桂枝挥发油对子宫腺肌病异位内膜细胞侵袭、迁移的研究

Effects of CINNAMOMI RAMULUS Volatile Oil on the Invasion and Migration of Adenomyosis Derived Cells

目的:探究桂枝挥发油对子宫腺肌病异位内膜细胞(AMDC)侵袭、迁移能力的影响并探讨其作用机制。方法:免疫荧光法鉴定提取细胞,检测波形纤维蛋白(Vimentin)、细胞角质蛋白7(CK7)、血管性血友病因子(Von Willebrand Factor)、上皮型钙粘蛋白(E-cadherin)、α-平滑肌肌动蛋白(Alpha smooth muscle)荧光表达能力。采用CCK-8法检测桂枝挥发油处理后细胞的活力与毒性,Transwell法检测细胞侵袭能力、损伤划痕法检测细胞迁移能力以及蛋白免疫印迹法(Western blotting)分析蛋白RhoA、RhoB、RhoC、ROCK1、ROCK2表达情况。结果:免疫荧光法鉴定细胞Vimentin强阳性,CK7、Von Willebrand Factor、E-cadherin、Alpha smooth muscle弱阳性;与模型对照组相比,桂枝挥发油1 mg/mL组细胞侵袭和迁移能力明显受到抑制,RhoA、RhoB、RhoC、ROCK1、ROCK2蛋白表达明显下调(P<0.05)。结论:所获细胞为AMDC,桂枝挥发油可抑制AMDC的活性,限制侵袭迁移能力,其机制可能与Ras同源基因/Rho相关螺旋卷曲蛋白激酶(Rho/ROCK)信号通路有关,参与子宫腺肌病的病理生理过程。

Objective:To explore the effects of CINNAMOMI RAMULUS volatile oil on the invasion and migration of adenomyosis-derived cells(AMDCs)and its mechanism.Methods:Immunofluorescence was used to identify and extract cells,detecting fluorescence expressions of vimentin,cytokeratin-7(CK7),von Willebrand Factor(vWF),E-cadherin,andα-smooth muscle actin(α-SMA).Cell activity and toxicity of AMDCs after treatment withCINNAMOMI RAMULUS volatile oil were detected by cell counting kit-8(CCK-8)assay.Transwell assay was used to detect the cell invasion ability,cell wound scratch assay was used to detect migration ability,and Western blotting was used to analyze the protein expressions of RhoA,RhoB,RhoC,Rho-associated kinase 1(ROCK1),and ROCK2.Results:Immunofluorescence results showed that vimentin cells were strongly positive,and CK7,vWF,E-cadherin,andα-SMA were weakly positive.As compared with the model control group,the cell invasion and migration ability was inhibited in the 1 mg/mLCINNAMOMI RAMULUS volatile oil group,and the protein expressions of RhoA,RhoB,RhoC,ROCK1,and ROCK2 were significantly down-regulated(P<0.05).Conclusion:The acquired cells are AMDCs.CINNAMOMI RAMULUS volatile oil can inhibit the activity of AMDCs and limit their invasion and migration ability,and the mechanism may be related to the Rho/ROCK signaling pathway,which is involved in the pathophysiology of adenomyosis.