Acute Oral Toxicity and Genotoxicity Test and Evaluation of Cinnamomum camphora Seed Kernel Oil.

Cinnamomum camphora seed kernel oil (CCSKO) is one of the important natural medium chain triglycerides (MCT) resources, with more than 95.00% of medium chain fatty acids found in the world, and has various physiological effects. However, CCSKO has not been generally recognized as a safe oil or new food resource yet. The acute oral toxicity test and a standard battery of genotoxicity tests (mammalian erythrocyte micronucleus test, Ames test, and in vitro mammalian cell TK gene mutation test) of CCSKO as a new edible plant oil were used in the study. The results of the acute oral toxicity test showed that CCSKO was preliminary non-toxic, with an LD50 value higher than 21.5 g/kg body weight. In the mammalian erythrocyte micronucleus test, there was no concentration-response relationship between the dose of CCSKO and micronucleus value in polychromatic erythrocytes compared to the negative control group. No genotoxicity was observed in the Ames test in the presence or absence of S9 at 5000 µg/mL. In vitro mammalian cell TK gene mutation test showed that CCSKO did not induce in vitro mammalian cell TK gene mutation in the presence or absence of S9 at 5000 µg/mL. These results indicated that CCSKO is a non-toxic natural medium-chain oil.

Identification and Safety Evaluation of Ochratoxin A Transformation Product in Rapeseed Oil Refining Process.

Ochratoxin A (OTA) is an important mycotoxin detected in edible oil, and it can be effectively removed by classical edible oil refining processes. However, the fate of OTA in the refining process has not been reported. In this study, we systematically tracked the OTA changes during the oil refining process by fortifying 100 µg/kg OTA in crude rapeseed oil. Results showed that about 10.57%, 88.85%, and 0.58% of OTA were removed during the degumming, deacidification, and decolorization processes. Among them, 16.25% OTA was transferred to the byproducts, including 9.85% in degumming wastewater, 5.68% in soap stock, 0.14% in deacidification wastewater, and 0.58% in the decolorizer; 83.75% OTA was found to transform into the lactone ring opened OTA (OP-OTA) during the deacidification stage, which is attributed to the hydrolysis of the lactone ring of OTA in the alkali refining. The OP-OTA was verified to distribute in the soap stock, and small amounts of OP-OTA could be transferred to deacidified wastewater when the OTA pollution level reached 500 µg/kg in crude rapeseed oil. The OP-OTA exhibited strong toxicity, especially nephrotoxicity, as reflected by the cell viability assay and in silico toxicity. Therefore, the safety of the soap stock processing products from OTA-contaminated rapeseed deserves attention.

Effects of medium chain triglycerides on lipid metabolism in high-fat diet induced obese rats.

This study aimed to compare the effects of three different medium chain triglycerides (MCTs) on lipid metabolism in obese rats. A high fat diet was fed to Sprague-Dawley rats to induce obesity, and then caprylic triglyceride (CYT), capric triglyceride (CT) and lauric triglyceride (LT) were synthesized and used to treat the obese rats for 12 weeks. The obesity phenotype and molecular changes related to lipid metabolism were determined. The results showed that all the three MCTs reduced the body weight (BW) and fat coefficient in obese rats, and the levels of plasma and liver lipids were also improved. Among the MCT groups, the LT group showed the lowest BW with the lowest food intake. Furthermore, three MCTs had different effects on the expression levels of lipid metabolism-related proteins (PPARs and SREBPs). The LT group performed the best among the three MCT groups in the protein expression levels. Interestingly, high-dose LT decreased the expression of the low density lipoprotein receptor (LDLR) in the liver, which impaired the transport of the very low density lipoprotein (VLDL)/low density lipoprotein (LDL) to the liver, resulting in high levels of total plasma cholesterol (TC) and LDL-c in the rats. We have for the first time found that different MCTs had different effects on the expression levels of triacylglycerol and cholesterol metabolism-related proteins in obese rats. These findings would help better understand the relationship between the health benefit and the type of MCT.

Effects of preheat treatment and polyphenol grafting on the structural, emulsifying and rheological properties of protein isolate from Cinnamomum camphora seed kernel.

In this study, protein isolate (PI) and purified polyphenol extract (PPE) were prepared from Cinnamomum camphora seed kernel (CCSK). The effects of preheat treatment (50-90 °C) combined with polyphenol grafting (5 % PPE, w/w) on the structural, emulsifying and rheological properties of PI were investigated. Results demonstrated the preheat treatments at 80 and 90 °C significantly increased the extent of protein aggregation of PI. Fluorescence spectra and thermal behavior analysis revealed that preheat-treated PI exhibited more compact structure and higher thermal stability. Moreover, the emulsifying stability and apparent viscosity of PI were enhanced after preheat treatments at 50, 60 and 70 °C. After modification by PPE, the secondary structural changes of preheat-treated PI were confirmed by FTIR. PPE modification improved the thermal stability and antioxidant activities of preheat-treated PI. These results provide a novel way to combine the advantages of preheat treatment and polyphenol grafting in developing a novel protein ingredient.

