gp and BCRP might be involved in the transportation of tedizolid.

目的 研究黄酮碳苷牡荆素在大鼠体内的代谢产物,并推测其代谢途径.方法 SD大鼠灌胃给予5 mg·kg-1牡荆素,收集0～3 h,3～6 h,6～12 h的尿液,采用UPLC-Q-TOF检测尿样中代谢产物.结果 采用Metabolynx XS代谢物分析软件,根据质谱碎片信息对代谢物进行结构鉴定,最终得到3个代谢产物.结论 牡荆素在大鼠尿液中检测得到1个一相代谢产物,2个二相代谢产物,推测牡荆素在大鼠体内主要发生氧化、甲基化和葡萄糖醛酸结合反应,其中葡萄糖醛酸化反应是较强的代谢种类.

Quzhou Fructus Aurantii (QFA) is an authentic herb of local varieties in Zhejiang, China, which is usually used to treat gastrointestinal illnesses, but its effects on respiratory inflammation have not been reported yet. In our study, the anti-inflammatory activity of QFA extract (QFAE) was evaluated on copper sulfate pentahydrate (CuSO 4 ·5H 2 O)-induced transgenic neutrophil fluorescent zebrafish model. QFAE showed a significant effect of anti-inflammation in CuSO 4 ·5H 2 O-induced zebrafish by reducing the neutrophil number in the inflammatory site. We investigated the anti-inflammatory activity of QFAE on lipopolysaccharide (LPS)-induced acute lung injury (ALI) mice models and RAW 264.7 cells. QFAE had an anti-inflammatory effect on reducing total cells, neutrophils, and macrophages in BALF and attenuated alveolus collapse, neutrophils infiltration, lung W/D ratio, myeloperoxidase (MPO) protein expression and other pulmonary histological changes in lung tissues, as well as hematological changes. Levels of pro-inflammatory cytokines, including TNF, IL-6, IFN-γ, MCP-1, and IL-12p70, were decreased, whereas anti-inflammatory cytokine IL-10 was increased after treatment with QFAE both in vivo and in vitro . In summary, our results suggested that QFAE had apparent anti-inflammatory effects on CuSO 4 ·5H 2 O-induced zebrafish, LPS-induced ALI mice, and RAW 264.7 cells. Furthermore, QFAE may be a therapeutic drug to treat ALI/ARDS and other respiratory inflammations.

Autophagy is an intracellular degradation pathway widely existing in eukaryotic cells. The degradation of intracellular protein complexes, damaged organelles and invading pathogens by autophagy after the package is then transported to the lysosomes and lysosomal acid hydrolase digestion, and released into the cytoplasm to maintain cell homeostasis. With the 2016 Nobel Prize in Physiology and Medicine awarded to researchers from the field of autophagy, autophagy research has attracted more and more attention. This review summarizes the processes of autophagy signal regulation, autophagy and lysosomal processes in autophagy, and discusses the molecular signaling pathways involved in autophagy.

课题组前期研究系统考察了参麦注射液在大鼠和犬等动物体内多种组分的毒代动力学.文章中提出了基于AUC自定义权重系数“中药多组分整合毒代动力学”思路,在参麦注射液犬和大鼠的毒代动力学数据基础上,根据各组分的AUC值计算各组分的权重系数,对血药浓度进行加权求和,获得基于AUC自定义权重系数的整合毒代动力学模型.参麦注射液的整合毒代模型能较为准确的反应参麦注射液在动物体内的暴露、蓄积等情况,并与相关生物标志物有高度的关联性.

参麦注射液是由红参和麦冬经提炼精制而成的中药复方注射剂,临床上主要用于病毒性心肌炎、冠心病等疾病的治疗.人参皂苷和麦冬皂苷是其主要活性成分,近年来围绕参麦注射液中皂苷类成分在体内的药代动力学研究不断丰富.本文就近年来参麦注射液中皂苷类化学成分的药代动力学研究进展进行综述,为相关研究提供参考.

