Safflower Extract Research Articles

Pharmacokinetic comparisons of hydroxysafflower yellow A in normal and blood stasis syndrome rats

Ethnopharmacological relevance Safflower is a popular Traditional Chinese Medicine (TCM) to invigorate the blood and dispel ‘blood stasis’, which arises from poor blood circulation. The differences of pharmacokinetic properties between normal and blood stasis syndrome rats were seldom reported. Aim of the study The present study was conducted to evaluate the pharmacokinetics of hydroxysafflower yellow A (HSYA) following oral administration of hydroxysafflower yellow A and safflower extract with approximately the same dose of HSYA 100 mg/kg in both normal and acute blood stasis rats. Materials and methods The animals were orally administered with HYSA monomer and safflower extract. The blood samples were collected according to the time schedule. The concentrations of HSYA in rat plasma were determined by HPLC. Various pharmacokinetic parameters were estimated from the plasma concentration versus time data using non-compartmental methods. Results It was found that AUC 0– t , C max, Vd and CL of HSYA in both HSYA monomer and safflower extract in acute blood stasis rats were with significant difference ( P < 0.05) comparing with that in normal rats. Conclusions The results indicated that HSYA was with high uptake and eliminated slowly in the animals with blood stasis syndrome, suggesting that the rate and extent of drug metabolism was altered in acute blood stasis animals.

Preparation of High Quality Safflower (Carthamus tinctorius L.) Seed Extract by High-Pressure Extraction Process

Safflower seed extract was prepared by a high-pressure extraction technology and its quality characteristics were compared to that of other conventional extraction techniques, such ultrasonic and reflux extractions. Safflower seeds were extracted with 80% aqueous ethanol by three above extraction methods, and further fractionated with Diaion HP-20 column chromatography to obtain a partially purified safflower seed extract (PPSSE). Among the three extraction techniques examined, the reflux extraction showed the higher yields of EtOH extract and PPE than the ultrasonic and high-pressure extractions. Levels of most phenolic compounds in the EtOH extract of safflower seed are higher in reflux and ultrasonic extractions than the high pressure extraction, but levels of two serotonin aglycones, N-(p-coumaroyl)serotonin (CS) and N-feruloylserotonin (FS), in PPSSE were higher in the high pressure extraction than the reflux and ultrasonic extractions. In addition, color values (L and a) of the PPSSE were higher in the high-pressure extraction than the reflux and ultrasonic extractions, although there were no significant differences in pH and UV maxima absorption spectra among three extraction techniques. These results indicate that the high-pressure extraction technology is a simple and effective extraction for preparation of a high quality of safflower seed extract containing CS and FS with anti-wrinkle activity.

Protective effects of purified safflower extract on myocardial ischemia in vivo and in vitro

Carthamus tinctorius L. (safflower) is one of the most commonly used Chinese herbal medicines to prevent and treat cardiac disease in clinical practice. However, the mechanisms responsible for such protective effects remain largely unknown. In this study, we investigated the anti-myocardial ischemia effects of a purified extract of C. tinctorius (ECT) both in vivo and in vitro. An animal model of myocardial ischemia injury was induced by left anterior descending coronary artery occlusion in adult rats. Pretreatment with ECT (100, 200, 400, 600 mg/kg body wt.) could protect the heart from ischemia injury by limiting infarct size and improving cardiac function. In the in vitro experiment, neonatal rat ventricular myocytes were incubated to test the direct cytoprotective effect of ECT against H(2)O(2) exposure. Pretreatment with 100-400 microg/ml ECT prior to H(2)O(2) exposure significantly increased cell viability as revealed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. ECT also markedly attenuated H(2)O(2)-induced cardiomyocyte apoptosis, as detected by Annexin V and PI double labeling with flow cytometry. The intracellular level of reactive oxygen species (ROS) was shown by 2',7'-dichlorofluorescin diacetate (DCFH-DA), and ECT pretreatment significantly inhibited H(2)O(2)-induced ROS increase. We made a preliminary examination of the signaling cascade involved in ECT mediated anti-apoptotic effects. Phosphatidylinositol 3 kinase (PI3K) inhibitor (LY294002) blocked the cytoprotective effect conferred by ECT. Taken together, our findings provide the first evidence that the cardioprotective effects of ECT in myocardial ischemia operate partially through reducing oxidative stress induced damage and apoptosis. The protection is achieved by scavenging of ROS and mediating the PI3K signaling pathway.

