雌激素对外周血T淋巴细胞间连接蛋白Cx40及Cx43的影响及机制研究

发布时间：2018-03-23 18:54

本文选题：缝隙连接　切入点：免疫　出处：《石河子大学》2017年硕士论文

【摘要】：目的:通过比较正常Sprague-Dawley(SD)大鼠和围绝经期模型的SD大鼠,正常育龄期女性和围绝经期女性外周血T淋巴细胞亚群、CD4+和CD8+T淋巴细胞上连接蛋白(Connexin,Cx)40和43表达、以及血清炎症因子IL-1、IL-2和IL-6的表达,探讨E_2(雌二醇)是否通过调节T淋巴细胞间缝隙连接通讯,抑制围绝经期低雌激素造成的T淋巴细胞亚群紊乱和促炎因子释放增加的慢性炎症过程,进而改善由慢性炎症引起的免疫功能异常。方法:动物血样实验:实验选用4月龄雌性SD大鼠,分为Control组(对照组)、OVX组(围绝经期动物模型组)及OVX+E_2(雌激素替代治疗组)。应用流式细胞术检测大鼠外周血T淋巴细胞亚群的比例以及CD4+和CD8+T淋巴细胞上CX40和CX43的表达频率;其次使用ELISA技术检测各组间雌二醇(estradiol,E_2)的差异以及各组炎症因子IL-1、IL-2、IL-6的表达。临床血样实验:分为育龄期组和围绝经期组,应用流式细胞术检测两组人群外周血T淋巴细胞亚群的比例以及CD4+和CD8+T淋巴细胞上CX40和CX43的表达频率;同时使用ELISA技术检测各组炎症因子IL-1、IL-2、IL-6的表达。结果:1.OVX组SD大鼠E_2水平低于Control组(P0.05),OVX+E_2组E_2水平比OVX组上升(P0.05)。2.OVX组CD4+CD25+T淋巴细胞比Control组上升(P0.05),OVX+E_2组比OVX组CD4+CD25+T淋巴细胞下降(P0.05);OVX组CD4+/CD8+比Control组下降(P0.05),OVX+E_2组CD4+/CD8+比OVX组上升(P0.05)。3.OVX组CD8+CX40、CD4+CX43均比Control组明显上升(P0.01),OVX+E_2组CD8+CX40、CD4+CX43均比Control组下降(P0.01),OVX+E_2组CD8+CX43比Control组及OVX组均下降(P0.01)。4.OVX组大鼠血浆IL-1β、IL-6浓度较Control均升高(P0.01),IL-2较Control组降低(P0.05),OVX+E_2组IL-1β、IL-6浓度较OVX组均降低(P0.01),IL-2较OVX组升高(P0.05)。5.Pearson相关性分析结果显示,大鼠IL-1β、IL-6与CD8+CX40、CD4+CX43和CD8+CX43均为正相关关系;IL-2与CD4+CX43为负相关关系。6.围绝经期组CD4+/CD8+比育龄组上升(P0.05)2.围绝经组女性外周血中CD4+CX40、CD4+CX43和CD8+CX43比育龄期组均上升(P0.05)。7.围绝经期女性外周血中IL-2及IL-6浓度均下降(P0.05)。结论:围绝经期是一种慢性低度炎症,伴随T淋巴细胞亚群比值异常,促炎因子释放增加。外源性给予E_2能够改善上述由围绝经期伴随的慢性炎症造成的损害,其可能机制是E_2通过下调T淋巴细胞上构成缝隙连接通道的CX40和CX43的表达,抑制T淋巴细胞间的信息通讯,进而减少促炎因子的释放。
[Abstract]:Objective: To compare the normal Sprague-Dawley (SD) rats and perimenopausal model of SD rats, normal women of childbearing age and menopausal women of peripheral blood T lymphocyte subsets, CD4+ lymphocytes and CD8+T junction protein (Connexin, Cx) 40 and 43 expression, and serum IL-1, the expression of IL-2 and IL-6 to investigate, E_2 (estradiol) whether by regulating the gap junctional intercellular communication of T lymphocytes, inhibit the disorder of T lymphocyte subsets in perimenopausal caused by low estrogen and proinflammatory cytokine release process of chronic inflammation increase, and improve the immune function caused by chronic inflammatory abnormalities. Methods: animal experiment: blood test selected 4 month old female SD rats were divided into Control group (control group), OVX group (perimenopausal animal model group) and OVX+E_2 (estrogen replacement therapy group). Rats were detected in peripheral blood T lymphocytes subsets by flow cytometry and CD4 CX40 + and CD8+T lymphocytes and the expression of CX43 frequency; secondly using ELISA technique to detect the estradiol (estradiol, E_2) as well as the differences between each group of inflammatory factors IL-1, IL-2, IL-6. The expression of clinical blood experiment: divided into age group and menopausal group, the proportion of peripheral blood T lymphocyte subsets in two group by flow cytometry and the expression frequency of CD4+ and CD8+T lymphocyte CX40 and CX43; at the same time using the detection of IL-1, each group of inflammatory factors ELISA IL-2, the expression of IL-6. Results: the E_2 level of 1.OVX group SD rats were lower than that of Control group (P0.05), the level of E_2 in group OVX+E_2 than in group OVX increased (P0.05) the.2.OVX group of CD4+CD25+T lymphocytes than in group Control increased (P0.05), OVX+E_2 group was lower than that of OVX group (P0.05); CD4+CD25+T lymphocyte in OVX group CD4+/CD8+ decreased than group Control (P0.05), OVX+E_2 group CD4+/CD8+ than in group OVX increased in group.3.OVX (P0.05) CD8+CX40, CD4+C X43 was higher than Control group significantly increased (P0.01), OVX+E_2 group CD8+CX40, CD4+CX43 were lower than group Control (P0.01), OVX+E_2 CD8+CX43 group than Control group and OVX group were significantly decreased (P0.01) in serum in.4.OVX group IL-1 beta, IL-6 concentration was Control increased (P0.01), IL-2 was lower than that of group Control (P0.05 OVX+E_2), group IL-1 beta, IL-6 concentration in OVX group were lower (P0.01), IL-2 increased compared with group OVX (P0.05).5.Pearson correlation analysis showed that rat IL-1 beta, IL-6 and CD8+CX40, CD4+CX43 and CD8+CX43 were positive correlation between IL-2 and CD4+CX43; negative correlation between.6. in perimenopausal group CD4+/CD8+ than childbearing age group increased (P0.05) 2. perimenopausal women group CD4+CX40 in the peripheral blood of CD4+CX43 and CD8+CX43 than the adult group were increased (P0.05.7.) in perimenopausal women and IL-2 IL-6 concentration in peripheral blood were decreased (P0.05). Conclusion: peri menopause is a chronic low-grade inflammation, with T lymphocyte subsets The ratio of abnormal proinflammatory cytokine release increase. Exogenous E_2 can improve the perimenopausal accompanied by chronic inflammatory damage, the possible mechanism is E_2 by inhibiting the expression of CX40 of gap junction channels and CX43 T lymphocytes, T lymphocytes in the suppression of information communication, thereby reducing the release of inflammatory factors.

【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R711.75

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