HPLC法测定丹参和丹参配方颗粒中丹参酮ⅡA的含量的研究

采用高效液相色谱法测定丹参和丹参配方颗粒中丹参酮ⅡA的含量方法。结果：该方法的线性范围为14～138μg／ml(r=1．0000)，平均回收率为98．39％，样品平均含量为1．33％和0．01％。提示本法简便、准确、灵敏度高、重现性好，可用于丹参和丹参配方颗粒的成分含量测定。     

丹参水溶性成分在慢性肾脏病中的研究进展

近年来,丹参总酚酸、丹参素、丹酚酸A、B等丹参水溶性成分因其肯定的抗氧化、抗血小板聚集等作用不仅广泛应用于肝病、心脑血管疾病和抗肿瘤等方面,而且其在慢性肾脏病防治中也取得了较大进步。本文就其在慢性肾脏病中的研究进展作一综述。     

丹参酮IIA磺酸钠注射液联合降脂方治疗非酒精性脂肪肝疗效观察

目的:观察丹参酮IIA磺酸钠注射液联合降脂方治疗非酒精性脂肪肝的临床疗效。方法:将60例患者随机分为2组各30例,试验组予丹参酮IIA磺酸钠注射液静脉滴注,并口服中药降脂方;对照组予阿托伐他汀钙口服,观察比较2组患者治疗前与治疗4周后肝/脾CT值、甘油三酯、总胆固醇及中医证候积分变化。结果:治疗后2组患者肝/脾CT值、甘油三酯、总胆固醇、中医证候积分均较治疗前明显改善(P0.05);治疗组各项指标改善较对照组更为显著(P0.05)。结论:丹参酮IIA磺酸钠注射液联合降脂方能改善患者临床症状及肝/脾CT值、甘油三酯、总胆固醇,疗效优于阿托伐他汀钙。     

Effects of tanshinone IIA, a major component of Salvia miltiorrhiza, on platelet aggregation in healthy newborn piglets

Ethnopharmacological relevance

Tanshinone IIA (STS), an active ingredient of the Chinese herb Danshen (Salvia miltiorrhiza) for angina and stroke in adults, has been reported to inhibit platelet function. However, its effect on platelet and underlying mechanism remain largely unknown, particularly in neonates.

Materials and Methods

To investigate the effect of STS on the platelet aggregation and its interaction with various platelet activation pathways, platelet aggregatory function was studied in whole blood stimulated by collagen (2-10 μg/ml) ex vivo in newborn piglets receiving intravenous STS (0.1-10 mg/kg, n = 8) and in vitro in whole blood from newborn piglets (n = 6) incubated with STS (0.1-100 μg/ml). The respective morphological changes of platelets were also examined by scanning electron microscopy. Plasma levels of nitrite/nitrate (NOx) and thromboxane B₂ (TxB₂), matrix metalloproteinase (MMP)-2 and -9 activities were also examined. To further delineate the mechanistic pathway, the effect of STS on endothelial microparticles release from cultured human umbilical vein endothelial cells (HUVECs) was quantified by flow cytometry.

Results

STS impaired the ex vivo, but not in vitro, collagen-stimulated platelet aggregation. Infusion of STS elevated the plasma level of TxB₂ at 10 mg/kg. However, STS had no effect on NOx level. Incubating cultured HUVECs with STS (1 and 10 μg/ml) caused a significant release of endothelial microparticles. Morphologically, STS elicited platelet activation in vivo, but not in vitro.

Conclusions

STS impairs the ex vivo whole blood platelet aggregatory function by activating platelet in vivo in healthy newborn piglets. It implies that STS may elicit its effects by stimulating endothelial microparticles production and eicosanoid metabolism pathway.     