Safflower extracts functionally regulate monoamine transporters

Safflower (HH), the dry flower of Carthamus tinctorius L., has long been used to empirically treat neuropsychological disorders such as stroke and major depression in traditional Chinese medicine, and recently been proven effective for regulating levels of dopamine and serotonin in new-born rat brain. The present study assessed whether HH would be bioactive for functionally regulating monoamine transporters using in vitro drug-screening cell lines. Our current results showed that all solvent-extracted HH fractions, in different degrees, markedly increased both dopamine uptake by Chinese hamster ovary (CHO) cells stably expressing dopamine transporter (DAT) and norepinephrine uptake by CHO cells expressing norepinephrine transporter (NET), and also showed that chloroform (HC), ethyl acetate (HE), and n-butyl alcohol extract strikingly depressed serotonin uptake by CHO cells expressing serotonin transporter (SERT); wherein, the potencies of ethanol extract, HC, HE, and aqueous extract to up-regulate dopamine/norepinephrine uptake and potency of HE to inhibit serotonin uptake were relatively stronger. Further investigation revealed that the enhancement of dopamine/norepinephrine uptake by HC and HE was dependent of DAT/NET activity, and the HE-induced inhibition of serotonin uptake was typical of competition. Thus, HH extracts are novel monoamine transporter modulators functioning as DAT/NET activators and/or SERT inhibitors, and would likely improve neuropsychological disorders through regulating monoamine-transporter activity.

Antioxidant and Neuroprotective Activities of Mogami-benibana (Safflower, Carthamus tinctorius Linne)

Free radical scavenging activity of the extracts of petals (bud, early stage, full blooming and ending stage), leaf, stem, root and seeds of Mogami-benibana (safflower, Carthamus tinctorius Linne), the contents of the major active components of carthamin and polyphenols, and neuroprotective effect of the petal extracts and carthamin in the brain of mice and rats were examined. Water extracts of Mogami-benibana petals scavenged superoxide, hydroxyl and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals and singlet oxygen. The scavenging activities of the extract of safflower petals with various colors showed the order of orange, yellow and white from high to low. This order is consistent with the contents of carthamin, which is a pigment of orange color and is found highest in orange petals and lowest in white petals. There was also a relationship between DPPH radical scavenging activity and carthamin content in the petal extracts of safflower. The neuroprotective effects were examined in cellular and animal models. Mogami-benibana petal extract inhibited glutamate-induced C6 glia cell death, significantly decreased the formation of malondialdehyde in mouse cerebrum, and inhibited the increase in thiobarbituric acid reactive substances and 8-hydroxy-2'-deoxyguanosine (8-OHdG) in the cerebral cortex of rats subjected to an injection of FeCl(3) solution into the sensory motor cortex. Carthamin showed similar effects in inhibiting 8-OHdG by the petal extract in rats. These results suggest that the petal extract of Mogami-benibana has free radical scavenging activity and neuroprotective effect and carthamin is one of the major active components.

Safflower extract: A novel renal fibrosis antagonist that functions by suppressing autocrine TGF‐beta

Progressive renal disease is characterized by the accumulation of extracellular matrix proteins in the renal interstitium. Hence, developing agents that antagonize fibrogenic signals is a critical issue facing researchers. The present study investigated the blood-circulation-promoting Chinese herb, safflower, on fibrosis status in NRK-49F cells, a normal rat kidney interstitial fibroblast, to evaluate the underlying signal transduction mechanism of transforming growth factor-beta (TGF-beta), a potent fibrogenic growth factor. Safflower was characterized and extracted using water. Renal fibrosis model was established both in vitro with fibroblast cells treated with beta-hydroxybutyrate and in vivo using rats undergone unilateral ureteral obstruction (UUO). Western blotting was used to examine protein expression in TGF-beta-related signal proteins such as type I and type II TGF-beta receptor, Smads2/3, pSmad2/3, Smads4, and Smads7. ELISA was used to analyze bioactive TGF-beta1 and fibronectin levels in the culture media. Safflower extract (SE) significantly inhibited beta-HB-induced fibrosis in NRK cells concomitantly with dose-dependent inhibition of the type I TGF-beta1 receptor and its down-stream signals (i.e., Smad). Moreover, SE dose-dependently enhanced inhibitory Smad7. Thus, SE can suppress renal cellular fibrosis by inhibiting the TGF-beta autocrine loop. Moreover, remarkably lower levels of tissue collagen were noted in the nephron and serum TGF-beta1 of UUO rats receiving oral SE (0.15 g/3 ml/0.25 kg/day) compared with the untreated controls. Hence, SE is a potential inhibitor of renal fibrosis. We suggest that safflower is a novel renal fibrosis antagonist that functions by down-regulating TGF-beta signals.

