本文選題：氟中毒 + 小鼠　； 參考：《山西農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：[目的]多年來(lái)對(duì)氟的研究表明,適量的氟有利于維持鈣磷正常代謝,預(yù)防齲齒,而過(guò)量的氟可導(dǎo)致多種組織器官的損傷,引起氟中毒。氟對(duì)雄性生殖系統(tǒng)的影響是多方面的,而氟對(duì)生殖內(nèi)分泌方面的研究還較少。本文主要通過(guò)real-time PCR、組織切片、透射電鏡等技術(shù),結(jié)合血清中生殖相關(guān)激素的含量、骨氟含量、精子畸形率等的變化,綜合分析氟暴露對(duì)小鼠下丘腦-垂體-睪丸軸的影響,并進(jìn)一步探討其對(duì)生殖系統(tǒng)損傷的分子機(jī)制。[方法]48只8周齡性成熟雄性昆明小鼠,隨機(jī)分為四組：對(duì)照組、低氟組、中氟組、高氟組,對(duì)照組給予去離子水,氟處理組分別給予含25、50、100mg/LNaF的去離子水,飼喂60天。飼養(yǎng)過(guò)程中記錄生長(zhǎng)狀況,并定期記錄體重。染毒結(jié)束后,采集血清、精子和下丘腦、垂體、睪丸、股骨等組織,進(jìn)行相關(guān)的指標(biāo)測(cè)定,包括使用氟離子選擇電極測(cè)量骨氟含量,使用放射免疫法測(cè)量血清中生殖相關(guān)激素的含量,使用HE染色和透射電子顯微鏡觀察組織形態(tài)結(jié)構(gòu)的變化,使用real-time PCR檢測(cè)生殖相關(guān)基因mRNA的表達(dá)等。[結(jié)果]①與對(duì)照組相比,各氟處理組骨氟含量明顯升高,在中氟組和高氟組差異顯著(p0.001)；而體重、體增重、主要臟器系數(shù)均無(wú)顯著性差異(p0.05)。②精子畸形率在各氟處理組明顯升高,與對(duì)照組相比,在高氟組差異顯著(p0.01)；而血清中卵泡刺激素(FSH)、黃體生成素(LH)、睪酮(T)激素含量與對(duì)照組相比無(wú)顯著性差異(p0.05)。③通過(guò)制作病理組織切片,HE染色后觀察發(fā)現(xiàn),下丘腦和垂體組織無(wú)明顯的病理變化,而睪丸組織在氟處理組出現(xiàn)了不同程度的損傷,高氟組尤為明顯,出現(xiàn)了管腔中成熟精子明顯減少,各級(jí)生精細(xì)胞排列紊亂,層數(shù)減少,生精小管周?chē)g隙變寬,間質(zhì)細(xì)胞出現(xiàn)了空泡。④透射電鏡結(jié)果顯示,下丘腦神經(jīng)元細(xì)胞核染色質(zhì)異常聚集,核膜腫脹,線粒體嵴斷裂,神經(jīng)髓鞘板層離散,呈蔥皮樣變；垂體中促性腺激素細(xì)胞染色質(zhì)聚集,線粒體腫脹、空泡化,內(nèi)質(zhì)網(wǎng)擴(kuò)張；睪丸組織中精原細(xì)胞、精母細(xì)胞、支持細(xì)胞染色質(zhì)濃縮,線粒體空泡化,精子細(xì)胞頂體膜破裂,間質(zhì)細(xì)胞核破裂,線粒體空泡化嚴(yán)重。⑤Real-time PCR檢測(cè)結(jié)果顯示,與對(duì)照組相比,下丘腦和垂體中的生殖相關(guān)激素的基因mRNA表達(dá)量無(wú)顯著差異(p0.05),而睪丸組織中的FSHR、LHR、INHα、INHβB和SHBG基因mRNA表達(dá)量都明顯下降,其中FSHR表達(dá)量只在高氟組差異顯著(p0.05),LHR、INHα和SHBG表達(dá)量在中氟組和高氟組差異顯著(p0.05),而INHβB表達(dá)量在低、中、高氟組均差異顯著(p0.05)。[結(jié)論]本次試驗(yàn)結(jié)果表明,60天氟暴露主要對(duì)睪丸的形態(tài)結(jié)構(gòu)和生殖相關(guān)的基因產(chǎn)生了影響,而通過(guò)血腦屏障對(duì)下丘腦和垂體產(chǎn)生的影響是其次的。
[Abstract]:[objective] the study on fluoride for many years showed that the proper amount of fluoride is beneficial to maintain the normal metabolism of calcium and phosphorus and prevent dental caries, and excessive fluoride can lead to the injury of various tissues and organs and lead to fluorosis. The effects of fluorine on male reproductive system are various, but there are few studies on reproductive endocrine. The effects of fluoride exposure on hypothalamus-pituitary-testicular axis in mice were analyzed by means of real-time, tissue section, transmission electron microscopy and the changes of serum reproductive hormones, bone fluorine content and sperm deformity rate. Furthermore, the molecular mechanism of its damage to reproductive system was discussed. [methods] Forty-eight 8-week-old male Kunming mice were randomly divided into four groups: control group, low fluorine group, middle fluorine group, high fluoride group, control group, deionized water, fluorine treatment group, and deionized water containing 2550 mg / L NAF for 60 days. During feeding, the growth status was recorded and the weight was recorded regularly. After the exposure, serum, sperm, hypothalamus, pituitary, testis, femur and other tissues were collected, and related indexes