丹参酮ⅡA对心血管系统药理作用的研究进展

丹参酮ⅡA是中药丹参中主要有效活性成分,能通过多种机制等发挥保护心血管系统的作用。文中总结了近年来科研工作者对丹参酮ⅡA对心血管系统的保护作用的基础研究,为丹参酮ⅡA更广泛应用于临床心脑血管等疾病的治疗提供保障。     

丹参酮IIA的抗肿瘤药理作用及其纳米给药系统研究进展＊

近年来，肿瘤发病率不断攀升，化疗是目前临床治疗肿瘤的主要策略，但因多数化疗药物毒副作用大且容易造成化学耐药性，导致化疗效果并不理想。而传统中药凭借其多途径、多靶点、多通路的特点在抗肿瘤方面具有治疗温和不剧烈、毒副作用小等独特的优势。丹参酮IIA（Tanshinone IIA，TSA）作为丹参中提取的主要菲醌类衍生物，具有明显的抗肿瘤作用，具备被开发为毒性小、疗效好的新型抗肿瘤药物的条件。但因其难溶于水、半衰期短、生物利用度低等缺点极大程度上限制了其临床应用。利用纳米技术，可以改善丹参酮IIA的不足；构建中药纳米递药系统可以提高肿瘤组织靶向性来改善半衰期短、稳定性差等实际临床应用中的问题。本文主要综述丹参酮IIA的抗肿瘤机理及其纳米递药系统的研究进展，以期为丹参酮IIA的深入研究和临床应用提供参考。     

丹参酮IIA对肾癌786-O细胞生长抑制作用及其分子机制

目的 研究丹参酮IIA对肾癌细胞生长抑制作用及其分子机制.方法 MTT法检测丹参酮IIA对肾癌细胞活力影响;流式细胞分析检测丹参酮IIA对肾癌细胞周期阻滞;免疫印迹检测细胞周期阻滞相关靶蛋白蛋白表达水平;激光共聚焦观察核仁蛋白Nucleophosmin (NPM)/B23定位的变化.结果 丹参酮能够呈浓度依赖方式显著抑制肾癌细胞生长活力(P＜0.05);细胞周期分析表明丹参酮IIA处理的肾癌细胞阻滞在S期;免疫印迹结果证明丹参酮IIA处理肾癌细胞,cyclin A,p53及其下游基因p21显著上调;激光共聚焦结果表明在丹参酮IIA作用下,NPM蛋白定位从核仁移位到核浆.结论 丹参酮IIA作用786-O细胞导致细胞周期S阻滞,其机制可能与NPM移位促使其相互作用靶蛋白p53和p21蛋白水平上调有关.     

右归丸治疗肾阳虚型慢性肾脏病2～3期的临床疗效研究

[目的]观察右归丸治疗肾阳虚型慢性肾脏病2~3期的临床疗效。[方法]选取80例肾阳虚型慢性肾脏病2~3期患者,随机分为治疗组和对照组,对照组(40例)予西医一体化治疗,治疗组(40例)在西医一体化治疗的基础上加用右归丸,观察两组患者在治疗前、治疗后的血清肌酐(Scr)、估算肾小球滤过率(e GFR)、24 h尿蛋白定量及中医证候积分的变化情况,观察周期为24周。[结果]在血清Scr、24 h尿蛋白定量、e GFR及中医证候改善方面,治疗组均优于对照组(P0.05)。[结论]右归丸可有效改善肾阳虚型慢性肾脏病2~3期患者的中医证候积分及其肾功能,降低24 h尿蛋白定量,具有一定的肾脏保护作用,并且临床运用安全。     

Neuroprotective effects of Tanshinone IIA on permanent focal cerebral ischemia in mice

The objective of this study was to evaluate whether Tanshinone IIA (TSA) was neuroprotective in permanent focal cerebral ischemia and to determine the possible mechanisms of its neuroprotection. Mice were subjected to permanent middle cerebral artery occlusion. The neuroprotection of TSA was investigated with respect to neurological deficit scores and infarct volume. Biochemical analyses for malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in serum, and nitric oxide (NO) content and the inducible nitric oxide synthase (iNOS) activity in brain tissue were performed at 24 h after ischemia. Immunohistochemistry was used to measure the expression of iNOS. In vitro, the effects of TSA were tested in the cultured astrocytes exposed to hydrogen dioxide (H2O2). TSA (5, 10 and 20 mg/kg, i.p.) significantly reduced the infarct volume and improve neurological deficit. TSA also significantly increased the activity of SOD after 24 h of ischemia and decreased the MDA level, NO content, and iNOS expression. In vitro, the translocation of NF-kappaB was inhibited by TSA and the survival rate of astrocytes was markedly increased and the NO production was decreased. In conclusion, these results illustrated that TSA protected the brain from ischemic injury by suppressing the oxidative stress and the radical-mediated inflammatory insult.     

