Detection Of The Apoptosis In Subclinical Mastitis Mammary Epithelial Cell And Analysis Of Related Differences

In order to observe the change of the cows with subclinical mastitis apoptosis of epithelial cells;In order to study the breast immune response mechanism with mastitis in cows, and provide scientific reference and basis for the mechanism for the pathogenesis and treatment of mastitis;Screening identified 18 subclinical mastitis Holstein cows with similar capacity, obtaining breast tissue respectively in no pregnancy, and after parturition(AP)8thd,20thd,120thd,240thd,300thd.Applying the immunohistochemical method to detect expression of apoptotic factor caspase3,bcl-2,bax,flow cytometry was used to analyze apoptotic cells quantitatively,while elected the healthy cows and subclinical mastitis cows and measured the quantity of lactation on after parturition on 2thd,8thd, 20thd, 40thd and so on,in 16 lactation milk time,each measure time elected 10 cows.The result of caspase3 by Immunohistochemistry and flow cytometry was analysesed differences and differences in milk yield between healthy cows and subclinical mastitis cows. healthy cow data was from own laboratory.Experiment results(:1)The expression of Caspase3 results:Positive cells were brownish yellow.The rate of positive expression at No pregnancy and AP 240thdrespectively disparity was exceedingly significant other stages(P<0.01);There were no significantly differences between 8thdand 300thd(P>0.05), the same with 20thdand 120thd(P>0.05),whille compared with other periods,both showed significant differences(P<0.01);Compared with healthy cows show:the rate of AP 8thd,20thdpositive cells compared to the same period,there were very significant differences(P<0.01);No pregnancy,AP 120thd,240thdand 300thd, positive cells over the same period were significantly differences(P<0.05).(2)Apoptotic factor bax expression:positive cells were brown.No pregnancy is the highest expression,AP 20thdis the lowest expression.There were no significant differences between AP 20thdand 120thdexpression (P>0.05),whille compared with other periods,showed very significantly differences(P<0.01);In no pregnant,AP 8thd,240thd,300thd,the rates of positive expression were very significantly different from each other (P<0.01).(3)Apoptotic factor bcl-2 expression:The rate of positive expression in no pregnancy and AP 300thdwere very significantly different from the other periods(P<0.01);AP 20thdand 120thdshowed no significant difference (P>0.05);There were no significant differences in AP 8thdand 240thd(P>0.05), which was significantly different from others(P<0.01).(4)Apoptosis by flow cytometry showed that: Subclinical mastitis cow mammary epithelial cell apoptosis in no pregnancy and AP 300thdwas very significantly different from the other periods respectively (P<0.01);There were no significant differences between AP 8thdand 240thd (P>0.05),but showed significantly differences compared with others(P<0.01);20thdand 120thdafter delivery were no significant difference(P>0.05).Compared with healthy cows show that:No pregnancy was no significant difference in the same period(P>0.05),AP 8thd,20thdshowed very significant differences(P<0.01),other periods were significantly different (P<0.05).(5)The quantify of lactation between subclinical mastitis cows and healthy was showed that:The quantify of subclinical mastitis cows lactation was significantly decreased than healthy,except milk 2th d ,8th d ,80th d ,100th d decreased no significant differences(P>0.05),at other time points showed very significant differences(P<0.01). Conclusion:The number apoptosis of breast epithelial cells of subclinical mastitis cow by Immuno -histochemistry and flow cytometry compared with healthy cows shows that:The occurrence of subclinical mastitis caused mammary epithelial cell damage,promote significantly increase of apoptosis of breast epithelial cells,and the number of different lactation periods were higher than health.Detection of apoptotic factor Bax and bcl-2 by Immunohistochemistry shows that,their changes with the coordination of inhibition in different lactation periods,that Bax and bcl-2 apoptotic factors involved in subclinical mastitis of bovine mammary cell apoptosis,and apoptosis in breast cells has a role in regulation.Subclinical mastitis cows milk yield compared with healthy cows show,subclinical mastitis during lactation milking was significantly lower than the amount of healthy,description subclinical mastitis caused substantial apoptosis of breast epithelial cells and the amount of cow milk has a direct impact, resulting in reduced milk yield, which led to economic losses.