电针对PCOS模型大鼠性激素及卵巢组织TSP-1表达的影响
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摘要
目的探讨电针对多囊卵巢综合征(polycystic ovary syndrome, PCOS)模型大鼠的治疗作用,以及对卵巢组织中血小板反应蛋白-1(thrombospondin-1, TSP-1)表达的影响。方法将32只雌性SD大鼠随机分为空白组10只、模型组12只、电针组10只,采用1 mg/kg来曲唑加l%羧甲基纤维素灌胃建立PCOS大鼠模型。治疗后检测大鼠血清睾酮(testosterone, T)、黄体生成素(luteinizing hormone, LH)、卵泡刺激素(follicle-stimulating hormone, FSH)水平,HE染色观察卵巢组织病理结构改变,免疫组化法检测卵巢组织中TSP-1的表达。结果治疗后,电针组大鼠体质量增量和卵巢相对质量较模型组降低(P<0.01),血清T、LH水平及LH/FSH均较模型组降低(P<0.01),卵巢切片可见小囊状扩张卵泡较模型组减少,颗粒细胞层增厚;TSP-1表达于卵巢组织颗粒细胞和卵泡膜细胞的胞浆,模型组TSP-1表达量较低空白组降低(P<0.01),电针组TSP-1表达较模型组升高(P<0.05)。结论电针治疗可改善PCOS大鼠性激素紊乱状态,提高卵巢组织中TSP-1的表达,通过调节血管抑制因子、减少卵巢组织中血管新生、改善微循环以达到治疗PCOS的目的。
Objective To observe the therapeutic effects of electroacupuncture(EA) on polycystic ovary syndrome(PCOS) model rats and its effects on the expression of thrombospondin-1(TSP-1) in ovarian tissues. Methods A total of 32 female SD rats were randomly divided into the blank group(n=10), the model group(n=12), and the EA group(n=10). PCOS rat model was established by intragastric administration of 1 mg/kg letrozole dissolved in 1% carboxymethylcellulose(CMC). After treatment, serum levels of testosterone(T), luteinizing hormone(LH), and follicle stimulating hormone(FSH) were measured by enzyme-linked immunosorbent assay. The pathological changes of the ovarian tissue were observed by HE staining. The expression of TSP-1 in ovarian tissues was detected by immunohistochemistry. Results Compared with the model group, the body weight gain and the relative ovarian mass of the EA group were decreased when compared with the model group(P<0.01), and the serum T, LH levels and LH/FSH were decreased when compared with the model group(P<0.01). The HE staining of the ovarian tissue showed decreased cystic expansion follicles, and the granular cell layer was obviously thicker. TSP-1 was expressed in the cytoplasm of ovarian tissue granulosa cells and ovarian membrane cells. The expression of TSP-1 was decreased in the model group than in the blank group(P<0.01). After treatment, the expression of TSP-1 in the EA group was increased when compared with model group(P<0.05). Conclusion Electroacupuncture can improve the state of sex hormone disorder in PCOS rats, increase the expression of TSP-1, reduce the angiogenesis in ovarian tissue by regulating angiogenesis inhibitors, and improve microcirculation to achieve the purpose of treating PCOS.
Objective To observe the therapeutic effects of electroacupuncture(EA) on polycystic ovary syndrome(PCOS) model rats and its effects on the expression of thrombospondin-1(TSP-1) in ovarian tissues. Methods A total of 32 female SD rats were randomly divided into the blank group(n=10), the model group(n=12), and the EA group(n=10). PCOS rat model was established by intragastric administration of 1 mg/kg letrozole dissolved in 1% carboxymethylcellulose(CMC). After treatment, serum levels of testosterone(T), luteinizing hormone(LH), and follicle stimulating hormone(FSH) were measured by enzyme-linked immunosorbent assay. The pathological changes of the ovarian tissue were observed by HE staining. The expression of TSP-1 in ovarian tissues was detected by immunohistochemistry. Results Compared with the model group, the body weight gain and the relative ovarian mass of the EA group were decreased when compared with the model group(P<0.01), and the serum T, LH levels and LH/FSH were decreased when compared with the model group(P<0.01). The HE staining of the ovarian tissue showed decreased cystic expansion follicles, and the granular cell layer was obviously thicker. TSP-1 was expressed in the cytoplasm of ovarian tissue granulosa cells and ovarian membrane cells. The expression of TSP-1 was decreased in the model group than in the blank group(P<0.01). After treatment, the expression of TSP-1 in the EA group was increased when compared with model group(P<0.05). Conclusion Electroacupuncture can improve the state of sex hormone disorder in PCOS rats, increase the expression of TSP-1, reduce the angiogenesis in ovarian tissue by regulating angiogenesis inhibitors, and improve microcirculation to achieve the purpose of treating PCOS.
引文
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[12]贺晓霞,王若光.平衡阴阳法对多囊卵巢综合征雌雄激素的调节[J].湖南中医药大学学报,2016,36(2):16-19.
[13]赵娜,杨正望,全春梅.多囊卵巢综合征临床分型与中医证型的相关性分析[J].湖南中医药大学学报,2017,37(9):966-968.
[14]施茵,虞莉青,尹小君.针灸治疗肥胖型多囊卵巢综合征的临床疗效观察[J].中华中医药学刊,2010,28(4):805-807.
[15]卓缘圆,吴家满,杨卓欣,等.“调任通督针刺法”治疗多囊卵巢综合征不孕症的临床疗效观察[J].中国针灸,2016,36(12):1237-1241.
[16] NOROOZZADEH M N M, BEHBOUDI-GANDEVANI S B S,ZADEH-VAKILI A Z A, et al. Hormone-induced rat model of polycystic ovary syndrome:A systematic review[J]. Life Sciences,2017,191:259-272.
