WO2021012065A1 - 一种牛黄清心丸在制备治疗抑郁症的药物中的应用 - Google Patents

一种牛黄清心丸在制备治疗抑郁症的药物中的应用 Download PDF

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WO2021012065A1
WO2021012065A1 PCT/CN2019/096628 CN2019096628W WO2021012065A1 WO 2021012065 A1 WO2021012065 A1 WO 2021012065A1 CN 2019096628 W CN2019096628 W CN 2019096628W WO 2021012065 A1 WO2021012065 A1 WO 2021012065A1
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tongrentang
pills
depression
niuhuang
qingxin
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PCT/CN2019/096628
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French (fr)
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沈剑刚
高充
李霞
吴炜
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北京同仁堂国药有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/24—Antidepressants

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  • the invention belongs to the field of biomedicine, and specifically relates to the application of Niuhuang Qingxin Pills in the preparation of drugs for treating depression
  • Depression severe depression or clinical depression
  • depression patients are often accompanied by sadness, anxiety, irritability, excessive guilt and shy emotions. Depression also severely affects patients' sleep, diet, and social life, and severe cases lead to suicide. Therefore, the development of antidepressant drugs has a profound impact on promoting social development and reducing the burden on the people.
  • ECT electroconvulsive stimulation
  • traditional Chinese medicine has been used in the treatment of emotional diseases for more than 2,000 years and has accumulated a large number of therapeutic prescriptions.
  • Clinical practice has shown that traditional Chinese medicine has unique advantages in the treatment of depression. Strong emotional regulation.
  • the theory of traditional Chinese medicine believes that depression is a symptom of deficiency and deficiency of the heart, spleen, liver and kidney, dysfunction of heart, spleen, liver and kidney, deficiency of heart, spleen, qi and blood, liver and kidney deficiency, stagnation of liver and qi, turbid phlegm and blindness.
  • Niuhuang Qingxin Pills is a representative prescription for regulating emotions.
  • It consists of bezoar, antelope horn, musk, ginseng, atractylodes macrocephala (stir-fried with bran), angelica, white peony, bupleurum, dried ginger, donkey-hide gelatin, platycodon, buffalo horn concentrated powder, etc. It is made up of 27 medicinal materials and has the functions of replenishing the heart and spleen, replenishing qi and nourishing blood, soothing the liver and relieving depression, calming the nerves, resolving phlegm and eliminating wind.
  • Niuhuang Qingxin Pills include: (1) Oral diseases, such as oral mucosal ulcers, mouth and tongue sores, toothache, heel swelling and pain, acute and chronic pharyngitis, etc.; (2) Inverse caused by disturbance of phlegm and fire; (3) ) Patients with intracranial hypertension and coma; (4) Cerebrovascular disease, Tianjin Heping District Traditional Chinese Medicine Hospital treats 281 cases of acute cerebrovascular disease. For those with impaired consciousness and impotence, they are given this product and Angong Niuhuang Pills, daily 1 ⁇ Take 2 pills alternately, and the effect is satisfactory. (5) Treatment of herpes neuralgia: 30 patients with postherpetic neuralgia were given 1 Tongren Niuhuang Qingxin Pill and 1 bag of Longdan Xiegan Pill, 18 cases were cured and 10 cases improved.
  • Oral diseases such as oral mucosal ulcers, mouth and tongue sores, toothache, heel swelling and pain, acute and chronic pha
  • Niuhuang Qingxin Pills can be used for depression.
  • toxicological studies showed that Tongrentang Niuhuang Qingxin Pills, 461 times the human dose, were administered to mice by gavage, and no obvious acute toxicity was seen.
  • Respectively equivalent to 50, 25 and 12.5 times the human daily dose SD rats were given intragastrically for 26 consecutive weeks. During the experiment (13, 26 weeks of administration and 4 weeks of discontinuation), animals in all dose groups did not die.
  • Tongrentang Niuhuang Qingxin Pills have emotional regulation effects and no obvious side effects after long-term use, there is no relevant research on whether Tongrentang Niuhuang Qingxin Pills can improve depression, so it is a blank application in the field of depression treatment.
  • the inventor provides an application of Niuhuang Qingxin Pills in the treatment of depression, wherein the Niuhuang Qingxin Pills are Tongrentang Niuhuang Qingxin Pills.
  • Tongrentang Niuhuang Qingxin Pills are the pharmaceutical form used in the experimental examples of this patent invention.
  • the composition and preparation methods of other domestic brands of Niuhuang Qingxin Pills in China are different from them.
  • the specific design is commercial secrets and it is inconvenient to discuss. Therefore, they are different from Niuhuang Qingxin Pills.
  • the functions possessed are significantly different, so the Niuhuang Qingxin Pills described in this patent application are preferably Tongrentang Niuhuang Qingxin Pills.
