Objective To Explore The Changes Of Cell Behavior In The Adenomyosis By Treated With The XiaoZheng Prescription

ObjectiveThis study was designed to explore whether the XiaoZheng Prescription could alleviate the migration, invasion and adhension abilities by treating the primary culture human adenomyosis eutopic endometrial stromal cells with the XiaoZheng Prescription medicated serum. Primarily, the detailed procedure of isolating primary culture cells from human adenomyosis eutopic endometrium tissues by collagenase I digestion, and the advantages of treating the primary culture cells with medicated serum are to discuss. Secondly, our research is to demonstrate that XiaoZheng Prescription could regulate the development and progression of adenomyosis through impairing the migration, invasion and adhension capacity of the cells. Finally, our study serves as an important experimental evidence and theoretical basis for XiaoZheng Prescription as an effective Traditonal Chinese Medicine therapy; and this study will put on a novel sight on the conservative treatment strategies for adenomyosis as well.Methods1. Endometrium tissues were excised from 8 patients who were under the panhysterectomy treatment in the department of gynaecology of the Second Affiliated Hospital of Guangzhou University of Chinese Medicine from January 2014 to May 2015. During the surgical excision, one part of the focus specimen was sent to pathology department and made a definite pathological diagnosis. The other part was preserved in the normal saline and transferred quickly to the clean bench. After collecting the endometrium tissues of adenomyosis by using an aseptic method, we used collagenase I to digest the tissues and then gathered the cells by a centrifuging method for culture. The cells were identified by immunocytochemical method.2.15 Specific-Pathogen-Free(SPF) female Spragne-Dawleg(SD) rats were chosed as the experimental animals for this study, which were randomly divided into 3 groups:XiaoZheng Prescription group, gestrinone group and the normal saline control group. Meanwhile, each group was intragastric administrated with different drug suspension in a dosage under conversion from human to experimental rats. After 1-week gavage, under 10%chloral hydrate anesthesia, the medicated serum of the rats was drawn from abdominal aorta by serum separation tube and subpackage for later application.3. The primary culture adenomyosis eutopic endometrial stromal cells were separately treated by 5%,10%, and 20%concentration medicated serum of XiaoZheng Prescription, gestrinone and the normal saline control serum for 24 hours. The alteration of the migration, invasion and adhesion capacity of the 3 groups primary culture cells were evaluated using Transwell migration assay, Matrigel invasion assay and Fibronectin adhesion assay.Results1. This study succeed to gain the human adenomyosis eutopic endometrial stromal cells from 7 cases of endometrium tissues of adenomyosis. And the primary culture cells were identified as immunopositive vimentin stromal cells.2. The effect of the XiaoZheng Prescription medicated serum on migration, invasion and adhesion of human adenomyosis eutopic endometrium stromal cells: Comparision of the primary cells treated with 5%,10%and 20%concentration normal saline control serum, has not seen remarkable differences (P>0.05) in the alteration of the migration, invasion and adhension capacity. Meanwhile, in comparision with the primary culture cells treated by 5%,10%or 20%normal saline control serum, both the gestrinone group and the XiaoZheng Prescription group counterpart could make an obvious effect (P＜0.05), on inhibiting the migration, invasion and adhesion capacity. When compared XiaoZheng Prescription group to the gestrinone group counterpart, the 20%concentration Xiao Zheng Prescription medicated serum has a significant (P＜0.05) inhibitive effect on the migration, invasion and adhesion function.Conelusion1. This study has clarified that the XiaoZheng Prescription could be able to make a significant effect on the treatment of adenomyosis by inhibiting the migration, invasion and adhesion capacity of human adenomyosis eutopic endometrium stromal cells.2. The inhibition of migration, invasion and adhesion of the human adenomyosis eutopic endometrial stromal cells, indicates that the XiaoZheng Prescription could suppress the genesis and development of the adenomyosis; and this study also laid a foundation of the research on the treatment of adenomyosis with a proper Traditional Chinese Medicine preparation, and put on a novel sight in researching the pathogenesis of adenomyosis as well.