本文選題：DBDPE + 甲狀腺激素�。� 參考：《廣西醫(yī)科大學(xué)》2013年碩士論文

【摘要】：研究目的 觀察十溴二苯乙烷(decabromodiphenyl ethane, DBDPE)經(jīng)口灌胃染毒后對(duì)青春期雄性大鼠甲狀腺組織的影響,通過測(cè)定染毒后甲狀腺激素3,5,3’三碘甲腺原氨酸(triiodothyronine, T3)、3,5,3',5'-四碘甲狀腺原氨酸(thyroxine, T4)和促甲狀腺激素(thyroid stimulating hormone, TSH)含量以探討其甲狀腺毒性；通過測(cè)定DBDPE染毒后肝臟微粒體代謝相關(guān)酶尿二磷酸葡萄糖醛酸轉(zhuǎn)移酶(uridine diphosphate glueuronsyl transferases, UDPGTs)活性及對(duì)甲狀腺和垂體特異性基因的影響以探討甲狀腺毒性作用機(jī)制；同時(shí)研究DBDPE對(duì)雄性大鼠青春期發(fā)育的影響,為全面了解DBDPE內(nèi)分泌干擾效應(yīng)和DBDPE的安全使用提供基礎(chǔ)毒理學(xué)資料。 研究方法 1.選取DBDPE制備染毒液,將出生后(postnatal day, PND)23天雄性大鼠按體重隨機(jī)分為組：對(duì)照組、100mg/kg組、300mg/kg組、600mg/kg組,每組9只。采用經(jīng)口灌胃染毒方式連續(xù)染毒至PND53,給藥期間每日稱量體重并根據(jù)體重適當(dāng)調(diào)整給藥劑量。同時(shí),每日觀察大鼠生殖器官發(fā)育狀況。 2.采用蘇木素-伊紅染色法觀察大鼠甲狀腺、垂體、肝臟、腎上腺、睪丸、前列腺組織病理學(xué)變化。 3.采用酶聯(lián)免疫吸附測(cè)定法(enzyme linked immunosorbent assay, ELISA)檢測(cè)大鼠血清甲狀腺激素T3、T4、TSH含量。 4.測(cè)定肝臟微粒體UDPGT活性觀察DBDPE對(duì)甲狀腺激素代謝相關(guān)酶活性的影響。 5.采用熒光定量PCR方法檢測(cè)大鼠甲狀腺和垂體相關(guān)基因表達(dá)。 研究結(jié)果 1. DBDPE染毒后青春期雄性大鼠甲狀腺、垂體、肝臟、腎上腺組織病理學(xué)與對(duì)照組相比沒有發(fā)生明顯改變。在300mg/kg和600mg/kg組,肝臟重量出現(xiàn)了顯著性降低,其它臟器重量沒有出現(xiàn)顯著性改變；染毒期間及解剖當(dāng)日染毒組大鼠體重隨劑量增高有下降趨勢(shì),但沒有出現(xiàn)統(tǒng)計(jì)學(xué)差異。 2.雄性大鼠經(jīng)DBDPE染毒后,解剖當(dāng)日,與對(duì)照組比較,甲狀腺激素T4、T3水平在600mg/kg劑量組都出現(xiàn)顯著性降低(p0.05),含量分別降低了10.3%和13.9%。同時(shí),血清TSH含量在染毒組均出現(xiàn)顯著性降低(p0.05),與對(duì)照組相比,100mg/kg劑量組TSH含量降低了43.1%,300mg/kg劑量組含量降低了49.3%,600mg/kg劑量組含量降低了35.0%。 3.測(cè)定了DBDPE染毒后肝臟微粒體代謝相關(guān)酶UDPGT活性,UDPGT活性在300mg/kg和600mg/kg劑量組出現(xiàn)顯著性升高(p0.05),并且隨著染毒劑量的增高各試驗(yàn)組UDPGT活性呈現(xiàn)出劑量-效應(yīng)關(guān)系,在600mg/kg劑量組肝臟微粒體UDPGT活性最高。 