Yuxin Chen

The Mechanism of Antifungal Action of Essential Oil from Dill (Anethum graveolens L.) on Aspergillus flavus

The essential oil extracted from the seeds of dill (Anethum graveolens L.) was demonstrated in this study as a potential source of an eco-friendly antifungal agent. To elucidate the mechanism of the antifungal action further, the effect of the essential oil on the plasma membrane and mitochondria of Aspergillus flavus was investigated. The lesion in the plasma membrane was detected through flow cytometry and further verified through the inhibition of ergosterol synthesis. The essential oil caused morphological changes in the cells of A. flavus and a reduction in the ergosterol quantity. Moreover, mitochondrial membrane potential (MMP), acidification of external medium, and mitochondrial ATPase and dehydrogenase activities were detected. The reactive oxygen species (ROS) accumulation was also examined through fluorometric assay. Exposure to dill oil resulted in an elevation of MMP, and in the suppression of the glucose-induced decrease in external pH at 4 µl/ml. Decreased ATPase and dehydrogenase activities in A. flavus cells were also observed in a dose-dependent manner. The above dysfunctions of the mitochondria caused ROS accumulation in A. flavus. A reduction in cell viability was prevented through the addition of L-cysteine, which indicates that ROS is an important mediator of the antifungal action of dill oil. In summary, the antifungal activity of dill oil results from its ability to disrupt the permeability barrier of the plasma membrane and from the mitochondrial dysfunction-induced ROS accumulation in A. flavus.

In vivo and in vitro antioxidant activity and α-glucosidase, α-amylase inhibitory effects of flavonoids from Cichorium glandulosum seeds.

The aim of this study was to investigate the antioxidant, anti-glucosidase and anti-amylase activities of total flavonoids (TFs) from Cichorium glandulosum seeds, and to analyse its chemical composition by HPLC-ESI/MS. In vitro study, radical scavenging IC50 values of TFs were 7.33±0.093, 9.24±0.100, 154.33±11.38 and 256.7±4.86 μg/ml for DPPH, ABTS, hydroxyl radicals, and superoxide anion, respectively. In the 8-64 mg/ml range, α-glucosidase and α-amylase were inhibited by TFs to a certain extent. In vivo, the treatment groups with TFs (100, 200, 400 mg/kg) showed a significant decrease in the malondialdehyde level, the superoxide dismutase and glutathione levels were restored to almost normal levels, and the catalase and glutathione peroxidase levels significantly increased compared to the CCl4-intoxicated group in rats. The present study suggests that C. glandulosum seeds should be given special attention because of their antioxidant and anti-glucosidase, anti-amylase activity.

Inhibition of pancreatic lipase, α-glucosidase, α-amylase, and hypolipidemic effects of the total flavonoids from Nelumbo nucifera leaves

ETHNOPHARMACOLOGICAL RELEVANCE Nelumbo nucifera Gaertn. leaves have been used as medicinal herbs in the past 1300 years, specifically utilized to cure hyperlipidemia, hyperglycemia, and obesity. It has been recorded in the most famous medicinal book in China for more than 400 years. The present study aims to identify the potential therapeutic activities of the flavonoids isolated from Nelumbo nucifera leaves. MATERIALS AND METHODS Nelumbo nucifera leaf flavonoids (NLF) were tested for the inhibition of lipase, α-glucosidase, and α-amylase activities in vitro. A single dose of NLF was administered by oral gavage in mice for acute toxicity. Wistar rats with high-fat diet-induced hyperlipidemia and two other animal models were used to evaluate the hypolipidemic effects of NLF. RESULTS Our in vitro biochemistry tests revealed that the NLF showed high inhibitory activity against porcine pancreatic lipase, α-amylase, and α-glucosidase with IC50 values of 0.38 ± 0.022, 2.20 ± 0.18, and 1.86 ± 0.018 mg/mL, respectively. Furthermore, the NLF significantly lowered the lipid components, such as the total cholesterol, triglycerides, low-density lipoprotein cholesterol, and malondialdehyde, in various established in vivo systems and raised the high-density lipoprotein cholesterol. Moreover, the NLF alleviated high-fat diet-induced lipid accumulation in the liver. CONCLUSIONS The results demonstrate that NLFs can effectively ameliorate hyperlipidemia and inhibit the key enzymes related to type 2 diabetes mellitus. Our findings may provide new pharmacological basis for the treatment of hyperlipidemia, hyperglycemia, and obesity using NLFs.