Effect of extracts from safflower seeds on osteoblast differentiation and intracellular calcium ion concentration in MC3T3-E1 cells

Although safflower seeds have long been used in Korea as herbal medicines, very little research has been published on the effects of safflower seed on bone formation or bone density. The study reported here therefore examined bone nodule formation, calcium uptake, alkaline phosphatase activity, and intracellular concentration of calcium ion [Ca2+]i in murine osteoblastic cells of the MC3T3-E1 line that were cultured on modified Eagle's minimal essential medium alone (controls) or with addition of 0.1% crude extract of safflower seed (experimental group I) or 0.1% aqueous fraction of safflower seed (experimental group II). Fluorescence spectrometry measurement of ([Ca2+]i) showed significantly accelerated rates of osteoblast differentiation in experimental group I (3 µL of crude extract in 8 × 104 cells) and experimental group II (2 µL of aqueous fraction in 8 × 104 cells) compared to the control group.

Determination of hydroxysafflor yellow A in rat plasma and tissues by high‐performance liquid chromatography after oral administration of safflower extract or safflor yellow

A simple and reproducible HPLC method for quantification of hydroxysafflor yellow A (HSYA) in rat plasma and tissues after oral administration of safflower extract or safflor yellow (SY) was developed. Sample preparation was achieved by protein precipitation of plasma and tissue homogenates with three volumes of methanol. p-Hydroxybenzaldehyde was used as the internal standard (IS). HSYA and IS were separated on a Hypersil BDS-C(18) column with a gradient elution system composed of acetonitrile and aqueous acetic acid. UV detection was used at 320 nm. The calibration curves were linear in all matrices (r(2) > 0.999) in the concentration ranges 0.51-101.36 microg/mL for plasma, 12.27-2454.46 microg/g for intestines and 0.96-192.20 microg/g for lung. The intra-day and inter-day precision were all less than 12.5%, and the extract recovery was in the range 64.1-103.7% with RSD less than 15.6% for HSYA in all matrices. The method was used successfully to quantify HSYA in rat plasma and tissue samples to support a pharmacokinectic study.

홍화씨와 한약재 흔합추출물이 Streptozotocin으로 유도한 흰쥐의 혈당과 혈액 성분에 미치는 영향

당뇨병은 생체내 당 이용 능력의 이상으로 일어나는 만성대사성 질환으로서 심혈관계 합병증이 일으키는 대표 증상인 고혈당과 지질대사를 치료하기 위하여 많은 환자들이 민간요법에 의존하고 있다. 이에 본 연구는 홍화씨와 한약재혼합 추출물이 혈당강하와 지질상태 개선에 대한 효능을 검증하고자 실시하였다. 당뇨는 streptozotocin(45 ㎎/㎏ B.W.)으로 S.D.계 흰쥐에게 근육주사로 유도시킨 다음 주재료로 볶은 홍화씨 50 g, 100 g 각각에 부재료로 맥문동, 감초, 상엽, 백복령, 숙지황, 비파엽, 독활, 대추, 산수유, 목단피, 천화분, 오미자를 섞어서 만든 한방 혼합 추출물을 당뇨 유도 쥐에게 체중 ㎏ 당 100 ㎎, 200 ㎎의 홍화씨를 섭취할 수 있도록 각각 경구 투여하였다. 당뇨쥐의 고혈당 증상은 100 g의 홍화씨를 넣고 추출한 한약재 혼합추출물의 투여로 정상수준은 아니었지만 34% 낮아졌다(p＜0.0.5). 혈장 내 지질과산화물 함량은 정상군과 당뇨 유도군 간에 유의한 차이는 없었으나, 간 마이크로좀의 지질과산화물 함량은 정상군은 6.20±2.25 n㏖/㎎ protein에 비하여 당뇨군은 7.74 n㏖/㎎ protein로 p＜0.05 수준에서 높았다. 그러나 100 g의 홍화씨를 첨가하여 만든 한약재 혼합추출물 투여했을 때 5.85 n㏖/㎎ protein 수준까지 낮추어 주었다. 혈액과 간 조직 내 중성지방 함량은 역시 100 g의 홍화씨를 첨가하여 만든 한약재 혼합추출물 투여로 p＜0.05 수준에서 저하효과가 현저하였다(p＜0.05). 그러나 혈액과 간 조직 내 유리지방산, 총콜레스테롤과 인지질함량은 실험 전 군에서 유의적인 차이를 볼 수 없었고, 정상군에 비하여 당뇨군에서 혈장내 LDL-콜레스테롤과 동맥경화지수가 유의적으로 높은 반면에 HDL-콜레스테롤은 낮았으며 홍화씨와 한약재 혼합추출물이 이들 함량 변화에 미치는 효과는 관찰되지 않았다. 이상의 본 실험결과는 당뇨에 의한 혈당, 지질과산화물 그리고 중성지방 증가는 홍화씨와 한약재 혼합추출물의 투여로 다소 완화되었다. 이러한 결과를 바탕으로 체내 혈당조절에 의한 당뇨병의 예방 및 치유효과가 있는 홍화씨와 같은 천연물질의 기능성 성분에 대하여 향후에 보다 심층 있는 연구를 통하여 건강 기능성 식품 소재로 활용하는 것이 매우 중요할 것으로 여겨진다.

