Pharmacokinetic properties of crocin (crocetin digentiobiose ester) following oral administration in rats

This study investigated the pharmacokinetic properties of crocin following oral administration in rats. After a single oral dose, crocin was undetected while crocetin, a metabolite of crocin, was found in plasma at low concentrations. Simultaneously, crocin was largely present in feces and intestinal contents within 24h. After repeated oral doses for 6 days, crocin remained undetected in plasma and plasma crocetin concentrations were comparable to the corresponding data obtained after the single oral dose. Furthermore, the absorption characteristics of crocin were evaluated in situ using an intestinal recirculation perfusion method. During recirculation, crocin was undetected and low concentrations of crocetin were detected in plasma. The concentrations of crocin in the perfusate were reduced through different intestinal segments, and the quantities of drug lost were greater throughout the colon. These results indicate that (1) orally administered crocin is not absorbed either after a single dose or repeated doses, (2) crocin is excreted largely through the intestinal tract following oral administration, (3) plasma crocetin concentrations do not tend to accumulate with repeated oral doses of crocin, and (4) the intestinal tract serves as an important site for crocin hydrolysis.     

Influence of crocetin on high-cholesterol diet induced atherosclerosis in rats via anti-oxidant activity together with inhibition of inflammatory response and p38 MAPK signaling pathway

The present study aimed to investigate the effect and possible mechanism of action of crocetin on the high cholesterol diet (HCD) induced atherosclerosis rat. The Wistar rats were used in the current investigation. The rats were divided into following group, Group I: control, Group II: HCD induced AS, Group III: AS + crocetin (25 mg/kg), Group IV: AS + crocetin (50 mg/kg) and Group V: AS + Simvastatin, respectively. AS was induced in the rats using the vitamin D₃ and HCD. The rats received the pre-determined treatment for the 10 weeks. After the study period, the level of lipid profile, malonaldehyde (MDA) and superoxide dismutase (SOD) were also estimated. The proinflammatory cytokines viz., tumor necrosis factor (TNF)-α and interleukin (IL)-6 were scrutinized using the ELISA kits. We also estimated the expression of phosphorylated p38 (p-p38) MAPK using the Western blot techniques. The results revealed that the AS was successfully induced in the rats. The AS control group rats showed the modulated level of lipid profile, and decreased the level of the SOD and boost the level of the MDA as compared with the normal control. However, crocetin thrived in enhancing the lipid profile toward the standard value in the normal control group rats. The crocetin and simvastatin group rats significantly inhibited the expression of the p-p38 MAPK as compared to the AS group rats. In conclusion, the current investigation revealed that the crocetin reduced the HCD induced dyslipidemia in the Wistar rats, the possible mechanism of action may be connected to the antioxidative, down regulating of p-p38 MAPK and antiinflammatory effect by crocetin.     

Metabolite and target transcript analyses during Crocussativus stigma development

Saffron, the desiccated stigmas of Crocussativus, is highly appreciated for its peculiar colour, flavour and aroma. Several studies have been conducted with the spice, but little is known about the evolution of volatile and non-volatile compounds generated during the development of the stigma. In this study, we have followed these compounds, with special attention to those of isoprenoid origin (carotenoids and monoterpenes), which are responsible for the organoleptic properties of saffron. The main compounds that accumulated throughout stigma development in C.sativus were crocetin, its glucoside derivatives and picrocrocin, all of which increased as stigmas reached a fully developed stage. The volatile composition of C.sativus stigmas changed notably as stigmas developed with each developmental stage being characterized by a different volatile combination. In red stigmas, β-cyclocitral, the 7,8 cleavage product of β-carotene, was highly produced, suggesting the implication of both β-carotene and zeaxanthin in crocetin formation. As stigmas matured, hydroxy-β-ionone and β-ionone were produced while safranal, the most typical aroma compound of the processed spice, was only detected at low levels. However, a safranal-related compound 2,2,2-trimethyl-2-cyclohexene-1,4-dione (4-oxoisophorone) increased rapidly at the anthesis stage and also in senescent stigmas. Monoterpenes were mainly emitted at the time of anthesis and the emission patterns followed the expression patterns of two putative terpene synthases CsTS1 and CsTS2. Fatty acid derivates, which predominated at the earlier developmental stages, were observed at low levels in later stages.     

