Compatibility art of traditional Chinese medicine: From the perspective of herb pairs

Ethnopharmacological relevance

Over the past decades, research of traditional Chinese medicine (TCM) mainly focused on developing potential candidates from Chinese medicinal herbs, while the wisdom of applying these traditional herbs has not been paid as much attention as it deserves. As is well-known, multi-herb therapy is one of the most important characteristics of TCM, but the modernization drive of this conventional wisdom has faced many obstacles due to its unimaginable complexity. Herb pairs, the most fundamental and the simplest form of multi-herb formulae, are a centralized representative of Chinese herbal compatibility. In light of their simplicity and the basic characteristics of complex formulae, herb pairs are of great importance in the studies of herb compatibility.

Materials and methods

A systematic search of herb pair related research was carried out using multiple online literature databases, books and monographs published in the past 20 years.

Results

A comprehensive introduction to the compatibility of TCM, the position of herb pairs in TCM and the progresses of several famous herb pairs were provided in this review. Furthermore, the clinical study and the future research trends of herb pairs were also discussed.

Conclusions

Herb pairs have played, and may continue to play a key role in full investigation of general herb compatibility for their indispensable position in TCM. Much more research is needed for the standardization, safety evaluation, and mechanism exploration of herb pairs.     

Identification of campesterol from Chrysanthemum coronarium L. and its antiangiogenic activities

Campesterol, a plant sterol in nature, is known to have cholesterol lowering and anticarcinogenic effects. Since angiogenesis is essential for cancer, it was surmised that an antiangiogenic effect may be involved in the anticancer action of this compound. This study investigated the effect of campesterol on basic fibroblast growth factor (bFGF)-induced angiogenesis in vitro in human umbilical vein endothelial cells (HUVECs) and an in vivo chorioallantoic membrane (CAM) model. Campesterol isolated from an ethylacetate fraction of Chrysanthemum coronarium L. showed a weak cytotoxicity in non-proliferating HUVECs. Within the non-cytotoxic concentration range, campesterol significantly inhibited the bFGF-induced proliferation and tube formation of HUVECs in a concentration-dependent manner, while it did not affect the motility of HUVECs. Furthermore, campesterol effectively disrupted the bFGF-induced neovascularization in chick chorioallantoic membrane (CAM) in vivo. Taken together, these results support a potential antiangiogenic action of campesterol via an inhibition of endothelial cell proliferation and capillary differentiation.     

Agkistrodon acutus-purified protein C activator protects human umbilical vein endothelial cells against H2O2-induced apoptosis

Context: Recent studies show that the Agkistrodon acutus (Viperidae) (syn. Deinagkistrodon acutus) protein C activator (PCA) treats acute myocardial infarction and ischaemia-reperfusion animal models effectively, while the underlying mechanism remains unknown.

Objective: To study the effect of PCA on the injury of human umbilical vein endothelial cells (HUVECs) induced by H₂O₂ and the underlying mechanism.

Materials and methods: Primary cultured HUVECs were pretreated with PCA (20, 40 and 80?μg/mL) for 1?h, then HUVEC apoptosis was induced by 300?μmol/mL H₂O₂. Apoptosis was analyzed by AnnexinV-FITC/PI, and reactive oxygen species (ROS) level was tested by flow cytometry. Colorimetric methods were used to detect the levels of NO and IL-1. In addition, real-time PCR and western blot analyses were used to detect the expression of eNOS and phospho-p38/MAPK.

Results: Morphological changes were induced by H₂O₂ in HUVECs. The cell survival rate was increased by 43.9, 64.0 and 80.6% in each PCA pretreated group (20, 40 and 80?μg/mL) compared to the model group. In each PCA pretreated group, oxidative stress level was also decreased to 54.7, 42.7 and 25.1%. Moreover, the level of IL-1 was decreased to 83.3, 62.2 and 30.7%. The level of NO was increased by 155.9, 232.4 and 317.6%. Apoptosis rate was decreased to 59.0, 47.7 and 32.7%. Phospho-p38 expression was downregulated, but eNOS expression was upregulated.

Discussion and conclusion: The results suggest that PCA can effectively protect the endothelial cells from injury induced by H₂O₂, which may be associated with antioxidation, upregulation of eNOS and downregulation of p38-MAPK.     