[17]宫静,程凯,张露芬.电针不同穴位对去卵巢大鼠HPO轴的影响[J].中华中医药杂志,2011,26(3):595-597.
[18]王君,张宝文,于新宇,等.疾徐捻转泻法针刺足三里对高血压家兔的降压作用[J].中国临床康复,2006,10(7):116-117.
[19]杨正望,文炯,张增玲.补肾化瘀方对PCOS大鼠卵巢颗粒细胞CYP450表达的影响[J].湖南中医药大学学报,2011,31(1):30-32,39.
[20]孙洁,赵继梦,季蓉,等.电针对多囊卵巢综合征大鼠卵巢高雄激素症的影响[J].针刺研究,2013,38(6):465-472.
[21]林鸾.穴位埋线对PCOS-IR模型大鼠生殖内分泌和胰岛素抵抗的影响[D].广州:广州中医药大学,2016.
[22]黄玉秀,郑秀,刘林芳,等.TSP-1在子宫内膜腺癌中的表达及与VEGF、CD105的关系[J].中国医师杂志,2009,11(10):1301-1303.
[23]叶璐,冯亦军,马奔,等.VEGF-C和TSP-1在卵巢上皮癌中的表达及意义[J].中华临床医师杂志,2008,2(10):1142-1147.
[2] QIU J J, LIU Y L, LIU M H, et al. Ovarian interstitial blood flow changes assessed by transvaginal colour Doppler sonography:predicting ovarian endometrioid cyst-induced injury to ovarian interstitial vessels[J]. Archives of Gynecology&Obstetrics,2012,285(2):427-433.
[3]任凤岩,李一宁,王丹波,等.血管生成与多囊卵巢综合征发病机制的关系[J].中国全科医学,2007,10(13):1121-1123.
[4]王凡,张正红,肖开转,等.下丘脑-垂体-肾上腺轴和下丘脑-垂体-卵巢轴在多囊卵巢综合征神经内分泌功能紊乱中的作用[J].中国医学科学院学报,2017,39(5):699-704.
[5]万赛英,谭峰,方美凤,等.电针干预高血压大鼠脑缺血血管内皮生长因子和促血管生成素1表达的研究[J].中华老年心脑血管病杂志,2010,12(2):173-176.
[6] RESHEF T A L, DAVID B S, RICHA RD V G, et al. Angiopoietin-1 and angiopoietin-2 are altered in polycystic ovarian syndrome during controlled ovarian stimulation[J]. Vascular cell,2013,5(1):18.
[7] OSZ K, ROSS M, PETRIK J.The thrombospondin-1 receptor CD36 is an important mediator of ovarian angiogenesis and folliculogenesis[J]. Reproductive Biology and Endocrinology, 2014(12):21.
[8] KAFALI H, IRIADAM M, OZARDAII I, et al. Letrozole-induced polycystic ovaries in the rat:a new model for cystic o varian disease[J]. Archives of Medecal Research, 2004,35(2):103-108.
[9]余曙光,徐斌.实验针灸学[M].北京:人民卫生出版社,2012:1.
[10]华兴邦,李辞蓉,周浩良,等.大鼠穴位图谱的研制[J].实验动物与动物实验,1991(1):1-5.
[11]曾仁宏,陈家旭.针灸辨证选穴之探讨[J].吉林中医药,2008,28(2):86-87.
[12]贺晓霞,王若光.平衡阴阳法对多囊卵巢综合征雌雄激素的调节[J].湖南中医药大学学报,2016,36(2):16-19.
[13]赵娜,杨正望,全春梅.多囊卵巢综合征临床分型与中医证型的相关性分析[J].湖南中医药大学学报,2017,37(9):966-968.
[14]施茵,虞莉青,尹小君.针灸治疗肥胖型多囊卵巢综合征的临床疗效观察[J].中华中医药学刊,2010,28(4):805-807.
[15]卓缘圆,吴家满,杨卓欣,等.“调任通督针刺法”治疗多囊卵巢综合征不孕症的临床疗效观察[J].中国针灸,2016,36(12):1237-1241.
[16] NOROOZZADEH M N M, BEHBOUDI-GANDEVANI S B S,ZADEH-VAKILI A Z A, et al. Hormone-induced rat model of polycystic ovary syndrome:A systematic review[J]. Life Sciences,2017,191:259-272.
[17]宫静,程凯,张露芬.电针不同穴位对去卵巢大鼠HPO轴的影响[J].中华中医药杂志,2011,26(3):595-597.
[18]王君,张宝文,于新宇,等.疾徐捻转泻法针刺足三里对高血压家兔的降压作用[J].中国临床康复,2006,10(7):116-117.
[19]杨正望,文炯,张增玲.补肾化瘀方对PCOS大鼠卵巢颗粒细胞CYP450表达的影响[J].湖南中医药大学学报,2011,31(1):30-32,39.
[20]孙洁,赵继梦,季蓉,等.电针对多囊卵巢综合征大鼠卵巢高雄激素症的影响[J].针刺研究,2013,38(6):465-472.
[21]林鸾.穴位埋线对PCOS-IR模型大鼠生殖内分泌和胰岛素抵抗的影响[D].广州:广州中医药大学,2016.
[22]黄玉秀,郑秀,刘林芳,等.TSP-1在子宫内膜腺癌中的表达及与VEGF、CD105的关系[J].中国医师杂志,2009,11(10):1301-1303.
[23]叶璐,冯亦军,马奔,等.VEGF-C和TSP-1在卵巢上皮癌中的表达及意义[J].中华临床医师杂志,2008,2(10):1142-1147.