多囊卵巢综合征-胰岛素抵抗大鼠子宫组织中PPARs异常表达与胰岛素抵抗的关系

doi:10.12300/j.issn.1674-5817.2020.206

摘要/Abstract

摘要： 目的探讨过氧化物酶体增殖物激活受体（peroxisome proliferator-activated receptors,PPARs）在多囊卵巢综合征（polycystic ovarian syndrome,PCOS）伴胰岛素抵抗大鼠模型子宫组织中异常表达与胰岛素抵抗的关系。方法将40只雌性SD大鼠随机分为正常对照组和PCOS-胰岛素抵抗模型组,采用胰岛素+人绒毛膜促性腺激素（human chorionic gonadotrophin,hCG）皮下注射法建立大鼠PCOS-胰岛素抵抗模型,正常对照组注射等量生理盐水（即0.9%NaCl溶液）。应用ELISA检测两组大鼠空腹胰岛素和空腹血糖水平,并计算胰岛素抵抗指数。取大鼠子宫组织行HE染色,免疫组织化学法检测PPARs表达水平,蛋白质印迹法检测PPARs、胰岛素受体底物（insulin receptor substrate,IRS）、葡萄糖转运蛋白4（glucose transporter 4,GLUT-4）和胰岛素样生长因子-1（insulin like growth factor-1,IGF-1）等蛋白表达。结果模型组大鼠血清空腹胰岛素、空腹血糖和稳态胰岛素评价指数显著高于正常对照组（P<0.05）。子宫组织HE染色提示,正常对照组大鼠子宫组织形态正常,模型组大鼠子宫内膜出现不同程度的增生性改变,腺腔变小,腺体数目减少,排列松散。免疫组织化学显示,PPARs的3种亚型PPARα、PPARβ及PPARγ在大鼠子宫组织中均有表达,模型组PPARα和PPARγ的表达下调（P<0.05）,PPARβ在两组间的表达差异无统计学意义（P>0.05）。蛋白质印迹法检测结果显示,模型组大鼠子宫组织PPARα、PPARγ、IRS和GLUT-4的表达显著低于正常对照组（P<0.05）,而IGF-1表达高于正常对照组（P<0.05）。结论在PCOS-胰岛素抵抗大鼠子宫组织中IRS、GLUT-4表达下调,而IGF-1表达上调,证实在子宫组织层面存在胰岛素抵抗,可能与PCOS大鼠子宫组织PPARα、PPARγ表达下调相关。

关键词: 过氧化物酶体增殖物激活受体, 多囊卵巢综合征, 胰岛素抵抗, 子宫组织, 大鼠

Abstract: Objective To investigate the relationship between abnormal expression of peroxisome proliferator-activated receptors (PPARs) and insulin resistance in the uterine tissues of rats with polycystic ovary syndrome and insulin resistance (PCOS-IR). Methods Forty female SD rats were randomly divided into normal control and PCOS-IR model groups. PCOS-IR models were established by subcutaneous injection of insulin and human chorionic gonadotropin (INS+hCG) method, and rats in the normal control group were injected with equal amounts of normal saline. Fasting insulin (FINS) and fasting plasma glucose (FPG) levels were measured by enzyme-linked immunosorbent assay (ELISA), and the insulin resistance index (HOMA-IR) was calculated. Uterine tissues of rats were stained with hematoxylin-eosin (HE), PPARs were detected by immunohistochemistry, and the expressions of PPARs, insulin receptors (IRS), glucose transporter 4 (GLUT-4), and insulin-like growth factor-1 (IGF-1) were detected by Western blotting. Results FPG and FINS levels and HOMA-IR in the model group were significantly higher than those in the normal control group (P < 0.05). HE staining of uterine tissues showed that the endometrium of the normal control group was normal, and the endometrium of the model group showed different degrees of proliferative changes, the glandular cavity became smaller, the number of glands decreased, and the arrangement was loose. Immunohistochemistry showed that three subtypes of PPARs (PPARα, PPARβ, and PPARγ) were expressed in the rat uterus. The expression of PPARα and PPARγ in the rat uterus was down-regulated in the model group (P < 0.05), and there was no significant difference in the expression of PPARβ between the two groups (P > 0.05). Western blotting results showed that the expression of PPARα, PPARγ, IRS, and GLUT-4 in the rat uterus of the model group was significantly lower than that of the normal control group (P < 0.05), while the expression of IGF-1 was higher than that of the normal control group (P < 0.05). Conclusion The expression of IRS and GLUT-4 is down-regulated and the expression of IGF-1 is up-regulated in the uterine tissue of PCOS-IR rats. The existence of IR is confirmed in the uterine tissue of PCOS-IR rats, and it may be related to the down-regulation of PPARα and PPARγ expression in the uterine tissues.