Assessment of the effect of ethanol extracts from Cinnamomum camphora seed kernel on intestinal inflammation using simulated gastrointestinal digestion and a Caco-2/RAW264.7 co-culture system.

Cinnamomum camphora seeds have multiple bioactivities. There were few studies on the effect of C. camphora seeds on intestinal inflammation in vitro and in vivo. The study aimed to investigate the effects of ethanol extracts from C. camphora seed kernel on intestinal inflammation using simulated gastrointestinal digestion and a Caco-2/RAW264.7 co-culture system. Results showed that the digested ethanol extracts (dEE) were rich in polyphenols, and a total of 17 compounds were tentatively identified using UPLC-LTQ-Orbitrap-MS/MS. dEE increased cell viability, while decreasing the production of reactive oxygen species, and the secretion and gene expression of inflammatory markers (NO, PGE2, TNF-α, IL-1ß and IL-6). dEE also down-regulated NF-κB/MAPK pathway activities by suppressing the phosphorylation of relevant signaling molecules (p65, IκBα, ERK and p38), as well as the expression of TLR4 receptor protein. Furthermore, dEE may improve intestinal barrier function by increasing the TEER value, and the expression of tight junction proteins (ZO-1, claudin-1 and occludin). The results suggest the ethanol extracts from C. camphora seed kernel may have strong anti-inflammatory activities, and a potential application in the prevention or treatment of intestinal inflammation and enhancement of intestinal barrier function in organisms.

High Dietary Intervention of Lauric Triglyceride Might be Harmful to Its Improvement of Cholesterol Metabolism in Obese Rats.

Hypercholesterolemia is often considered to be a major risk factor for atherosclerosis, and medium-chain fatty acids have been found to reduce the total cholesterol (TC) level and maintain low-density lipoprotein cholesterol (LDL-c) stability. However, we unexpectedly found that the levels of TC and LDL-c were increased in obese rats treated with high-dose lauric triglycerides (LT). The study aimed to investigate the effect and mechanism of LT on cholesterol metabolism in obese rats. Our results showed that LT intervention could reduce cholesterol biosynthesis by downregulating the expression of HMG-CoA reductase in obese rats. LT increased the expression levels of PPARγ1, LXRα, ABCA1, and ABCG8 in the liver. These results indicated that LT could improve the lipid transfer and bile acid efflux. However, LT significantly increased the expression of PCSK 9, resulting in accelerated degradation of LDLR, thus reducing the transport of very LDL (VLDL) and LDL to the liver. Together with the increased expression of NPC1L1 protein, LT impaired the uptake of VLDL/LDL by the liver and increased the reabsorption of sterols, leading to an increase in the levels of TC and LDL-c in obese rats.

Covalent modification by phenolic extract improves the structural properties and antioxidant activities of the protein isolate from Cinnamomum camphora seed kernel.

In this study, protein isolate (PI) and purified phenolic extract (PPE) were prepared from Cinnamomum camphora seed kernel (CCSK). The effects of covalent modification of PI by PPE at different concentrations (1, 2, 3, 4 and 5%, w/w) were investigated with respect to structural properties and antioxidant activities of protein. Fifteen bioactive compounds in PPE were tentatively identified by UPLC-ESI-MSn. With the increase of PPE concentration, the turbidity, covalent binding rate, phenolic content and color intensity of the PI-PPE complexes were gradually increased. Fourier transform infrared spectroscopy and circular dichroism spectroscopy analysis showed that the secondary and tertiary structures of the complexes were changed and became greater order than PI. Furthermore, the complexes exhibited stronger thermal stability and antioxidant activities than those of PI. These results suggested that the protein-phenolic covalent complexes obtained from CCSK may have great potential to be used in food formulations as functional ingredients.

Ethanol extracts from Cinnamomum camphora seed kernel: Potential bioactivities as affected by alkaline hydrolysis and simulated gastrointestinal digestion.

