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以临床解热镇痛药基本成分对乙酰氨基酚(APAP)为模药,调查APAP在家蚕体内的代谢和对蚕体主要组织(系统)的损伤以及蚕体主要解毒酶基因的表达变化,探讨家蚕作为肝毒性药物毒理研究替代实验动物的可能性。家蚕5龄2 d幼虫经口给予APAP后0.5 h,在消化管、脂肪体和血淋巴中均能够检测到原药,APAP主要糖基化解毒代谢途径中的关键酶——尿苷二磷酸-葡萄糖醛酸基转移酶(UGT)活性也随即快速上升,其中脂肪体中的UGT酶活性上升早于血淋巴和消化管。实时荧光定量PCR(qRT-PCR)分析显示,幼虫消化管和脂肪体细胞中UGT家族基因Ugt30、Ugt86和Ugt89的转录水平在给予APAP后0.5 h开始显著上调。HE染色显示,高剂量APAP对蚕体消化管和脂肪体组织具有损伤作用,但AO/PI染色未发现血淋巴中死细胞增多。研究结果表明,家蚕的消化管、脂肪体和血淋巴都有通过糖基化代谢APAP的解毒功能,过量的APAP对消化管和脂肪体组织都有损伤作用。
To investigate the metabolism of APAP in silkworm (Bombyx mori L.) and its damage to the main tissue (system) of silkworm as well as the changes of the main detoxifying enzyme genes in silkworms by using acetaminophen (APAP) as the basic component of clinical antipyretic and analgesic drugs Bombyx mori as hepatotoxicity toxicology study to replace the possibility of experimental animals. The 2nd instar larvae of the 5th instar larvae of the silkworm were able to detect the original drug 0.5 h after oral administration of APAP in the digestive tract, fat body and hemolymph. The key enzyme in the major glycosylation and detoxification pathway of APAP, uridine diphosphate- Glucurbituryltransferase (UGT) activity was also rapidly increased, UGT enzyme activity in fat body rose earlier than the hemolymph and digestive tract. Real-time quantitative PCR (qRT-PCR) analysis showed that transcriptional level of UGT family genes Ugt30, Ugt86 and Ugt89 in larval digestive tract and fat body cells were significantly up-regulated from 0.5 h after APAP administration. HE staining showed that high dose APAP had damaging effects on digestive duct and fat body of silkworm, but no increase of dead cells in hemolymph was found by AO / PI staining. The results showed that the digestive duct, fat body and hemolymph of silkworm had the detoxification function of APAP through glycosylation metabolism. Excessive APAP had damaging effects on digestive duct and fat body tissues.