The pharmacokinetic profile of crocetin in healthy adult human volunteers after a single oral administration

Crocetin, a unique carotenoid with a short carbon chain length, is an active compound of saffron and Gardenia jasminoides Ellis used as traditional herbal medicine. The present study was undertaken to investigate the pharmacokinetic profiles of crocetin in healthy adult subjects. The study was conducted as an open-label, single dose escalation with 10 Filipino volunteers (5 men and 5 women). The subjects received a single dose of crocetin at three doses (7.5, 15 and 22.5 mg) in one week interval. Blood samples were collected from the brachial vein before and at 1, 2, 4, 6, 8, 10 and 24 h after administration. Plasma concentrations of crocetin were determined by high-performance liquid chromatography (HPLC). Crocetin was rapidly absorbed and detected within an hour of administration with a mean time to reach maximum concentration (T_max) of crocetin ranging from 4.0 to 4.8 h. The mean values of C_max and AUC_0-24 h ranged from 100.9 to 279.7 ng/ml and 556.5 to 1720.8 ng^.h/ml respectively. C_max and AUC values increased with dose proportional manner. Crocetin was eliminated from human plasma with a mean elimination half life (T_1/2) of 6.1 to 7.5 h.In summary, there were no serious adverse events up to 22.5 mg dose of crocetin while crocetin was found to be absorbed more quickly than the other carotenoids such as β-carotene, lutein and lycopene.     

转基因烟草表达西红花CSzCD基因产生西红花酸

为研究转基因烟草中产生西红花酸的可行性,在本研究中,西红花玉米黄素裂解酶（CSzCD）基因插入到pBI121载体的花椰菜花病毒（CaMV）35S启动子下游,通过农杆菌介导整合到烟草基因组中。通过Southern blotting 分析得到21株转基因烟草植株系; 转基因烟草叶片提取物Western blotting 和HPLC分析显示有西红花酸的产生,然而阴性对照中并没有发现西红花酸的存在。     

Interaction of tRNA with Safranal,Crocetin, and Dimethylcrocetin

Abstract

Saffron is the red dried stigmas of Crocus sativus L. flowers and used both as a spice and as a drug in traditional therapeutic. The biological activity of saffron in modern medicine is in development. Its numerous applications as an anti-oxidant and anti-cancer agent are due to its secondary metabolites and their derivatives (safranal, crocins, crocetin, dimethylcrocetin). The aim of this study was to examine the interaction of transfer RNA with safranal, crocetin, and dimethylcrocetin in aqueous solution at physiological conditions. Constant tRNA concentration (6.25 mM) and various drug/tRNA (phosphate) molar ratios of 1/48 to 1/8 were used. FT-IR and UV-Visible difference spectroscopic methods have been applied to determine the drug binding mode, the binding constants and the effects of drug complexation on the stability and conformation of tRNA duplex. External binding mode was observed for safranal crocetin and dimethylcrocetin, with overall binding constants K_safranal = 6.8 (± 0.34) × 10³ M^?1, K_CRT = 1.4 (± 0.31) × 10⁴ M^?1, and K_DMCRT = 3.4 (± 0.30) × 10⁴ M^?1. Transfer RNA remains in the A-family structure, upon safranal, crocetin and dimethylcrocetin complexation.     