Background: Ginsenosides have been widely used clinically for many years and were regarded as very safe. However, a few researches on the toxicities of these kinds of agents showed that some ginsenosides may have side-effect on the rats or dogs. So it is extremely necessary to further clarify the potential toxicity of ginsenosides. This study was carried out to investigate long-term toxicity and genotoxicity of 25-methoxydammarane-3, 12, 20-triol (25-OCH3-PPD), a new derivative of ginsenoside, in beagle dogs. Methods: Twenty-four beagle dogs were divided randomly into four treatment groups and repeatedly orally administered with 25-OCH3-PPD capsule at 60, 120, and 240 mg/kg/day for 91 consecutive days. Ames, micronucleus, and chromosomal aberration tests were established to analyze the possible genotoxicity of 25-OCH3-PPD. Results: There was no 25-OCH3-PPDeinduced systemic toxicity in beagle dogs at any doses. The level of 25-OCH3-PPD at which no adverse effects were observed was found to be 240 mg/kg/day. The result of Ames test showed that there was no significant increase in the number of revertant colonies of 25-OCH3-PPD administrated groups compared to the vehicle control group. There were also no significant differences between 25-OCH3-PPD administrated groups at all dose levels and negative group in the micronucleus test and chromosomal aberration assay. Conclusion: The highest dose level of 25-OCH3-PPD at which no adverse effects were observed was found to be 240 mg/kg per day, and it is not a genotoxic agent either in somatic cells or germs cells. 25-OCH3-PPD is an extremely safe candidate compound for antitumor treatment. (C) 2018 The Korean Society of Ginseng, Published by Elsevier Korea LLC.

1 毒代动力学的历史和含义 临床前药物安全性评价研究中的毒代动力学(Toxicokinetics,TK)研究受试物在毒性试验中不同剂量水平下的全身暴露程度和持续时间,预测受试物在人体暴露时的潜在风险,是非临床毒性试验的重要研究内容之一[1].药物TK是一门较新的学科,人用药品注册技术要求国际协调会议(ICH)首次提出TK研究的指导原则(ICH,S3,a、b部分)[24].国内药物TK研究起步更晚,2004年修订的指导原则中才首次提出药物TK的相关内容[5],并鼓励创新药物进行TK研究,相关的技术性指导文件主要参照ICH关于TK的相关内容.2014年我国颁布了《药物毒代动力学研究技术指导原则》.在最新颁布的TK指导原则中,鼓励扩大开展TK研究的药物范围,而不仅限于创新药物,并将伴随TK从长期毒性试验推荐扩展至单次/重复给药毒性试验、遗传/生殖毒性试验和致癌性试验,可见TK在整个药物安全性评价体系中的地位大大提升,已成为药物安全性评价一个及其重要的组成部分[6-9].

目的 建立大鼠多次采集脑脊液的动物模型,以进行药物在大鼠脑部的动力学研究.方法 将大鼠麻醉后,利用脑立体定位仪,通过大鼠双侧耳道与上方门齿固定大鼠头部,切开大鼠头部皮肤,暴露颅骨与颈部肌肉交界处,将采样针对准大鼠颅骨与颈部肌肉交界处的中心位置,利用脑立体定位仪的上下轴将采样针竖直缓慢向下刺入,刺入约0.6～0.9 cm(视大鼠体质量而定)后,脑脊液即可被采出.结果 脑脊液采集成功率(脑脊液成功采样1次及以上)100％(26例),动力学采样成功率(脑脊液成功采样6次及以上)80％(10例).结论 本模型操作简单、结果稳定、成功率高、重复性高,适合脑部作用药物进行脑脊液动力学研究.

目的 探讨新型抗氧化剂依达拉奉(edaravone,EDA)拮抗阿霉素(doxorubicin,DOX)诱导的H9C2心肌细胞损伤中的自噬机制.方法 H9C2心肌细胞加EDA和不同浓度DOX共同孵育后,MTT法检测细胞存活率并用试剂盒测定过氧化物岐化酶(SOD)活性和氧化代谢产物丙二醛(MDA)水平,DAPI染色法观察细胞凋亡情况,Western blot测定自噬相关蛋白LC3B表达水平.结果 在实验设置细胞评价体系下,浓度为1μg/ml的EDA可以显著提高DOX作用下H9C2心肌的存活率,同时能显著降低DOX(浓度为1uμmol/L)引起细胞中MDA水平(P ＜0.05,P＜0.01)的升高,且使DOX诱导的SOD活性的降低得到显著提升(P＜0.05),上述差异有统计学意义.DAPI染色结果显示EDA能够减少DOX导致的心肌细胞凋亡,并显著抑制DOX诱导的自噬相关蛋白LC3B的增加,差异有统计学意义(P ＜0.05,P＜0.01).结论 DOX能引起H9C2细胞产生氧化应激,并诱导细胞自噬进而使细胞凋亡.而EDA对DOX诱导的心肌细胞凋亡具有保护作用,这种作用可能是通过调节自噬并降低细胞内氧化应激损伤而保护的心肌细胞.