were measured, including the use of fluorine ion selective electrode to measure the fluorine content in bone. The contents of reproductive related hormones in serum were measured by radioimmunoassay, the morphological changes of tissues were observed by HE staining and transmission electron microscope, and the expression of reproductive related genes was detected by real-time PCR. [results] 1Compared with the control group, the bone fluorine content in each fluorine treatment group was significantly higher than that in the middle fluorine group and the high fluorine group (p0.001), while the weight, body weight, weight gain, There was no significant difference in the main organ coefficients (p0.05) .2 the sperm deformity rate was significantly higher in each fluorine treatment group than in the control group (p0.01). However, the levels of FSH, LH and T in serum were not significantly different from those in the control group (p0.05). The results showed that there were no obvious pathological changes in hypothalamus and pituitary after HE staining. However, testicular tissue was damaged in different degrees in fluoride treatment group, especially in high fluoride group, in which mature spermatozoa decreased obviously, spermatogenic cells of all levels were arranged in disorder, the number of layers decreased, and the space around seminiferous tubules became wider. The results of transmission electron microscope showed that hypothalamic neurons showed abnormal aggregation of nuclear chromatin, swelling of nuclear membrane, rupture of mitochondrial ridge, scattered lamellar layer of nerve myelin, and scallion dermatosis. Pituitary gonadotropin cell chromatin aggregation, mitochondria swelling, vacuolation, endoplasmic reticulum dilatation; testicular tissue spermatogonium, spermatocyte, Sertoli cell chromatin concentration, mitochondrial vacuolation, sperm cell acrosome membrane rupture, When the interstitial nucleus was ruptured and the mitochondria vacuolated seriously, the results of .5Real-time PCR showed that, compared with the control group, There was no significant difference in mRNA expression of reproductive hormone between hypothalamus and pituitary (p0.05), while the mRNA expression of FSHR-LHR-INH 偽 -INH 尾 B and SHBG in testicular tissue decreased significantly. The expression of FSHR was significantly different only in the high fluorine group (p0.05) and in the middle fluorine group and the high fluorine group (p0.05), while the expression of INH 尾 B was lower in the middle fluorine group than in the high fluoride group (p0.05). [conclusion] the results showed that 60 days of fluoride exposure mainly affected the morphological structure and reproductive genes of testis, while the effect of blood-brain barrier on hypothalamus and pituitary was the second.
【學(xué)位授予單位】：山西農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：X503.22

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 鐘先玖,吳鑫,周元陵,黃簡(jiǎn)抒,金泰^，

本文編號(hào)：2116315