Key words: Peroxisomal proliferator-activated receptors, Polycystic ovary syndrome, Insulin resistance, Uterine tissue, Rat

中图分类号:

魏巍, 陈艺华, 张秀智, 冷义福, 李纯, 尹天晓. 多囊卵巢综合征-胰岛素抵抗大鼠子宫组织中PPARs异常表达与胰岛素抵抗的关系[J]. 实验动物与比较医学, 2021, 41(5): 435-442.

WEI Wei, CHEN Yihua, ZHANG Xiuzhi, LENG Yifu, LI Chun, YIN Tianxiao. Relationship Between the Abnormal Expression of PPARs and Insulin Resistance in Uterus of Rats with Polycystic Ovary Syndrome and Insulin Resistance[J]. Laboratory Animal and Comparative Medicine, 2021, 41(5): 435-442.

图/表 4

参考文献

[1] 中华医学会妇产科学分会内分泌学组及指南专家组. 多囊卵巢综合征中国诊疗指南[J]. 中华妇产科杂志, 2018, 53(1):2-6. DOI:10.3760/cma.j.issn.0529-567x.2018.01.002.
[2] 张春梅, 赵越, 乔杰. 多囊卵巢综合征合并代谢综合征临床特点及治疗进展[J]. 中国实用妇科与产科杂志, 2016, 32(9):915-918.
[3] BANU J, FATIMA P, SULTANA P, et al.Association of infertile patients having polycystic ovarian syndrome with recurrent miscarriage[J]. Mymensingh Med J, 2014, 23(4):770-773.
[4] HAOULA Z, SALMAN M, ATIOMO W.Evaluating the association between endometrial cancer and polycystic ovary syndrome[J]. Hum Reprod, 2012, 27(5):1327-1331. DOI:10.1093/humrep/des042.
[5] SZCZEPAŃSKA A A, ŁUPICKA M, SOCHA B M, et al. The influence of arachidonic acid metabolites on PPAR and RXR expression in bovine uterine cells[J]. Gen Comp Endocrinol, 2018, 262:27-35. DOI:10.1016/j.ygcen.2018.03.009.
[6] 魏巍, 徐泰安, 王秋实, 等. 两种多囊卵巢综合征伴胰岛素抵抗大鼠模型的比较研究[J]. 哈尔滨医科大学学报, 2019, 53(1):12-16. DOI:CNKI:SUN:HYDX.0.2019-01-003.
[7] 李轶, 梁晓燕. 多囊卵巢动物模型的研究进展[J]. 国际生殖健康/计划生育杂志, 2008, 27(6):380-383. DOI: 10.3969/j.issn.1674-1889.2008.06.014.
[8] 陈如枫, 刘新敏. 多囊卵巢综合征胰岛素抵抗动物模型构建方法的研究概况[J]. 世界中西医结合杂志, 2018, 13(5):733-736. DOI:10.13935/j.cnki.sjzx.180536.
[9] PORETSKY L, PIPER B.Insulin resistance, hyper-secretion of LH, and a dual-defect hypothesis for the pathogenesis of polycystic ovary syndrome[J]. Obstet Gynecol, 1994, 84(4):613-621.
[10] 李轶, 梁晓燕, 杨星, 等. 胰岛素和人绒毛膜促性腺激素建立高雄激素多囊卵巢大鼠模型[J]. 国际生殖健康/计划生育杂志, 2008, 27(1):54-57. DOI:10.3969/j.issn.1674-1889.2008.01.019.
[11] 谢来娣, 张伊娜, 龚丽芬, 等. 多囊卵巢综合征伴胰岛素抵抗相关发病机制研究[J]. 中国妇幼保健, 2019, 34(8):1926-1929. DOI:10.7620/zgfybj.j.issn.1001-4411. 2019.08.75.
[12] 许良智, 单丹. 多囊卵巢综合征——一种代谢障碍性全身性疾病在卵巢的表现[J]. 中华妇幼临床医学杂志(电子版), 2011, 7(5):321-325.
