The Effect Of Sulfated Polysaccharide From Pine Pollen On The Calcium Concentration And Proliferation In Rat Arterial Smooth Muscle Cells

In order to enhance the biological activity of polysaccharides, structural modified polysaccharides appeared great significance. There are a lot of modification methods, such as sulfation, carboxymethylation, selenylation, and so on. Sulfated derivation of polysaccharide, which is rich in SO₄^2-, is easier to bind the specific domain of protein. Sulfated modification can change polysaccharides orignal activity.Polysaccharide was extracted and precipitated by 60% alcohol from Pinus massoniana pollen, naming PPM60. Protein was removed by Sevage method. Four components PPM60-A,PPM60-B,PPM60-C,PPM60-D were obtained separately by molecular sieve Sephacryl S-400HR. These four components were chemically modified by chlorosulfonic acid- pyridine method, respectively naming SPPM60-A,SPPM60-B,SPPM60-C,SPPM60-D, and their substituted degree were1.28,1.63,1.31,1.49.The original generation smooth muscle cells (VSMCs) were obtained by tissue-piece inoculation and Enzyme digestion with Collagenase.We use 0.25% trypsin digestion to obtain passage cells. The cells were long spindle shape, the membranes were integrated, and their fringes were smooth, which were smooth muscle cells identified with immunohistochemical staining and living cells detected by trypan blue staining were more than 90%. A lot of smooth muscle cells can be obtained by these two methods for further research.In this experiment, these four components can reduce the calcium concentration of smooth muscle cells, and the component of PPM60-A and SPPM60-A have signifcant difference effects on the VSMCs.High K⁺ and NE can increase the calcium concentration of VSMCs. PPM60-A and SPPM60-A affect VSMCs 5mins can reduce the role of these agonists.We speculate PPM60-A and SPPM60-A can reduce the calcium concentration of VSMCs by intention on the voltage-dependent calcium channel and receptor-operated calcium channel. PPM60-A and SPPM60-A also can reduce the high calcium concentration of VSMCs increased by K⁺ beforehand.Norepinephrin (NE) can promote the proliferation of VSMCs.We use NE to set up a proliferation model to test the effects of PPM60-A and SPPM60-A on the VSMCs.We found that these two components can inhibite the promotion of NE in 24h or 48h, and they performance a bound of dose-dependent.