| As one of the most common diseases of female reproductive system,endometritis is usually occult and the pathological changes of uterus are slight in the early stage,and there is no obvious clinical manifestation in the dog.When it develops to the middle or late stage of the course of disease,the pathological changes of uterus worsen and even appear systemic inflammatory response syndrome,which will endanger the life of the dog.Therefore,in-depth study on the characteristics of uterine lesions is helpful to understand the pathogenesis.Furthermore,the study on the expression differences of inflammatory factors and aging-related factors in different pathological types of uterine tissues can reveal the pathological characteristics of endometritis.In this experiment,30 cases of endometritis dogs with relevant clinical manifestations were collected and counted in an animal hospital in Harbin,and uterine tissue samples were collected for anatomical and pathological observation and classification.In addition,uterine tissue samples of 10 healthy female dogs who were subjected to physiological sterilization were taken as control observation.According to the clinical manifestations of 40 female dogs and the observation results of uterine anatomy and histology,40 dogs were divided into the following categories:(1)Group without clinical manifestations: 1)Healthy,n=7;2)Endometrial cystic hyperplasia,n=3.(2)Group with clinical symptoms: 1)No inflammation,n=1;2)Mild inflammatory infiltration,n=9;3)Endometrial hyperplasia type severe inflammatory infiltration type,n=12;4)Endometrial atrophy type severe inflammatory infiltration type,n=8.Subsequently,RT-q PCR was used to detect the m RNA content of IL-1?,IL-6 and IL-8 genes in healthy,mild inflammatory infiltration and two types of severe inflammatory infiltration of uterine tissue In addition,according to the relationship between dog weight and relative age of human beings,the classification standard for young and old dogs is established.Western Blot technique was used to detect the expression levels of aging-related proteins(p53,p21)in uterine tissues of six groups of young or old health,mild inflammatory infiltration of young or old and severe inflammatory infiltration of young or old.The experimental results: Statistical results of basic information of affected dogs: The youngest was 1 year and 5 months,and the oldest was 13 years with an average age of 8.10±3.03.In addition,of the 30 dogs affected,only 2 dogs had a history of giving birth,and none of the other dogs were bred.All cases had no history of exogenous sex hormone drugs.Statistical results of clinical investigation: 3 cases had symptoms within 30 days to 55 days after mating,27 cases had clinical manifestations between 50 days to 85 days after ending of last oestrus.When seeing a doctor,the affected dog may have different degrees of clinical symptoms,including depression,fever,loss of appetite or even disuse,increased desire to drink,frequent urination,abnormal secretion of vulva,increased abdominal circumference,gastrointestinal disorders,abnormal estrus cycle and other manifestations.B-ultrasonic images showed that 30 dogs had enlarged uterine horn,dark liquid area in uterine cavity and hyperplasia or atrophy of uterine wall.Blood routine test showed that the dogs had different degrees of inflammatory infection and anemia.Biochemical test results showed that some dogs had some degree of liver and kidney damage.Results of uterine anatomy and pathology:Of the 10 clinically asymptomatic female dogs undergoing physiological sterilization,3 cases had normal uterine tissue samples for necropsy,but the endometrium was observed to be cystic proliferative lesions under the microscope.Of the 30 cases with clinical symptoms,1 case had histologically no inflammatory lesion but its endometrium was atrophic,while the other 29 cases had inflammatory cell infiltration in varying degrees,of which 9 cases had slight inflammatory cell infiltration but no significant intimal hyperplasia.The other 20 cases showed diffuse and severe inflammatory infiltration,and were divided into 12 endometrial hyperplastic and 8 endometrial atrophic types according to the thickness ratio of healthy endometrium to myometrium.In addition,compared with healthy uterine horn,the radius of endometrial hyperplastic and atrophic severe inflammatory infiltrative uterine horn increased significantly(p<0.001)and no significant change in other groups.In addition,the ratio of endometrial thickness to myometrial thickness of endometrial hyperplasia with severe inflammatory infiltration was significantly higher than that of healthy uterus(p<0.01).The ratio of endometrial thickness to myometrial thickness was significantly lower in endometrial atrophic type than in hyperplastic type.Detection Results of Expression of Inflammatory Factors and Aging Factors in Uterine Tissue: Compared with healthy uterine tissue samples,the m RNA contents of IL-1?,IL-6 and IL-8 genes in uterine samples with inflammatory histology were significantly increased.Among them,the m RNA contents of three genes in endometrial hyperplastic and atrophic severe inflammatory infiltrating uterine tissues were significantly increased compared with the other two groups(p<0.001).In the uterus with severe inflammatory infiltration,the m RNA content of IL-1? and IL-6 genes in endometrial atrophic type was significantly higher than that in hyperplastic type(p<0.05 and p<0.001),but the change of IL-8 was not significant.Compared with the healthy uterus of the young and the old,the expression of p53 and p21 proteins in the uterus tissues with inflammatory characteristics of the young and the old are significantly increased.At the same time,the contents of two proteins in the uterine tissue of the severe inflammatory infiltration group were significantly higher than those of the mild inflammatory infiltration group(p<0.01 or p<0.001).Combined with the above research,it is found that the uterus of healthy dogs and affected dogs can have pathological changes in different degrees in anatomy and histology.Affected dogs can be divided into four types: non-inflammatory,mild inflammatory infiltration,endometrial hyperplastic severe inflammatory infiltration and atrophic severe inflammatory infiltration.In addition,endometrial cystic hyperplasia appeared in healthy female dogs.The expression of inflammatory factors(IL-1?,IL-6 and IL-8)is positively correlated with the degree of inflammatory infiltration in uterine tissue.In addition,the expression levels of p53 and p21 proteins in uterine tissue indicate that the degree of uterine aging is positively correlated with the degree of inflammatory infiltration. |
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