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胡桃醌对大鼠血清中与脑组织损伤相关生物标志物的影响
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| 篇名： | 胡桃醌对大鼠血清中与脑组织损伤相关生物标志物的影响 |
| TITLE： | Effects of Juglone on Serum Biomarkers Related to Brain Tissue Injury in Rats |
| 摘要： | 目的：考察胡桃醌对大鼠脑组织的影响及其与脑组织损伤相关生物标志物的关系。方法：将40只大鼠分为空白组和胡桃醌高、中、低剂量组（34.832、17.416、8.708mg/kg），每组10只，每天灌胃给药1次，连续给药4周。末次给药后，考察各组大鼠的一般行为学、脑指数及脑组织病理形态。采用超高效液相色谱-多反应检测质谱（UPLC-MRM-MS）法检测各组大鼠血清中左旋多巴（L-Dopa）、L-酪氨酸（L-Tyr）、L-色氨酸（L-Trp）的含量。色谱条件为色谱柱WatersAcquityUPLCBEHC18，流动相为乙酸铵水溶液-乙腈，梯度洗脱，流速为0.3mL/min，进样量为5μL，柱温为30℃；质谱条件为电喷雾离子源，正离子模式下检测，毛细管电压为3500V，去溶剂气流量为650L/h，去溶剂化温度为350℃，离子源温度为110℃。结果：与空白组比较，胡桃醌各剂量组大鼠出现倦怠、四肢无力的行为；脑组织可见大、小脑皮质内血管充血，锥体细胞层部分神经元固缩，核深染，形态不规则，边界不清晰等病理性改变；胡桃醌高剂量组脑指数显著增加（P＜0.05）。建立的UPLC-MRM-MS法专属性良好，L-Dopa、L-Tyr、L-Trp的检测质量浓度线性范围分别为31.25～32000、31.25～32000、15.625～16000ng/mL（r＝0.9991～0.9999），检测限分别为6.250、5.625、3.125ng/mL，定量限分别为15.625、18.75、10.00ng/mL，精密度、准确度、稳定性（24h）试验RSD均小于2.6％（n＝6），基质效应为95.1％～100.1％（RSD均未超过3.25％，n＝3）。动物实验结果显示，与空白组比较，胡桃醌中、高剂量组L-Dopa含量显著升高（P＜0.01）；胡桃醌低、中、高剂量组L-Tyr含量显著升高，L-Trp含量显著降低（P＜0.05或P＜0.01）。结论：胡桃醌对大鼠的行为学、脑指数、脑组织形态均有明显影响，可能通过升高血清中L-Dopa、L-Tyr的含量，降低L-Trp的含量影响大鼠的脑组织功能。 |
| ABSTRACT： | OBJECTIVE：To investigate th e effects of juglone on brain tissu e of rats and its relationship with the biomarkers related to brain tissue injury. METHODS ：Totally 40 rats were divided into blank group ，juglone high-dose ，medium-dose and low-dose groups （34.832，17.416，8.708 mg/kg），with 10 rats in each group. They were given medicine intragastrically once a day ， for consecutive 4 weeks.After last administration ，the general behavior ，brain index and brain tissue morphology were investigated. UPLC-MRM-MS method was used to determine the serum contents of L-dopa（L-Dopa），L-tyrosine（L-Tyr）and L-tryptophan（L-Trp） in rats. The chromatographic condition included Waters Acquity UPLC BEH C 18 column，mobile phase consisted of ammonium acetate aqueous solution-acetonitrile ，gradient elution ，at the flow rate of 0.3 mL/min，sample size of 5 μL，column temperature of 30 ℃；MS condition include electrospray ion source ，in positive ion mode ，capillary voltage of 3 500 V，desolvent gas flow of 650 L/h，desolvent temperature of 350 ℃，ion source temperature of 110 ℃. RESULTS ：Compared with blank group ，the rats in each dose group showed the behavior of tiredness and weakness of limbs. The brain tissue morphology showed pathological changes ， which contained blood vessel congestion in the cerebral and cerebellar cortex ，partial cell nucleus pyknosis in the pyramidal cell layer，deep staining of nuclei ，irregular shape and unclear boundary and other pathological changes ；the brain index of juglone high-dose group increased significantly （P＜0.05）. The established UPLC-MRM-MS method showed good specificity and the range of L-Dopa，L-Tyr and L-Trp were 31.25-32 000，31.25-32 000，15.625-16 000 ng/mL（r＝0.999 1-0.999 9），respectively. The limits of detection were 6.250，5.625，3.125 ng/mL，respectively. The limits of quantitation were 15.625，18.75，10.00 ng/mL，respectively. RSDs of precision ，accuracy and stability （24 h）tests were all Matrix effects were 95.1％-100.1％ （RSD are not more than 3.25％，n＝3）. Compared with blank group，the contents of L-Dopa were increased significantly injuglone medium-dose and high-dose groups （P＜0.01）. The contents of L-Tyr were increased significantly in juglone lowdose，medium-dose and high-dose gro ups，while the contents of L-Trp were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：Juglone has an effect on the general behavior，brain index and brain tissue morphology of rats. It may affect the brain function of rats by increasing the contents of L-Dopa and L-Tyr in serum and decreasing the contents of L-Trp. |
| 期刊： | 2020年第31卷第03期 |
| 作者： | 董文婷，霍金海，孙国东，王伟明 |
| AUTHORS： | DONG Wenting ，HUO Jinhai，SUN Guodong ，WANG Weiming |
| 关键字： | 胡桃醌；脑组织；生物标志物；超高效液相色谱-多反应检测质谱法；含量测定 |
| KEYWORDS： | Juglone；Brain tissue ；Biomarker；UPLC-MRM-MS；Content determination |
| 阅读数： | 274 次 |
| 本月下载数： | 5 次 |
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