홍화(Carthamus tinctorius L.)씨의 sterol 및 Phytoestrogen 분석

홍화씨가 골절, 골다공증 및 골형성부전증 등의 골 질환 치료에 효과가 있다는 것이 알려져 민간요법으로 사용되고 있으나 홍화씨의 어떤 성분이 이러한 작용을 하는지에 관해서는 거의 밝혀져 있지 않다. 본 연구에서는 흥화씨의 sterol 조성을 분석하고 폐경기 여성들의 골다공증 치료에 사용되고 있는 합성 estrogen 대체물질로써 그 기능이 확인된 phytoestrogen 활성을 측정하였다. Cholesterol에서 성호르몬인 estrogen이 합성되는 것처럼 식물 sterol 도 phytoestrogen 합성을 위한 전구체 작용을 하는 것으로 알려져 있다. 홍화씨에 존재하는 식물성 sterol은 stigmast-5-en-3-ol (3<TEX>$\beta$</TEX>, 24S)-form인 <TEX>${\gamma}$</TEX>-sitosterol (clionasterol)로 확인되었으며 홍화에 존재하는 식물성 sterol의 함량은 총지질의 약 4%를 함유하였다. 홍화씨의 sterol 분석은 GC-MS와 Wiley MS spectrum library를 사용하여 분석하였으며 홍화씨에 존재하는 중요한 4-desmethyl sterols 에는 sitosterol 만이 확인되었다. 홍화씨의 식물성 estrogen 효과는 <TEX>$\beta$</TEX>-estradiol (<TEX>$10^{-8}$</TEX> M)의 활성을 1로 했을 때 홍화씨의 에탄을 층(5 mg/ml) 및 메탄올 층(5 mg/ml)에서 각각 0.37과 0.43으로 estrogen 유사효과를 나타내었다(ANOVA p<0.001). 홍화씨에 포함된 estrogen에 의한 전사활성 촉진은 MCF7/pDS-CAT-ERE119-Ad2MLP system을 사용하여 estrogen수용체와 reporter gene 발현을 이용하는 방법으로 세포에 직접 처리함으로써 phytoestrogen 활성을 측정할 수 있었다. This study was done to investigated the phytosterol compositions of safflower (Carthamus tinctorius L.) seed. The phytoestrogen activity was also determined using CAT-ELISA Kit in ethanol extract of safflower seed. The phytosterol of safflower seeds was identified using gas chromatography-mass spectrometry after saponification of the oils. The phytosterol content and composition of safflower seed oils were 4% and identified stigmast-5-en-3-ol (3<TEX>$\beta$</TEX>, 24S)-form, <TEX>${\gamma}$</TEX>-sitosterol (clionasterol) with Wiley MS spectrum library. The synergistic effect of human estrogen receptor (hER) has been investigated using a minimal chimeric promoters composed of the TATA region of the adenovirus-2 major late promoter (A22MLP) and two consensus perfectly polindromic Xenopus vitellogenin A2 gene estrogen responsive elements (XVEREl19). Transient transfection experiments in tile human breast adenocarcinoma cell line MCF-7, which is known to express the estrogen receptor endogenously, revealed that phytoestrogen from Carthamus tinctorius L. acts as estrogen. We have observed the transcriptional activities stimulated methanol and ethanol extract of safflower seed in MCF-7, were 0.43 and 0.37 respectively, compared to that by <TEX>$\beta$</TEX>-estradiol as 1.0. Our data showed that safflower seeds have estrogenic activity methanol and ethanol extracts and ethanol lower than that of <TEX>$\beta$</TEX>-estradiol. This result provides the first evidence that the beneficial effect of safflower seeds may be mediated, at least in part, by the stimulating effect of phytoestrogen ell bone-protecting.