Cytotoxicity of Tanshinone IIA combined with Taxol on drug‐resist breast cancer cells MCF‐7 through inhibition of Tau

Drug resistance represents a major obstacle to improving the overall response and survival of cancer patients. Taxol is one of the most commonly used chemotherapy agents in breast cancer. As with many cancer therapeutic agents, resistance remains a significant problem when using Taxol to treat malignancies. In this study, estrogen receptor positive breast cancer cells MCF‐7 were induced Taxol resistance. And Tanshinone IIA combined with Taxol was chosen to treat it. The drugs combination showed additive effect in most drug concentrations. Drug resistance cancer cells showed a higher microtubule associated protein (Tau) expression, which was considered as one of the reasons for Taxol resistance. Tanshinone IIA inhibited the expression of Tau in MCF‐7 cells and resulted in higher sensibility of Taxol. Moreover, Tanshinone IIA also showed cytotoxicity to MCF‐7, which might be related to its estrogenicity effect. In conclusion, the combination of Tanshinone IIA and Taxol showed higher cytotoxicity to Taxol resistant MCF‐7 cells, which might be related to the inhibition of Tau.     

冠脉通片防治慢性肾脏病大鼠血管钙化的作用研究

[目的] 观察冠脉通片对大鼠慢性肾脏病（CKD）血管钙化的防治作用。[方法] 采用灌胃给予腺嘌呤加髙磷饮水的方法制备大鼠CKD血管钙化模型。造模同时灌胃给予不同剂量冠脉通片,连续10周。腹主动脉取血检测血清尿素、肌酐、钙、磷水平,试剂盒检测腹主动脉钙含量,苏木精-伊红（HE）染色观察肾和血管组织变化,硝酸银染色观察血管钙化情况。[结果] 冠脉通片0.46、0.23 g/kg组能显著降低模型大鼠血清尿素、肌酐及磷水平,升高钙水平;明显降低腹主动脉钙含量;肾脏HE染色结果提示,与正常组相比,模型组大鼠肾小球结构紊乱,肾小管代偿性扩张,局部见棕褐色结晶沉积,肾脏间质弥漫淋巴和单核细胞浸润,大量成纤维细胞增生,形成广泛纤维化,肾脏血管明显减少。与模型组比较,冠脉通片组正常肾小球数目增多,肾小管扩张程度降低,炎性细胞数量下降,腺嘌呤结晶减少。其中,冠脉通片0.46 g/kg组效果更好。HE与硝酸银染色显示,与模型组相比,冠脉通片0.46、0.23 g/kg组大鼠胸主动脉钙化灶面积明显缩小,其中冠脉通片0.46 g/kg组效果更好。[结论] 冠脉通片对CKD血管钙化大鼠有明显防治作用,可改善肾功能,纠正钙、磷代谢紊乱,降低动脉钙含量,缩小钙化面积。     

CVVHDF联合乌司他丁治疗慢性肾脏病并发急性胰腺炎疗效观察

目的观察连续静脉静脉血液透析滤过(CVVHDF)联合乌司他丁治疗慢性肾脏病并发急性胰腺炎患者的临床效果。方法 30例慢性肾脏病并发急性胰腺炎患者随机分为3组,每组10例。A组:予以血液透析(HD)+常规治疗;B组:予以CVVHDF+常规治疗;C组:予以CVVHDF+乌司他丁+常规治疗。观察比较3组患者治疗后腹部体征改善情况、血尿淀粉酶、肝功能、白细胞计数、C反应蛋白、APACHEⅡ评分、低血压发生次数及心律失常发生次数。结果治疗后3组患者血尿淀粉酶、白细胞计数、肝功能等指标均有改善。与A组相比,B组、C组患者腹部体征改善时间均明显缩短,低血压及心律失常发生次数明显减少。且B组、C组在第7天APACHEⅡ评分及第3天C反应蛋白均明显下降,而C组下降水平较B组更为明显。结论 CVVHDF具有血流动力学稳定及清除溶质更为彻底的优点,在慢性肾脏病合并急性胰腺炎患者的治疗中效果明显优于HD,且联合应用乌司他丁可以进一步改善患者全身炎症状态,进一步提高治疗效果。     

