Pure Artemisia capillaris oil for candle and soap making wholesale diffuser essential oil new for reed burner diffusers

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Rodent model design

The animals were randomly divided into five groups of fifteen mice each. The control group and model group mice were gavaged with sesame oil for 6 days. Positive control group mice were gavaged with bifendate tablets (BT, 10 mg/kg) for 6 days. The experimental groups were treated with 100 mg/kg and 50 mg/kg AEO dissolved in sesame oil for 6 days. On day 6, the control group was treated with sesame oil, and all of the other groups were treated with a single dose of 0.2% CCl4 in sesame oil (10 ml/kg) by intraperitoneal injection. The mice were then fasted free of water, and blood samples were collected from the retrobulbar vessels; collected blood was centrifuged at 3000 × g for 10 min to separate the serum. Cervical dislocation was performed immediately after withdrawal of blood, and liver samples were promptly removed. One part of the liver sample was immediately stored at −20 °C until analysis, and another part was excised and fixed in a 10% formalin solution; the remaining tissues were stored at −80 °C for histopathological analysis (Wang et al., 2008, Hsu et al., 2009, Nie et al., 2015).

Measurement of the biochemical parameters in the serum

Liver injury was assessed by estimating the enzymatic activities of serum ALT and AST using the corresponding commercial kits according to the instructions for the kits (Nanjing, Jiangsu Province, China). The enzymatic activities were expressed as units per liter (U/l).

Measurement of MDA, SOD, GSH and GSH-Px in liver homogenates

Liver tissues were homogenized with cold physiological saline at a 1:9 ratio (w/v, liver:saline). The homogenates were centrifuged (2500 × g for 10 min) to collect the supernatants for the subsequent determinations. Liver damage was assessed according to the hepatic measurements of the MDA and GSH levels as well as the SOD and GSH-Px activities. All of these were determined following the instructions on the kit (Nanjing, Jiangsu Province, China). The results for MDA and GSH were expressed as nmol per mg protein (nmol/mg prot), and the activities of SOD and GSH-Px were expressed as U per mg protein (U/mg prot).

Histopathological analysis

Portions of freshly obtained liver were fixed in a 10% buffered paraformaldehyde phosphate solution. The sample was then embedded in paraffin, sliced into 3–5 μm sections, stained with hematoxylin and eosin (H&E) according to a standard procedure, and finally analyzed by light microscopy (Tian et al., 2012).

Statistical analysis

The results were expressed as the mean ± standard deviation (SD). The results were analyzed using the statistical program SPSS Statistics, version 19.0. The data were subjected to an analysis of variance (ANOVA, p < 0.05) followed by Dunnett’s test and Dunnett’s T3 test to determine the statistically significant differences between the values of various experimental groups. A significant difference was considered at a level of p < 0.05.

Results and discussion

Constituents of AEO

Upon GC/MS analysis, the AEO was found to contain 25 constituents eluted from 10 to 35 min, and 21 constituents accounting for 84% of the essential oil were identified (Table 1). The volatile oil contained monoterpenoids (80.9%), sesquiterpenoids (9.5%), saturated unbranched hydrocarbons (4.86%) and miscellaneous acetylene (4.86%). Compared with other studies (Guo et al., 2004), we found abundant monoterpenoids (80.90%) in the AEO. The results showed that the most abundant constituent of AEO is β-citronellol (16.23%). Other major components of AEO include 1,8-cineole (13.9%), camphor (12.59%), linalool (11.33%), α-pinene (7.21%), β-pinene (3.99%), thymol (3.22%), and myrcene (2.02%). The variation in the chemical composition may be related to the environmental conditions that the plant was exposed to, such as mineral water, sunlight, the stage of development and nutrition.

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Liver disease, a common disorder caused by viral hepatitis, alcoholism, liver-toxic chemicals, unhealthy dietary habits and environmental pollution, is a global concern (Papay et al., 2009). However, medical treatment for this disease is often hard to administer and has a confined effect. Traditional Chinese herbal medicines, which underlie numerous prescriptions used to treat liver diseases, are still widely used by the Chinese (Zhao et al., 2014). Artemisia capillaris Thunb., Asteraceae, according to the Bencao Gangmu, the most famous records of Chinese Traditional Medicine, has been widely used as a medicine to clear heat, promote diuresis and remove jaundice and has also been used as a flavor in beverages, vegetables, and pastries because of its particular fragrance. A. capillaris has been regarded as a type of Chinese folk medicine and food by a growing number of people. Therefore, there have been considerable efforts to develop useful herbal medicines, such as A. capillaris, for the treatment of liver disease.

In recent years, herbal medicines have gained more attention and popularity for the treatment of liver disease because of their safety and efficacy (Ding et al., 2012). A. capillaris has been proven to possess good hepatoprotective activity based on modern pharmacological methods (Han et al., 2006). It is also an important medicinal material in China and is a popular anti-inflammatory (Cha et al., 2009a), choleretic (Yoon and Kim, 2011), and anti-tumor (Feng et al., 2013) herbal remedy.

Phytochemical studies have revealed a number of volatile essential oils, coumarins, and flavonol glycosides as well as a group of unidentified aglycones from A. capillaris (Komiya et al., 1976, Yamahara et al., 1989). The essential oil of A. capillaris (AEO) is one of the main pharmacological active compounds and confers anti-inflammatory (Cha et al., 2009a) and anti-apoptotic properties (Cha et al., 2009b). However, as AEO is one of the main compounds of A. capillaris, the potential hepatoprotective activities of the major constituents from A. capillaris should be explored.

In this study, the protective effect of AEO on carbon tetrachloride (CCl4)-induced hepatotoxicity was evaluated by biochemical methods, such as hepatic reduced glutathione (GSH), malondialdehyde (MDA) levels, superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) activity, as well as the activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the serum. The extent of CCl4-induced liver injury was also analyzed through histopathological observations, accompanied with phytochemical analysis by GC–MS to identify the constituents of AEO.