  • Tongrentang Niuhuang Qingxin Pills is preferred, that is, all components in it are natural components (of which natural ingredients are preferably used for bezoar, musk, and borneol), and its antidepressant effect is better than the artificial formula of Niuhuang Qingxin Pills . See Example 2.
  • the cause of the depression is increased psychological pressure or endocrine disorders under normal circumstances.
  • Tongrentang Niuhuang Qingxin Pill Due to the different causes and manifestations of depression, Tongrentang Niuhuang Qingxin Pill has a significant effect on depression caused by increased psychological stress or endocrine disorders in normal environments in the experimental cases.
  • the adult administration dose of the Tongrentang Niuhuang Qingxin Pill is 1-20 g/day.
  • the adult administration dose of the Tongrentang Niuhuang Qingxin Pill is 3-15 g/day.
  • the adult administration dose of the Tongrentang Niuhuang Qingxin Pill is 6-12 g/day.
  • the use method of the Tongrentang Niuhuang Qingxin Pills is oral.
  • the use dose of the Tongrentang Niuhuang Qingxin Pills in a mouse test is 20 mg/kg, which significantly promotes the occurrence of hippocampal nerves.
  • Figure 1 Tongrentang Niuhuang Qingxin Pills effectively prevented the depression and anxiety behavior caused by the restraint experiment;
  • Figures 1a-1f correspond to the sugar water spray experiment, tail suspension test, forced swimming, open field experiment, and new things eating in the restraint group. Incubation period and food intake study);
  • Figures 2a-2b correspond to the number of newborn cells (BrdU+) in the hippocampus of mice in the restraint group and the number of newborn neurons (BrdU+) /DCX+));
  • Figure 3 Tongrentang Niuhuang Qingxin Pills effectively prevented the symptoms of depression and anxiety caused by corticosterone feeding;
  • Figures 3a-3f correspond to the sugar water spray test, tail suspension test, forced swimming, open field test, and new things eating in corticosterone group mice. (Study on food incubation period and food intake);
  • Figure 4 Tongrentang Niuhuang Qingxin Pills improve hippocampal neurogenesis suppressed by chronic corticosterone feeding.
  • Figures 4a-4c correspond to the number of newborn cells (BrdU+) in the hippocampus of mice, the number of newborn neurons (BrdU+/NeuN+) in the hippocampus, and the number of newborn neural precursor cells (BrdU+/ The influence of the number of DCX+)).
  • Tongrentang Niuhuang Qingxin Pills the specification is 3g/per pill (HKC-18084, Tongrentang Niuhuang Qingxin Pills are approved for listing in Hong Kong, please visit the following website: http://www.cmchk.org.hk/pcm/chi/ #main_listpcm_2018.htm), the adult dosage is 1 to 2 pills once; twice a day, the children should reduce it.
  • This experiment used conventional methods such as Splash Test, Tail Suspension Test, Forced swimming Test, Open Field Test, Novelty Suppressed Feeding and other conventional methods of restraint group mice. Indicators for research.
  • Depression animal model This study uses a long-term behavioral restraint model and a corticosterone feeding model to simulate depression caused by two factors, external pressure and endocrine disorders.
  • control group normally reared, no treatment, 0.1ml saline is given daily during the experiment
  • depression model group 50ml plastic restraint tube with 11 stomata, mice are placed inside, Tighten the lid and make it unable to move freely, and restraint for 6 hours every day from 9 am for 30 days; during the experiment, 0.1ml of normal saline was given to the stomach before restraint
  • treatment group before restraint was given daily, Gavage mice with Tongrentang Niuhuang Qingxin Pills normal saline suspension (20mg/kg, equivalent to one pill for adults
  • Western medicine control group using the classic antidepressant fluoxetine as a control, before daily restraint, give And mouse fluoxetine physiological saline solution (18mg/kg)).
  • Corticosterone feeding experiment the control group (normal feeding, no treatment; from the 15th day of the modeling process, 0.1ml of normal saline was administered daily); corticosterone feeding model (corticosterone was fed at 70 ⁇ g/ml Dissolved in 0.45% beta-cyclodextrin in drinking water for 30 days; from the 15th day of the modeling process, 0.1ml of normal saline was administered daily during the process); treatment group (the drinking water treatment is the same as the model group; From the 15th day of the model process, the Niuhuang Qingxin Pill Suspension 20mg/kg was given intragastrically every day during the process); Western medicine control group (using the classic antidepressant fluoxetine as the control, the drinking water treatment was the same as the model group; From the 15th day of the model process, the fluoxetine physiological saline solution was given intragastrically at 18 mg/kg daily).
  • BrdU labeling BrdU was injected intraperitoneally (50 mg/kg/d) for 5 consecutive days to label newborn cells. Seven days after the last labeling, the mice were sacrificed and the brain tissues were fixed with 4% paraformaldehyde. And the frozen section has been immunofluorescent staining.
