Expression of transforming growth factor-β and Smad4 in ovarian granulosa cells in menopausal transitional rat

doi:10.16098/j.issn.0529-1356.2018.01.018

Abstract

Abstract:

Objective To investigate the expression of transforming growth factor beta(TGF-β) and Smad4 in ovarian granulosa cells of rats during menopausal transition and to analyze their relationship with the decline of ovarian function. Methods Female SD rats were divided into control group (group C, 6 months old, vaginal smear screening, n=9) and menopausal transition(MT) group (MT group, 12-14 months old, vaginal smear screening, n=8), rats were sacrificed quickly after ovariectomy. The ovarian granulosa cells were released by mechanical separation method, and cultured in CO₂ incubator. The expression of follicle-stimulating hormone receptor(FSHR) protein was observed by immunocytochemistry to identify ovarian granulosa cells. Immunohistochemistry was used to detect the expression of TGF-β and Smad4 protein in ovarian granulosa cells and images were collected and analyzed; The expression of TGF-β and Smad4 mRNA in granulosa cells was detected by Real-time PCR. Results Immunocytochemistry showed that the purity of granulosa cells was > 95%. The intensities of immunocytochemical positive staining for TGF-β and Smad4 in MT group was lower than those in normal group (P<0.05).Real-time PCR showed that both TGF-β and Smad4 mRNA were expressed. The amount of Real-time PCR for TGF-β and Smad4 mRNA was significantly lower than those of the control group (P<0.05). Conclusion TGF-β and Smad4 may be involved in the decline of ovarian function in rats with menopausal transition. The causes of the functional decline in postmenopausal rats may be partly related to the decrease of TGF-β and Smad4 expression in ovarian granulosa cells.

Key words: Menopausal transitional, Ovarian granulosa cells, Transforming growth factor-β, Smad4, Immunohis tochemistry, Real-time , PCR, Rat

YUAN Li-juan REN Jun-xu JI Hong-yu ZHANG Jing ZHANG Juan DING Yan. Expression of transforming growth factor-β and Smad4 in ovarian granulosa cells in menopausal transitional rat[J]. Acta Anatomica Sinica, 2018, 49(1): 108-112.

References

［1］Devine K, Mumford SL,Wu M, et al. Diminished ovarian reserve in the United States assisted reproductive technonlogy poplulation:diagnosite trend among 181,536 cycles from the Socitey for Assisted Reproductive Technology Clinic Outcomes Reporting System［J］. Fertil Steril, 2015, 104(3): 612-619.

［2］Nie ChF, Lu FM, Huang QY. Serum anti-Mullerian hormone and sex hormone test the clinical value of diagnosis of premature ovarian failure［J］.Shenzhen Journal of Integrated Traditional Chinese and Western Medicine, 2016, 26(4):87-88. (in Chinese)

聂超凤，卢芳梅，黄秋云. 血清抗苗勒管激素及性激素检测对卵巢早衰诊断预测的临床价值［J］.深圳中西医结合杂志， 2016, 26(4):87-88.

［3］Li Q, Zhang JH, Tian XW, et al.Ovarian function decline in patients with serum anti-Mullerian hormone level change and significance［J］.Shandong Medical Journal, 2016, 56(21):63-64. (in Chinese)

李强，张建海，田向文，等.卵巢功能衰退患者血清抗苗勒管激素的水平变化及意义［J］. 山东医药，2016, 56(21):63-64.

［4］Ye J, Ding T, Wang ShX. Advances in clinical diagnosis and treatment of ovarian aging［J］.Progress in Obstetetrics and Gynecology, 2013, 22(5):419-421.

［5］Shen ChY, Dong JX, Xu J. Smad3 promotes the autophagy of granulosa cells in rats［J］. Acta Anatomica Sinica, 2014, 45(1):109-113. (in Chinese)

申春艳，董静霞，徐建. Smad3促进大鼠卵巢颗粒细胞自噬［J］.解剖学报,2014,45(1):109-113

［6] Wan M， Li C， Zhen G， et al. Injury-activated transforming growth factor beta controls mobilization of mesenchymal stem cells for tissue remodeling［J］. Stem Cells，2012，30（11）：2498-2511.

［7］Liu Y，Chen X, Xue X, et al. Effects of Smad3 on the proliferation and steroidogenesis in human ovarian luteinized granulosa cells［J］. IUBMB,2014, 66(6):424-437.

［8］Miao X, Luo Q, Zhao H, et al. Co-expression analysis and identification of fecundity-related long non-coding RNAs in sheep ovaries［J］.SCI Rep, 2016, 6(1):393-398.

［9］Muhammad WS, Lena K, Kostas S. Genetic alterations in pancreaticcancer［J］. Gastroenterology, 2007, 13(33): 4423-4430.

［10］Wang X, Gao JL, Zhao MM, et al. Inhibition of bone marrow mesenchymal stem cells on epithelial mesenchymal transformation induced by transforming growth factor-β1［J］. Acta Anatomica Sinica, 2016, 47(6):765-762. (in Chinese)

王鑫, 高俊玲, 赵曼曼，等. 骨髓间充质干细胞抑制转化生长因子TGF-β1 诱导的肺泡Ⅱ型上皮细胞间质化［J］.解剖学报, 2016, 47(6):756-762.

［11］Huang S, Zhang F, Miao L, et al Lentiviral-mediated Smad4 RNAi induced anti-proliferation by pl6 up-regulation and apoptosis by caspase 3 down-regulation in hepatoma SMMC-7721 cells［J］. Oncol Rep, 2008, 20(5): 1053-1059.

