敲低KIF3B基因通过抑制Shh信号通路对小鼠胚胎腭突间充质细胞自噬的促进作用

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[中图分类号] R782.2 [文献标志码] A

ABSTRACT Objective：To discuss the effct of knock down of gene of kinesin family member 3B （KIF3B），an important component of primary cilia（PC） of in mouse embryonic palatal mesenchymal on the autophagy level of cels （mEPMCs）cels，and to clarify its mechanism. Methods：The mEPMCs from gestational day 14.5 C57BL/6J mice cultured in vitro were collected and divided into control group （administered normal saline），empty lentivirus transfected cel group （sh-NC group）（administered lentivirus transfection），KIF3B knockdown group （sh-KIF3B group）（administered KIF3B gene knockdown），and KIF3B knockdown plus Smoothened receptor agonist（SAG） group （sh-KIF3B + SAG group）（administered KIF3B gene knockdown followed by SAG addition），based on whether the KIF3B gene was knocked down and whether the SAG was used to activate the sonic hedgehog（Shh） signaling pathway and its downstream coreceptor Smo，with 5 rats in each group.Transmisson electron microscope wasused to observe the morphology and the number of autophagosomes/autolysosomes in the mEPMCs in various groups;Western bloting method was used to detect the expression levels of autophagy-related proteins Beclin-l and p62， and the Shh signaling pathway proteins Shh and Smo in the mEPMCs in various groups. Results： The transmission electron microscope observation results showed that compared with control group，the number of autophagosomes/autolysosomes in sh-KIF3B group was significantly increased （ P<0.05） ；compared with sh-KIF3B group，the number of autophagosomes/autolysosomes in the mEPMCs in sh-KIF3B+SAG group was significantly decreased （ ⋅P<0.05 ）．The Western blotting results showed that compared with control group，the Beclin-1 protein expression level in the mEPMCs in sh-KIF3B group was significantly increased （ .P<0.05 ），and the KIF3B，p62，Shh，and Smo protein expression levels were significantly decreased ΔP<0.01 ）；compared with sh-KIF3B group， the Shh ，Smo，and p62 protein expression levels in the mEPMCs in sh-KIF3B + SAG group were significantly increased （ P<0.01 ），and the Beclin-1 protein expression level was significantly decreased （ P<0.01 ）. Conclusion：Knockdown of KIF3B gene can promote autophagy of the mEPMCs，and the mechanism may be related to its inhibition of the Shh signaling pathway.

KEYWORDsCleft lip and palate；Kinesin family member 3B； Primary cilia；Sonic hedgehog signaling pathway; Autophagy

唇腭裂（cleftlipandpalate，CL/P）是最常见的颌面部畸形之一，CL/P在全世界的发病率约为1/700，而在中国的发病率可达 1.82/1 000[1.2] 。（剩余10375字）