CVVHDF联合乌司他丁治疗慢性肾脏病并发急性胰腺炎疗效观察

目的观察连续静脉静脉血液透析滤过（CVVHDF）联合乌司他丁治疗慢性肾脏病并发急性胰腺炎患者的临床效果。方法 30例慢性肾脏病并发急性胰腺炎患者随机分为3组,每组10例。A组：予以血液透析（HD）＋常规治疗;B组：予以CVVHDF＋常规治疗;C组：予以CVVHDF＋乌司他丁＋常规治疗。观察比较3组患者治疗后腹部体征改善情况、血尿淀粉酶、肝功能、白细胞计数、C反应蛋白、APACHEⅡ评分、低血压发生次数及心律失常发生次数。结果治疗后3组患者血尿淀粉酶、白细胞计数、肝功能等指标均有改善。与A组相比,B组、C组患者腹部体征改善时间均明显缩短,低血压及心律失常发生次数明显减少。且B组、C组在第7天APACHEⅡ评分及第3天C反应蛋白均明显下降,而C组下降水平较B组更为明显。结论 CVVHDF具有血流动力学稳定及清除溶质更为彻底的优点,在慢性肾脏病合并急性胰腺炎患者的治疗中效果明显优于HD,且联合应用乌司他丁可以进一步改善患者全身炎症状态,进一步提高治疗效果。     

基于网络药理学探讨丹参酮ⅡA治疗脊髓损伤的可能机制及相关通路

目的:基于网络药理学分析丹参酮ⅡA治疗脊髓损伤的分子生物学机制及相关通路。方法:利用TCMSP、BATMAN-TCM、DTPS、STITCHI、Swiss Target Prediction 5个数据库筛选出丹参酮ⅡA的作用靶点,利用OMIM、GeneCards、CTD数据库检索脊髓损伤相关基因,利用韦恩图筛选丹参酮ⅡA治疗脊髓损伤的作用靶点。利用STRING数据库进行蛋白互作网络分析,利用Cytoscape软件构建蛋白相互作用网络,利用Metascape数据库进行GO分析和KEGG信号通路富集分析。结果:丹参酮ⅡA共有潜在靶点182个,与脊髓损伤相关的靶点157个,其中肿瘤蛋白p53(TP53)、Myc原癌基因蛋白(MYC)、G1/S-特异性周期蛋白-D1(CCND1)、半胱氨酸天冬氨酸蛋白酶3(CASP3)、JUN原癌基因蛋白(JUN)5个靶点在蛋白互作网络中具有较为重要的作用。丹参酮ⅡA治疗脊髓损伤涉及的生物学过程主要包括药物反应、脂多糖反应、细胞对DNA损伤刺激的反应等;涉及的分子功能主要包括酶结合、转录因子结合、蛋白结合等。其作用机制可能与cAMP signaling pathway、MAPK signaling pathway、PI3K-Akt signaling pathway、HIF-1 signaling pathway、TNF signaling pathway、Apoptosis等通路有关。结论:丹参酮ⅡA治疗脊髓损伤具有多靶点、多通路作用的特点,其可能通过促进神经元存活、促进轴突再生、抑制神经细胞凋亡、抗炎等过程对脊髓损伤过程进行干预。     

Tanshinone IIA,a constituent of Danshen,inhibits the release of glutamate in rat cerebrocortical nerve terminals

Ethnopharmacological relevance

Danshen is a commonly used traditional Chinese medicine and has received considerable attention due to their beneficial effects on the health, including prevention of cardiovascular disease, and cancer. Tanshinone IIA, a major active constituent of Danshen, has been reported to have a neuroprotective profile.

Aim of the study

An excessive release of glutamate is considered to be related to neuropathology of several neurological diseases. In this study, we investigated whether tanshinone IIA could affect endogenous glutamate release and explored the possible mechanism.

Materials and methods

The experimental model was the isolated nerve terminals (synaptosomes) purified from the rat cerebral cortex. The release of glutamate was evoked by the K⁺ channel blocker 4-aminopyridine (4-AP) and measured by one-line enzyme-coupled fluorometric assay. We also used a membrane potential-sensitive dye to assay nerve terminal excitability and depolarization, and a Ca²⁺ indicator, Fura-2-acetoxymethyl ester, to monitor cytosolic Ca²⁺ concentrations ([Ca²⁺]_C).

