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雷公藤多苷片对Ⅱ型胶原诱导性关节炎雌性大鼠生殖毒性的影响

[Effect of Tripterygium Glycosides Tablets on reproductive toxicity in female rats with Ⅱ type collagen induced arthritis].

作者信息

Fan Yuan-Fang, Xu Ying, Su Xiao-Hui, Liu Li-Ling, Tian Ya-Ge, Zhao Yuan, Kong Xiang-Ying, Lin Na

机构信息

Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences Beijing 100700,China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2019 Aug;44(16):3486-3493. doi: 10.19540/j.cnki.cjcmm.20190523.402.

Abstract

The aim of this paper was to observe the toxic effect of Tripterygium Glycosides Tablets( TG) on the reproductive system of Ⅱ type collagen induced arthritis( CIA) male rats,and to explore the toxic mechanism preliminarily. Fifty SD rats were randomly divided into normal control group( Con),model group( CIA),Tripterygium Glycosides Tablets clinical equivalent dose groups of 1,2,4 times( 9,18,36 mg·kg-1),10 rats in each group,and were given by gavage once a day for 42 days after the first immunization.The organ indexes of uterine and ovarian were calculated on days 21 and 42. Histopathological and morphological changes of uterine and ovarian were observed under optical microscope. The concentration of estradiol( E2),follicle-stimulating hormone( FSH),luteinizing hormone( LH),17α-hydroxylase( CYP17 A1) and cytochrome P450 19 A1( CYP19 A1) in serum were detected by ELISA. Immunohistochemistry was used to observe the expression of Bax and Bcl-2 related proteins in the apoptosis pathway of uterus and ovary. The results showed that compared with the Con group,CIA group could reduce the number of uterine glands( P<0.05),but no significant changes were observed in other groups. Compared with the CIA group,there were no significant changes in the coefficients of uterus and ovary in the Tripterygium Glycosides Tablets groups. The number of uterine glands,total follicles in the ovary,mature follicles and corpus luteum,the distribution of blood vessels and mitochondria had a certain inhibitory trend,and also slightly increased the number of atresia follicles,but the histopathological quantitative indicators were not statistically different. Except that 2 times clinical dose of Tripterygium Glycosides Tablets could significantly reduce the content of CYP19 A1( P<0. 05) after 42 d administration,there were no significant changes in serum estrogen E2,FSH,LH and estrogen synthesis key enzymes CYP17 A1 in each administration group. Medium and high doses of Tripterygium Glycosides Tablets could increase the expression of apoptotic protein Bax in uterine and ovarian tissues( P<0. 05,P<0. 01),and all the administration groups could inhibit the expression of apoptotic inhibiting protein Bcl-2( P <0. 05,P<0. 01,P<0.001),42 d was more obvious than 21 d. In conclusion,4 times and less than 4 times Tripterygium Glycosides Tablets did not cause obvious toxicity and histopathological changes in the reproductive organs of CIA rats,but it could reduce the level of serum estrogen synthesis key enzyme CYP19 A1 and affect the content of apoptosis-related proteins Bax and Bcl-2 in uterus and ovary tissues. The relevant mechanism needs further study.

摘要

本文旨在观察雷公藤多苷片(TG)对Ⅱ型胶原诱导性关节炎(CIA)雄性大鼠生殖系统的毒性作用,并初步探讨其毒性机制。将50只SD大鼠随机分为正常对照组(Con)、模型组(CIA)、雷公藤多苷片临床等效剂量1倍、2倍、4倍组(9、18、36 mg·kg-1),每组10只,首次免疫后每天灌胃给药1次,连续给药42天。于第21天和第42天计算子宫和卵巢的脏器指数。光镜下观察子宫和卵巢的组织病理学及形态学变化。采用酶联免疫吸附测定法(ELISA)检测血清雌二醇(E2)、卵泡刺激素(FSH)、黄体生成素(LH)、17α-羟化酶(CYP17 A1)和细胞色素P450 19 A1(CYP19 A1)的浓度。采用免疫组织化学法观察子宫和卵巢凋亡途径中Bax和Bcl-2相关蛋白的表达。结果显示,与Con组比较,CIA组子宫腺数量减少(P<0.05),其他组未见明显变化。与CIA组比较,雷公藤多苷片各剂量组子宫和卵巢系数未见明显变化。子宫腺数量、卵巢中卵泡总数、成熟卵泡及黄体数量、血管及线粒体分布有一定抑制趋势,闭锁卵泡数量略有增加,但组织病理学定量指标差异无统计学意义。各给药组血清雌激素E2、FSH、LH及雌激素合成关键酶CYP17 A1除雷公藤多苷片2倍临床剂量给药42天后可使CYP19 A1含量显著降低(P<0.05)外,未见明显变化。中、高剂量雷公藤多苷片可使子宫和卵巢组织凋亡蛋白Bax表达增加(P<0.05,P<0.01),各给药组均可抑制凋亡抑制蛋白Bcl-2表达(P<0.05,P<0.01,P<0.001),第42天较第21天更明显。综上所述,4倍及以下剂量雷公藤多苷片对CIA大鼠生殖器官未引起明显毒性及组织病理学改变,但可降低血清雌激素合成关键酶CYP19 A1水平,影响子宫和卵巢组织凋亡相关蛋白Bax和Bcl-2含量,其相关机制有待进一步研究。

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