  • Tongrentang Niuhuang Qingxin Pill can effectively relieve the behavioral symptoms of depression and anxiety caused by depression.
  • the inventors evaluated the neurogenesis in the hippocampal DG area of each group of mice.
  • Niuhuang Qingxin Pill treatment significantly increased the density of newborn cells and neurons in the hippocampal DG area, and this effect was the same as that of the fluoxetine positive control group ( Figure 2a-2b, one- way ANOVA, #: p ⁇ 0.05 compared with model group).
  • Tongrentang Niuhuang Qingxin Pills can significantly promote hippocampal neurogenesis damaged by long-term external pressure. This result provides a neurobiological basis for Tongrentang Niuhuang Qingxin Pill to prevent depression and anxiety caused by depression.
  • Tongrentang Niuhuang Qingxin Pill can effectively combat depression and anxiety symptoms caused by behavioral restraint and promote hippocampal neurogenesis. It suggests that Tongrentang Niuhuang Qingxin Pills can have a significant therapeutic effect in fighting depression and anxiety caused by external pressure.
  • glucocorticoids are significantly increased in patients with stress and depression.
  • Glucocorticoids can play a series of effects such as decreased neurogenesis levels by acting on the glucocorticoid receptors of neurons in the brain. These neurological changes are ultimately reflected in depression and anxiety behavior. Therefore, the inventor designed a long-term feeding model with glucocorticoids to simulate depression caused by endocrine changes, and to understand the therapeutic effect of Tongrentang Niuhuang Qingxin Pills on this depression model.
  • Figure 3 shows the depressive behavior of mice with different treatments. It can be concluded from the figure that long-term feeding of corticosteroids (CORT) caused significant depression-like behavior in mice.
  • CORT corticosteroids
  • the specific performance is that the self-organization time in the sugar water spraying experiment was significantly lower than that of the control group (Figure 3a, one-way ANOVA, **: p ⁇ 0.01 compared with the control group); and activities in the tail suspension test and the forced swimming experiment Time reduction (Figure 3b-3c, one-way ANOVA, ***: p ⁇ 0.001 compared with the control group).
  • mice showed more time to self-organize in the sugar water spray experiment (Figure 3a, one-way ANOVA, #: p ⁇ 0.05 compared with the model group); At the same time, Niuhuang Qingxin Pills in the tail suspension experiment and the forced swimming experiment also significantly restored the activity time of the mice ( Figure 3b-3c, one-way ANOVA, #: p ⁇ 0.05 compared with the model group); this result is consistent with the positive The control drug fluoxetine has been (one-way ANOVA, ###: p ⁇ 0.001 compared with the model group).
  • Tongrentang Niuhuang Qingxin Pills significantly reversed the prolongation of foraging latency and the reduction of food intake in mice caused by long-term feeding of corticosterone (Figure 3e-3f, one-way ANOVA) , ##: p ⁇ 0.05 compared with the model group). And this kind of effect is consistent with the fluoxetine effect.
  • Tongrentang Niuhuang Qingxin Pills can effectively treat depression and anxiety models caused by elevated corticosterone.
  • the inventors in order to detect the effect of Tongrentang Niuhuang Qingxin Pills on hippocampal neurogenesis in a corticosterone-fed model, the inventors also used immunofluorescence to detect the cell density of BrdU-labeled nerve cells and precursor cells in the hippocampal DG area.
  • the corticosterone feeding model resulted in a significant decrease in the number of BrdU positive cells in the DG zone (4a, one-way ANOVA, ***: p ⁇ 0.001), and the inventor found that Tongrentang Niuhuang Qingxin Pills
  • the treatment significantly increased the number of BrdU cells in the hippocampal DG area and the effect was consistent with the positive control drug fluoxetine (4a, one-way ANOVA, ##: p ⁇ 0.01). This shows that Tongrentang Niuhuang Qingxin Pills can promote the proliferation of hippocampal DG cells under depression.
  • the inventors co-stained BrdU and the mature neuron marker NeuN and used statistics of BrdU/NeuN double-positive cells to identify the neurogenesis effect of Tongrentang Niuhuang Qingxin Pills.
  • the corticosterone feeding model also significantly reduced the level of BrdU/NeuN positive cells in the DG zone (D, one-way ANOVA, **: p ⁇ 0.01 compared with the control group).
  • the inventor’s results showed that Tongrentang Niuhuang Qingxin Pills can significantly improve the number of newborn neurons in the DG area.
  • Tongrentang Niuhuang Qingxin Pills can also improve the number of BrdU/DCX double-positive cells suppressed by corticosterone, and the effect is consistent with the positive control fluoxetine ( Figure 4c, one-way ANOVA, #: p ⁇ 0.05, ##: p ⁇ 0.01 compared with the model group).
