發(fā)布時間：2018-03-13 04:38

  本文選題：高雄激素血癥　切入點：雄激素受體　出處：《醫(yī)學研究生學報》2017年05期 　論文類型：期刊論文

【摘要】：目的雄激素信號通路參與調(diào)控卵泡早期生長發(fā)育以及卵泡閉鎖過程,而性激素結(jié)合球蛋白是調(diào)控卵巢局部雄激素水平的重要因素。文中探討雄激素對卵巢顆粒細胞中性激素結(jié)合球蛋白(SHBG)表達的影響。方法 1培養(yǎng)人卵巢顆粒癌細胞株(KGN),分別給予含0 nmol/L DHT,500 nmol/L DHT,500 nmol/L DHT+60μmol/L flutamide的細胞培養(yǎng)液,提取細胞蛋白。用Western blot、免疫熒光等方法檢測SHBG的表達。2脫氫表雄酮(DHEA)誘導高雄激素血癥模型,12只雌性SD大鼠隨機數(shù)字表法分為HA組和對照組,每組6只。HA組每只皮下注射DHEA 6 mg/(100 g·d)溶于0.2 m L實驗級大豆油,對照組給予等體積的大豆油。連續(xù)注射35 d后腹腔注射5%水合氯醛麻醉各組大鼠,下腔靜脈取各組大鼠血液,ELISA方法檢測各組血清中SHBG的水平;剪切各組大鼠卵巢和部分肝,免疫組化方法檢測卵巢顆粒細胞中SHBG的表達,Western blot方法檢測肝雄激素受體(AR)及SHBG表達情況。結(jié)果與0nmol/L DHT處理24h AR表達(1.06±0.03)相比,300、400、500nmol/L DHT處理后(1.06±0.02、1.61±0.11、2.38±0.14)均升高(P0.05);SHBG與AR表達變化一致,隨DHT濃度梯度增加表達升高,但在500 nmol/L DHT處理24 h后SHBG表達升高顯著(P0.01)。與0 nmol/L DHT比較,500 nmol/L DHT處理后AR蛋白、SHBG蛋白表達均升高(P0.01);與500 nmol/L DHT比較,500 nmol/L DHT+60μmol/L flutamide處理后AR蛋白、SHBG蛋白表達均降低(P0.05)。免疫熒光結(jié)果表明,與0 nmol/L DHT相比,DHT促進AR的表達,同時加入flutamide可明顯抑制AR的表達。SHBG的表達與AR相一致。卵巢組織HE染色結(jié)果表明HA組與對照組相比卵巢形態(tài)發(fā)生改變,表現(xiàn)為多囊卵巢;卵巢免疫組化結(jié)果表明SHBG在HA組中表達高于對照組。HA組血清SHBG表達低于對照組[(2.41±0.14)vs(4.80±0.35),P0.01]。HA組中肝SHBG表達表達低于對照組,而AR蛋白表達高于對照組(P0.05)。結(jié)論雄激素信號通路激活后SHBG在卵巢顆粒細胞癌細胞以及大鼠卵巢顆粒細胞中表達升高。
[Abstract]:Objective androgen signaling pathway is involved in early follicular growth and development and follicular atresia. The effect of androgen on the expression of SHBG in ovarian granulosa cells was studied. Methods 1. The cell culture medium containing 0 nmol/L DHT 500 nmol/L nmol/L DHT 60 渭 mol/L flutamide was used. Cell proteins were extracted. 12 female SD rats were randomly divided into HA group and control group by Western blot2 and immunofluorescence. 2. Dehydroepiandrosterone (DHEA) induced hyperandrogenemia model. 6 rats in each group were injected subcutaneously with DHEA 6 mg/(100 g 路d) dissolved in 0.2ml soybean oil. The control group was given soybean oil of the same volume. After 35 days of continuous injection, 5% chloral hydrate was injected intraperitoneally to anesthetized rats in each group. The levels of SHBG in serum of each group were detected by Elisa, and the ovary and partial liver of each group were cut off. The expression of SHBG and SHBG in granulosa cells were detected by immunohistochemical method. Results compared with 0 nmol / L DHT treatment for 24 h, the expression of AR was 1.06 鹵0.03). Compared with 0 nmol / L DHT, the expression of AR was 1.06 鹵0.02nmol / L DHT (1.06 鹵0.02nmol / L) and 1.06 鹵0.02nmol / L DHT + 0.112.38 鹵0.14). The expression increased with the increase of DHT concentration gradient. However, after treatment with 500 nmol/L DHT for 24 h, the expression of SHBG increased significantly (P0.01G). Compared with 0 nmol/L DHT, the expression of AR protein of nmol/L DHT was significantly higher than that of 500 nmol/L DHT, and the expression of AR protein was significantly decreased after treatment of 500 nmol/L DHT with 500 nmol/L DHT 60 渭 mol/L flutamide. The results of immunofluorescence showed that, after treatment with 500 nmol/L DHT, the expression of SHBG was significantly higher than that of 0 nmol/L DHT. Compared with 0 nmol/L DHT, the expression of AR was enhanced by flutamide, and the expression of SHBG was significantly inhibited by flutamide. The results of HE staining in ovarian tissue showed that the morphology of ovary in HA group was changed compared with that in control group, which showed polycystic ovary. The expression of SHBG in HA group was higher than that in control group. The expression of serum SHBG in HA group was lower than that in control group [2.41 鹵0.14 vs 4.80 鹵0.35 P0.01] .The expression of liver SHBG in HA group was lower than that in control group. The expression of AR protein was higher than that of the control group P0.050.Conclusion the expression of SHBG in ovarian granulosa cell carcinoma and rat ovarian granulosa cells is increased after androgen signaling pathway activation.
【作者單位】： 南京大學醫(yī)學院江蘇省醫(yī)學分子技術(shù)重點實驗室;
【基金】：國家自然科學基金(81471422)
【分類號】：R711.75

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