西红花酸治疗DHT诱导的PCOS小鼠的机制研究

目的:观察西红花酸对双氢睾酮(Dihydrotestosterone,DHT)诱导的多囊卵巢综合症(Polycystic ovarian syndrome,PCOS)小鼠的疗效并探讨其作用机制。方法:妊娠15-18天给予孕鼠皮下注射DHT诱导子代雌鼠PCOS模型。待子鼠8周龄后,随机选择一半数量的PCOS小鼠连续4周西红花酸灌胃,作为西红花酸给药组(n=18)。给药期间检测体重和动情周期,待小鼠16周龄左右,通过眼球取血后处死,取出下丘脑、卵巢。采用HE染色观察卵巢组织的病理改变;ELISA试剂盒检测血清中雌二醇(Estradiol,E2)、睾酮(Testosterone,T)、孕酮(Progesterone,P4)、促黄体生成素(Luteinizing hormone,LH)、卵泡刺激素(Follicle-stimulating hormone,FSH);采用免疫组化、Western blot和实时荧光定量PCR法检测下丘脑的前腹侧视旁核(Anteroventral periventricular,AVPV)、弓状核(Arcuate,ARC)的kisspeptin以及视前区(Preoptic area,POA)里Gn RH的表达水平。结果:与对照组相比,PCOS小鼠卵巢与体重的比值上升了22.56%±6.77%,动情周期延长,卵巢内出现大的空泡,闭锁卵泡数量增加了138.74%±33.22%,窦状卵泡、成熟窦状卵泡和黄体数量分别减少了38.80%±4.69%、56.35%±7.32%和63.77%±7.25%,血清中E2、P4和FSH水平分别降低了40.99%±2.69%、56.91%±5.25%、和38.80%±4.69%,而T、LH水平分别升高了43.23%±4.70%和148.46%±28.16%,下丘脑中AVPV中kisspeptin神经元表达减少,ARC中kisspeptin神经元表达增多,POA中Gn RH神经元减少,而以上情况能够被西红花酸改善。结论:西红花酸分别通过促进和抑制下丘脑AVPV核和ARC核团的kisspeptin表达改善PCOS的病理变化。     

藏红花酸抗小鼠肝纤维化的实验研究

目的:通过观察藏红花酸对肝纤维化小鼠肝组织病理学改变及p38MAPK蛋白表达的影响,探讨其抗肝纤维化作用及可能的机制。方法:36只6周龄雄性昆明小鼠随机分为3组:藏红花酸组、模型组、正常组。除正常对照组外,其余两组小鼠给予30%四氯化碳橄榄油溶液腹腔注射,首次注射5 m L/Kg,以后每次3 m L/Kg,每3日1次,连续12周。同时藏红花酸组给予藏红花酸5mg/Kg灌胃,每日1次,连续12周。实验过程中,观察各组小鼠的一般状态。12周末处死小鼠,肝组织切片行HE、Masson染色,显微镜下观察病理改变,免疫组化检测小鼠肝组织p38MAPK蛋白的表达。结果:实验过程中,模型组小鼠一般状态较差,体重增长缓慢;藏红花酸组小鼠一般状态尚可,体重较模型组增长明显(P0.05)。肝组织HE、Masson染色结果显示:模型组小鼠肝纤维化程度较重(造模成功),与其相比,藏红花酸组小鼠肝纤维化程度有所改善,差异有统计学意义。与模型组相比,藏红花酸组肝组织p38MAPK表达显著下降(P0.05)。结论:藏红花酸能有效减轻小鼠肝损伤及纤肝维化程度,其抗肝纤维化的机制可能与下调p38MAPK蛋白的表达有关。     

Molecular dynamic simulation and DFT study on the Drug-DNA interaction; Crocetin as an anti-cancer and DNA nanostructure model

In this research, the interaction of Crocetin as an anti-cancer drug and a Dickerson DNA has been investigated. 25 ns molecular dynamic simulations of Crocetin and DNA composed of 12 base pairs and a sequence of d(CGCGAATTCGCG)₂ were done in water. Three definite parts of the B-DNA were considered in analyzing the best interactive site from the thermodynamic point of view. Binding energy analysis showed that van der Waals interaction is the most important part related to the reciprocal O and H atoms of the Crocetin and DNA. Stabilizing interactions, obtained by ΔG calculations, showed that maximum and minimum interactions are related to the S1 and S3 regions, respectively. This means that the most probable van der Waals interaction site of the Dickerson B-DNA and Crocetin is located in the minor groove of DNA. Two sharp peaks at 2.55 and 1.75 Å in radial distribution functions of the PO?HO and NH?OC parts are related to new hydrogen bonds between the Crocetin and DNA in the complex which can be considered as the driving force of the anti-cancer mechanism of the Crocetin. Average values of 0.3 au and zero for the electron densities of the H?O bonds for DNA and complex, obtained by Quantum theory of atoms in molecules (QTAIM), means that the origin of DNA instability after complexation may be related to the H-bond denaturation by Crocetin. Finally, the evaluation of the dispersion interactions using the dispersion functional, -148.76 kcal.mol^?1, confirmed the importance of the dispersion interaction in drug-DNA complex.