α-细辛醚对秦岭滑蜥断尾后芽基形成时期细胞增殖的影响

Effect of α-Asarone on Cell Proliferation During the Stage of Blastema Formation in Tail Regeneration of Scincella tsinlingensis

蜥蜴再生芽基是研究羊膜类附肢替换的良好模型。以秦岭滑蜥为对象,运用挤压法建立断尾再生模型,设原尾空白组、再生尾对照组和注射α-细辛醚的再生尾实验组进行研究,在其分别生长15 d、18 d和20 d后取芽基并进行冰冻切片免疫组织化学染色,检测PCNA性细胞平均灰度值,积累α-细辛醚影响断尾后芽基形成时期细胞增殖的基础资料。结果显示:PCNA在不同实验分组和不同阶段尾组织中均有表达,在空白组18 d的脊髓中表达最弱,灰度值为188.09±1.98,而实验组20 d的脊髓中表达最强,灰度值为123.10±2.91。实验组的再生尾皮肤组织、肌肉组织和脊髓中PCNA的表达强度均高于空白组和对照组,且对照组与实验组的脊髓中的表达强度具有显著差异(P<0.05)。上述结果表明α-细辛醚能够促进秦岭滑蜥断尾后芽基形成时期的细胞增殖,促进再生脊髓细胞增殖的效应最为显著。

The regeneration blastema of lizards is an amniote model for the study of appendage replacement.To provide basic insight for further research on reptile tail regeneration,we studied the effect ofα-asarone on cell proliferation during the process of regeneration of Scincella tsinlingensis.We created a tail regeneration model by gently pinching to initiate autotomy.The lizards were assigned to three groups,viz.blank group(lizards with original tail),control group(lizards with regenerated tail),and experimental group(lizards with regeneration tail and injection ofα-asarone).We collected the regeneration blastema of three groups at 15 d,18 d,and 20 d after the start of regeneration,and used these cells to prepare immunohistochemistry stained frozen slices.We tested the average gray-scale value of proliferating cell nuclear antigen(PCNA)positive cells.PCNA positive cells were distributed in all experimental groups and all in all developmental stages of tail tissue.The weakest expression in the spinal cord at 18 d was found in the blank group,for which the gray-scale value was 188.09±1.98.The strongest expression in the spinal cord at 20 d was found in the experimental group,for which the gray scale value was 123.10±2.91.The expression intensity of PCNA in regenerative tail skin,muscle and spinal cord was higher in the experimental group than that in either blank group or control group.The intensity of expression in the spinal cord differed between controls and the experimental group(P<0.05).These results suggest thatα-asarone can promote cell proliferation during the stage of blastema formation in S.tsinlingensis tail regeneration,and the most significant effect of promoting the proliferation can be detected in regenerated spinal cord cells.