Therapeutic Effect Of Photolon Mediated Photodynamic Therapy For Endometriosis In Rat Model

OBJECTIVE To investigate the effect of photodynamic therapy(PDT)mediated by photolon on the treatment of Balb/c endometriosis mouse model and the killing effect on ectopic endometrial cells of Balb/c female mouse.METHODS Measure the absorbance of photosensitizer.Balb/c female mice with an average weight of 20g in 6-7 weeks were treated with autologous endometrial transplantation under hydrated chloral hydrant anesthesia to construct endometriotic model.Hormone was given after surgery to promote the growth of endometriosis model,When the average size of the lesion is 6 mm,it means that the model is established successfully.Eutopic and ectopic endometrial cells of Balb/c female mice were primary cultured,after the success of the passaging,we observed the differences in eutopic and ectopic endometrial cell morphology of mice.Ectopic endometrial cells that were primary cultured were divided into control group,simple light group,photosensitizer group and Photolon-mediated PDT group.Morphological changes after treatment were observed under microscope.MTT assay was used to analyze the inhibition rate and half lethal dose of Photolon mediated PDT treatment on ectopic endometrial cells in mice.Fourteen mice were randomly divided into two groups:control group and Photolon-mediated PDT treatment group.(The control group:mice injected with 200ul saline,after 3 hours of exposure to natural light 10min;PDT treatment group:mice injected with Photolon solution of 200ul(2mg/ml),after 3 hours of exposure to PDT treatment,the irradiation dose was 2.5J/cm2(660nm,1W,distance from the lesion surface to 7min).At the end of treatment,the size of the ectopic lesion was measured and recorded every 7 days.Each group of mice were sacrificed 5 days after treatment.The pathological sections of mouse endometriosis were prepared,HE staining was used to observe the pathological changes of ectopic endometrium in each group.RESULTS 1.The leydig cell of the ectopic endometrium cell cultured in vitro is smaller than the eutopic leydig cell,and grows dispersedly.With clear edge and less patina,the cell is in lathy fusiform and stretch and straight form.Its nucleus,which is relatively larger,is in oval and grows fast.Its growth model is similar to that of cancer cell.2.After employing PDT treatment of Photolon mediation to treat the ectopic endometrium cell cultured in vitro,the cells are massive dead.The original lathy fusiform of the cell is damaged.And its.edge is blurry,its patina is broken,and its cell nucleus is pyknosis,even is deformed,crazed and disappeared.3.The inhibition ratios to ectopic endometrium cell of rat of different concentrations Photolon(0.015ug/ml,0.03ug/ml,0.06ug/ml,0.125ug/ml,0.25ug/ml,and 0.5ug/ml)are all higher than the control group.And the half lethal amount of cell is reached when the concentration is 0.160ug/ml.4.The control group rat's ectopic endometrium focus increases consistently,while the focus of Photolon+PDT group stops increase and gradually decreases.5.HE staining results are shown:Compared with the control group,the successive arrangement of cells in the Photolon+PDT group was disrupted.The cell shape was distroyed and the proportion of cytoplasm decreased.Some nuclei were pyknosis,and these could be seen with vesicular nuclei and perinuclear halo.CONCLUSION Photolon-mediated PDT therapy can effectively inhibit the growth of mouse endometriosis.