Results

Tanshinone IIA inhibited the release of glutamate evoked by 4-AP in a concentration-dependent manner. Inhibition of glutamate release by tanshinone IIA was prevented by the chelating the extracellular Ca²⁺ ions, and by the vesicular transporter inhibitor bafilomycin A1. However, the glutamate transporter inhibitor DL-threo-beta-benzyl-oxyaspartate did not have any effect on the action of tanshinone IIA. Tanshinone IIA decreased the depolarization-induced increase in [Ca²⁺]_C, whereas it did not alter the resting synaptosomal membrane potential or 4-AP-mediated depolarization. Furthermore, the effect of tanshinone IIA on evoked glutamate release was prevented by the Ca_v2.2 (N-type) and Ca_v2.1 (P/Q-type) channel blocker ω-conotoxin MVIIC, but not by the ryanodine receptor blocker dantrolene or the mitochondrial Na⁺/Ca²⁺ exchanger blocker CGP37157. Mitogen-activated protein kinase (MEK) inhibition also prevented the inhibitory effect of tanshinone IIA on evoked glutamate release.

Conclusion

These results show that tanshinone IIA inhibits glutamate release from cortical synaptosomes in rats through the suppression of presynaptic voltage-dependent Ca²⁺ entry and MEK signaling cascade.     

雷公藤多甙治疗慢性肾脏病Ⅰ期患者的研究

目的观察雷公藤多甙治疗慢性肾脏病(CKD)Ⅰ期患者的临床疗效。方法将CKDⅠ期患者随机分成2组,雷公藤组予雷公藤多甙片60 mg/d,对照组予肌苷片0.6 g/d。2组均治疗6个月,停药观察6个月。观察指标为尿常规2、4 h尿蛋白定量、肌酐清除率(Ccr)、血压、肝功能、血常规、血脂、临床症状等。结果治疗6个月后,雷公藤组尿蛋白减少,且停药6个月后尿蛋白仍稳定。尿蛋白定性由治疗前(+)~(),减为治疗后(-)~(+);镜下血尿也减轻。结论雷公藤多甙能显著降低CKDⅠ期患者蛋白尿与血尿,保护肾功能,有利于患者长期病情稳定。     

丹参酮ⅡA对心肌梗死模型大鼠心肌钙调蛋白及心功能的影响

目的：观察丹参酮ⅡA对心肌梗死模型大鼠心肌钙调蛋白(cardiac muscle, CaM)及钙调蛋白依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependentproteinkinase-Ⅱ, CaMKⅡ)mRNA 表达和心功能的影响。方法100只大鼠按随机数字表法分为假手术组、模型组、丹参酮ⅡA组。模型组、丹参酮ⅡA组采用结扎左侧冠状动脉前降支45 min再灌注45 min,并反复3次阻断与再灌注的方法建立大鼠心肌梗死模型。丹参酮ⅡA组尾静脉注射丹参酮ⅡA磺酸钠注射液10 mg/kg,假手术组、模型组尾静脉注射等体积生理盐水。连续给药7 d后断头处死,取心脏用Langendorff离体心脏灌流器灌流离体心脏,测定各组大鼠给药前后离体心脏左室最大上升/下降速率(±LVdp/dtmax)、心肌收缩张力、心率等心功能指标;TTC染色观察心肌梗死面积;检测缺血区心肌组织CaM及CaMKII mRNA表达。结果与模型组比较,丹参酮ⅡA 组 CaM mRNA[(1.29±0.19)比(2.31±0.21)]及 CaMKⅡ mRNA[(1.10±0.07)比(2.13±0.18)]表达下调(P＜0.05),心肌梗死范围[(25.12±0.43)%比(35.15±0.64)%]缩小(P＜0.05),心肌收缩张力[(2.03±0.14)g比(1.06±0.12)g]及±LVdp/dtmax[(4701.2±135.3)mmHg/s比(3214.7±110.2)mmHg/s;(2518.7±65.4)mmHg/s比(1960.3±62.5)mmHg/s]增高(P＜0.05)。结论丹参酮ⅡA可下调急性心肌梗死模型大鼠心肌细胞CaM、CaMKⅡ mRNA表达,对心肌缺血有保护作用。     