  • the results of the inventors indicate that Tongrentang Niuhuang Qingxin Pills have a significant effect on promoting neurogenesis in the treatment of depression caused by corticosterone, and the effect on the brain area is more extensive than that of the traditional antidepressant western drug fluoxetine.
  • Tongrentang Niuhuang Qingxin Pills can effectively combat depression and anxiety symptoms caused by corticosterone feeding and promote hippocampal neurogenesis. It is suggested that Tongrentang Niuhuang Qingxin Pill has a significant therapeutic effect in combating depression and anxiety caused by endocrine disorders.
  • Tongrentang Niuhuang Qingxin Pill has a very significant effect in combating anxiety and depression caused by depression.
  • Tongrentang Niuhuang Qingxin Pill is a representative prescription for regulating emotions. It has the effects of replenishing the heart and spleen, nourishing qi and blood, soothing the liver and relieving depression, calming the nerves, resolving phlegm and eliminating wind.
  • This research project confirmed the therapeutic effect of Tongrentang Niuhuang Qingxin Pills on depression from the perspectives of behavior and neurobiology.
  • Tongrentang Niuhuang Qingxin Pills can relieve depression and anxiety symptoms caused by two different reasons.
  • the inventors discovered the promoting effect of Tongrentang Niuhuang Qingxin Pill on hippocampal neurogenesis. Based on this, the inventors more clearly proved that Tongrentang Niuhuang Qingxin Pill has a neurobiological mechanism against depression, and it may be Tongrentang Niuhuang Qingxin Pill.
  • Treatment of other neurodegenerative diseases such as senile dementia, brain damage after stroke, etc. provide neurobiological evidence.
  • Tongrentang Niuhuang Qingxin Pills can effectively treat depression and anxiety symptoms caused by behavioral restraint and corticosterone feeding, and can improve neurological function in depression by promoting hippocampal neurogenesis.