In vitro and in vivo hepatoprotective and antioxidant activity of ethanolic extract from Meconopsis integrifolia (Maxim.) Franch

ETHNOPHARMACOLOGICAL RELEVANCE Meconopsis integrifolia (Maxim.) Franch is a high mountain endemic species used as a traditional Tibetan and Mongolian herb to treat hepatitis, pneumonia, and edema. This study aims to investigate the hepatoprotective and antioxidant effects of Meconopsis integrifolia ethanolic extract (MIE) in vitro and in vivo. MATERIALS AND METHODS The in vitro antioxidant property of MIE was investigated by employing various established systems. Rats with carbon tetrachloride (CCl4)-induced liver injury were used to assess the hepatoprotective and antioxidant effect of MIE in vivo. The level or activity of alkaline phosphatase (ALP), glutamate pyruvate transaminase (ALT), aspartate aminotransferase (AST), and total bilirubin (TB) in the blood serum and thiobarbituric acid reactive substances (TBARS), superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) in the liver and kidney of the rats were assayed using standard procedures. RESULTS MIE exhibited strong antioxidant ability in vitro. In the rats with CCl4-induced liver injury, the groups treated with MIE and silymarin showed significantly lower levels of ALT, AST, ALP, and TB. MIE demonstrated good antioxidant activities in both the liver and kidney of the rats in vivo. CONCLUSIONS MIE exhibits excellent hepatoprotective effects and antioxidant activities in vitro and in vivo, supporting the traditional use of Meconopsis integrifolia in the treatment of hepatitis.

Antifungal mechanism of essential oil from Anethum graveolens seeds against Candida albicans

This work studied the antifungal mechanism of dill seed essential oil (DSEO) against Candida albicans. Flow cytometric analysis and inhibition of ergosterol synthesis were performed to clarify the mechanism of action of DSEO on C. albicans. Upon treatment of cells with DSEO, propidium iodide penetrated C. albicans through a lesion in its plasma membrane. DSEO also significantly reduced the amount of ergosterol. These findings indicate that the plasma membrane of C. albicans was damaged by DSEO. The effect of DSEO on the functions of the mitochondria in C. albicans was also studied. We assayed the mitochondrial membrane potential (mtΔψ) using rhodamine 123 and determined the production of mitochondrial dysfunction-induced reactive oxygen species (ROS) via flow cytometry. The effects of the antioxidant l-cysteine (Cys) on DSEO-induced ROS production and the antifungal effect of DSEO on C. albicans were investigated. Exposure to DSEO increased mtΔψ. Dysfunctions in the mitochondria caused ROS accumulation in C. albicans. This increase in the level of ROS production and DSEO-induced decrease in cell viability were prevented by the addition of Cys, indicating that ROS are an important mediator of the antifungal action of DSEO. These findings indicate that the cytoplasmic membrane and mitochondria are the main anti-Candida targets of DSEO.

Hepatoprotective activity of flavonoids from Cichorium glandulosum seeds in vitro and in vivo carbon tetrachloride-induced hepatotoxicity.

ETHNOPHARMACOLOGICAL RELEVANCE Cichorium glandulosum Boiss. et Huet was used historically in Uyghur folk medicine. Its roots, seeds, and aerial parts are extensively used by Uyghur residents in Xinjiang to eliminate savda typhoid, dredge and cure obstructive jaundice variety liver disorders. AIM OF THE STUDY To evaluate the hepatoprotective activity of total flavonoids (TFs) obtained from C. glandulosum seeds against carbon tetrachloride (CCl4)-induced liver damage in vitro and in vivo. To investigate the mechanisms of hepatoprotective effects for TFs. MATERIALS AND METHODS The dried seeds of C. glandulosum were extracted with 70% aqueous ethanol, and the extract was chromatographed with D101 macroporous resin. In vitro the antioxidant capacity against lipid peroxidation (LPO) was evaluated using ferrothiocyanate, thiobarbituric acid, β-carotene bleaching, and LPO inhibition assay. The cytotoxicity and hepatoprotective activity of TFs were evaluated in human liver hepatoma cells (HepG2). MTT assay, hepatic injury markers aspartate aminotransferase (AST), alanine aminotransaminase (ALT), lactate dehydrogenase (LDH) leakage, malondialdehyde (MDA), and glutathione (GSH) were performed. In vivo the hepatoprotective activity of TFs against CCl4-induced acute liver injury was evaluated in rats. A series of biochemical and antioxidant parameter levels were measured in liver homogenate. The suppressive effect on pancreatic lipase activity was determined. RESULTS Results indicated that TFs showed antioxidant capacity against lipid peroxidation (LPO). Administrating CCl4 (1%, v/v) caused a significant decrease in HepG2 viability. Treatment with TFs at doses (62.5, 125, and 250 μg/ml) could significantly ameliorate the cytotoxicity and decline the levels of AST, ALT, and LDH induced by CCl4. The markers including MDA and GSH, which were close to oxidative damage, were restored. Oral treatment with TFs in vivo at doses of 100, 200, and 400 mg/kg significantly reduced the levels of AST, ALT, alkaline phosphatase (ALP), total bilirubin (TB), and thiobarbituric acid reactive substances (TBARs) in the serum compared with CCl4-induced acute liver injury in rats. TFs showed dose-dependent suppressive effects on pancreatic lipase activity, and the IC50 was 1.318 ± 0.164 mg/ml. CONCLUSION TFs from C. glandulosum seeds demonstrated significant hepatoprotection against CCl4-induced hepatotoxicity. TFs exhibited significant suppression of LPO and pancreatic lipase capacity, which may be the mechanisms of hepatoprotective effects against CCl4. Our results contribute towards validation of the traditional use of C. glandulosum seeds in the treatment of liver disorders.

