雌激素及其受体调节剂对APOE基因敲除小鼠胸主动脉CD40/CD40L的影响

Effects of estrogen and its receptor modulators on CD40 / Cd40L expression in thoracic aorta tissue of castrated APOE(-/-) mice

目的探讨雌激素及其受体拮抗剂对去势APOE基因敲除小鼠主动脉血管中CD40和CD40L表达的影响。方法 24只去势APOE(-/-)小鼠随机均分为三组:第一组给予生理盐水(对照组);第二组给予非特异雌激素受体(estrogen receptor,ER)拮抗剂ICI和雌激素(estrogen,E2)(ICI+E2组);第三组给予特异性雌激素β受体拮抗剂PHTTP和雌激素(PHTTP+E2组)。喂养8周后麻醉处死,获取动脉粥样硬化(atherosclerosis,AS)的组织标本,提取总RNA。应用反转录,定量PCR方法检测CD40/CD40L mRNA表达量。石蜡切片行Masson染色、CD40/CD40L免疫组化和免疫荧光染色。结果三组小鼠CD40/CD40L的表达量有显著不同(H=10.38,P=0.03)。与对照组相比,PHTTP+E2组CD40/CD40L表达量明显下降(q=12.46,P=0.021),ICI+E2组CD40/CD40L虽有下降,但差异无统计意义。Masson染色结果显示:与对照组相比ICI+E2组和PHTTP+E2组胶原纤维降解少,斑块稳定。结论雌激素可能通过雌激素受体α(ERα)发挥预防或治疗妇女AS的作用,ERα可能是妇女心血管疾病的保护性因素。

Objective To explore the effects of estrogen and its receptor antagonists on the expression of CD40 and CD40 ligand（ CD40L） in the thoracic aorta tissues of the castrated APOE（-/-） mice. Methods Twenty-four castrated APOE（-/-） mice were randomly divided into three groups： control group（ fed by saline）,ICI ＋ E2 group[fed by non-specific estrogen receptor（ ER） antagonists ICI and estrogen（ E2） ],PHTTP ＋ E2 group（ fed by specific estrogen receptor β antagonist PHTTP and E2）. At 8 week,the mice were anesthetized and sacrificed to get artery atherosclerosis（ AS） tissue samples for extracting the total RNA. The reverse transcription and quantitative PCR methods were used to detect CD40 / CD40 L mRNA expression. The paraffin sections were used for Masson staining,CD40 / CD40 L immunohistochemistry and immunofluorescence staining. Results The expression of CD40 / CD40 L was significantly different among three groups（ H = 10. 38,P = 0. 03）. Compared with control group,the expression of CD40 / CD40 L significantly decreased in PHTTP ＋ E2 group（ q = 12. 46,P = 0. 02）,and it also decreased in in ICI ＋ E2 group,but there was no statistical difference. Masson staining results showed that the collagen fibers degradated less in ICI ＋ E2 group and HTTP ＋ E2 group than in control group,which displayed the plaque stable. Conclusion Estrogen may play a role in prevention or treatment of women AS by estrogen receptor（ ERα）,which might be a protective factor for cardiovascular disease in women.