Study On Rhodiola Sachalinensis Saccharide And Herba Leonuri Polysaccharide For Cryopreservation Of Bull And Boar Spermatozoa

Finding the optimal cryoprotectants was still one of the main and important problems atpresent. There were many reports on the pharmacological researches of Herba Leonuripolysaccharide, apple polyphenol, and grape seed procyanidins, but so far no studies on theireffects on semen cryoprotection had been reported. This research used semen samples frombulls and boars as the investigation target, to explore the effects of Rhodiola sachalinensissaccharide (RSS) and Laminaria japonica polysaccharide (LJP) on bull cryo-sperm, and theeffects of Herba leonuri polysaccharide, apple polyphenol and grape seed procyanidin onboar cryo-sperm. The sperm viability was evaluated by the Computer Assisted SpermAnalysis System (CASA). Propidium iodide (PI) and rhodamin123(Rh123) staining,hypoosmotic swelling test (HOST) and fluorescein isothiocyanate-peanut agglutinin(FITC-PNA) staining were used to test sperm mitochondrial activity, plasma membraneintegrity and acrosomal integrity. In addition, the antioxidase activities (SOD, LDH et al.)were evalueated by assay kits. The results of this research were shown below.(1) The sperm viability, curvilinear line velocity (VCL), mitochondrial activity, plasmamembrane integrity and acrosomal integrity of bull cryo-spermatozoa were significantlyimproved with the supplement of RSS and LJP, compared to that of the control (P <0.05).The optimal concentrations of RSS and LJP added into basic extenders was0.06mg/mL and1.00mg/mL.(2) Herba leonuri polysaccharide, apple polyphenol, grape seed procyanidin added to theextenders protected the sperm well during cryopreservation. The sperm viability, curvilinearline velocity (VCL), mitochondrial activity, plasma membrane integrity and acrosomalintegrity of boar cryo-spermatozoa were significantly improved when adding0.12mg/mL ofHerba Leonuri polysaccharide,0.06mg/mL of apple polyphenol and0.04mg/mL of grapeseed procyanidin to the freezing extender, compared with the control group (P <0.05).(3) The activities of the super oxide dismutase (SOD), lactate dehydrogenase (LDH),aspartate transaminase (AST) in bull cryo-sperm, and the super oxide dismutase (SOD),catalase (CAT), total antioxidant capacity (T-AOC) in boar cryo-sperm were lower than thatof the corresponding fresh sperm. The activities of SOD, LDH and AST in the bull sperm wasraised significantly after adding RSS and LJP to the freezing extender (P <0.05). The optimaladded concentrations of RSS and LJP were0.06and1.00mg/mL, respectively. (4)The activities of SOD, CAT and T-AOC in the boar sperm were fallen after thecryopreservative process. Compared with the control group, the enzyme activities of boarcryo-sperm were increased significantly with Herba Leonuri polysaccharide, applepolyphenol, grape seed procyanidin added in the freezing extender (P <0.05). The optimalconcentrations of Herba Leonuri polysaccharide, apple polyphenol and grape seedprocyanidin were0.12,0.06and0.04mg/mL, respectively.