4. DBDPE染毒后,甲狀腺中Tpo(thyroid peroxidase, Tpo)mRNA表達(dá)在300mg/kg及600mg/kg劑量組中出現(xiàn)顯著性增高,與對(duì)照組相比,分別相對(duì)增高了2.00倍及2.37倍；DBDPE染毒后,與對(duì)照組相比,Tg(thyroglobulin, Tg)mRNA表達(dá)在300mg/kg劑量組出現(xiàn)顯著性增高,相對(duì)增高了1.8倍,在600mg/kg劑量組相對(duì)增高了1.6倍；Tshr (thyrotropin receptor, TSHR) mRNA表達(dá)在300mg/kg劑量組相對(duì)顯著性上調(diào)了1.33倍；Dio2(iodothyronine deiodinase, Dio) mRNA表達(dá)在300mg/kg劑量組出現(xiàn)顯著性增高,相比增高了1.72倍；Diol基因和Dio3基因與對(duì)照組相比,染毒組沒有出現(xiàn)顯著性差異；同時(shí),染毒后Slc5a5(solute carrier family5, member5, Slc5a5) mRNA表達(dá)也沒有出現(xiàn)顯著性差異(p0.05)。垂體中,DBDPE染毒后Tshβ基因表達(dá)在各染毒組均出現(xiàn)顯著性降低,與對(duì)照組相比,在100mg/kg、300mg/kg和600mg/kg組分別下調(diào)了1.17倍、1.15倍、1.07倍。在mRNA水平,100mg/kg組Dio2基因顯著增高了1.46倍。Tra (thyroid hormone receptor α, Trα)、Trhr (TRH receptor, Trhr)、Dio1、 Dio3基因均未出現(xiàn)顯著性差異(p0.05)。 5.觀察DBDPE對(duì)雄性大鼠青春期發(fā)育的影響：結(jié)果表明,青春期雄性大鼠在染毒期間,包皮完全分離時(shí)間及完全分離時(shí)的體重,與對(duì)照組相比,各染毒組均無顯著差異性(p0.05)。各染毒組大鼠雙側(cè)睪丸、前列腺、精囊重量與對(duì)照組比較也沒有出現(xiàn)統(tǒng)計(jì)學(xué)改變。同時(shí),染毒劑量至600mg/kg時(shí)大鼠睪丸、前列腺未見顯著的組織病理學(xué)改變。 研究結(jié)論 DBDPE可干擾青春期雄性大鼠甲狀腺激素T3、T4、TSH在大鼠機(jī)體中的穩(wěn)態(tài),使青春期雄性大鼠血清T3、T4、TSH激素含量有所下降。其甲狀腺毒性作用機(jī)制可能是通過影響肝臟微粒體激素代謝相關(guān)酶UDPGT使血清甲狀腺激素T3、T4含量降低,體循環(huán)中甲狀腺激素T3、T4含量的降低可引起甲狀腺中特異性基因Tpo、Tg mRNA水平的增高,促使甲狀腺自身合成更多的甲狀腺激素T3、T4以補(bǔ)償體循環(huán)中甲狀腺激素的下降。DBDPE還可通過影響垂體中Dio2基因的表達(dá),使垂體中的T4更多地轉(zhuǎn)化為T3增多,在垂體局部,相對(duì)增多的T3負(fù)反饋引起Tsh mRNA水平下調(diào),使垂體TSH分泌減少,從而導(dǎo)致體循環(huán)中TSH含量下降。此外,DBDPE對(duì)雄性大鼠的青春期發(fā)育影響不明顯。
[Abstract]:Purpose of study


