| Rheumatoid Arthritis(RA) is a painful and often crippling disease that initially results in swollen and imflammed joints, but often progresses to deformed or completely destroyed joints. Recent genetic research strongly suggests that this is a result of the body mistakenly attacking type II collagen(CII), which is the major portion of cartilage tissue. In theory, oral administration of highly purified type II collagen (CII) represents a cure treatment for Rheumatoid Arthritis. Chick sternal xiphoid cartilage primarily comprises abundant type II collagen and proteoglycan and can be applied to produce an edible supplement to prevent or alleviate the symptoms of RA. The present research studied the preparation of cartilage powder which contains abundant type II collagen from chick sternal cartilage.In addition, the composition and biological activities of the product were determined. Major content of this paper as following:Firstly, The fresh cartilage was digested by pepsin to eliminate chicken and cartilage membrane from cartilage and the optimum enzymatic conditions were as follows. The enzymatic concentration was 400 mg·L-1, the temperature was 28℃, the weight proportion was 1:5(material: liquid), the reaction time was 24 h. We applied chloroform: methanol=2:1 to remove grease from cartilage and the resulting lipid content was 0.05%. Freeze-drying could eliminate 97%water from cartilage. The cartilage was ground with the presentation of liquid nitrogen and the average granule diameter was 73.37μm.Then, Hydrochloric carbamidine was used to remove non-collagen components, then pepsin was added to extract type II collagen. The results of SDS-PAGE electrophoresis and RP-HPLC showed the prepared Type II collagen is a high purity product. The structure and composition of type II collagen were studied. Type II collagen had a high content of glycine, hydroxyproline and proline residues, with 313, 118 and 94 residues per 1000 amino acids residues respectively, and small amounts of tyrosine, cysteine, histidine and methionine residues with 5, 17 and 2 residues per 1000 amino acids residues respectively. The denaturing temperature was determined by differential scanning calorimeter to be about 44°C for purified type II collagen. The minimal difference between denaturation temperature of purified type II collagen from cartilage powder and intact cartilage indicated that purification process has little influence for stability of collagen. Analysis results of UV absorbance spectrogram,infrared spectrum and CD spectrogram proved that type II collagen of cartilage powder remained natural structure. All the results indicated that the type II collagen in chick cartilage powder retained most intermolecular crosslinks during the preparation process.Lastly, the influence on rat rheumatoid arthritis (RA) by oral administration of chick sternal cartilage and Glucosamine Sulfate was studied. The collagen-induced rat rheumatoid arthritis model (CIA) was established in male Sprague-Dawley (SD) rats with intradermal injection of type II collagen in Freund's complete adjuvant. CIA rats were treated with oral administration of chick sternal cartilage and Glucosamine Sulfate. The development of RA was observed in CIA rats and some relevant characteristics were determined. The results indicated that oral administration of chick sternal cartilage obviously relieved the arthritis symptoms and markly suppressed the lymphocyte proliferation. In addition, the levels of TNF-α, IL-1 and IL-2 in the experimental rats were markedly lowered versus vehicle-treated rats. Hence, our studies demonstrated that chick sternal cartilage and Glucosamine Sulfate could be applied to prevent and cure the RA as a functional healthy food which are safe and lack of side-effects. |
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