Study On The Expression Of Fractalkine,CX3CR1 And Their Correlation In Endometriosis

Objective: To study the expression of Fractalkine and it's receptor CX3CR1 in eutopic endometrium,ectopic endometrium in endometruosis and the expression in peritoneal fluid and blood serum ,and a thinPrep cytological test and immunocychemistry detection on the peritoneal fluid cells to get a better understanding of the roles FKN and CX3CR1 play in the occurrence and development of endometriosis and to provide a theoretical guideline for the clinical diagnosis and treatment.Methods:1 The study subject:The study group:Patients with ovarian endometriosis who were operated in the gynaecology of the Second Hospital of HeBei Medical University, selected from June 2007 to October 2008(congfirmed by the pathology after operated),and divided into four groups: one group was endometrium group, this group was further divided into two small groups: eutopic endometrium group and ectopic endometrium group ,each of which included 30 cases (16 patients in the proliferative stage and 14 patients in the secretory stage ). The median age of patients in this group was 35 years old (22 to 43 years old). The second and the third group were peritoneal fluid group and blood serum group, each of which included 24 cases (12 patients in the proliferative stage and 12 patients in the secretory stage), and the median age of the patients in this group was 27.5 years old (21 to 35 years old); the fourth group was peritoneal fluid cells group including 30 cases (15 patients in the proliferative stage and 15 patients in the secretory stage), and the median age of the patients in this group was 25.5 years old ( 21 to 34 years old).The control group: patients with infertility who were operated with peritoneoscopy in the same hospital and the same periods were selected and divided into four groups: one group was normal endometrium including 20 cases which were confirmed through pathology (11 patients in the proliferative stage and 9 patients in the secretory stage), and the median age of the patients in this group was 29.5 years old(24 to 36 years old); the second and the third group were peritoneal fluid group and blood serum group, each of which included 20 cases (10 patients in the proliferative stage and 10 patients in the secretory stage), and the median age of the patients in this group was 30.5 years old (21 to 37 years old); the fourth group was peritoneal fluid cells group, including 20 cases (10 patients in the proliferative stage and 10 patients in the secretory stage), and the median age of the patients in this group was 28 years old ( 21 to 37 years old). We distinguished the menstrual cycle on the basis of last menstrual period and the pathology of the endometrium . All of the women in the two groups had normal menstrual cycles and no case history of irregularial vaginal bleeding in recent six months ; no case history of taking oral esteroid hormone and fixing IUD in recent six months ; no case history of hypertension,heart disease , diabete , renopathy ,urgent and chronic pelvic inflammatory disease.2 Sample collecting :2.1 Endometrium group: The endometrium was collected from the study and control group on their laparoscopic operatings, bathed by PS again and again and then fixed by PFA.2.2 Peritoneal fluid group : The peritoneal fluid was taken suction from recto-uterine fossa on their laparoscopic operat- ings , then centrifuged 10 mintues in common temperature 3000r/min and lastly put into freezer in -70℃to be measured .2.3 Blood serum group : The blood samples were taken from the patients'ulnar vein when their stomach were empty , and then centrifuged 10 minutes in common temperature 3000r/min and lastly put into the freezer in -70℃to be measured .2.4 Peritoneal fluid cells group:The cells from precipitation of peritoneal fluid was processed by ThinPrep cytological test .3 The experiment methods : To determine the expression of FKN and it's receptor CX3CR1 in the eutopic endometrium,ectopic endometrium in endometriosis and normal endometrium by immuno-histechemical staining ; To determine the expression of FKN and it's receptor CX3CR1 in the PF and blood serum of the study group and control group by ELISA . To determine the expression of FKN and it's receptor CX3CR1 in peritoneal fluid cells by ThinPrep cytological test and immunocychemistry . To analyze the results by the means of the SPSS13.0.Result:1 FKN was lacalized on cytoplasm and plasmalemma of glandular epithelial cells,interstitial cells,blood vessel endothelium cells in the ectopic(Fig.1) and eutopic (Fig.2)endometrium .There were so many positive particles on the surface of glandular cavity ,but little expressing on normal (Fig.3)endometrium . CX3CR1 was lacalized on cytoplasm and plasmalemma of