[13] 杨嘉琦, 郭雪桃, 周梦, 等. 育龄期伴高雄激素多囊卵巢综合征临床特征及代谢异常相关因素分析[J]. 中国药物与临床, 2017, 17(6):789-792. DOI:10.11655/zgywylc2017.06.004.
[14] CHANG E M, HAN J E, SEOK H H, et al.Insulin resistance does not affect early embryo development but lowers implantation rate in in vitro maturation-in vitro fertilization-embryo transfer cycle[J]. Clin Endocrinol, 2013, 79(1):93-99. DOI:10.1111/cen.12099.
[15] 宋学茹, 张慧英, 张艳芳, 等. 细胞外信号调节激酶在多囊卵巢综合征患者子宫内膜中的活化及意义[J]. 中华妇产科杂志, 2010, 45(10):767-771. DOI:10.3760/cma.j.issn.0529-567x.2010.10.011.
[16] 王菊. 胰岛素抵抗与多囊卵巢综合征育龄妇女生育功能的关系[J]. 中国实用医药, 2018, 13(26):70-71. DOI:10.14163/j.cnki.11-5547/r.2018.26.038.
[17] 李扬璐, 阮祥燕, MUECK A O.多囊卵巢综合征对妊娠结局的影响研究进展[J]. 首都医科大学学报, 2016, 37(4):449-453. DOI:10.3969/j.issn.1006-7795.2016. 04.007.
[18] PATEL R, SHAH G.Insulin sensitizers modulate GnRH receptor expression in PCOS rats[J]. Arch Med Res, 2018, 49(3):154-163. DOI:10.1016/j.arcmed.2018. 08.001.
[19] VITTI M, DI EMIDIO G, DI CARLO M, et al.Peroxisome proliferator-activated receptors in female reproduction and fertility[J]. PPAR Res, 2016, 2016:4612306. DOI:10.1155/2016/4612306.
[20] ALTSHULER D, HIRSCHHORN J N, KLANNEMARK M, et al.The common PPARgamma Pro12Ala polymor-phism is associated with decreased risk of type 2 diabetes[J]. Nat Genet, 2000, 26(1):76-80. DOI:10.1038/79216.
[21] UNLUTURK U, HARMANCI A, KOCAEFE C, et al.The genetic basis of the polycystic ovary syndrome: a literature review including discussion of PPAR-gamma[J]. PPAR Res, 2007, 2007:49109. DOI:10.1155/2007/49109.
[22] WANG M H, TAFURI S.Modulation of PPARgamma activity with pharmaceutical agents: treatment of insulin resistance and atherosclerosis[J]. J Cell Biochem, 2003, 89(1):38-47. DOI:10.1002/jcb.10492.
[23] 梁秀文, 季新梅, 马娟. 多囊卵巢综合征患者外周血单个核细胞中RPS 26及PPARγ基因的表达[J]. 中国计划生育和妇产科, 2017, 9(1):55-58. DOI:10.3969/j.issn.1674-4020.2017.01.14.
[24] MINGE C E, BENNETT B D, NORMAN R J, et al.Peroxisome proliferator-activated receptor-γ agonist rosiglitazone reverses the adverse effects of diet-induced obesity on oocyte quality[J]. Endocrinology, 2008, 149(5):2646-2656. DOI:10.1210/en.2007-1570.
[25] CUI Y, MIYOSHI K, CLAUDIO E, et al.Loss of the peroxisome proliferation-activated receptor gamma (PPARgamma) does not affect mammary development and propensity for tumor formation but leads to reduced fertility[J]. J Biol Chem, 2002, 277(20):17830-17835. DOI:10.1074/jbc.m200186200.