  • Tongrentang Niuhuang Qingxin Pill (Natural Recipe) (20mg/kg/d) was given to male C57BL/6N mice by gavage for one week, and the same dose of Tongren Niuhuang Qingxin Pill (Artificial Recipe) was administered for one week.
  • the basic nerve regeneration in the hippocampus Observed.
  • Tongrentang Niuhuang Qingxin Pills natural recipe, bezoar, musk, and borneol are all made of natural sources
  • Tongren Niuhuang Qingxin Pills artificial recipe, bezoar, musk, and borneol are all made of artificial ingredients
  • DCX is a neuroblast marker, and its density represents the differentiation ability of hippocampal neural stem cells.

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Abstract

牛黄清心丸，特别是同仁堂制的牛黄清心丸在制备治疗抑郁症的药物中的应用。

Description

一种牛黄清心丸在制备治疗抑郁症的药物中的应用 技术领域
本发明属于生物医药领域，具体涉及一种牛黄清心丸在制备治疗抑郁症的药物中的应用
技术背景
抑郁症(重度抑郁或临床抑郁症)已经成为了严重威胁当今人类精神健康的心理疾病。抑郁症病人常常伴随悲伤、焦虑、易怒、过分的负罪感以及羞涩的情绪，抑郁症还严重影响患者睡眠、饮食、以及社会生活，严重者导致患者自杀。因此，抗抑郁药物的开发对促进社会发展及减轻人民负担具有深远的影响。
早期心理干预对抑郁症患者确有疗效，但是心理干预并不能改变患者长期所处的生活环境，心理干预对抑郁症尤其是重度抑郁的改善作用有限。药物治疗是目前抑郁症主要的治疗途径，通用的抗抑郁西药包括SSRI(选择性5-羟色胺再摄取抑制剂)、SNRIs(5-羟色胺去甲肾上腺素再摄取抑制剂)、NDRIs(去甲肾上腺素多巴胺再摄取抑制剂)、Tricyclic antidepressants(三环类抗抑郁药物)以及MAOIs(单胺氧化酶抑制剂)。西药治疗抑郁症有短期疗效同时副作用也十分明显，除药物成瘾外，上述药物还具有其他副作用，常见例如口干、睡眠异常、尿潴留、视力模糊、头痛、焦虑等。其他治疗方法如电休克刺激(ECT)方法也无法满足目前抑郁症人群的通用治疗需求。
作为替代疗法，中医药治疗情志病的历史上已有2000多年，积累了大量的治疗方剂，临床实践表明，中医药在治疗抑郁症方面有独到的优势，多种经典传统成方都具有很强的情绪调节作用。中医药理论认为，抑郁症为本虚标实之证，与心脾肝肾功能失调，心脾气血亏虚，肝肾不足，肝郁气滞，痰浊蒙闭心神有关。牛黄清心丸就是一个代表性的调节情志的方剂，由牛黄、羚羊角、麝香、人参、白术(麸炒)、当归、白芍、柴胡、干姜、阿胶、桔梗、水牛角浓缩粉等27味药材配置而成，具有补益心脾、益气养血、疏肝解郁、镇静安神、化痰熄风功效。适用于气血不足、痰热上扰引起；也用于胸中郁热、惊悸虚烦、头目眩晕、中风不语、口眼歪斜、半身不遂、言语不清、神志昏迷等痰涎壅盛病证。
目前为止牛黄清心丸疗效报道包括：(1)口腔疾病，如口腔粘膜溃疡、口舌生疮、 牙痛、牙跟肿痛、急慢性咽炎等；(2)痰火上扰所致呢逆；(3)颅内高压昏迷患者；(4)脑血管病，天津市和平区中医医院治疗急性脑血管病281例，凡遇有意识障碍属阳闭者，给予本品及安宫牛黄丸，每日1～2丸交替服用，效果满意。(5)治疗疱疹神经痛：30例带状疱疹后遗神经痛患者，给予同仁牛黄清心丸1丸与龙胆泻肝丸1袋，治愈18例，好转10例。
直至目前，尚未有研究显示牛黄清心丸可以用于抑郁症的研究或报道。同时毒理学研究显示，以相当于人用剂量的461倍的同仁堂牛黄清心丸对小鼠灌胃给药，未见明显急性毒性反应。分别相当于人日用剂量的50、25和12.5倍，连续给SD大鼠灌胃26周，实验期间(给药13、26周以及停药4周)所有剂量组动物均未出现死亡，对3个剂量组的大鼠一般状况、体重及摄食量均无明显影响；尿糖、尿酮、尿胆素原、尿蛋白等11项尿液指标与正常对照组比较未见明显异常；凝血时间、血常规指标、血液生化指标均在正常范围，同对照组比较均无明显异常；给药组大鼠受检脏器无明显改变，胸、腹等体腔无异常积液；脏器指数均在正常范围，同对照组比较均无明显差异；病理学检测显示主要脏器组织均未见因药物引起的形态学改变，也未见明显药物蓄积性或药物延迟性毒性。说明同仁堂牛黄清心丸可以作为长期使用安全的药物，如果其抗抑郁症疗效可以证实，将有重大意义。
