Study On Velvet Antler Ossification And The Relationship Between The Calcification And Its Related Factors In Tarim Red Deer (Cervus Elaphus)

Velvet antler is a rare traditional Chinese medicine,has a warm kidney yang,raw lean blood,strong reinforcement of marrow effect.It is a special tissue of the deer family with a strong ability to regenerate in the case of natural growth each year to undergo a process of occurrence,growth,ossification,shedding and so on.Is the unit weight of Tarim red deer antler yield the highest deer subspecies in antler antler growth period of antler growth rapidly,in essence,is the process of bone formation in the cartilaginous skeleton,in the process of antler ossification,especially a large number of mineral elements calcium deposition in antler,deposition of calcium the needs of a variety of factors of ion regulation factors,relatively close relationship with the calcium sensing receptor?CaSR?,parathyroid hormone?PTH?,androgen?ADG?,calcitonin?CT?,bone isoenzyme alkaline phosphatase?BALP?,bone sialoprotein?BSP?and bone gla protein?BGP?and other hormones.In order to clear the relationship between deposition,calcium ions in the antler and antler ossification and regulation factors,contents and changes of Antlerogenic period determination of calcium,calcium,antler bone calcium,blood ADG,PTH,CT,BALP,BGP and BSP hormone from the test and training of Tarim red deer antler.Mesenchymal cells,chondroplast cells,cartilage cells and periosteal cells in vitro,using laser scanning confocal microscopy observation of extracellular calcium ions to flow and determination of intracellular calcium levels by Real-time PCR,Western Blot,CaSR,BSP,Runx2 analysis,GAPDH gene expression,analysis of pilose antler angle of ossification.Through experimental study,the following results were obtained:1.The Tarim red deer antler and antler growth,calcium,calcium,bone calcium changes.The growth rate of the youth group in deer antlers of the tower began on the eighty-fourth day growth reached the maximum value in the antler blood Ca²⁺ content,beginning seventieth days of growth decreased,then increased in antler,the first eighty-fourth days of growth reached the highest value in antler.The middle-aged and the elderly group of red deer began to grow on the seventieth day of the antler growthrate reached the maximum value in antler,blood Ca²⁺ content began in fifty-seventh days of antler growth reached the highest value,appear in the antler growth began seventieth days after the content of Ca²⁺ and reduce the trend,rising slowly.The Tarim red deer,antler growth period of about 100 days,from the beginning to the growth of antler ossification period,Tarim red deer antler groups are calcium increasing,the main body of Tarim red deer were decalcified bone,including rib decalcification is most serious,followed by pelvic bone,and bone and decalcification of the lightest degree of scapular.2.The relationship between the hormones in the blood,and antler ossification.In the different growth stages of antler,the content of blood serum ADG,PTH,CT,BALP,BGP and BSP of the Tarim red deer exist outside of the unceasing change,especially the stage of ossification of the most obvious changes.The most obvious changes of serum ADG level is in antler,before this has been at a low level,then increased rapidly,reaching significant difference?P<0.01?.The serum PTH in antler growth phase change is not obvious,until the ossification period in the low level,but in ossification late and serum levels of PTH were significantly higher?P<0.01?red deer.Serum levels of BALP were in the red deer antler to the early rapid growth period showed a downward trend,the rapid growth period to ossification late and serum levels of BALP were increased in red deer antler,after the harvest,decreased significantly.The serum BGP and BSP were with antler growth in elevated into the ossification period began to decline after the antler harvest has been lowered,and the antler growth rate positively.The concentration of serum CT and antler growth rate is the same,but lagged behind changes in the antler growth rate,in the early stage,velvet antler rapid growth period were increased slowly,to reach the highest stage of ossification,and then began to decline.3.The connection of cultivation antler cells and calcium regulation associated genes.We collected the tip of antler in the growing and cultivated the antler cells,experimental results showed that:ACC,MSC was easy to cultivate,and CC was slower growth.It was detected that the [Ca²⁺]i in the MSC was higher than in the ACC,CC by laser scanning confocal method.It was longer that the time required of the CC Ca²⁺inflow from extracellular to intracellular than the MSC,ACC When added calcium.With the advent of ossification stage,the antler cartilage cells number was increasing,the amount of calcium needed was more and more.The levels was significantly higher in ACC which genes expression of BSP,CaSR,Runx2 mRNA than which in MSC by real-time PCR amplification,and the levels was higher in CC which genes expression only of Runx2 mRNA than which in MSC.We used the Western Blot method to detected and get the same result.These experiments illustrate that the BSP,CaSR,and Runx2 genes was closely related with the rapid growth of antler,and this genes play an important role in antler ossification beginning.