Protective Effects of Hot Water Extract of Safflower Leaves and Its Component Luteolin-7-O-Glucoside on Paraquat-Induced Oxidative Stress in Rats.

Suppressive effects of hot water extract of safflower leaves (Ex) and its major component luteolin-7-O-glucoside (LuGlc) on oxidative stress were determined in rats in which oxidative stress had been induced by paraquat (PQ). Both Ex and LuGlc suppressed lowering of food intake, and body weight gain, increases in lung weight, liver thiobarbituric acid reactive substances (TBARS), and a decrease in the liver triacylglycerol concentration were all induced by PQ-feeding. Rises in the liver total glutathione and oxidized glutathione concentrations in the PQ-fed rats were suppressed by adding Ex and LuGlc to the PQ diet. Moreover, Ex and LuGlc tended to suppress decreases in catalase activity in the liver mitochondrial fraction and liver α-tocopherol concentration which were induced by PQ-feeding. These results showed that both Ex and LuGlc, which is a major component of Ex, may be available to suppress oxidative stress in vivo.

A study on the teratogenic and cytotoxic effects of safflower extract

In recent years, the use of carthami flos (the flowers of Carthamus tinctorius L.) as a coloring and flavoring agent has increased as a food additive in Iran. In order to evaluate its safety, the teratogenic effects of carthami flos on the central nervous system development in mice was investigated. Furthermore, its cytotoxic effect on the rat nervous cell culture was studied to complete safety evaluations. For teratogenic studies, after natural mating, pregnant mice were divided into test and control groups. The groups were treated with different dosage regimens of aqueous carthami flos extract during 0–8 days of gestation. Embryos were then isolated at the 13th gestation day and evaluated for macroscopic, microscopic and morphometric characteristics. The results showed that in higher doses (1.6 and 2 mg/kg/day) the embryos were absorbed, whereas with lower dose (1.2 mg/kg/day) changes in external, internal and longitudinal diameters, open neuropore, changes in cellular orientation and cellular degeneration were observed. The results obtained from cytotoxic assay also demonstrated a concentration-dependent cytotoxic effect of carthami flos extract. It is concluded that the use of carthami flos as a food additive should be reconsidered.

Immunomodulating activities of polysaccharide fractions from dried safflower petals.

In the course of screening for immunomodulators, we found a significant blastogenic activity specific for splenic B cells in the extracts of safflower (Carthamus tinctorius L.). Active fractions termed SF1 and SF2 were purified from dried petals of safflower by boiling water extraction, ethanol precipitation and Sepharose CL-2B column chromatography. The elution profiles of the gel filtration indicated that the molecular weight of SF1 and SF2 was estimated to be more than 100 kD. Major components of SF1 and SF2 seem to be polysaccharides, and structural analysis of alditol acetate derivatives by GC-MS revealed some differences between SF1 and SF2 in the sugar component. Biological activities of SF1 and SF2 on B cells and macrophages were examined in comparison with lipopolysaccharides (LPS). SF1 and SF2 induced both the proliferation and the IgM production of B cells to the equivalent level as those induced by LPS. In macrophages, SF1 and SF2 effectively stimulated the production of NO. However, SF1 stimulated the production of IL-1, IL-6, and TNF as much as LPS, while SF2 induced them only weakly or not at all. Thus, these results suggest that SF1 and SF2 activate B cells and macrophages in different mechanisms.

Microbiology of siljo, a traditional Ethiopian fermented legume product

Siljo was prepared by thoroughly cooking powdered horsebean (Vicia faba) in safflower (Carthamus tinctorius) extract into a semi-solid slurry and then adding to it untreated black mustard (Brassica nigra) powder after cooling to 50°C. The black mustard powder was the source of starter microorganisms, with Lactobacillus acidophilus, L. plantarum and L. delbruekii initiating and later dominating the fermentation process. The pH of the fermenting mass dropped to 4.5 within 36 h and reached 4.0 at 168 h. Aerobic mesophilic bacteria and lactic acid bacteria were each present at about 1×10(10) c.f.u./ml after 36 h of fermentation but Enterobacteriaceae were never detected. The dominant aerobic mesophilic flora consisted of Micrococcus, Bacillus and Lactobacillus spp. Crude protein, crude fat and ash increased slightly during the fermentation, with final values of around 28%, 25% and 7%, respectively, but there was a marked increase in protein availability and concentration during the fermentation.