Application of mir-509-3p expression inhibitor in preparation of medicine for treating polycystic ovary syndrome

A technology for inhibitors and synthesizing drugs, applied in the field of biomedicine

Inactive Publication Date: 2016-09-14
YANTAI YUHUANGDING HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are no related studies on miRNAs in granulosa cells of PCOS patients

Method used

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  • Application of mir-509-3p expression inhibitor in preparation of medicine for treating polycystic ovary syndrome
  • Application of mir-509-3p expression inhibitor in preparation of medicine for treating polycystic ovary syndrome
  • Application of mir-509-3p expression inhibitor in preparation of medicine for treating polycystic ovary syndrome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Morphological differences in the development of granulosa cells and oocytes between PCOS patients and women of normal childbearing age in Example 1

[0030] PCOS patients, especially PCOS patients with obesity, have abnormal oocyte morphology, thick cytoplasmic granules, and uneven thickness of the zona pellucida, resulting in low fertilization rate, uneven blastomere size and fragmented embryos after fertilization. More, slow cleavage speed or stagnant embryonic development, high-quality embryos cannot be obtained, which seriously affects the treatment outcome of these patients ( figure 1 ).

Embodiment 2

[0031] Example 2 Differential expression of miR-509-3p in granulosa cells of PCOS patients and normal women.

[0032] Total RNA was extracted by Trizol method, and after reverse transcription, real-time quantitative PCR was used to amplify and measure miRNA expression. The TaqMan probe method was used, and the reagents were primers and kits commercialized by Applied Biosystems (ABI) (reverse kit: name FG, Taqman (r) microRNA RT Kit200rxns, article number 4366596; PCR kit: name TaqMan GeneExpression Master Mix, cat. no. 4369016; reverse primer and PCR primer: name MicroRNA Assays50RT+150PCR, Cat. No. 4427975).

[0033] Eight cases of newly diagnosed PCOS patients and eight normal women were collected, and the expression of miR-509-3p in granulosa cells was detected. It was found that the expression of miR-509-3p in PCOS patients was significantly higher than that in normal women, with a statistical difference (4.6±0.13vs1. 0±0.03, p figure 2 ).

Embodiment 3

[0034] Example 3 Dual-luciferase reporter gene system verification of the predicted target gene of miR-7

[0035] The target genes of miR-7 were predicted by Targetscan, PicTar and miRanda software. According to whether the biological functions of these target genes were related to hormone synthesis and ovulation mechanism, PTGS1 was selected as the target gene of miR-509-3p for verification.

[0036] Clone the 3'-UTR sequence of the predicted target gene PTGS1 of miR-509-3p, and add restriction sites XbaI (5'-TCTAGA-3') and NotI (5'-GCGGCCGC-3') at both ends of the primer . The primer sequences are as follows:

[0037]

[0038] The pmiRGLO vector (purchased from Promega) and the amplified sequence were subjected to XbaI and NotI double enzyme digestion, respectively. After the target fragments were recovered, the respective amplified fragments were connected to the vector to construct a dual luciferase reporter gene vector carrying the PTGS1 3'UTR region.

[0039] The c...

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Abstract

The invention discloses the application of miR-509-3p expression inhibitor in the preparation of medicine for treating polycystic ovary syndrome. The present invention also provides a polycystic ovary syndrome drug containing an miR-509-3p expression inhibitor. After inhibiting the expression of miR‑509‑3p in granulosa cells, mRNA levels of prostaglandin endoperoxide synthase 1 (PTGS1) were upregulated, prostaglandin synthesis was increased, and tissue plasminogen activator activity was increased in the ovary, thereby Trigger ovulation. It shows that miR-509-3p expression inhibitor can affect the synthesis of prostaglandins and ovulation by regulating the expression of PTGS1 gene, so as to play a role in the treatment of abnormal ovulation in PCOS.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the application of miR-509-3p expression inhibitors in the preparation of medicines for treating PCOS. Background technique [0002] Polycystic ovary syndrome (PCOS) is a common endocrine disorder in women of childbearing age. For more than 70 years, it has been one of the diseases concerned by reproductive endocrinologists at home and abroad. In 2003, ESHRE / ASRM established diagnostic criteria for PCOS in Rotterdam, the Netherlands: ① Anovulation or rare ovulation; ② Clinical and biochemical features of hyperandrogenism; ③ Ultrasound examination showed polycystic changes in the ovaries and excluded other androgen abnormalities. Multiple related diseases, such as congenital adrenal hyperplasia (CAH), Cushing's syndrome, hyperprolactinemia, severe insulin resistance syndrome, androgen-secreting tumors and (or) thyroid dysfunction, etc. According to this standard, we conduc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61P15/08
Inventor 黄鑫郝翠芳包洪初丛建香
Owner YANTAI YUHUANGDING HOSPITAL
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