虽然同仁堂牛黄清心丸具有情志调节作用，且长期服用下无明显副作用，但同仁堂牛黄清心丸是否对抑郁症具有改善作用，尚未存在相关研究，因此其在抑郁症治疗领域属于应用空白。
发明内容
基于上述技术问题，发明人提供了一种牛黄清心丸在治疗抑郁症的药物中的应用，其中，所述牛黄清心丸为同仁堂牛黄清心丸。
同仁堂牛黄清心丸是本专利发明中实验例所采用的药剂形式，而中国国内的其他品牌牛黄清心丸的组成和制备方法均与其具有区别，具体区别设计商业机密，不便论述，因此不同牛黄清心丸所具有的功能具有显著区别，因此本专利申请中所述牛黄清心丸优选同仁堂牛黄清心丸。
特别是优选同仁堂牛黄清心丸的天然方，即其中的所有组分采用天然组分(其中优选牛黄、麝香、冰片均选用天然成分)，其抗抑郁的药效要优于牛黄清心丸的人工方。 见实施例2。
优选地，在上述的同仁堂牛黄清心丸在治疗抑郁症的药物中的应用，所述抑郁症的致病原因为心理压力升高或正常环境下内分泌失调。
由于抑郁症的成因不同，表现形式也各异，同仁堂牛黄清心丸在实验例中对于心理压力升高或正常环境下的内分泌失调引起的抑郁症具有显著效果。
优选地，在上述的同仁堂牛黄清心丸在治疗抑郁症的药物中的应用，所述同仁堂牛黄清心丸的成人给药剂量1-20g/日。
优选地，在上述的牛黄清心丸在治疗抑郁症的药物中的应用，所述同仁堂牛黄清心丸的成人给药剂量为3-15g/日。
优选地，在上述的牛黄清心丸在治疗抑郁症的药物中的应用，所述同仁堂牛黄清心丸的成人给药剂量为6-12g/日。
优选地，在上述的牛黄清心丸在治疗抑郁症的药物中的应用，所述同仁堂牛黄清心丸的使用方法为口服。
优选地，在上述的同仁堂牛黄清心丸在治疗抑郁症的药物中的应用，所述同仁堂牛黄清心丸在小鼠试验中，使用剂量为20mg/kg，显著促进海马神经的发生。
附图说明
图1同仁堂牛黄清心丸有效阻止了拘束实验引起的抑郁焦虑行为；图1a-1f分别对应拘束组小鼠的糖水喷涂实验、悬尾试验、强制游泳、旷场实验、新事物进食中的(进食潜伏期和进食摄入量研究)；
图2同仁堂牛黄清心丸促进了抑郁症压抑的海马神经发生；图2a-2b分别对应同仁堂牛黄清心丸对拘束组的小鼠海马DG区新生细胞(BrdU+)的数量和DG区新生神经元(BrdU+/DCX+)的数目的影响)；
图3同仁堂牛黄清心丸有效阻止了皮质酮饲喂引起的抑郁焦虑症状；图3a-3f分别对应皮质酮组小鼠的糖水喷涂实验、悬尾试验、强制游泳、旷场实验、新事物进食中的(进食潜伏期和进食摄入量研究)；
图4同仁堂牛黄清心丸改善了慢性皮质酮饲喂压抑的海马神经发生。(图4a-4c分别对应同仁堂牛黄清心丸对皮质酮的小鼠海马区新生细胞(BrdU+)的数量、海马区新生神经元(BrdU+/NeuN+)的数目和海马区新生神经前体细胞(BrdU+/DCX+)数量的影 响)。
具体实施方式
实验例1
药物：同仁堂牛黄清心丸，规格为3g/每丸(HKC-18084，同仁堂牛黄清心丸在香港获批上市信息可登陆下列网址查询：http://www.cmchk.org.hk/pcm/chi/#main_listpcm_2018.htm)，成人用量为一次1～2丸；一日2次，小儿酌减。
本实验采用了拘束组小鼠的糖水喷涂实验Splash Test、悬尾试验Tail Suspension Test、强制游泳Forced Swim Test、旷场实验Open Field Test、新事物进食Novelty Suppressed Feeding等常规方法对小鼠的各项指标进行研究。
1抑郁症动物模型：本研究采用了长期行为拘束模型与皮质酮饲喂模型分别模拟外界压力与体内内分泌紊乱两种因素所导致的抑郁症。
2动物分组：选择8周雄性C57BL/6N小鼠作为实验动物，具体操作如下：
2.1拘束实验组：对照组(正常饲养，无处理，实验过程中每日给予0.1ml生理盐水灌胃)；抑郁症模型组(50ml塑料拘束管，带有11个气孔，将小鼠放置入内，拧紧盖子并使其无法自由活动，每天从上午九点开始进行6小时拘束，持续30天；实验过程中每日拘束前给予0.1ml生理盐水灌胃)；治疗组(在每日给予拘束前，于小鼠灌胃同仁堂牛黄清心丸生理盐水混悬液(20mg/kg，相当于成人一丸计量))；西药对照组(采用经典的抗抑郁药物氟西汀作为对照，在每日拘束前，给与小鼠氟西汀生理盐水溶液(18mg/kg))。
2.2皮质酮饲喂实验：对照组(正常饲养，无处理；从造模过程第15日起过程中每日给予0.1ml生理盐水灌胃)；皮质酮饲喂模型(将皮质酮以70μg/ml溶解于0.45％beta-环糊精的饮水中，持续30天；从造模过程第15日起过程中每日给予0.1ml生理盐水灌胃)；治疗组(饮水处理与模型组相同；从造模过程第15日起过程中每日给予牛黄清心丸混悬液20mg/kg灌胃处理)；西药对照组(采用经典的抗抑郁药物氟西汀作为对照，饮水处理与模型组相同；从造模过程第15日起过程中每日给氟西汀生理盐水溶液18mg/kg灌胃处理)。
