Study On The Volatile Constituents And Bioactivity Of Black Coral

Chaoshan area is located on the southeast coast of China in the subtropical zone. It has distinctive monsoon climates. The advantageous geographical location indicates the gestation of various species from sea to land. Some of them are used as traditional medicines in folk for treating some diseases.Hailiu (Antipathes japonica Brook), one genera of black corals (BC), has some pharmaceutical functions used in Chinese folk, especially used as marine biotical materials for cigarette holders in Chaoshan area. At present, there was little known about the chemical constituents extracted of Hailiu. This study aimed to investigate the bioactivities of volatile constituents in BC and to explore the possible benefits of black corals cigarette holders (BCCHs).Method:1. The volatile constituents of BC were extracted by supercritical fluid extraction (SFE) with carbon dioxide (CO₂–SFE).2. The BC extract (BCE) identified and analyzed the constituents by gas chromatography–mass spectrometry (GC–MS) combined with retention index.3. Inorganic elements of BC were determined by inductively coupled plasma spectrometer (ICPS).4. 1, 1-Diphenyl-2-picrylhydrazyl (DPPH) method and Phenanthroline-Fe2+ oxidative assay of hydroxyl radical were applied to evaluate in vitro antioxidation of BCE.5. Antimicrobial susceptibility testing for BCE involved the Kirby–Bauer (KB) disk diffusion method. And a broth microdilution method was used to determine the minimal inhibitory concentration (MIC) against the susceptible microorganisms.6. 60 mice were randomized into 6 groups. Before they were exposed to cigarette smoke (CS) in treatment groups mice were given BCE ultrasonic atomizing inhalation of 0.3mg/ml, 1.5mg/ml and 3mg/ml in normal saline (NS), respectively, for 10 minutes. While control group and CS group mice were treated NS,positive control group was administrated 6mg/ml of Vitamin C (Vc). All animals were euthanized via anesthetic overdose ip. The content of malondialdehyde (MDA), the activity of superoxide dismutase (SOD) and myeloperoxidase (MPO) were analyzed. Histological and morphological studies of right upper lung were performed.Result:1. In total, 15 components were reliably identified in BC and found to be biologically active. These included triethyl phosphate, butylated hydroxytoluene, cedrol, n-hexadecanoic acid, squalene, and cholesterol.2. 13 inorganic elements P, Ca, Mg, S, B, Si, Fe, Cu, Zn, Ba ect were determined by ICPS.3. In the bioactivities tests, BCE showed a scavenging activity of DPPH and hydroxyl radicals.4. The BCE exhibited moderate antimicrobial activity against the Gram-positive microorganisms. The results of the MIC determination indicated the BCE inhibited the microorganisms tested. S. aureus and S. epidermidis had a MIC of 73 and 37μg/μl, respectively.5. Histopathological evaluation: Typical pathological inflammation lesions in the lung were observed in acute lung injury group,including alveolar congestion,hemorrhage,infiltration of neutrophils in the airspace or vessel wall,thickness of the alveolar wall. Pathological changes were relieved obviously in BCE treated groups. Compared with normal group,MPO and MDA in acute lung injury group were all higher(P<0.01),SOD was lower;MPO and MDA in BCE groups were all lower than acute lung injury model group (P<0.05 or P<0.01),SOD was higher.Meanwhile the BCE showed the dose dependent.Conclution:It is the first time to present some volatile constituents of BC. The functions of BC cigarette holders (BCCHs) are related to the active chemical compositions. The antioxidant and antimicrobial activities of BC from in vitro and in vivo may be protective for cigarette smokers who use BCCHs, especially for acute lung injury.