拟青霉代谢物提取物对小鼠体内单胺氧化酶和慢性应激大鼠海马单胺类神经递质的影响

Effect of bioactive compounds from paecilomyces tenuipes on monoaminoxidase activity in mice and levels of monoamine neurotransmitter in hippocampus of rats with chronic unpredictable stress

目的:观察一种拟青霉代谢物提取物对小鼠体内组织单胺氧化酶活性和慢性应激大鼠海马单胺类神经递质的影响。方法:实验于2004-08在安徽医科大学药理学教研室进行。①将180只小鼠随机分为吗氯贝胺组和拟青霉代谢物提取物组,雌雄各半。吗氯贝胺组一次性灌胃吗氯贝胺40mg/kg,拟青霉代谢物提取物组均灌胃拟青霉代谢物提取物100mg/kg。按照取材时间分为0,0.25,0.5,1,2,4,8,l6,24h9个时间点,每个时间点10只大鼠。采用紫外分光光度计在每个时间点测定小鼠脑、肝组织单胺氧化酶活性。②选用成年雄性SD大鼠48只,随机分为6组:对照组、模型组、吗氯贝胺组、拟青霉代谢物提取物80,40,20mg/kg组,每组8只。采用慢性应激法造成大鼠抑郁模型,用药组及模型组大鼠共接受20d各种不同的应激(包括电击足底、4℃冷水刺激5min、45℃热刺激5min、高速水平摇晃10min、夹尾60s、禁水24h、禁食48h、悬吊5min和昼夜颠倒,平均每种刺激各2次)。刺激同时用药组灌胃给药,1次/d,共22d。模型组及对照组灌胃蒸馏水,吗氯贝胺组灌胃剂量为40mg/kg、单胺氧化酶剂量组灌胃剂量分别为20,40,80mg/kg。采用高效液相-电化学法检测应激大鼠海马肾上腺素、5-羟色胺、多巴胺及其代谢产物5-羟基吲哚乙酸、3,4-二羟基苯乙酸的含量。结果:所用实验动物全部进入结果分析。①拟青霉代谢物提取物100mg/kg1次性给药对小鼠脑组织单胺氧化酶A具有一定的选择性抑制作用,拟青霉代谢物提取物给药后4h,对小鼠脑组织单胺氧化酶A的抑制率达到66.7%。②与对照组比较,模型组大鼠海马肾上腺素、多巴胺、5-羟色胺的含量降低,3,4-二羟基苯乙酸升高,5-羟基吲哚乙酸无明显改变,多巴胺/3,4-二羟基苯乙酸,5-羟色胺/5-羟基吲哚乙酸的比值降低;与模型组比较,拟青霉代谢物提取物80,40mg/kg组肾上腺素、多巴胺、5-羟色胺的含量均升高,3,4-二羟基苯乙酸、5-羟基吲哚乙酸的含量均降低,拟青霉代谢物提取物20mg/kg组3,4-二羟基苯乙酸含量降低。拟青霉代谢物提取物80,40mg/kg可提高多巴胺/3,4-二羟基苯乙酸、5-羟色胺/5-羟基吲哚乙酸的比值;拟青霉代谢物提取物20mg/kg可提高多巴胺/3,4-二羟基苯乙酸的比值。结论:拟青霉代谢物提取物可能通过选择性抑制单胺氧化酶A和调节海马单胺类神经递质发挥抗抑郁作用。

AIM： To study the effects of bioactive compounds from paecilomyces tenuipes （BCPT） on monoaminoxidase （Mao） activity in brain and liver organs of mice and the levels of monoamine neurotransmitters in hippocampus of chronic unpredictable stress rats. METHODS： The experiment was conducted in the Department of Pharmacology of Anhui Medical University in August 2004. ①180 mice were randomly divided into moclobemide group and BCPT group with half females and half males. Mice in the modobemide group were intragastrically administrated with moclobemide 40 mg/kg, and mice in the BCPT group were given intragastric administration of BCPT 100 mg/kg. The ultraviolet spectrophotometer was adopted to measure the Mao activity in brain and liver tissues of mice respectively at 0, 0.25, 0.5, 1, 2, 4, 8, 16 and 24 hours with 10 mice tested in each time-point.②Forty-eight adult SD rats were selected and randomly divided into six groups： control group, model group, moclobemide group, BCPT 80,40,20 mg/kg groups with 8 rats in each group. The rat models of depression were established by chronic stress, and rats in the model group and administration group were given different stress for 20 days （including sole-shocking, stimulating at 4 ℃ in water for 5 minutes, 5- minute thermal stimulus, 10-minute weaving at high speed, 60-second tail- clamping, 24-hour water-prohibition, 48-hour fasting, 5-minute suspending and reversed time feeling for twice each）. Gastric perfusion was given to rats of the administration group at the same time of stimulating once a day for 22 days. Rats in the model group and control group were given intragastric administration of distilled water, while rats in the moclobemide group received intragastric administration at 40 mg/kg. Rats in BCPT large dose group were treated by intragastric administration respectively at 20,40, 80 mg/kg. The levels of adnephrin, 5-hydroxyptamine （HT）, dopamine and its metabolic products 5-hydroxyindoleacetic acid, 3, 4-dihydroxybenzene in hippocampus of rats with unpredictable stress were measured by high performance liquid chromatography with electrochemical detector. RESULTS： All experimental animals were involved in the analysis of results.①Once administration of 100 mg/kg BCPT had certain inhibition effect on MAO-A in brain tissues of mice, while 4 hours after administration with BCPT, the inhibition ratio of MAO-A was 66.7%.②Compared with the control group, levels of adnephrin, 5-HT and dopamine in rats of the model group were decreased, while the level of 3, 4-dihydroxybenzene was increased, and there was no change in level of 5-hydroxyindoleacetic acid. The ratios of dopamine/3, 4-dihydroxybenzene and 5-HT/5-hydroxyindoleacetic acid decreased. Compared with the model group, the levels of adnephrin, 5-HT and dopamine were increased, while levels of 3, 4-dihydroxybenzene, 5-hydroxyindoleacetic acid were decreased. The level of 3, 4-dihydroxybenzene in 20 mg/kg BCPT group was reduced. BCPT could enhance the ratios of dopamine/3, 4-dihydmxybenzene, 5-HT, 5-hydroxyindoleacetic acid at 80mg/kg and 40 mg/kg, and BCPT could enhance the ratio of dopamine/3, 4-dihydroxybenzene at 20 mg/kg. CONCLUSION： BCPT has anti-effect on depression, which may works by selectively inhibit MAO-A and regulate the monoamine neurotransminer in hippocampus of chronic unpredictable stress rats.