Masson Pine Pollen Polysaccharide Extraction, Esterification And Intracellular Calcium Ion Flow In The Primary

The old researches of polysaccharide and its sulfate mostly focused on its functions of antivirus, antitumor, antioxidation, anticoagulant and improve immunity, but there are few reports about its functions on heart and such myocytes. Here, we extracted polysaccharide from Pinus massoniana pollen and then sulfated; observed the effect of its 60% component on cardiomyocytes and primary discussed the possible mechanism.1. Polysaccharide was extracted from Piuns massoniana pollen with hot water and precipitated by 40%, 60% alcohol. The efficiency of extraction is separately 0.328% and 3.071%. We chose 60% alcohol precipitated polysaccharide (PPM60) to become sulfated (SPPM60) by chlorosulfonic axid-pyridine method, the efficiency is 147%.2. There is a typical absorption of polysaccharide and no nucleic acid and protein absorption through UV spectrum scan; Polysaccharide content of PPM60 is 88.69% and the degree of its sulfate is 1.762; Infrared ray spectrum indicates there are typical characteristic absorption of PPM60 and characteristic absorptions of sulfate ester bond of SPPM60.3. By means of perfusing apparatus of isolated toad heart, we observed the effects of PPM60 and its sulfate SPPM60 on the contractility of toad cardiac muscle. By adding drugs like propranolol (Pro), CdCl2 and verapamil (Ver) to block the specific receptor on cardiomyocytes; compared its effect with other types of polysaccharide and cardiac glycoside like digoxin, we discussed the probable effecting mechanism of SPPM60 on cardiomyocytes. The results showed that PPM60 could inhibit myocardial contractility; SPPM60 could significantly strengthen the myocardial contractility. Both of them had no effect on heart rate. SPPM60 could reverse the inhibition activity of Pro and CdCl2. But in the presence of Ver, the positive inotropic effect of SPPM60 was inhibited completely. It proved that the positive inotropic effect of SPPM60 may work through L-type Ca2+ channel, and be independent of theβ1-receptor. Besides, the positive inotropic effect of SPPM60 is its peculiar function and can be enhanced by its degree of sulfate.4. We observed the effects of PPM60 and SPPM60 on the contractile ability of toad skeletal muscle. The results showed that PPM60 could strengthen its contractility; however, SPPM60 had no effect on skeletal muscle. So, PPM60 and SPPM60 had different effects on different musculature, the probable mechanism relates to the distribution of different types of L-Ca2+ channel.5. We used the peroxides and superoxides sensitnve fluorescence probe DCFH-DA and confocal microscopes to observe the effects of PPM60, SPPM60 on the production of oxygen free radicals (OFR) in cardiomyocytes. The results showed that PPM60 could slightly increase OFR, while SPPM60 could reduce the concentration of OFR. It indicated that SPPM60 didn't cause Ca2+ overloading when increasing the concentration of Ca2+.