Ischemic stroke leads to severe brain damage and high mortality. Chlorogenic acid is a phenolic compound known to have neuroprotective properties. Bcl-2 family protein plays an important role in the regulation of apoptosis. We investigated whether chlorogenic acid exerts neuroprotective effects against ischemic injury by modulating Bcl-2 and Bax proteins. Middle cerebral artery occlusion (MCAO) was performed to induce cerebral ischemia and rats were injected intraperitoneally with phosphate buffered saline or chlorogenic acid (30 mg/kg) for 2 h after MCAO. Cortical tissues were collected 24 h after MCAO injury and reverse transcription-quantitative real time polymerase chain reaction and Western blot analyses were performed to investigate the expression of Bcl-2 and Bax. The regulation of Bcl-2 and Bax proteins by chlorogenic acid during glutamateinduced cell damage were examined. Cells were collected at 24 h after administration of glutamate (5 mM) and chlorogenic acid (10, 30, 50 μM). These results showed a decrease in Bcl-2 expression and an increase in Bax expression in MCAO animals, but chlorogenic acid treatment alleviated these changes by MCAO damage. Glutamate significantly reduced cell viability, and chlorogenic acid treatment alleviated this reduction in a dose-dependent manner. Glutamate induced a decrease in Bcl-2 expression and an increase in Bax expression, but chlorogenic acid treatment alleviated these changes. We found that chlorogenic acid alleviates changes in the expression of Bcl-2 and Bax proteins induced by brain injury. Therefore, our findings provide an evidence that chlorogenic acid has neuroprotective effects against MCAO damage by modulating Bcl-2 and Bax proteins.

Ischemic stroke causes severe neuronal damage. Chlorogenic acid is a phenolic substance present in fruits and coffee. It also exerts neuroprotective effects against various brain injuries. The 14-3-3 family protein perform a variety of functions including metabolism, signal transduction, cell differentiation, and apoptosis. The purpose of this study is to investigate whether chlorogenic acid regulates the expression of 14-3-3 protein in stroke animal models. Ischemic stroke was induced by middle cerebral artery occlusion (MCAO) surgery. Phosphate buffered saline (PBS) or chlorogenic acid (30 mg/kg) were intraperitoneally injected to adult male rats 2 h before MCAO surgery. Adhesive-removal test was performed 24 h after MCAO surgery and cerebral cortical tissues were collected for further study. MCAO damage caused severe neurological impairment and chlorogenic acid treatment ameliorated this disorder. Our proteomic approach showed a decrease in 14-3-3 expression in MCAO animals with PBS. The decrease in 14-3-3 expression alleviated in MCAO animal with chlorogenic acid. We confirmed changes in various 14-3-3 protein isoforms, including beta/alpha, zeta/delta, gamma, epsilon, eta, and tau through reverse transcription-PCR. These results explained that chlorogenic acid regulates the expression of 14-3-3 protein in MCAO-induced cerebral ischemia. 14-3-3 is considered to be an important protein for cell survival through binding to pro-apoptotic proteins. The maintenance of 14-3-3 levels is an important event in neuroprotection against ischemic injury. Therefore, we can demonstrate that the 14-3-3 protein contributes to the neuroprotective effect of chlorogenic acid in stroke animal models.

본 연구에서는 chlorogenic acid 이성질체 9종과 arbutin을 분석하기 위하여, 전처리 과정과 UHPLC-MS/MS 동시 분석 조건을 확립하였다. UHPLC-MS/MS에서 C18 column을 사용하여 15분 동안 분석할 수 있는 용매 조성을 만들고, 10가지 물질에 대한 정량, 정성 이온을 선택하여 negative 모드에서 분석하는 동시분석법을 확립하였고 과일류를 대상으로 추출, 진탕 추출, 초음파 추출, 원심분리, 농축 등의 과정을 거치는 전처리법을 개발하였다. 기기분석과 전처리법에 대한 유효성은 특이성, 직선성, 정확성, 정밀성 그리고 검출한계 및 정량한계 등을 통하여 확인하였다. 회수율은 48.1-120.3%, intraday precision (RSD)는 0.4-7.9%, interday precision (RSD)는 0.0-7.2%로 확인되었다. 따라서, 본 연구에서 확립한 동시분석방법은 과일류 중 chlorogenic acid와 arbutin을 효과적으로 분석하는데 유용할 것으로 사료된다.

A greening phenomenon has been observed in some plant foods such as chestnut, sweet potato,burdock, and others during processing. The formation of the pigments was postulated as reactions of primary amino compounds with chlorogenic acid or caffeic acid ester, yielding acridine derivatives. Acridine derivatives have been regarded as mutagenetic agents. For the reason, the bacterial reverse mutation test was carried out to evaluate the genotoxicity of green pigment using Salmonella typhimurium TA98 and TA100. Alanine, arginine, aspartic acid, glycine,lysine, and phenylalanine were reacted repectively with chlorogenic acid to synthesize model compound. Green pigment was extracted from sweet potato. Maximum concentration of 2 and 50 mg/plate was tested for the synthetic green pigments and extracted green pigment respectively, taking bacterial survival, solubility, and color intensity into consideration. There was no signigicant increase in the reverse mutation either with or without S9 activation system by any test material. Though further studies with other genotoxicity test system are necessary, both synthetic and sweet potato green pigments seemed not to cause mutation despite the acridine moiety in their structures.