肝素钠对氟尿嘧啶诱导的人脐静脉内皮细胞损伤的影响探讨

目的通过观察不同浓度肝素钠对氟尿嘧啶诱导的人脐静脉内皮细胞(HUVECs)损伤的影响,探讨不同浓度肝素钠稀释液及不同作用时相点对氟尿嘧啶致血管内皮细胞损伤的影响及可能的作用机制。方法将体外培养的2~3代HUVECs随机分为对照组、氟尿嘧啶组、肝素钠组,其中对照组只用培养基培养;氟尿嘧啶组用终浓度为25mg/L的氟尿嘧啶为刺激物;肝素钠组分别以终浓度为12.5U/mL、25U/mL、50U/mL肝素钠稀释液联合终浓度为25mg/L的氟尿嘧啶为刺激物。酶联免疫吸附实验(ELISA)检测细胞白介素-6(IL-6)、白介素-1(IL-1)及组织纤维溶酶原激活物(t-PA)含量。结果氟尿嘧啶组IL-6、IL-1及t-PA蛋白表达较对照组升高(P0.05);与氟尿嘧啶组相比,肝素钠组可抑制氟尿嘧啶诱导的HUVECs释放IL-6、IL-1及t-PA(P0.05),以50U/mL肝素钠组作用最为显著,但IL-6、IL-1及t-PA蛋白表达仍高于对照组(P0.05)。不同浓度肝素钠组间差异无统计学意义(P0.05)。同一药物浓度组,与药物作用1d后相比,第4天、第7天、第14天均有统计学差异(P0.05),其中IL-6蛋白表达呈时间依赖性升高。结论肝素钠可抑制内皮细胞释放IL-1、IL-6及t-PA蛋白,对内皮细胞具有一定保护作用。     

干扰人乳腺癌MDA-MB-231细胞 EPCR表达对内皮细胞增殖､迁移及脉管形成的影响

目的 :通过观察干扰乳腺癌MDA-MB-231细胞内皮蛋白C受体(endothelial protein C receptor,EPCR)表达对内皮细胞的增殖、迁移、脉管形成能力的变化,分析EPCR在肿瘤血管形成中的作用。方法:采用si RNA方法降低人乳腺癌MDA-MB-231细胞EPCR的表达,通过RT-PCR及Western blot技术检测EPCR干扰效果,实验分为EPCR干扰组、无关序列组及未处理组。制备肿瘤条件培养基模拟肿瘤微环境培养HUVECs细胞,通过CCK-8法检测各组内皮细胞增殖能力,Transwell小室检测内皮细胞迁移能力,Matrigel检测内皮细胞脉管形成能力。结果:与未处理组及无关列序组相比,EPCR干扰组细胞的增殖、迁移及脉管形成能力均明显降低(P<0.05)。结论:干扰乳腺癌MDA-MB-231细胞EPCR的表达能够抑制内皮细胞增殖、迁移及脉管形成能力,提示EPCR可能在肿瘤血管形成中起重要作用。     

丹酚酸A对过氧化氢诱导的人脐静脉内皮细胞损伤的保护作用

目的研究丹酚酸A对人脐静脉内皮细胞(HUVECs)氧化损伤的保护作用。方法采用过氧化氢(H2O2)诱导的HUVECs氧化损伤模型,观察丹酚酸A(10,20和40μg·mL-1)对血管内皮细胞损伤的保护作用,MTT法检测细胞存活率,试剂盒检测细胞上清液中的超氧化物歧化酶(SOD)、丙二醛(MDA)、乳酸脱氢酶(LDH)、一氧化氮(NO),比色法测定Caspase-3的活性。结果丹酚酸A可抑制H2O2诱导的血管内皮细胞的凋亡,同时增加SOD的活性,降低MDA的水平,减少LDH的漏出量,增加NO的释放,降低Caspase-3的活性。且丹酚酸A的上述作用均呈质量浓度依赖性。结论丹酚酸A可以有效保护内皮细胞免受氧化损伤。     

流体切应力和Ox—LDL地内皮细胞表面ICAM—1表达的影响

目的研究动脉粥样硬化(AS)的危险因素高脂血症、流体切应力及二者共同作用对血管内皮细胞细胞间粘附分子-1(ICAM-1)表达的影响。方法以人脐静脉内皮细胞(HUVECs)为研究对象,应用免疫组化ACB方法,观测Ox-LDL、流体切应力及二者共同作用对HUVECs的粘附分子ICAM-1表达的影响。结果与对照组相比,(1)Ox-LDL显著增加HUVECs表面ICAM-1表达;(2)流体切应力使HUVECs表面ICAM-1表达增加,但ICAM-1的表达随时间依赖性增加的趋势不明显;(3)共同作用后,HUVECsICAM-1的表达较基础表达显著增加。结论Ox-LDL、流体切应力的改变及二者共同作用显著增加H-UVECs表面ICAM-1表达。     

汉防己甲素抑制血管生成的作用

背景与目的:血管生成在肿瘤从良性向恶性转变、癌细胞进入血液循环、转移灶发展和破裂中都起着重要作用.本研究探讨汉防己甲素对体外、体内血管生成的抑制作用及其可能的机制.方法:采用M1_r法观察汉防己甲素对人脐静脉血管内皮细胞(human umbilical vein endothelial cell HUVEC)和人肠癌LoVo细胞增殖的影响.通过Transwell小室趋化实验、体外小管形成实验观察汉防己甲素对HUVEC迁移、成血管能力的影响.建立裸鼠kIVo细胞皮下移植瘤模型.给予汉防己甲素灌胃,观察用药对肿瘤微血管密度的影响.结果:2-8 μg/mL的汉防己甲素作用48 h时对HUVEC的细胞增殖抑制率为24.6%-76.9%,对LoVo细胞增殖抑制率为11.6%～14.0%;体外小管形成实验发现.2～8 μg/mL的汉防己甲素作用24 h时HUVEC小管形成数目减少,且管腔不完整,与对照组比较差异有统计学意义(P值均＜0.001).经2-8 μg/mL的汉防己甲素处理12 h后HUVEC迁移数明显少于对照组(P值均＜O.001).在裸鼠皮下移植瘤体内实验中,80 mg/kg汉防己甲素作用于裸鼠LoVo细胞皮下移植瘤后其微血管密度与生理盐水对照组比较.差异具有统计学意义(p0.035).结论:汉防己甲素在体外能有效抑制血管生成,其机制可能与抑制HUVEC增生、迁移和小管形成,诱导HUVEC凋亡,抑制HUVEC DNA的合成有关.汉防己甲素在体内对裸鼠LoVo移植瘤具有抗血管生成作用.     

