Study On The Mechanism Of The Muliti-Ways Effect Of Laminaria Japonica On Atheroselerosis

With the development of social economy and the transformation of human way of life, cardiovascular and cerebrovascular diseases have become the major killer of human health, while atherosclerosis is the arch-criminal which induced cardiovascular and cerebrovascular diseases. Therefore, the screening of the anti atherosclerosis drugs or health products has became the hotspot at present. Because of the disadvantages such as obvious adverse reaction of chemical synthetic drugs, the screening of anti atherosclerosis activity ingredients from the traditional Chinese medicine or the homology of medicine and food plants has gradually attracted people’s attention. Laminaria japonica is one kind of edible algae with high nutrition value, modern pharmacological research shows that the Laminaria japonica has anti atherosclerosis effects to a certain extent, but the effect of different components of Laminaria japonica in the role of anti atherosclerosis has not been systematically researched and reported. In this experiment, the Laminaria japonica was taken as the research object, on the basis of the extraction, separation and purification of polysaccharides and polyphenols, further research on the biological activity of each component of Laminaria japonica was studied to explor the multi-ways effect of Laminaria japonica on atherosclerosis from different views. Main experimental contents and the results are as follows:A comprehensive extraction plan of Laminaria japonica was designed to get the polyunsaturated fatty acids, total polyphenols and total polysaccharides with appropriate solvents according to the different polar of the components in Laminaria japonica. Further research was done on the extraction, separation and purification of the polyphenols and polysaccharides.With the extraction yields of total polyphenol as the index, The ultrasonic extraction process was optimized by single factor experiment and orthogonal test, the result shows that the best extraction condition is the materials liquid ratio of 1: 15(g:mL),70% ethanol as the solvent, extracting temperature of 30 ℃, the extracting time of 40 min and the average extraction yield of total polyphenol is 2.55% with good repeatability. Total polyphenols of Laminaria japonica were separated and purified by the adsorption of macroporous resin (AB-8) and gradient elution acodding to the different dissolution characteristics of polyphenol components, with the UV spectroscopy and thin layer chromatography, the polyphenols are divided into 5 groups:Frl, Fr2, Fr3, Fr4, Fr5 respectively, which is an ideal result.With the extraction yields of polysaccharide as the index, the process of hot water extraction was optimized by single factor experiment and orthogonal test, the result shows that the best extraction parameters are as follows:extraction temperature of 100 ℃, the material liquid ratio of 1:40(g:mL), the extraction time of 1 h,2 times of extraction, under this condition,the average extraction yield of polysaccharide is 7.24% with good repeatability. The deproteinization of the total polysaccharide of Laminaria japonica obtained by hot water extraction and ethanol precipitation was carried out using papaya proteinase-Sevag method,and with the method of precipitation of CaCl2 and sixteen alkyl three methyl bromide(CTAB), the total polysaccharides in Laminaria japonica are separated into 3 three components:laminaran, fucose and alginate calcium.With the mechanisms of atherogenesis as the starting point of the study, the antioxidant, hypolipidemic and anti-inflammatory activities of each component in Laminaria japonica obtained by separation and purification were studied.The investigation of antioxidant activity was done based on the scavenging capacity of DPPH’and’OH together with the anti-lipid peroxidation ability. Expect for alginate calcium and Frl, Fr2, all the components in Laminaria japonica have the DPPH·and·OH cavenging activities, and the cavenging rate is in line with the sample concentration. The total polysaccharide of Laminaria japonica shows the strongest DPPH·nd·OH scavenging capacity for at the concentration of 5.0 mg/mL, the scavenging rate reached to 19.85% and 25.93%. The capacities of Laminaria japonica components’inhibition of the linoleic oxidation was determined by the thiobarbituric acid reactive substances(TBARs) method, the result shows that the fucoses have the strongest ability to inhibit the oxidation of linoleic acid, the inhibition rate reached to 53.1% and the effect time lasting for over 10 days.The result of the investigation of the hypolipidemic ability by combinding activities to sodium taurocholate in vitro shows that all the components in Laminaria japonica have hypolipidemic ability, except for alginate calcium and Fr1, Fr2. The total polysaccharide has the highest combinding rate of sodium taurocholate in vitro which is 26.89%, shows the best hypolipidemic ability.The result of the inhibiting activities to capillary permeability of mouse peritoneal shows that polysaccharides and unsaturated fatty acid show a certain pencent of anti-inflammatory activity, the inhibition rate is41.39% andl6.52% respectively, there is no anti-inflammatory activity detected in the polyphenols. All the polysaccharides, unsaturated fatty acid and polyphenols have weak inhibiting ability of the malondialdehyde (MDA) and the inhibiting rate is less than 10%.In summary, there is a certain and different percent of hypolipidemic, antioxidant and anti-inflammatory ability in the polysaccharide, polyphenols and unsaturated fatty acids of Laminaria japonica, the results explains the mechanism of Laminaria japonica in the role of multi-ways anti atherosclerosis from different views, provides scientific reference to the further research on the medical value of Laminaria japonica and the development of natural drugs for anti atherosclerosis.