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Mechanism of MiR-296-3p Regulating Granulosa Cell Apoptosis in Rats with Polycystic Ovary Syndrome |
BAI Xue1,YU Xin-yue1,*(),ZHEN Chun-yang2,HOU Cong3,LI Min4 |
1. General Hospital of Northern Theater Command, Shenyang 110052, China 2. Shenyang Jinghua Hospital Co., Ltd., Shenyang 110001, China 3. Liaoning Huizhi Tongda Biological Science Co., Ltd., Shenyang 110034, China 4. Beijing Zhongyuan Co., Ltd., Beijing 100016, China |
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Abstract Objective: To investigate the effect of miR-296-3p on granulosa cell apoptosis in rats with polycystic ovary syndrome (PCOS). Methods: The levels of estradiol (E2), progesterone (P) and testosterone (T) in serum of PCOS rats were detected by automatic chemiluminescence immunoassay; HE staining was used to detect the pathological changes of the ovary; TUNEL was used to detect apoptosis in the ovary; The expression level of miR-296-3p and PRKCA in the ovary was detected by qPCR; Ovarian granulosa cells (GCs) were divided into miR-NC, miR-296-3p mimic, miR-296-3p inhibitor, miR-296-3p mimic+Vector, miR-296-3p mimic+PRKCA and miR-296-3p mimic+PRKCA+NF-κB groups after transfection of various plasmids, and untreated GCs were used as control group. The expression level of p38, p-p38 and NF-κB was detected by Western blot. StarBase website and double luciferase report test predicted and verified the targeting relationship between miR-296-3p and PRKCA, respectively. Flow cytometry was used to detect cell apoptosis rate. Results: Compared with the control group, the levels of E2, P and T in the ovaries of PCOS rats increased; The proportion of apoptosis increased; The expression of miR-296-3p increased. miR-296-3p has a targeting relationship with PRKCA, and there are binding sites in the 3'UTR region. Compared with miR-NC group, the expression level of PRKCA mRNA in miR-296-3p mimic group was significantly decreased, and p-p38/p38 and NF-κB expression was up-regulated. Compared with miR-NC group, the expression level of PRKCA mRNA in miR-296-3p inhibitor group was significantly increased, and p-p38/p38 and NF-κB expression was down-regulated. Compared with the Mir-NC group, the percentage of apoptosis of miR-296-3p mimic and miR-296-3p mimic+Vector groups was significantly up-regulated. Compared with miR-296-3p mimic and miR-29-3p mimic+Vector, the percentage of apoptosis of ovarian granulocyte cells in miR-296-3p mimic+PRKCA group was significantly down-regulated. Compared with miR-296-3p mimic+PRKCA+NF-κB group, the percentage of apoptosis of ovarian granulosa cells in miR-296-3p mimic+PRKCA+NF-κB group was significantly up-regulated. Conclusion: MiR-296-3p activates p38 MAPK/NF-κB signal pathway by targeting PRKCA and mediating granulosa cell apoptosis in rats with polycystic ovary syndrome.
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Received: 02 March 2023
Published: 04 May 2023
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