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异虎耳草素对PCPA致松果体损伤模型大鼠的改善作用及生物钟基因表达的影响
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| 篇名： | 异虎耳草素对PCPA致松果体损伤模型大鼠的改善作用及生物钟基因表达的影响 |
| TITLE： | Improvement Effects of Isopimpinelline on PCPA-induced Pineal Gland Injury Model Rats and Its Effects on Expression of Biological Clock Gene |
| 摘要： | 目的：研究异虎耳草素对对氯苯丙氨酸（PCPA）致松果体损伤模型大鼠的改善作用及其对生物钟基因表达的影响。方法：将60只大鼠随机分为空白对照组（2％聚山梨酯80溶液）、模型对照组（2％聚山梨酯80溶液）、阳性对照组（褪黑素，10mg/kg）和异虎耳草素高、中、低剂量组（3、1.5、0.75mg/kg），每组10只。除空白对照组外，其余各组大鼠均采用腹腔注射PCPA（450mg/kg）的方法构建松果体损伤模型。造模结束后，各组大鼠灌胃给药，每天1次，连续7d。在给药第6天，通过戊巴比妥钠协同睡眠实验考察各组大鼠的入睡潜伏期和睡眠持续时间；末次给药后，采用酶联免疫吸附法测定大鼠血清中褪黑素水平，分别在荧光显微镜和电子显微镜下观察大鼠松果体组织的病理变化和细胞超微结构改变，并采用实时荧光定量-聚合酶链式反应法测定大鼠松果体中生物钟基因Clock、Bmal1、Per1、Per2、Per3、Cry1、Cry2mRNA表达水平。结果：与空白对照组比较，模型对照组大鼠的入睡潜伏期显著延长（P＜0.05）；血清中褪黑素水平和松果体中Bmal1、Per1mRNA表达水平显著降低（P＜0.05或P＜0.01），Per3mRNA表达水平显著升高（P＜0.05）；松果体细胞排列紊乱、核固缩，空泡变性明显增多，数目明显减少，线粒体多见肿胀、嵴断裂、固缩。与模型对照组比较，异虎耳草素高剂量组大鼠的入睡潜伏期显著缩短（P＜0.05）、睡眠持续时间显著延长（P＜0.05），血清中褪黑素水平和松果体中Clock、Bmal1、Per1、Cry1、Cry2mRNA表达水平显著升高（P＜0.05或P＜0.01）；异虎耳草素中剂量组大鼠的入睡潜伏期显著缩短（P＜0.05），血清中褪黑素水平和松果体中Clock、Bmal1、Per1、Cry1、Cry2mRNA表达水平显著升高（P＜0.05或P＜0.01），Per3mRNA表达水平显著降低（P＜0.05）；异虎耳草素低剂量组大鼠Clock、Bmal1、Per2、Cry2mRNA表达水平显著升高（P＜0.05），Per3mRNA表达水平显著降低（P＜0.05）；异虎耳草素各剂量组大鼠松果体细胞排列紊乱均有一定程度改善、核固缩空泡变性减少，线粒体肿胀、嵴断裂、固缩均有一定程度减少。结论：异虎耳草素对PCPA致大鼠松果体损伤具有一定的改善作用；其可上调松果体正向调节因子Clock、Bmal1和负向调节因子Per1、Per2、Cry1、Cry2的表达，下调负向调节因子Per3的表达。 |
| ABSTRACT： | OBJECTIVE：To study the improvement effects of isopimpinelline on p-chlorophenylalanine（PCPA）-induced pineal injury model rats and its effect on expression of biological clock gene. METHODS ：Totally 60 rats were divided into blank control group（2％ polysorbate solution），model control group （2％ polysorbate solution），positive control group （melatonin，10 mg/kg） and isopimpinelline high-dose ，medium-dose and low-dose groups （3，1.5，0.75 mg/kg）. Except for blank control group ，rats in other groups were given PCPA intraperitoneally （450 mg/kg）to establish pineal injury model. After modeling finished ，they were given relevant medicine intragastrically ，once a day ，for consecutive 7 d. On the 6th day of administration ，the sleep latency and sleep duration of rats in each group were investigated by pentobarbital sodium coordination sleep test ；after last administration ， ELISA assay was used to determine the serum level of melatonin in rats. Fluorescence microscope and electron microscope were used to observe the pathological tissue and cell ultrastructure changes of the pineal gland. RT-qPCR was used to detect the mRNA expressions of biological clock gene Clock，Bmal1，Per1，Per2，Per3，Cry1，Cry2 in pineal gland of rats. RESULTS ：Compared with blank control group ，model control group had significantly longer sleep latency （P＜0.05）；serum melatonin ，mRNA expressions of Bmal1 and Per1 in pineal gland were significantly decreased （P＜0.05 or P＜0.01）while mRNA expression of Per3 was increased significantly （P＜0.05）. The pineal gland cell arrangement disorder ，nuclear pyknosis ，vacuolar degeneration increased and cell number decreased significantly ；mitochondria swollen ，cristae broken and pyknosis were observed. Compared with model control group ，the sleep latency of isopimpinelline high-dose group was shortened significantly （P＜0.05），sleep duration time was prolonged significantly （P＜0.05）；the levels of melatonin in serum ，mRNA expressions of Clock，Bmal1， Per1，Cry1 and Cry2 in pineal gland of rats were increased significantly （P＜0.05 or P＜0.01）. In isopimpinelline medium-dose group，the sleep latency was shortened significantly （P＜0.05）；the levels of melatonin in serum and mRNA expressions of Clock， Bmal1，Per1，Cry1，Cry2 in pineal gland were increased significantly （P＜0.05 or P＜0.01），while mRNA expression of Per3 was decreased significantly （P＜0.05）. In isopimpinelline low-dose group ，the levels of mRNA expressions of Clock，Bmal1，Per2 and Cry2 were increased significantly （P＜0.05），while mRNA expression of Per3 was decreased significantly （P＜0.05）. Cell arrangement disorder was improved and nuclear pyknosis vacuole degeneration was decreased to some extent in isopimpinelline groups；mitochondria swelled ，cristae fractured ，and pyknosis decreased to some extent. CONCLUSIONS ：Isopimpinelline can improve PCPA-induced pineal gland injury in rats ；it can up-regulate the expressions of positive regulators Clock，Bmal1 and negative regulators Per1，Per2，Cry1，Cry2，while down-regulate the expression of negative regulator Per3. |
| 期刊： | 2020年第31卷第17期 |
| 作者： | 王丹妮，杨钤，仲启明，宋美卿，仝立国，贾力莉，牛艳艳，冯玛莉 |
| AUTHORS： | WANG Danni，YANG Qian，ZHONG Qiming ，SONG Meiqing ，TONG Liguo，JIA Lili，NIU Yanyan ，FENG Mali |
| 关键字： | 异虎耳草素；松果体；生物钟基因；大鼠 |
| KEYWORDS： | Isopimpinelline；Pineal gland ；Biological clock gene ；Rats |
| 阅读数： | 394 次 |
| 本月下载数： | 3 次 |
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