PM2.5中不同组分对人脐静脉内皮细胞的联合毒性作用研究

目的：探讨PM_2.5中各种组分单独以及联合暴露对于人脐静脉内皮细胞（HUVECs）的毒性作用,并探讨不同组分之间是否存在交互作用。方法：以永生化的HUVECs细胞为研究对象,分别给予0、5、10、20、40、80、160 μg/mL纳米炭黑颗粒（下称炭黑）,0、2.5、5、10、20、40、80、160 μg/mL尘土颗粒（下称尘土）,0、6.25、12.5、25、50、100、200、400 μmol/L醋酸铅,0、5、10、20、40、80、160 μmol/L亚砷酸钠和氯化镉,染毒24 h,采用CCK-8法测定不同受试物对细胞存活率的影响,并计算不同受试物对HUVECs细胞的半数抑制浓度（IC₅₀）。然后进行联合染毒实验,选用细胞存活率为80%时的剂量,炭黑（或尘土）分别与铅、砷、镉对HUVECs进行联合染毒24 h,用CCK-8测定细胞存活率;另外根据PM_2.5中各组分实际占比,按m_铅：m_{炭黑/尘土}=1：50,m_砷：m_{炭黑/尘土}=1：100,m_镉：m_{炭黑/尘土}=1：500的比例进行两两联合染毒,以及按m_铅：m_砷：m_镉：m_{炭黑/尘土}=10：5：1：500的比例进行四者联合染毒。染毒24 h后,用CCK-8法测定细胞存活率。结果：HUVEC细胞存活率随着炭黑、尘土、醋酸铅、亚砷酸钠、氯化镉染毒浓度的增加而下降（P<0.05）,以上各受试物对HUVECs细胞的IC₅₀分别为16 μg/mL、110 μg/mL、184 μmol/L、15 μmol/L、14 μmol/L。炭黑与醋酸铅、亚砷酸钠、氯化镉联合染毒时,细胞存活率比炭黑单独染毒时分别下降了17.8%、43.8%、41.2%;尘土与醋酸铅、亚砷酸钠、氯化镉联合染毒时,细胞存活率比尘土单独染毒时分别下降了11.6%、27.8%、28.3%。联合染毒时细胞存活率比单独染毒时明显下降（P<0.05）。炭黑与亚砷酸钠、炭黑与氯化镉之间存在交互作用（均为P<0.05）。结论：PM_2.5中不同组分均可以对血管内皮细胞产生毒性作用,单独暴露时炭黑的细胞毒性作用比尘土大;但与3种金属联合暴露时,尘土比炭黑的联合暴露细胞毒性更大。铅、砷、镉对细胞的毒性作用大小排序为氯化镉>亚砷酸钠>醋酸铅,且炭黑颗粒与亚砷酸钠、炭黑颗粒与氯化镉之间存在交互作用,表现为协同作用。     

Low levels of the air pollutant 1-nitropyrene induce DNA damage,increased levels of reactive oxygen species and endoplasmic reticulum stress in human endothelial cells

Both epidemiological and experimental studies suggest that exposure to high levels of air pollution is a risk factor associated with cardiovascular disease. Traffic emission is a major source of exposure to persistent air pollutants such as nitrated polycyclic aromatic hydrocarbons (nitro-PAHs). 1-Nitropyrene (1-NP), one of the most abundant nitro-PAHs in diesel exhausts, was selected as a model nitro-PAH for the present study. The aim of the study was to investigate the effects of 1-NP in human umbilical vein endothelial cells (HUVECs) and the metabolic pathways involved. The nitroreductase inhibitor dicoumarol and the coplanar aryl hydrocarbon receptor (AhR) ligand PCB 126 were used to modulate the metabolism of 1-NP. The results revealed that low levels (≤10 μM) of 1-NP induced DNA damage, increased levels of reactive oxygen species (ROS) and increased protein expression of the endoplasmic reticulum (ER) stress chaperone GRP78. A decrease in cell viability was only observed following exposure to a higher level of 1-NP (15 μM). Inhibition of nitroreductive metabolism by dicoumarol attenuated the induction of DNA damage, intracellular ROS levels and GRP78 expression. This suggests that the effects of 1-NP on HUVEC were mediated by metabolites mainly formed at nitroreduction. Our findings suggest that the human blood vessel endothelium is a sensitive target tissue for the major nitro-PAH constituent in diesel exhaust.