發(fā)布時(shí)間：2018-06-01 14:07

  本文選題：丙烯酰胺 + 睪丸�。� 參考：《重慶醫(yī)科大學(xué)》2013年碩士論文

【摘要】：目的 研究丙烯酰胺（acrylamide,AA）對(duì)大鼠睪丸組織的亞急性毒性作用，及其對(duì)增殖細(xì)胞核抗原（proliferating cell nuclear antigen，PCNA）和表皮生長(zhǎng)因子受體（epidermal growth factor receptor，，EGFR）蛋白表達(dá)的影響，初步探討丙烯酰胺生殖毒性的可能作用機(jī)制。 方法 將32只5～6周齡雄性SD大鼠隨機(jī)分為4組，每組8只，分別為空白對(duì)照組，4mg/kg、12mg/kg和36mg/kg劑量丙烯酰胺染毒組，連續(xù)灌胃染毒30天。處理結(jié)束后，心臟取血，化學(xué)發(fā)光法檢測(cè)血清中睪酮（testosterone，T）和雌二醇（estradiol，E2）的含量,取雙側(cè)睪丸組織，HE染色觀察睪丸組織病理學(xué)變化，TUNEL檢測(cè)睪丸細(xì)胞凋亡情況，RT-PCR檢測(cè)睪丸組織PCNA、EGFR mRNA的表達(dá),免疫組化和Westernblot檢測(cè)睪丸組織PCNA、EGFR蛋白的表達(dá)。 結(jié)果 與對(duì)照組比較，12、36mg/kg染毒組大鼠體重明顯下降（P0.05）,36mg/kg染毒組大鼠睪丸臟器系數(shù)升高（P0.05）；HE染色顯示，12、36mg/kg染毒組大鼠睪丸組織出現(xiàn)了明顯的病理形態(tài)學(xué)改變，具體表現(xiàn)為生精細(xì)胞減少、生精上皮萎縮、管腔出現(xiàn)空泡等；TUNEL顯示12、36mg/kg染毒組睪丸凋亡細(xì)胞顯著增多（P0.05）；化學(xué)發(fā)光法顯示，各染毒組大鼠血清T濃度呈劑量依賴型下降（P0.05），12、36mg/kg染毒組大鼠血清E2濃度下降（P0.05）；RT-PCR顯示，12、36mg/kg染毒組大鼠睪丸組織PCNA和EGFR的mRNA表達(dá)均明顯降低（P0.05）；免疫組化和Western blot顯示，12、36mg/kg染毒組大鼠睪丸組織PCNA的蛋白表達(dá)明顯降低（P0.05），各染毒組大鼠睪丸組織EGFR的蛋白表達(dá)明顯降低（P0.05），差異具有統(tǒng)計(jì)學(xué)意義。 結(jié)論 一定劑量的丙烯酰胺染毒可導(dǎo)致大鼠睪丸組織發(fā)生病理學(xué)變化，凋亡細(xì)胞增多。PCNA、EGFR蛋白在大鼠睪丸組織中的表達(dá)減少，在一定程度上可抑制生精細(xì)胞DNA的合成，打亂睪丸生精細(xì)胞的分裂、增殖與凋亡之間的動(dòng)態(tài)平衡，擾亂性激素水平，最終導(dǎo)致大鼠不同程度的生殖毒性，為丙烯酰胺生殖毒性的機(jī)制研究提供實(shí)驗(yàn)依據(jù)。
[Abstract]:Purpose To study the subacute toxicity of acrylamide acrylic acid (AA) to testis of rats and its effect on the expression of proliferating cell nuclear antigen-PCNA (PCNA) and epidermal growth factor receptor (EGFR) protein of proliferating cell nuclear antigen (PCNA) and epidermal growth factor receptor (EGFR). To explore the possible mechanism of reproductive toxicity of acrylamide. Method Thirty-two 5-week-old male SD rats were randomly divided into 4 groups (n = 8 in each group). The rats in the control group (n = 8) were treated with 4 mg 路kg ~ (-1) of 36mg/kg and 12 mg / kg / kg of 36mg/kg respectively, and the rats were given intragastric administration for 30 days. After the treatment, the blood was taken from the heart, and the contents of testosterone Tand estradiolone E2) in serum were detected by chemiluminescence method. The histopathological changes of testis were observed by HE staining in bilateral testis. The apoptosis of testicular cells was detected by Tunel. The expression of PCNA- EGFR mRNA in testicular tissues was detected by RT-PCR, and the expression of PCNA- EGFR protein was detected by immunohistochemistry and Westernblot. Result Compared with the control group, the weight of the rats in the 125mg / kg group decreased significantly (P 0.05 mg / kg) and the testicular organ coefficient increased. The HE staining showed that the testicular tissues of the rats exposed to 123mg / kg P0.05mg / kg showed obvious pathological morphology changes, and the spermatocyte decreased. Testicular apoptotic cells were significantly increased in 12 ~ 36mg / kg group with atrophy of spermatogenic epithelium and vacuole in the lumen, and showed by chemiluminescent method. Serum T concentration decreased in a dose-dependent manner in each exposure group (P 0.05 mg / kg). Serum E _ 2 concentration decreased in the rats of each group. RT-PCR showed that the mRNA expression of PCNA and EGFR in testis of rats exposed to 12 ~ 36mg / kg was significantly lower than that of the control group, and the expression of mRNA in testis was significantly decreased by immunohistochemistry and Western blot, and the results of Western blot showed that the rats were exposed to 120.36 mg / kg of P0. 05 mg / kg. The expression of PCNA protein in testis of the rats in the control group decreased significantly, while the expression of EGFR protein in the testis of each exposed group was significantly lower than that in the control group (P 0.05). Conclusion A certain dose of acrylamide could induce pathological changes in rat testis, decrease the expression of DNA protein in testis and inhibit the synthesis of DNA in spermatogenic cells. Disrupting the division of testicular spermatogenic cells, the dynamic balance between proliferation and apoptosis, and disturbing the level of sex hormones, eventually lead to different degrees of reproductive toxicity in rats, which provides experimental basis for the study of the mechanism of acrylamide reproductive toxicity.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類(lèi)號(hào)】：R114

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