PGD_(2)/DP_(1)途径对细菌感染奶牛子宫内膜组织中HMGB-1和PAFR表达的影响

Effect of PGD_(2)/DP_(1) Pathway on Expression of HMGB-1 and PAFR in Endometrial Tissue of Cows Infected with Bacteria

为了研究前列腺素D2(prostaglandin D2,PGD2)/DP1受体途径对大肠杆菌(Escherichia coli)和金黄色葡萄球菌(Staphylococcus aureus)感染奶牛子宫内膜组织中炎症介质HMGB-1和PAFR的表达及对组织损伤程度的影响,试验以体外培养奶牛子宫内膜组织为研究对象,应用1×10-6 mol/L DP1受体激动剂(BW-245C和15d-PGJ2)和等量(1×106 CFU/mL)大肠杆菌、金黄色葡萄球菌共同处理奶牛子宫内膜组织,采用实时荧光定量PCR、免疫组化和HE染色法检测奶牛子宫内膜组织中HMGB-1和PAFR的表达并评价组织损伤情况。结果显示,与空白对照组相比,大肠杆菌和金黄色葡萄球菌感染奶牛子宫内膜组织中HMGB-1和PAFR表达量显著升高(P<0.05),而DP1受体激动剂与大肠杆菌和金黄色葡萄球菌共处理奶牛子宫内膜组织中DP1受体激动剂显著抑制奶牛子宫组织中HMGB-1和PAFR的表达(P<0.05)。HE染色结果显示,大肠杆菌与金黄色葡萄球菌感染的奶牛子宫内膜组织上皮细胞和腺上皮细胞完全脱落、坏死、崩解;而DP1受体激动剂、大肠杆菌、金黄色葡萄球菌共同处理奶牛子宫内膜组织中,DP1受体激动剂的加入显著减轻奶牛子宫内膜组织的损伤程度(P<0.05)。免疫组化染色法结果与以上两种方法结果一致。结果表明,PGD2能够抑制大肠杆菌与金黄色葡萄球菌感染的奶牛子宫内膜组织中损伤相关因子HMGB-1、PAFR的表达,减轻组织损伤程度,这一作用可能是由DP1受体所介导的。

The aim of this study was to investigate the effects of PGD2/DP1 receptor pathway on the expression of inflammatory mediators HMGB-1 and PAFR,and the degree of tissue injury in the endometrium of dairy cows infected with Escherichia coli(E.coli)and Staphylococcus aureus(S.aureus).The endometrial tissues of cows were treated with 1×10-6 mol/L DP1 receptor agonists(BW-245C and 15d-PGJ2)and infected with E.coli and S.aureus at a concentration of 1×106 CFU/mL.The expression of HMGB-1 and PAFR was detected by Real-time quantitative PCR,immunohistochemistry,and the tissue injury was evaluated by HE staining.The results showed that the expression of HMGB-1 and PAFR of endometrium tissue in dairy cows was significantly higher than that in blank control group after E.coli and S.aureus infection at a concentration of 1×106 CFU/mL(P<0.05),while the expression of HMGB-1 and PAFR in bovine endometrium was significantly inhibited by DP1 receptor agonists(P<0.05).HE staining showed that the endometrial epithelial cells and glandular epithelial cells was completely exfoliated,necrotic and disintegrated after E.coli and S.aureus infection.While the DP1 receptor agonists treated bovine endometrium,it significantly reduced the degree of injury of bovine endometrium(P<0.05).The results of immunohistochemical staining were consistent with those of the above two methods.The results showed that PGD2 could inhibit the expression of HMGB-1 and PAFR and reduce the degree of tissue damage in the bovine endometrium infected with E.coli and S.aureus,which might be mediated by DP1 receptor.