吡格列酮、二甲双胍治疗PCOS雌鼠后卵巢组织炎性因子以及超微结构改变的研究

目的:探讨吡格列酮、二甲双胍治疗后多囊卵巢综合征(PCOS)雌鼠卵巢组织匀浆中促炎因子、抑炎因子以及卵巢超微结构的改变。方法:将50只成年雌性SD大鼠随机分为5组,其中40只建立PCOS雌鼠模型。A组为正常雌鼠,B组为PCOS雌鼠,C组为经吡格列酮灌胃干预的PCOS雌鼠,D组为经二甲双胍灌胃干预的PCOS雌鼠,E组为吡格列酮联合二甲双胍灌胃干预的PCOS雌鼠。各组雌鼠干预处理后32h取卵巢组织,将一侧卵巢制备成组织匀浆液行固相夹心法酶联免疫吸附实验,检测促炎因子超敏C反应蛋白(hs-CRP)、降钙素原(PCT)与抑炎因子白介素10(IL-10)、脂联素(ADP)含量变化。将另一侧卵巢在透射电镜下观察超微结构的变化。结果:A、B、C、D、E组hsCRP水平分别为(541.62±74.26)、(820.81±46.84)、(686.33±99.12)、(779.02±38.46)、(501.92±56.18)ng/ml,B、C、D组hs-CRP水平均高于A组及E组(P均0.05);A、B、E组PCT水平分别为(42.57±2.42)、(93.31±7.17)、(47.21±11.10)pg/ml,B组PCT水平均高于A、E两组(P均0.05)。A、B、C、D、E组IL-10水平分别为(6.68±0.83)、(3.41±0.88)、(5.25±1.38)、(7.84±1.00)、(7.87±0.26)pg/ml,B、C组IL-10水平均低于A、D、E组(P均0.01)。ADP水平各组两两比较差异均无统计学意义(P均0.05)。A、E组卵巢颗粒细胞超微结构均为正常表现,B组颗粒细胞呈凋亡改变,C、D组颗粒细胞超微结构较B组有改善。结论:PCOS雌鼠卵巢组织匀浆中促炎因子与抑炎因子失衡,颗粒细胞呈凋亡变化。二甲双胍、吡格列酮联合干预可能改善这种慢性轻度炎症状态。

Effects of metformin and pioglitazone on inflammatory cytokine concentration and ultrastructure of ovarian tissue in female rats with polycystic ovary syndrome

Objective： To explore changes of inflammatory cytokine level in ovarian tissue homogenates and ovarian ultrastructure female rats suffering from polycystic ovary syndrome （PCOS） after taking metformin and pioglitazone. Methods： Models of female rats suffering from PCOS were established. Fifty female rats were divided into 5 groups randomly and evenly with 10 per group, in which there were two control groups named as group A and group B. Rat in group A included normal matured female rats with no intervention, while those in group B were matured female rats with PCOS. Both groups were fed 0.9% of normal saline. The rest three groups of C, D and E were experimental groups with matured female rats with PCOS. Rats in group C, D and E were fed metformin, pioglitazone, both metformin and pioglitazone, respectively. The rats were sacrified and their ovaries were taken. ELISA was conducted on one side of ovarian tissue homogenates and levels of high--sensitivity C-reactive protein （hs-CRP）, proealcitonin （PCT）, interleukin （IL）-10 and adiponeetin （ADP） were measured. Meanwhile, the uhrastructural changes of the other side of the ovary were observed under the TEM. Results： Concentrations of hs--CRP of five rouns were 541.62±74.26, 820.81±46.84, 686.33±99.12, 779.02±38.46, 501.92±56.18 ng/ml, respectively. The levels of hs--CRP in groups B, C and D were significantly higher than those in group A and E （P all〈0.05）. Concentrations of PCT of group A, B and E were 42.57±2.42, 93.31±7.17 and 47.21±11.10 pg/ml, respectively. PCT level in groups B was higher than those in group A and E （P all〈0.05）. Concentrations of IL--10 of five groups were 6.68±0.83, 3.41±0. 88, 5.25±1.38, 7.84±1.00, and 7.87±0.26 pg/ml, respectively. IL-10 levels in group B and C were lower than those in group A , D and E （P all 〈0.01）. However, no statistically significant differences were found in multiple comparisons of the five groups for ADP levels （P all〉0.05）. The ultrastructure of granulosa cells of group A and E was normal, while that of group B was approaching apoptosis. Whereas, those of group C and D were better than that of group B. Conclusion： The proinflammatory and inhibitory inflammatory cytokine in ovarian tissue homogenates of female rats suffering from PCOS are in an unbalanced status with granulosa cells are approaching apoptosis. The coordinated intervention of metformin and pioglitazone may alleviate chronic low--grade inflammation.