3焦虑抑郁行为学测定：通过糖水喷涂实验(Splash)、强制游泳(FST)、悬尾实验(TST)、旷场试验(OFT)以及新环境进食抑制(NSF)这五种行为学测定手段评价 动物的抑郁症程度(各组动物均需12只放可以达到统计标准)。(所述实验方法均可参照相关实验教程，属于本领域常用方法)
4海马神经发生研究：本研究通过目前最直接的激光共聚焦显微镜技术(confocal microscope)结合免疫荧光染色(immunofluorescence)直接对脑内神经干细胞的增殖，分化，以及成熟过程进行观察。通过各处理海马DG区不同标记物阳性的神经干细胞/前提细胞数平均牛黄清心丸的抗抑郁治疗效果。
BrdU标记：连续5天进行BrdU腹腔注射(50mg/kg/d)以标记新生细胞。在最后一次标记后七天将小鼠处死并将脑组织以4％多聚甲醛固定。并进行冰冻切片，已经免疫荧光染色。
实验结果
1同仁堂牛黄清心丸有效阻止了拘束实验引起的抑郁焦虑行为
如图1所示，行为拘束实验导致了小鼠在不同行为学测试中表现了明显的抑郁与焦虑现象。在糖水喷涂实验(Splash，图1a)中，小鼠整理毛发时间显著下降，说明了小鼠失去了主观躲避不适体验的能动性，提示了其抑郁症状。在悬尾试验(TST，图1b)与强制游泳实验(FST，图1c)中，小鼠在模拟的绝望环境下的活动时间明显减少。三组行为学均证明，拘束造成了显著的抑郁情绪。同时，旷场实验(OFT，图1d)提示小鼠在中心区域的运动时间显著减少；而新环境进食抑制(NSF，图1e-1f)结果表明小鼠在经历过饥饿后，其在旷场中进食潜伏期明显延长。这两组数据均提示了小鼠的焦虑行为。而从数据上可以看出，同仁堂牛黄清心丸与氟西汀效果一致，均逆转了小鼠的焦虑与抑郁行为。统计学分析，单因素方差分析(one-way ANOVA，n＝12per group，*：p<0.05，***：p<0.001与对照相比；#：p<0.05，##：p<0.01，###：p<0.001与模型组相比)。从六种行为学数据可以得出结论，同仁堂牛黄清心丸可以有效缓解抑郁症引起的抑郁焦虑的行为症状。
2同仁堂牛黄清心丸促进了抑郁症压抑的海马神经发生
通过免疫荧光共染色神经前体细胞标记物DCX与新生细胞标记BrdU，发明人对各组小鼠海马DG区的神经发生进行了评价。发明人的实验结果显示，行为拘束模型显著降低了小鼠海马DG区新生细胞(BrdU+)的数量(图2a；n＝6per group，one-way ANOVA，***：p<0.001与对照组相比)；同时，该模型也显著降低了DG区新生神经元(BrdU+/DCX+)的数目(图2b；n＝6per group，one-way ANOVA，***：p<0.001 与对照组相比)。与拘束模型组相比，牛黄清心丸处理(NHQXW)显著提高了海马DG区新生细胞与新生神经元的密度，并且这种效应与氟西汀阳性对照组结果相同(图2a-2b，one-way ANOVA，#：p<0.05与模型组对比)。这一结果提示了，同仁堂牛黄清心丸能够显著的促进由外界长期压力所损伤的海马神经发生。这一结果对同仁堂牛黄清心丸防止抑郁症引起的抑郁焦虑行为提供了神经生物学依据。
基于1和2部分结果，发明人能够证明，同仁堂牛黄清心丸可以有效对抗行为拘束引起的抑郁焦虑症状并促进海马神经发生。提示了在对抗外界压力引起的抑郁焦虑情绪中，同仁堂牛黄清心丸能够起到显著的治疗效果。
3同仁堂牛黄清心丸有效阻止了皮质酮饲喂引起的抑郁焦虑症状
糖皮质激素作为一种应激激素，其在压力应激以及抑郁症病人中会显著升高。而糖皮质激素通过作用脑内神经元的糖皮质激素受体会起到一系列的作用例如神经发生水平下降等。这些神经功能改变最终反映在抑郁焦虑行为上。所以，发明人设计了以糖皮质激素长期饲喂模型从而模拟内分泌改变引发的抑郁症，并以此了解同仁堂牛黄清心丸对该抑郁症模型的治疗效果。
如图所示，图3显示了不同处理小鼠的抑郁行为。从图中可以得出长期饲喂糖皮质激素(CORT)导致了小鼠出现显著的抑郁样行为。其具体表现为在糖水喷涂实验中自我整理时间较对照组显著下降(图3a，one-way ANOVA，**：p<0.01与对照组相比)；以及在悬尾试验与强制游泳实验中活动时间的降低(图3b-3c，one-way ANOVA，***：p<0.001与对照组相比)。与模型组相比，同仁堂牛黄清心丸治疗后，小鼠在糖水喷涂实验中表现出了更多的自我整理时间(图3a，one-way ANOVA，#：p<0.05与模型组相比)；同时，悬尾实验与强制游泳实验中牛黄清心丸也显著恢复了小鼠的活动时间(图3b-3c，one-way ANOVA，#：p<0.05与模型组相比)；这一结果与阳性对照药物氟西汀一直(one-way ANOVA，###：p<0.001与模型组相比)。