Proliferation and migration molecular mechanisms induce by earthworms (Pheretima aspergilum) in Schwann cells.

• Main Authors: Yung-Ming Chang, 張永明
• Other Authors: Tung-Yuan Lai
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/46326954062690804683

博士 === 中國醫藥大學 === 中國醫學研究所博士班 === 98 === The earthworm, which has stasis removal and wound healing functions, is a widely used Chinese herbal medicine in China. Schwann cell migration and proliferation are critical for the regeneration of injured nerves. Schwann cells provide an essentially supportive activity for neuron regeneration. However, the molecular migration and proliferation mechanisms induced by earthworms in Schwann cells remain unclear. Here, we investigate the molecular mechanisms, which includes : (1) migration signaling, MAPKs (Mitogen-activated protein kinases) mediated PAs and MMP2/9 pathway ; (2) survival and proliferative signaling, IGF-I (Insulin-like growth factor-I) mediated PI3K/Akt pathways and cell cycle regulation. Using MTT and migration assay, we confirmed that treatment with 125 μg/ml earthworm for 24 h appears to stimulate cell proliferation and migration. In addition , examined by western blotting, found that earthworm extract induced phosphorylation of ERK1/2 and p38, but not JNK, activate the downstream signaling expression of PAs (Plasminogen activators) and MMPs (Matrix metalloproteinases) in a time-dependent manner. Earthworm stimulated ERK1/2 and p38 phosphorylation was attenuated by pretreatment with chemical inhibitors (U0126 and SB203580), and small interfering ERK1/2 and p38 RNA, resulting in migration and uPA related signal pathway inhibition。 On the other hand, earthworm extract treatment also induces the phosphorylation of IGF-I mediated PI3K/Akt pathway, activate protein expression of PCNA (proliferating cell nuclear antigen) and cell cycle regulatory proteins (cyclin D1, cyclin E and cyclin A) in a time dependent manner. And accelerated G1 phase progression with earlier S phase entry and that significant numbers of cells entered the S phase.siRNA mediated knockdown of PI3K, that significantly reduces PI3K protein expression levels, resulting in Bcl2 survival factor reduction, revealing a marked blockage of G1 to S transition in proliferating cells. Taken together, our data reveals the unknown RSC96 cell migration and proliferation mechanism induced by earthworm extract, which find use as a new medicine for nerve regeneration.