方法：对贴壁生长的人角膜缘来源的基质干细胞(LNC)进行分离培养及传代，对骨髓间充质干细胞系(BMMSC)进行培养传代。通过将BMMSC作为阳性对照，采用Western blot技术，免疫荧光技术，real-time PCR技术从蛋白水平及基因水平验证Hedgehog信号通路成员SHH，patched，SMO，Gli-1在LNC中的表达情况。最后采用Cell Count Kit-8 检测不同浓度Gli抑制剂GANT61(空白，1，5，10，15，20，25，30μmol/L)处理LNC和BMMSC48h后的增殖抑制效应反向验证Hedgehog信号通路在LNC中的存在。

结果：Western blot技术，免疫荧光技术，real-time PCR技术均证明LNC细胞表达Hedgehog信号通路中的Gli-1，patched， SMO，且Gli蛋白抑制剂GANT61可以显著抑制LNC细胞增殖(P<0.05)。

结论：Hedgehog信号通路在LNC细胞的增殖过程中发挥重要作用。

METHODS: Culture and passage the mesenchymal stem cells derived from LNCs and bone marrow-derived mesenchymal stem cells(BMMSCs)in vitro. Using BMMSCs as a positive control, the expression of Hedgehog signaling pathway in LNCs was evaluated by Western blot, immunofluorescence and real-time PCR. Furthermore, different doses(1, 5, 10, 15, 20, 25, 30μmol/L)of GANT61(the Gli inhibitor)effect on the proliferation of LNCs and BMMSCs was detected by Cell Count Kit-8.

RESULTS:The results of Western blot, immunofluorescence and real-time PCR proved that LNCs expressed Gli-1, patched and SMO, which are the members of Hedgehog signaling. In addition, GANT61(the Gli inhibitor)inhibited the proliferation of LNCs significantly(P<0.05).

CONCLUSION:Hedgehog signaling pathway plays an important role in the proliferation of LNCs.