日本汉方药治疗慢性肾脏病研究的进展

慢性肾脏病(chronic kidney disease, CKD)已成为世界范围内危害公众健康的疾病。在日本，柴苓汤和温脾汤是治疗CKD的常用汉方药。文章综述了柴苓汤和温脾汤等汉方药对CKD进展过程中各种肾损害的保护作用，包括改善系膜组织损伤，减轻炎症细胞浸润，减少炎症介质表达，清除氧化应激产物，调节醛固酮代谢等作用，并且，在分子水平阐明了这些药物对各种损伤因子的作用机制。柴苓汤和温脾汤都是延缓CKD进展的有效药物。     

尿毒清对慢性肾脏病3~4期患者肠道微生态影响的临床研究

[目的] 探讨尿毒清颗粒对慢性肾脏病（CKD）3~4期未透析患者肠道微生态的影响。[方法] 选取就诊的慢性肾脏病3~4期未透析患者60例,随机分为CKD组30例和尿毒清组30例,尿毒清组在CKD组常规治疗的基础上加用尿毒清颗粒,另外征集健康志愿者10名作为对照组,分别于试验前及试验后3个月观察各组肠道菌群结构及多样性、血清尿毒素及炎症因子水平、肾功能指标的变化。[结果] CKD组和尿毒清组分别有2例和3例失访,最终分别纳入28例和27例。试验前,与对照组相比,CKD组及尿毒清组肠道菌群均发生了显著变化,且血清尿毒素及炎症因子水平、肾功能指标均明显升高,具有统计学差异（P<0.05）;试验后3个月,与CKD组相比,尿毒清组有肠道菌群多样性增加的趋势,且菌群结构更加接近对照组的菌群分布。此外,尿毒清组的硫酸吲哚酚（IS）及部分肾功能指标血清肌酐（Scr）、尿白蛋白肌酐比值（ACR）显著下降,具有统计学差异（P<0.05）。[结论] CKD患者的肠道菌群发生了显著变化,经尿毒清干预可在一定程度上调节肠道菌群失调,减轻机体炎症反应状态以延缓慢性肾脏病进展,而肠道菌群失调与肾功能改变之间的相关性尚需进一步研究。     

丹参酮ⅡA磺酸钠治疗糖尿病肾病的Meta分析

目的系统评价在随机对照试验中丹参提取物丹参酮ⅡA磺酸钠治疗糖尿病肾病(Diabetic kidney disease,DKD)的疗效及安全性。方法利用计算机检索PubMed、Embase、The Cochrane library、中国知网(CNKI)、维普(VIP)、中国生物医学文献数据库(CBM)及万方数字化期刊全文数据库等数据库,检索时限为各数据库最早收录时间至2018年11月,所有纳入的文献都属于随机对照试验(RCT)。根据Cochrane Handbook评价文献质量,并采用Review Manager 5.3软件对纳入的数据进行统计分析。结果总共纳入15个随机对照试验,共1 176例受试者。Meta分析结果显示在改善肾功能方面治疗组优于对照组,其中主要结局指标尿白蛋白排泄率(UAER)[MD=-19.77,95%CI(-32.74,-6.80),Z=2.99,P=0.003]及24 h尿蛋白定量(24 h-UP)[SMD=-2.13,95%CI(-2.91,-1.35),Z=5.35,P<0.000 01]具有统计学意义。在次要结局指标方面,治疗组与对照组相比,丹参酮ⅡA磺酸钠对血肌酐(SCr)、尿素氮(BUN)等肾功能指标具有统计学意义(P<0.05);对降低总胆固醇(TC)、甘油三酯(TG)及空腹血糖(FBG)等糖、脂代谢指标,治疗组优于对照组,差异具有统计学意义(P<0.05)。本次研究的15篇文献均未报导严重的不良反应。结论西医常规治疗基础上加用丹参酮ⅡA磺酸钠(治疗组)能提高DKD的治疗效果,这种联合用药可能具有防治DKD的潜力,并在降低糖、脂方面具有一定的作用,为临床应用提供了参考。