To investigate the effect of decabromodiphenyl ethane ( DBDPE ) on thyroid tissue of adolescent male rats after oral gavage , and determine the thyroid toxicity by measuring the contents of 3 , 5 , 3 ' triiodomethane ( T3 ) , 3 , 5 , 3 ' , 5 ' - tetraiodothyroxin ( TSH ) and thyroid stimulating hormone ( TSH ) .
To explore the mechanism of thyroid toxicity by measuring the activity of glucuronosyltransferase ( UDPGTs ) activity of liver microsomes after exposure to DBDPE and its effects on thyroid and pituitary - specific genes .
At the same time , the effects of DBDPE on the puberty development of male rats were studied to provide basic toxicological data for the comprehensive understanding of DBDPE endocrine disruption effect and safe use of DBDPE .


Research Methods


1 . The rats were randomly divided into two groups : control group , 100mg / kg group , 300 mg / kg group and 600mg / kg group .


2 . The pathological changes of the thyroid , pituitary , liver , adrenal gland , testis and prostate in rats were observed by the method of Su - lignin - eosin staining .


3 . Serum thyroid hormones T3 , T4 and TSH were measured by enzyme linked immunosorbent assay ( ELISA ) .


4 . The effect of DBDPE on the activity of thyroid hormone - related enzymes was observed by measuring the activity of UDPGT in liver microsomes .


5 . The expression of thyroid and pituitary - related genes in rats was detected by fluorescence quantitative PCR .


Results of the study


1 . The pathology of thyroid , pituitary , liver and adrenal gland of male rats after DBDPE had no significant change compared with control group . There was a significant decrease in liver weight in 300 mg / kg and 600 mg / kg group .
The body weight of the rats exposed to the virus during the exposure period and the anatomic day decreased with increasing dose , but there was no statistical difference .


2 . Compared with the control group , the levels of thyroid hormones T4 and T3 decreased by 10.3 % and 13.9 % in the dose group of 600 mg / kg , respectively , and the content of TSH in the dosage group decreased by 43.1 % and 49.3 % in the dosage group of 300 mg / kg , and the content of 600 mg / kg group was decreased by 33.0 % .


3 . The activity of UDPGT , UDPGT , UDPGT , UDPGT and UDPGT activity in liver microsomes were significantly increased after DBDPE exposure ( P < 0.05 ) , and the activity of UDPGT increased with the increase of the dosage of the dye . The activity of UDPGT in liver microsomes was the highest at the dosage of 600 mg / kg .


4 . After the DBDPE was poisoned , the mRNA expression of Tpo ( thyroid peroxidase , Tpo ) mRNA in thyroid was significantly higher in the dosage group of 300 mg / kg and 600 mg / kg , which was 2.00 times and 2.37 times higher than that in the control group , respectively .
Compared with the control group , the expression of Tg ( Tg ) mRNA increased significantly in the 300 mg / kg dose group compared with the control group , which was 1.8 times higher than that of the control group and 1.6 times higher in the 600 mg / kg group .
The mRNA expression of Tshr ( TSHR ) was up - regulated by 1.33 times in the 300 mg / kg dose group .
Dio mRNA expression was significantly higher in the 300 mg / kg dose group than that in the 300 mg / kg dose group , which was increased by 1 . 72 times .
Compared with the control group , the Diol gene and the Dio3 gene had no significant difference compared with the control group .
At the same time , there was no significant difference in mRNA expression of Slc5a5 ( P0.05 ) . In the pituitary , the expression of Tsh尾 gene was significantly decreased at 100 mg / kg , 300 mg / kg and 600 mg / kg in the pituitary . There was no significant difference in the mRNA level at 100 mg / kg , 300 mg / kg and 600 mg / kg , respectively .


5 . The effects of DBDPE on the puberty development of male rats were observed : the results showed that the total separation time and the weight of the male rats were not significantly different from the control group ( p < 0.05 ) .


Conclusions of the study


DBDPE can interfere with the steady state of thyroid hormones T3 , T4 and TSH in the rat body during puberty , and decrease the content of serum T3 , T4 and TSH in the rat body .
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R114

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