interstitial cells in the ectopic(Fig.4) and eutopic(Fig.5) endometrium , but little expressing on normal (Fig.6)endometrium .2 The expression of FKN,CX3CR1 in ectopic,eutopic and normal endometrium(Tab.1 Tab.2)The positive ratio of FKN on ectopic,eutopic and normal endometrium were 93.33%,70.00%,15.00% .The expression level of FKN in ectopic endometrium and eutopic endometrium of endometriosis were obviously higher than that in normal endometrium (X2=31.99,P<0.01; X2=15.15,P<0.01); The expression level of FKN in ectopic endometrium were obviously higher than that in eutopic endometrium (X2=16.49,P<0.01);FKN was found no differences between different menstrual cycle(P>0.05). The positive ratio of CX3CR1 on ectopic,eutopic and normal endometrium were 86.67%,63.33%,25.00% .The expression level of CX3CR1 in ectopic endometrium and eutopic endometrium of endometriosis were marked higher than that in normal endometrium (X2=23.37,P<0.01; X~2=8.26,P<0.05); The expression level of CX3CR1 in ectopic endometrium was obviously higher than that in eutopic endometrium (X~2=21.97,P<0.01);CX3CR1 was found no differences between different menstrual cycle(P>0.05).There was a positive relationship between the expression of FKN and CX3CR1 in ectopic endometrium (r=0.65, P<0.01); and in eutopic endometrium (r=0.63, P<0.05); and in normal endometrium (r=0.62, P<0.05) .3 The expression of FKN,CX3CR1 in peritoneal fluid and blood serum.The level of FKN in the PF in the study group was higher than that in the control group (2438.50±662.70 pg/ml,1396.03±677.09pg/ml) and both had significant difference (P<0.05),The difference was not significance(P>0.05)when comparing the blood serum (2671.56±1156.63pg/ml,3411.06±864.02pg/ ml)of these two groups . The difference was not significant(P> 0.05)when comparing the blood serum of different menstrual cycle .The level of CX3CR1 in the PF in the study group was higher than that in the control group (20.40±8.30 ng/ml,7.04±3.849ng/ml) and both had significant difference ( P< 0.05),The difference was not significant(P>0.05)when comparing the blood serum (32.14±12.29ng/ml,31.16±12.21ng/ml)of these two groups . The difference was not significant when comparing the blood serum of different menstrual cycle (P>0.05).4 FKN(Fig.7) was lacalized on cytoplasm and plasmalemma of mononuclear macrophage,peritoneum mesothelial cells . CX3CR1(Fig.9)was lacalized on cytoplasm and plasmalemma of mononuclear macrophage,natural killer cells,lymphocyte .5 The expression of FKN,CX3CR1 in peritoneal fluid cellsThe positive ratio of FKN on study group and control group were 100%,75.00% . The level of FKN in the study group was higher than the control group (X~2=26.53,P<0.01), The difference was not significant when comparing the different menstrual cycle of these two groups(P>0.05).The positive ratio of CX3CR1 on study group and control group were 100%,75.00% . The level of CX3CR1 in the study group was higher than the control group (X~2=26.53,P<0.01), The difference was not significant when comparing the different menstrual cycle of these two groups(P>0.05).There was a positive relationship between the expression of FKN and CX3CR1 in the study group(r=0.54, P<0.05), and the control group(r=0.69, P<0.05).Conclusion:1 The expression levels of FKN and CX3CR1 in normal endometrium, eutopic endometrial, ectopic endometrium have the tendency to increase, which shows that both of them play an important role in the formation of EMs2 The content of FKN and CX3CR1 in the peritoneal fluid of the study group were higher than that of the control group, which shows that they are involved in the intra-abdominal environmental changes of the endometriosis patients and the promotion of the adhesion and plantation of the ectopic endometrium, the formation of blood vessel in ectopic focus.3 FKN and CX3CR1 are the valid indicator to reflect the invasive growth of EMs, but the content of FKN and CX3CR1 in the serum of the study group and control group was not obviously different, which indicates that they are not involved in the systemic immune disorders of the EMs patients4 The expression of FKN and CX3CR1 in the group of endometrium and the group of peritoneal fluid cells has nothing to do with the menstrual cycle.5 The expression of FKN and CX3CR1 in the endometrium group and the peritoneal fluid cells group are relevant to each other, which shows that they interact with each other and promote the development of Ems together.6 Statistical analysis shows that the content of FKN and CX3CR1 in the endometrium, peritoneal fluid, peritoneal fluid cells of the EMs patients were higher than those in the control group and they play an important role in the occurrence and development of Ems. So the study of FKN and CX3CR1 provides a new way for the treatment of Ems.