目的:筛选大鼠体内碱性彗星试验中阳性药环磷酰胺(CP)和甲基磺酸乙酯(EMS)的给药周期.方法:将SD大鼠随机分为空白对照组、CP组(40 mg/kg,腹腔注射)和EMS组(100 mg/kg,灌胃),每组9只,每24 h给药1次,连续给药4次.分别于第2、3、4次给药后3 h(即首次给药后27、51、75 h)各组随机选取3只大鼠,处死并收集肝和胃样本,经过制备单细胞悬液、制片、裂解、解链、电泳及染色后,使用Comet AssayⅣ彗星试验数据分析系统进行分析,以彗星尾部DNA百分比(Tail DNA％)、尾长(TL)和尾矩(TM)为评价指标,筛选最佳给药周期.结果:空白对照组大鼠3个检测时间点肝和胃的Tail DNA％、TL、TM差异均无统计学意义(P>0.05),表明彗星试验体系比较稳定.与空白对照组比较,CP组和EMS组大鼠3个检测时间点肝和胃的Tail DNA％、TL(27 h的胃除外)、TM均明显升高(P<0.05).其中,CP组大鼠3个检测时间点间肝的Tail DNA％、TL、TM差异均无统计学意义(P>0.05),胃的Tail DNA％、TL均在给药51 h时最大,TM随给药时间的延长而升高;EMS组大鼠肝的Tail DNA％、胃和肝的TM均随给药时间的延长而升高.EMS组大鼠胃的Tail DNA％、肝和胃的TL均在给药51 h时最大.结论:在进行体内碱性彗星试验时,建议CP和EMS的给药周期是每24 h给药1次,连续给药3次.

Objective:To study the influence of PTD-CHMP1A (protein transduction domain-chromatin modifying protein 1A) fusion proteins on proliferation,migration and invasion of renal cell carcinoma (RCC) cells in vitro and the growth of transplanted rumor in vivo.Methods:RCC A498 and 786-0 cells were treated with PTD-CHMP1A (1,5,and 20 μg/ml),CHMP1A (5 μg/ml) and PBS respectively.The effect of PTD-CHMP1A on the proliferation ofA498 cells was detected by MTT assay.The effect of PTD-CHMP1A on the migration ofA498 cells was detected by scratch assay.The effect of PTD-CHMP 1A on the invasion ability ofA498 and 786-0 cells was detected by Transwell invasion assay.Finally,the transplanted tumor models were constructed by injecting 5 × 105 A498 cells into nude mice,and the effect of 20 μg PTD-CHMP1A on tumor growth was observed.Results:Compared with CHMP1A and PBS groups,PTD-CHMP1A fusion proteins (1,5,and 20 μg/ml) showed a significantly restrained effect on the proliferation ofA498 tumor cells,which was in dose-dependent and time-dependent manners.5 μg/ml of PTD-CHMP1A fusion proteins could effectively inhibit the migration ofA498 cells (P＜0.01 compared with CHMP1A group),and the invasion ofA498 and 786-0 cells (compared with Chmp1A group,P＜ 0.05),besides,PTD-CHMP1A inhibited the invasion ofA498 cells in a dose-dependent manner.Furthermore,PTD-CHMP1A could also observably inhibit tumor growth in vivo.Conclusion:PTD-CHMP1A fusion proteins effectively inhibited the proliferation,migration and invasion of RCC cells and the growth of transplanted tumor.

Doxorubicin (Dox) is an effective wide-spectrum antitumor drug. However, its clinical application may be hampered by dose-dependent cardiotoxicity. The mechanisms of cardiotoxicity have not been clearly elucidated, but are known to involve oxidative stress, mitochondrial dysfunction and apoptosis. Autophagy is a lysosome-dependent protein degradation pathway. More recently, many studies have suggested that autophagy plays an important role in Dox-induced cardiotoxicity. This paper gives a systematic review of the role of autophagy in Dox-induced cardiotoxicity.