In all living organisms, respiration may lead to oxidative stress, a state where increased formation of reactive oxygen species overwhelms host protection and subsequently induces DNA damage, lipid peroxidation, and protein denaturation. As a phenolic acid, chlorogenic acid occurs ubiquitously in food. It has been proven to have a number of biological effects in vitro and in vivo, including antioxidant, anti-inflammatory, and anticarcinogenic properties. Therefore, to some extent, chlorogenic acid can promote human health, and hopefully provide new methods for treatment of chronic disease. Recent studies have focused on the antioxidant properties of dietary polyphenols. The in vitro data often conflict with results obtained from in vivo studies on the antioxidant capacity of plasma or the resistance of plasma and lipoproteins to oxidation ex vivo after consumption of polyphenol-rich foods by human subjects. These inconsistencies are likely explained by the limited bioavailability of dietary polyphenols and their extensive metabolism in humans. Polyphenols exert multifaceted actions, and any clinical application using these substances should be based on a precise understanding of the physiologically relevant mechanisms.

Background : Campanula takesimana is a herb(Jabanpungnyeoncho) used traditionally in the korea private and we tried the development on a medicinal material. It was known that it has chlorogenic acid, as a immunoadjuvant activity. In this study we investigated the contents of the chlorogenic acid in Companula takesimana at different harversting time by the high performance liquid chromatographic(HPLC)-PDA. Campanula takesimana was collected on May maddle (Flower buds farmed), June middle (flowers opened) and July middle (seeds were mature). Methods and Results : HPLC analysis was carried out using X-bridge C18 column (5um, 250*4.6mm) and a mobile phase consisting of acetonitrile and water containing 0.1% formic acid at a flow rate of 1.0 ml/min. The optimum wavelength for the detection of the Chlorogenic acid was 330 nm using Photo Diode Array Detector. The contents of the chlorogenic acid in the Campanula takesimana extrat were 1.00% (May middle), 0.84% (June middle), 0.05% (July middle), respectively. As the different parts of Campanula takesimana, chlorogenic acid contents in May middle extrat were 2.73% (aerial part), 0.05% (root) and in June middle extrat were 1.61% (aerial part), 0.03% (root), 0.70% (flower) and in July middle were 0.13% (aerial part), 0.03% (root), 0.00% (seed), respectively. Conclusion : From the above results, the highes content of chlorogenic acid was observed in aerial part and May middle extract of Campanula takesimana.

Chlorogenic acid, formed of an ester of caffeic acid and quinic acid, which is naturally abundant in many plant species, was used as a model O-dihydoxy phenolic compound. In the previous study, we have reported that the isolated constituent from Apocynum venetum leaves has an inhibitory effect on Cu2+-induced oxidative modification of low-density lipoprotein (LDL). Among them, chlorogenic acid showed the most potent anti-LDL oxidative activity than other compounds. For the reason, we investigated the inhibitory effect of the chlorogenic acid on Cu2+-induced oxidative modification of LDL, monitored a lag time in the conjugated-diene formation and TBARS formation, and measured TNBS free amino acid group, and form cell formation in vitro system. The TBARS- and diene- formation were strongly inhibited by chlorogenic acid (0~100 μg/ml) with dose dependent manner. On the other hand, TNBS reactive lysine amino groups on LDL oxidation were protected by chlorogenic acid- treated cell group. Therefore, chlorogenic acid inhibited to cholesterol accumulation in the isolated peritoneal macrophage.

A series of aqueous or methanol extracts from four different Korean salad plants were assayed to determine their allelopathy and antioxidant activity. The extracts applied on filter paper in a Petri-dish bioassay significantly inhibited root growth of against alfalfa (Medicago sativa) seedlings. Leaf extracts from 40 g dry tissue L-1 of Aster yomena was most phytotoxic to alfalfa root growth, and followed by that of Cirsium japonicum, Taraxacum officinale, and Ixeris dentate. Methanol extracts of plants dose-dependently increased DPPH free radical scavenging activity in vitro. Antioxidant activity of methanol extracts from the same plant species was investigated, and the result showed high DPPH free radical scavenging activity in Cirsium japonicum, Aster yomena, and Ixeris dentate, however, in Taraxacum officinale was least activity. By means of HPLC analysis, chlorogenic acid, p-coumaric acid, and total phenolics with 7.68, 17.47 and 18.64 mg, 100g-1, respectively, showed the highest amounts in methanol extracts from Cirsium japonicum leaves. These results suggest that Compositae salad plants contain water-soluble substances with allelopathic potential as well as antioxidant activity.

Lettuce (Lactuca sativa L.) is known to contain water-soluble substances that improve antioxidant status due to the richness in antioxidants. Greenhouse experiment was carried out under different shading conditions during spring lettuce growing season. Shade significantly reduced shoot weight, number of leaves and chlorophyll content, while it increased shoot length of lettuce plants. The antioxidant potential of the individual fraction was in order of n-butanol 〉 ethyl acetate 〉 water 〉 n-hexane fraction, although was less than that of commonly used antioxidants, BHT and ascorbic acid. Fractions from lettuce plants dose-dependently increased DPPH free radical scavenging activity, in vitro test. By means of HPLC analysis, BuOH fraction of cultivar 'Hwahyang' (57.93 mg 100g-1) had the highest amount of antioxidant chi orogenic acid. Shading treatment increased average amount of chlorogenic acid of all cultivars in BuoH, EtOAc, hexane and water fractions by 33, 120, 144, and 58%, respectively. These results suggest that lettuce plants had potent antioxidant activity, and their activities were differently exhibited depending on cultivar and fraction.