Dill (Anethum graveolens L.) seed essential oil induces Candida albicans apoptosis in a metacaspase-dependent manner.

Dill (Anethum graveolens L.) has been used in traditional Uighur medicine for its various pharmacological activities. Previous studies have suggested that dill seed essential oil (DSEO) has anti-Candida potential and the mechanism of its action also has been studied. Our study examined whether DSEO induces apoptosis in the human pathogen Candida albicans ATCC 64550. Our results indicate that C. albicans ATCC 64550 cells treated with DSEO show some typical apoptosis characters, such as decrease in adenosine triphosphatase (ATPase) activity, chromatin condensation, nuclear fragmentation, and phosphatidylserine (PS) exposure. The DSEO promoted cytochrome c (cyt c) release and metacaspase activation, which resulted in C. albicans ATCC 64550 apoptosis. L-cysteine prevented the DSEO-induced nuclear fragmentation, PS externalization, and metacaspase activation, thus indicating that the reactive oxygen species (ROS) is an important mediator of DSEO-induced apoptosis. To our knowledge, this study is the first to report the induction of apoptosis of this pathogen with concomitant metacaspase activation by DSEO.

In vitro and in vivo evaluation of inhibition activity of lotus (Nelumbo nucifera Gaertn.) leaves against ultraviolet B-induced phototoxicity

Lotus (Nelumbo nucifera Gaertn.), an aquatic vegetable, is extensively cultivated in eastern Asia, particularly in China. Our previous study showed that lotus leaf extracts (LLEs) have strong antioxidant effects in vitro and in vivo. The main antioxidants in lotus leaf have been identified via liquid chromatography-mass spectrometry. Ultraviolet B (UVB) protective effects have been associated with plant extracts rich in antioxidants. The current study focuses on the mitochondria model to evaluate the potent inhibition activity of LLE against UVB-induced phototoxicity. The level of thiobarbituric acid reactive substances, glutathione, lipid hydroperoxide, conjugated diene, and 4-hydroxynonenal were measured. The in vivo activity of LLE was also investigated in mice model. The results showed that all concentrations of LLE (10, 100, and 1000μg/ml) possessed strong protective effect against UVB-induced phototoxicity in the mitochondria model. The in vivo test showed that LLE have significant protective effects on the level of superoxide dismutase, catalase, and glutathione peroxidase, as well as the contents of hydroxyproline and malondialdehyde in the skin samples. This study would provide a foundation for broadening the applications of lotus leaf in both the medical and food industries.

Coptis chinensis inflorescence extract protection against ultraviolet-B-induced phototoxicity, and HPLC-MS analysis of its chemical composition

Cultivated Coptis chinensis inflorescence has been highly valued in Chinese tea production for many years. The main alkaloid compounds in C. chinensis inflorescence ethanolic extracts (CE) were identified by high-performance liquid chromatography-mass spectrometry. The detected compounds included jatrorrhizine (4.87 mg/g), coptisine (17.18 mg/g), palmatine (3.32 mg/g), and berberine (31.81 mg/g), as well as columbamine and epiberberine (tentatively identified). CE protective activity against ultraviolet-B (UVB)-induced phototoxicity in a mitochondria model was determined by measuring thiobarbituric acid-reactive substrates, lipid hydroperoxide, conjugated diene, 4-hydroxynonenal, and glutathione. The results showed that CE excellently inhibited UVB-induced lipid peroxidation and glutathione reduction in vitro. This photoprotective effect of CE may be caused by the presence of the abovementioned alkaloid compounds and phenolic compounds that enhances CE antioxidant activity. Therefore, CE possesses potent photoprotective property that may find valuable applications in food industries and in anti-phototoxicity formulations.

In vitro and in vivo antioxidant properties of extracts from Coptis chinensis inflorescence.

The inflorescence of cultivated Coptis chinensis has been valued for tea production for many years in China. The antioxidant activities of C. chinensis inflorescence extracts prepared by various solvents were investigated by using several established in vitro systems: 2,2′-azinobis(3-ethylbenzthiazoline-6-sulphonic acid (ABTS), α,α-diphenyl-β-picrylhydrazyl (DPPH) and superoxide radical scavenging assays, reducing power assay, and ferrothiocyanate (FTC) and thiobarbituric acid (TBA) assays. The results showed that the 70% ethanol extract (EE) had the strongest antioxidant activity in vitro among the various extracts. Based on the in vitro results, EE was used to evaluate the antioxidant activity of C. chinensis inflorescence in vivo. The liver and kidney of intoxicated animals showed a significant decrease in superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) levels, while the malondialdehyde (MDA) level showed a significant increase. These changes were significantly reversed after treatment with EE and the standard vitamin E. Thus, the C. chinensis inflorescence may be a valuable natural source that can be applicable to food industries.