Screening of anti allergic active components from Cornus officinalis and their effects on degranulation of RBL-2H3 cells
Allergic disease is one of the diseases that the World Health Organization has listed as one of the key research and prevention diseases in the 21st century. The incidence rate in the world has reached 20%~30%, so finding effective anti allergic drugs is the current research hotspot. At present, there are many anti allergic chemical drugs, but most of them have the characteristics of single action steps and significant side effects. Plant extracts are complex mixtures of natural compounds with different structures and sources, with multi-target anti allergic ability, affecting different key steps of allergic reactions, and have mild effects and minimal side effects. China has abundant resources of traditional Chinese medicine, which provides a good foundation for the research of anti allergic natural substances.

Cornus officinalis Sieb. et Zucc. is a medicinal and edible plant in China, which contains cyclohexene ether terpenes and their glycosides, triterpenoids, flavonoids and other substances. In traditional Chinese medicine theory, it has good nourishing effects and is commonly used to treat deficiency syndrome, liver and kidney diseases, and reproductive system diseases. Cornus officinalis has a variety of biological activities, such as anti allergy, anti-inflammatory, antioxidant, anti diabetes, anti osteoporosis, vasodilation and other activities.

Kim et al. found that the water extract of Cornus officinalis can inhibit lung allergic reactions in OVA induced asthma models and has certain anti allergic effects. Kao et al. found that a traditional Chinese medicine formula containing Cornus officinalis can effectively alleviate allergic asthma symptoms induced by allergen Der-p and reduce serum IgE levels. Li et al. found that traditional Chinese medicine preparations containing Cornus officinalis (Cornus officinalis accounts for 11%) have significant inhibitory effects on skin itching, PCA response, and skin dysfunction. Atopic dermatitis is an allergic skin disease. Mechesso et al. reported that C2RTPL extract (a formula consisting of Cornus officinalis: Rose multiflora: Lespedeza: Platycodon grandiflorus: Chestnut in a ratio of 4:1:1:1) can improve the symptoms of atopic dermatitis and reduce the release of β – aminoglucosidase in a concentration dependent manner, indicating its anti allergic activity. Quah et al. found that 70% ethanol extract of Cornus officinalis can reduce the release of anti DNP IgE induced β – aminoglucosidase, indicating its anti allergic effect.

At present, there are no reports on the specific components of Cornus officinalis extract that have anti allergic effects. Therefore, based on previous research, this article further explores the anti allergic active ingredients in Cornus officinalis, aiming to discover the exact components with anti allergic activity in Cornus officinalis. Allergic reactions are classified into types I, II, III, and IV based on their mechanisms, induced by chemical mediators such as histamine, prostaglandins, leukotrienes, inflammatory factors, etc. Studies have shown that the vast majority of type I allergic diseases are related to the activity of hyaluronidase (HAase) in the body, and HAase is one of the target enzymes directly controlling mast cell degranulation (related to type IV allergic reactions). Therefore, hyaluronidase inhibition experiments and mast cell degranulation inhibition experiments are commonly used in in vitro anti allergic research.

This article uses hyaluronidase inhibition activity as an indicator to screen for anti allergic components in Cornus officinalis. The anti allergic activity of the components was further investigated through MTT assay and cell degranulation inhibition experiment, and the component with the highest anti allergic activity was obtained. Finally, gas chromatography-mass spectrometry (GC-MS) was used to analyze and identify the components with anti allergic activity in Cornus officinalis, providing a scientific basis for the comprehensive development and utilization of anti allergic active components in Cornus officinalis.

 

 

This article compares the hyaluronidase inhibitory activity of different polar parts of Cornus officinalis and finds that the petroleum ether part of Cornus officinalis has significantly higher activity than other parts, indicating that substances with lower polarity in Cornus officinalis have higher anti allergic activity. According to Li et al., the extract of a traditional Chinese medicine formula containing Cornus officinalis showed a 70% inhibition rate of hyaluronidase at 3mg/mL, indicating good anti allergic activity. However, the component COPE-8 isolated by silica gel column chromatography in this study showed a hyaluronidase inhibition rate of over 70% at only 2.5mg/mL, indicating higher anti allergic activity.

After activation, mast cells secrete and release mediators and inflammatory factors, which trigger allergic reactions. This process is called mast cell degranulation and is an important link in the occurrence of allergies in the body. Among the released pro-inflammatory mediators, histamine is the most characteristic and effective vasoactive mediator, and is considered a marker of degranulation. Therefore, in this experiment, the anti allergic activity of four active components was evaluated by assessing their effects on the release of histamine and β – glucosidase. The COPE-8 component with the best activity was further screened, which reduced the release levels of histamine and β – glucosidase by 46% and 39% respectively at 20 μ g/mL, while the C2RLP extract mainly composed of Cornus officinalis only reduced the release level of β – glucosidase by 24% at 30 μ g/mL. In addition, the 70% ethanol extract of Cornus officinalis only reduced the release level of β – glucosidase by 18% at 30 μ g/mL, further demonstrating the advantages of component COPE-8 in anti allergic activity.

This study is the first to explore the anti allergic active ingredients in Cornus officinalis by focusing on hyaluronidase inhibition activity. Through systematic separation and purification of Cornus officinalis extract, COPE-8 component with significant inhibition of hyaluronidase activity and cell degranulation was obtained. It can significantly reduce the release of histamine and β – aminoglucosidase induced by calcium ion carrier A23187, and is superior to the reported crude or formula extracts of Cornus officinalis, indicating its strong anti allergic activity. Through GCMS analysis, it was determined that the anti allergic active substances in COPE-8 are mainly terpenoids, including α – coumarinol, β – coumarinol, and perilla alcohol, which also have anti-inflammatory and antioxidant effects, indicating their wide range of biological activities.

In summary, the components with anti allergic activity in Cornus officinalis are alpha cinnamyl alcohol, beta cinnamyl alcohol, and perilla alcohol, which have strong anti allergic activity and both anti-inflammatory and antioxidant biological activities, and have broad application prospects.