The Establishment Of Starfish Quality Control And Effect Of Extracts On Osteoblasts

Starfish is a traditional Chinese medicine, first recorded in Compendium of Materia Medica. Research on the quality control of starfish varies, and limited in character identification. There is no microscopic identification, general test and determination of active ingredient content. Starfish has been used as Shang-ke Bone-knitting Tab.ts for the treatment of orthopedic. But the activity on the proliferation and differentiation of osteoblasts on starfish is less, also there is no effect of active components of starfish on proliferation and differentiation of osteoblasts. Therefore, this paper has carried on the following several aspects of the experiment:1) The standard of starfish quality was set up by the 2010 edition of Chinese Pharmacopoeia. The characteristics were observed by microscopic identification:showed that striped-muscle fibers with dense transverse striations alternated bright with dark, straight or sinuous; long ovoid in cross section view, having spot-or cleft-shape voids. Fragments of bone are irregular. Cell boundary of fragments of skin indistinct in surface containing brown pigment granules. Collagenous fibers twisted each other into masses. Through the gengral tests,determination of moisture, total ash, extract content inspection and starfish polysaccharides, saponins content, the esTab.lishment of quality control for water content not higher than 8.0%, total ash content is not more than 60%, the water soluble extract content of not less than 16% and 70% ethanol soluble extract content of not less than 10%. The content of polysaccharides and saponins in starfish was not less than 2.45% and 1.33%.2) In this study, acid and enzyme formulation was combined to separated polysaccharides from starfish, after removal of protein, dialysis and hydrogen peroxide bleaching, and then made as crude polysaccharides which appeared as a yellow powder. The total sugar content was determined to be 37.62% and protein content was 3.22%. The polysaccharides also contained sulfuric acid base which content was 14.70%. Molish and Seliwanoff reaction showed it contained aldehyde sugar residue, excluding ketone sugar.3) After repeated use of 70% ethanol extraction, petroleum ether degreasing, water-saturated n-butanol extraction, acetone precipitation total and macroporous resin chromatography and then made as crude asterosaponins from starfish. After foam reaction, Molish reaction, Salkowski reaction, Lieberman-Burchaard reaction, Resen-Heimer reaction and sulfuric acid group identification, indicated the asterosaponins was proved to be steroidal saponins which containing sulfuric acid base. Determine the optimal absorption wavelength by vanillin perchloric acid method is 547nm, and its content was 32.32%.4) The original generation of osteoblasts of 24 h old S.D rats were isolated and cultured in vitro with modified enzymatic digestion method. Different concentrations of polysaccharides and asterosaponins extract acting on the primary osteoblasts, with a series experiments such as cell proliferation assay, ALP activity determination, Alizarin red-S staining, quantitative detection of mineralized nodule formation. Experiments showed that starfish polysaccharides, asterosaponins extracts not only stimulated the growth and proliferation of osteoblasts, but also promoted the differentiation of osteoblasts. And starfish polysaccharides, asterosaponins extracts promoted the bone matrix mineralization by stimulating the deposition of calcium.Above all, this thesis focuses on the improvement of starfish quality control, provides the basis and methods for controlling the quality of starfish. The extraction of polysaccharides and saponins extract from starfish has effect on proliferation and differentiation of osteoblasts, also provides a scientific basis for the exploitation and utilization of starfish.