［12］Deckers M, Dinther M, Buijs J.The tumor suppreessor Smad4 is required for transforming frowth factor beta-induced epithelial to mesenchymal transition and bone metastasis of breast cancer cells［J］. Cancer Res, 2006, 66(4): 2202-2209.

［13］Wang ZP，Mu YX, Guo M, et al.Transforming growth factor-signaling participates in the maintenance of primordial follicle pool in the mouse ovary［J］.J Biol Chem, 2014（3）21：8299-8311.

［14］Pangas SA, Li X, Robertson EJ, et al. Premature luteinization and cumulus cell defects in ovarian-specific Smad4 knockout mice［J］. Molecular Endocrinology, 2006, 20(6): 1406-1422.

[1]	YANG Meng-yi NIU Shi-bo ZHANG Jing DANG Jie MA Zhan-bing LU Hong HUO Zheng-hao. Association between index finger and ring finger length ratios and polymorphism of homeobox A11 gene locus among Ningxia college students [J]. Acta Anatomica Sinica, 2024, 55(1): 62-66.
[2]	XIE Hui-lan FANG Fang LIN Yi. Mechanism of Chir99021 regulating Wnt/β-catenin signaling pathway inhibiting osteogenic differentiation of rat dental pulp stem cells [J]. Acta Anatomica Sinica, 2024, 55(1): 67-72.
[3]	WEI Xi-xi LI Chao-hong ZHAO Chen-lu TANG Jia-ping LIU Yu-zhen. Characterization of group Ⅰ metabotropic glutamate receptors in rat superior cervical ganglion and their changes following chronic intermittent hypoxia [J]. Acta Anatomica Sinica, 2024, 55(1): 3-9.
[4]	MA Ting-ting CHEN Jian-hong LIU Ai-cui LI Hai-ning. Role of inhibiting lncRNA TUG1 to down-regulate nucleotide binding oligomerization domain like receptor protein 1 inflammasome in delaying the progression of Alzheimer’s disease [J]. Acta Anatomica Sinica, 2024, 55(1): 32-42.
[5]	ZHENG Li-ping LIU Xia DU Xiao-hua WU Ya-juan LIU Shan-shan . Expression and distribution of brain-derived neurotrophic factor in different cerebrum regions of yak and cattle [J]. Acta Anatomica Sinica, 2024, 55(1): 10-16.
[6]	SU Peng WANG Xing-yi LIANG Jing-yan XIONG Tian-qing. Optimization and identification of a low density and high purity method for primary hippocampal neuron culture from fetal rats [J]. Acta Anatomica Sinica, 2024, 55(1): 113-119.
[7]	HAO Yong-ming GUO Lei ZHOU Cheng-hui PEI Han-jun LI Li ZHAO Zhe . Establishment of a rat model of myocardial hypertrophy by a modified abdominal aortic coarctation method [J]. Acta Anatomica Sinica, 2024, 55(1): 120-124.
[8]	XU Ya-ping YU Yue-xin CHEN Jin-fu WANG Ke-ke GUO Zhikun . Structural distribution and mechanism of elastic fibers and collagen fibers in ventricular interstitium of aged rats [J]. Acta Anatomica Sinica, 2023, 54(6): 716-721.
[9]	LIU Li-ping ZHU Meng-ni XU Hui . Effect of interleukin-6 on nucleated erythrocytes in lipopolysaccharide induced preeclampsia rats [J]. Acta Anatomica Sinica, 2023, 54(6): 722-729.
[10]	ZHAO Wei-ming LI Xiao YU Guo-ying WANG Gai-ping CHANG Cui-fang . Effect of serine protease inhibitor Kazaltype 1 on the proliferation of hepatocellular carcinoma cell and its mechanism [J]. Acta Anatomica Sinica, 2023, 54(6): 695-702.
[11]	WANG Wen-juan DING Yu-tong SU Hao REN Jie SUN Yan-rong WANG Han-fei LU Jia-li ZHANG Lin-qian BAI Yu QIN Li-hua. Changes of Nogo-A expression in hippocampus and striatum of rats under low estrogen condition [J]. Acta Anatomica Sinica, 2023, 54(6): 620-627.
[12]	ZHANG Qin YANG Jun-xia JIANG Qing-song. Effect of fosinopril on angiotensin converting enzyme 2 and vascular endothelial growth factor in diabetic retinopathy mice [J]. Acta Anatomica Sinica, 2023, 54(6): 628-634.
[13]	LIANG Pan-pan YU Ai-mei DU Jing KOU Wen-hui WANG Huan-huan SONG Ai-xia. Inhibitory effect of sodium ferulate on inflammatory response in migraine rats based on c-Jun N-terminal kinase/p38 mitogen-activated protein kinase signaling pathway [J]. Acta Anatomica Sinica, 2023, 54(6): 652-659.
[14]	YAO Su-hua YAN Jun XIE Fang LIN Chun-hua HUANG Zheng-chun. Nerve repair effect of olanzapine-mediated cyclic AMP response element binding protein/brain-derived neurotrophic factor/receptor tyrosine kinase receptors B pathway on schizophrenic rats [J]. Acta Anatomica Sinica, 2023, 54(6): 660-667.
[15]	XU Ying LIU Bin ZHENG Xing HE Zong-zhao. Role and mechanism of complement C3a receptor in the cognitive impairment of septic rats induced by cecal ligation and perforation [J]. Acta Anatomica Sinica, 2023, 54(6): 668-675.