[26] KADAM L, KOHAN-GHADR H R, DREWLO S. The balancing act - PPAR-γ's roles at the maternal-fetal interface[J]. Syst Biol Reprod Med, 2015, 61(2):65-71. DOI:10.3109/19396368.2014.991881.
[27] NADRA K, ANGHEL S I, JOYE E, et al.Differentiation of trophoblast giant cells and their metabolic functions are dependent on peroxisome proliferator-activated receptor beta/delta[J]. Mol Cell Biol, 2006, 26(8):3266-3281. DOI:10.1128/MCB.26.8.3266-3281.2006.
[28] ASAMI-MIYAGISHI R, ISEKI S, USUI M, et al.Expression and function of PPARgamma in rat placental development[J]. Biochem Biophys Res Commun, 2004, 315(2):497-501. DOI:10.1016/j.bbrc.2004.01.074.
[29] FOURNIER T, TSATSARIS V, HANDSCHUH K, et al.PPARs and the placenta[J]. Placenta, 2007, 28(2-3):65-76. DOI:10.1016/j.placenta.2006.04.009.
[30] PARAST M M, YU H, CIRIC A, et al.PPARgamma regulates trophoblast proliferation and promotes labyrinthine trilineage differentiation[J]. PLoS One, 2009, 4(11): e8055. DOI:10.1371/journal.pone. 0008055.
[31] 李巍巍, 贾莉婷, 张展. PAI-1在多囊卵巢综合征大鼠脂肪组织中的表达及意义[J]. 中国优生与遗传杂志, 2008, 16(12):28-31. DOI:CNKI:SUN:ZYYA.0.2008-12-012.
[32] HARA M, ALCOSER S Y, QAADIR A, et al.Insulin resistance is attenuated in women with polycystic ovary syndrome with the Pro(12)Ala polymorphism in the PPARgamma gene[J]. J Clin Endocrinol Metab, 2002, 87(2):772-775. DOI:10.1210/jcem.87.2.8255.
[33] 胡卫红, 陈琳, 同军,等. PPARγmRNA在卵巢颗粒细胞的表达调节及与多囊卵巢综合征的相关性[J]. 北京大学学报(医学版), 2013, 45(6):859-863. DOI:10.3969/j.issn.1671-167X.2013.06.007.
[34] 姜慧洁, 张晓静, 张慧, 等. 基于PPARs靶标改善胰岛素抵抗的中药活性成分研究进展[J]. 中国中药杂志,2015, 40(22):4355-4358. DOI:10.4268/cjcmm 20152207.
[35] 赵博文, 陈艳昆, 张栩, 等. 基于PPARγ-LXRα-ABCA1通路的中药降脂成分研究[J]. 北京中医药大学学报, 2018, 41(2):131-139. DOI:10.3969/j.issn. 1006-2157.2018.02.007.
[36] 张秀红, 宣姣, 亓志刚. PPARα, γ和δ: 胰岛素抵抗治疗的靶点[J]. 中国生物化学与分子生物学报, 2014, 30(6):543-548. DOI:10.13865/j.cnki.cjbmb.2014.06.004.
[37] WARREN J D, BLUMBERGS P C, THOMPSON P D.Rhabdomyolysis: a review[J]. Muscle Nerve, 2002, 25(3):332-347. DOI:10.1002/mus.10053.
[38] 秦春焕, 曹衡, 李红莉. PPARβ/δ在兔动脉粥样硬化模型中的表达及作用[J]. 中国分子心脏病学杂志, 2014, 14(5):1080-1084. DOI:10.16563/j.cnki.1671-6272. 2014.05.001.