The aim of the study was to evaluate the changes of potential bioactivities of ethanol extracts (EE) from Cinnamomum camphora seed kernel (CCSK) after alkaline hydrolysis and simulated gastrointestinal digestion. A total of 13 compounds in EE, mainly phenolics and saponins were tentatively identified using HPLC-ESI-QTOF-MS2 analysis. The total phenolic and total flavonoid contents in EE decreased by 30.6%, 1%, 33% and 11.8% after hydrolysis and digestion, respectively. The total saponins content decreased by 17% after hydrolysis while increased by 48% after digestion. The total condensed tannin contents increased by 70.3% and 17.2% after hydrolysis and digestion, respectively. The 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), cupric ion reducing activity (CUPRAC), ferric reducing antioxidant power (FRAP) and metal chelating activity (MCA) were used to determine antioxidant activity. Overall, the changes of antioxidant activities by hydrolysis and digestion were consistent with the changes of their total phenolic and flavonoid contents. The α-amylase and α-glucosidase inhibitory activities in EE increased by 817% and 33.3% after digestion while decreased by 12.3% and 19% after hydrolysis, respectively. Although the inhibitory activities of cholinesterase, tyrosinase and xanthine oxidase were decreased by hydrolysis and digestion, most of these activities were retained. These results showed that CCSK ethanol extracts had strong bioactivities and were reasonably stable to alkali and digestive enzymes.

Cinnamomum camphora Seed Kernel Oil Improves Lipid Metabolism and Enhances ß3-Adrenergic Receptor Expression in Diet-Induced Obese Rats.

The effects of dietary Cinnamomum camphora seed kernel oil (CCSKO) containing medium-chain triacylglycerols on lipid metabolism and mRNA and protein expression of ß-3 adrenergic receptor in adipose tissue were studied in diet-induced obese rats. High fat food-induced obese rats were randomly divided into CCSKO group, Lard group, Soybean oil (SOY) group and naturally restoring group (n = 10). Rats fed with low fat food were used as a normal control group. Significant decreases in body mass and abdominal fat mass/body mass after 12 weeks were found in CCSKO group as compared with Lard and SOY groups (p < 0.05). Levels of blood total cholesterol (TC), triglyceride, free fatty acid, fasting insulin and insulin resistance in the CCSKO group were decreased significantly, and noradrenaline level and insulin sensitivity index in the CCSKO group were significantly higher than other groups. Meanwhile liver TC and triglyceride levels in the CCSKO group were also decreased markedly. Expression levels of ß3-adrenergic receptor mRNA and protein were higher in CCSKO group than in Lard and SOY groups. These results suggest that CCSKO may contribute to reduction of the body fat mass, promote lipid metabolism and up-regulate ß3-adrenergic receptor expression in high fat diet-induced obese rats.

Cinnamomum camphora Seed Kernel Oil Ameliorates Oxidative Stress and Inflammation in Diet-Induced Obese Rats.

Cinnamomum camphora seed kernel oil (CCSKO) was found to reduce body fat deposition and improve blood lipid in both healthy and obese rats. The study was aimed to investigate the antioxidative stress and anti-inflammatory effects of CCSKO in high-fat-diet-induced obese rats. The obese rats were treated with CCSKO, lard, and soybean oil, respectively, for 12 wk. The level of total antioxidant capacity (T-AOC), activities of superoxide dismutase (SOD), glutathione peroxidase, and catalase, and levels of malondialdehyde (MDA), tumor necrosis factor (TNF)-α, peroxisome proliferator-activated receptor (PPAR)-γ, interleukin (IL)-6, and P65 were compared among CCSKO, lard, and soybean oil groups. Our results showed that the level of T-AOC and activities of SOD and catalase were significantly increased and the level of MDA was significantly decreased in CCSKO group. In addition, CCSKO treatment reduced the activities of serum glutamic oxaloacetic transaminase and glutamate-pyruvate transaminase, and levels of serum TNF-α, IL-6, and P65 through raising the level of PPAR-γ. In conclusion, CCSKO has, for the first time, been found to ameliorate oxidative stress and inflammation in high-fat-diet-induced obese rats.

Camphor Tree Seed Kernel Oil Reduces Body Fat Deposition and Improves Blood Lipids in Rats.

The total and positional fatty acid composition in camphor tree (Cinnamomum camphora) seed kernel oil (CKO) were analyzed, and for the first time, the effect of CKO on body fat deposition and blood lipids in rats was studied. The major fatty acids in CKO were determined to be decanoic acid (C10:0, 51.49%) and dodecanoic acid (C12:0, 40.08%), and uniformly distributed at Sn-1, 3, and Sn-2 positions in triglyceride (TG). Rats were randomly divided into control, CKO, lard, and soybean oil groups. At the end of the experiment, levels of blood lipids and the fats of abdomen in the rats were measured. The main organ were weighted and used for the histological examination. The results showed that body weight and fat deposition in CKO group were significantly lower than the lard and soybean groups. Moderate consumption of CKO was found to improve the levels of blood TG and low density lipoprotein cholesterol.