Med J Chin PLA, Vol. 41, No. 3, March 1, 2016 175 [ ] 30 SD 5 (A) (B) (C) (D) (E) 0.206 0.514 1.028mg/(kg d) ELISA HE Masson (TGF- ) 9(MMP-9) C D E A B (P<0.05) (P<0.05) B C D E (P<0.05) D E B C D E TGF- (P<0.05) MMP-9(P<0.05) D E [ ] 9 [ ] R711 [ ] A [ ] 0577-7402(2016)03-0175-05 [DOI] 10.11855/j.issn.0577-7402.2016.03.01 Preventive effect of estradiol valerate on intrauterine adhesion formation in rats WANG Xian, HE Yuan-li *, Cai Hui-hua, LI Hui-juan, REN Xu-wen, HU Hai-yan, CHEN Si-ping, ZHANG Dong-mei Department of Obstetrics and Gynecology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China * Corresponding author, E-mail: heyuanli310@163.com This work was supported by the National Natural Science Foundation of China (81270658) [Abstract] Objective To investigate the preventive effect and the mechanism of estradiol on the formation of intrauterine adhesion in rats. Methods Thirty female SD rats were randomly assigned into 5 groups (6 each), and rats intrauterine adhesion model was reproduced by curettage and infection. Estradiol valerate was gavaged for prevention of adhesion. Rats in control group (A) were not given intrauterine injury or estradiol. In physiological estrogen group (B) intrauterine injury was reproduced, but no estradiol valerate was given. The rats in low dose estrogen group (C), moderate dose estrogen group (D) and high dose estrogen group (E) were treated with estradiol valerate of 0.206, 0.514 or 1.028mg/(kg d) respectively after reproduction of the model. ELISA was used to determine the serum concentrations of estradiol. HE staining was performed to observe the number of endometrial glands in rats. Masson staining was used to calculate the area of endometrial fibrosis, and immunohistochemical method was employed to detect the expressions of matrix metalloproteinase 9 (MMP-9) and transforming growth factor beta (TGF- ). Results On account of administration of estradiol valerate, the serum estradiol levels were significantly higher in groupc,dandethan in group A and B, and the serum estradiol levels increased along with the increase of dosage of estradiol (P<0.05). Compared with group B, rats in group C, D and E presented increased number of endometrial glands and decreased ratio of endometrial fibrosis area (P<0.05), but no significant difference was found between group D and group E. Compare to group B, rats in Group C, DandE showed higher MMP-9 levels and lower TGF- levels (P<0.05), but no significant difference was found between group D and group E. Conclusion Estradiol may reduce the formation of intrauterine adhesion, and the effect is positively correlated with the dosage of estradiol used. [Key words] intrauterine adhesion; estradiol valerate; transforming growth factor beta; matrix metalloproteinase 9; fibrosis [ ] (81270658) [ ] [ ] 510282 ( ) [ ] E-mail heyuanli310@163.com
176 2016 3 1 41 3 [1] [2] 1000 19% [1,3] [4] [1] (TGF- ) 9(MMP-9) 1 1.1 (LPS) Sigma ELISA Masson TGF- Bio-Rad MMP-9 Abcam 1.2 10 6mg/L 4 24h 10% 0.35ml/100g 5mm 4mm 24h 7d [5] 1.3 8~10 SD 30 [ SCXK( )2011-0015] 250~280g 5 ( 6 Y 12 ) (A ) (B ) [5] (C ) 0.206mg/ (kg d) (D ) 0.514mg/(kg d) (E ) 1.028mg/(kg d) [6] 60kg 2 5 10mg/d 24d HE Masson TGF- MMP-9 1.4 24d 1500r/min 10min ELISA 1.5 HE Masson HE5 Masson 5 = / 3 Image Pro Plus 6.0 1.6 TGF- MMP-9 5 3Image Pro Plus 6.0 1.7 SPSS 20.0 x±s
Med J Chin PLA, Vol. 41, No. 3, March 1, 2016 177 (One-Way ANOVA) LSD-t P<0.05 (6.33±0.58) E (5.67±0.58) A B (P<0.01) D EC D E( 1) 2 2.1 C (15.9436±0.5221ng/L) D (20.5099±0.6250ng/L) E (24.8740±0.8988ng/L) A (12.6367±0.5319ng/L) B (13.3493±0.8767ng/L) C D E (P<0.05) A B (P=0.954) 2.2 A (0.2422±0.0191) B (0.9519±0.0319) C 2.3 TGF- MMP-9 A (0.0046± 0.0006) TGF- (P<0.001) B (0.1018±0.0189)TGF- C (0.0684±0.0062) D (0.0457±0.0030) E (0.0444±0.0022) ( P <0.001) D E TGF- (P=0.864) C (0.0448±0.0036) D (0.0896±0.060) E (0.0938±0.0066) MMP-9 A (0.0220±0.0093) B (0.0200±0.0032) (P<0.001) D E A B (P=0.698 2) (0.6237±0.0178) D (0.4207±0.0069) E (0.4449±0.0190) (P<0.01) C D E B ( P <0.01) D E ( P =0.180) A (8.67±0.58) B (1.67±0.58) (P<0.01) C (4.33±0.58) D 3 25%~30% Group A Group B Group C Group D Group E Masson HE 1 Masson HE ( 400) Fig.1 Masson and HE staining of endometrium in each group of rats ( 400) Group A Group B Group C Group D Group E TGF- MMP-9 2 TGF- MMP-9 ( 400) Fig.2 Expressions of TGF- and MMP-9 in endometrium of each group of rats (Immunohistochemistry 400)
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