这部分结果说明了同仁堂牛黄清心丸在调节内分泌引起的抑郁症方面的治疗效果。另外，图3结果显示了小鼠的焦虑行为实验。同仁堂牛黄清洗丸同样可以逆转皮质酮慢性饲喂引起的小鼠在旷场实验中中心区域活动的减少(图3d，one-way ANOVA，##：p<0.01与模型组相比)。与此同时，而新环境进食抑制(NSF)结果也表明同仁堂牛黄清心丸显著逆转了长期饲喂皮质酮导致的小鼠觅食潜伏期的延长以及食量的降低(图3e-3f，one-way ANOVA，##：p<0.05与模型组相比)。并且这类效应与氟西汀效果一致。综上显示，同仁堂牛黄清心 丸能够有效治疗皮质酮升高引起的抑郁与焦虑模型。
4同仁堂牛黄清心丸改善了慢性皮质酮饲喂压抑模型中的海马神经发生状态
如图所示，为了检测同仁堂牛黄清心丸对皮质酮饲喂模型中海马神经发生的作用，发明人同样通过免疫荧光方法检测了海马DG区BrdU标记的神经细胞以及前体细胞的细胞密度。如图4a所示，皮质酮饲喂模型导致了BrdU在DG区阳性细胞数目的显著下降(4a，one-way ANOVA，***：p<0.001)，并且，发明人发现了同仁堂牛黄清心丸治疗显著升高了BrdU在海马DG区的细胞数目并且效果与阳性对照药物氟西汀一致(4a，one-way ANOVA，##：p<0.01)。这说明了同仁堂牛黄清心丸在抑郁症状态下能够促进海马DG区细胞的增殖。为了鉴定增殖的细胞类型，发明人共染了BrdU与成熟神经元标记物NeuN并通过统计BrdU/NeuN双阳性细胞鉴定同仁堂牛黄清心丸的神经发生效应。如图4b所示，皮质酮饲喂模型也显著降低了BrdU/NeuN阳性细胞在DG区的水平(D，one-way ANOVA，**：p<0.01与对照组相比)。而发明人的结果显示了同仁堂牛黄清心丸能够显著改善DG区的新生神经元数目，与模型相比，同仁堂牛黄清心丸治疗组海马内BrdU/NeuN双阳性细胞数目显著升高并于氟西汀组一致(图4b，one-way ANOVA，#：p<0.05，##：p<0.01与模型组相比)。同样，通过对比BrdU与新生非成熟神经元标记物DCX共染水平发明人发现同仁堂牛黄清心丸也能够改善皮质酮压抑的BrdU/DCX双阳性细胞的数量并且效果与阳性对照组氟西汀一致(图4c，one-way ANOVA，#：p<0.05，##：p<0.01与模型组相比)。发明人的结果说明，同仁堂牛黄清心丸在对抗皮质酮引起的抑郁症中具有显著的促进神经发生的功效并且作用的脑区比传统抗抑郁西药氟西汀更为广泛。
基于3和4部分结果，发明人能够证明，同仁堂牛黄清心丸可以有效对抗皮质酮饲喂引起的抑郁焦虑症状并促进海马神经发生。提示了其在对抗内分泌失调引起的抑郁焦虑情绪中，同仁堂牛黄清心丸具有显著的治疗效果。
总结与讨论
通过两种不同病机为基础的抑郁症模型，发明人发现了同仁堂牛黄清心丸在对抗抑郁症引起的焦虑抑郁行为具有十分显著的疗效。同仁堂牛黄清心丸就是一个代表性的调节情志的方剂，具有补益心脾，益气养血，疏肝解郁，镇静安神，化痰熄风功效。本研究项目从行为学和神经生物学角度，确认了同仁堂牛黄清心丸对抑郁症的治疗效果。以行为拘束和皮质酮饲喂模型分别模拟心理压力升高以及正常环境下内分泌失调两种原 因导致的抑郁症状；这两种病因也是当今社会抑郁症最重要的两大原因。通过两种不同的模型，发明人确认了同仁堂牛黄清心丸能够缓解两种不同原因导致的抑郁焦虑症状。同时，发明人发现了同仁堂牛黄清心丸对海马神经发生的促进作用，以此为依据，发明人更加明确证明同仁堂牛黄清心丸具有对抗抑郁症的神经生物学机制，并且有可能为同仁堂牛黄清心丸治疗其他神经退行性疾病例如老年性痴呆，中风后脑损伤等提供神经生物学依据。
综上所述，同仁堂牛黄清心丸能够有效治疗行为拘束和皮质酮饲喂引起的抑郁焦虑症状并能够通过促进海马神经发生改善抑郁症中的神经功能。
实施例2
同仁堂牛黄清心丸(天然方)(20mg/kg/d)进行8周雄性C57BL/6N小鼠灌胃处理一周，与同仁牛黄清心丸(人工方)相同剂量灌胃处理一周后海马基础神经再生状况观察。
经对比发现同仁堂牛黄清心丸(天然方，牛黄、麝香、冰片均采用天然来源)与人工方的同仁牛黄清心丸(人工方，牛黄、麝香、冰片均采用人工成分)对海马神经干细胞增值水平并无显著差别。DCX为成神经细胞标记物，其密度代表海马神经干细胞分化能力。经比较发现相同剂量的同仁堂牛黄清心丸(天然方)DCX阳性细胞数显著高于人工方，证明天然方具有比人工方更好的促进海马神经分化的潜能，提高小鼠抗抑郁能力。

Claims (8)

  1. 一种牛黄清心丸在制备治疗抑郁症的药物中的应用，其特征在于，所述牛黄清心丸为同仁堂牛黄清心丸。
  2. 根据权利要求1所述的应用，其特征在于，所述抑郁症的致病原因为心理压力升高或正常环境下内分泌失调。
  3. 根据权利要求1所述的应用，其特征在于，所述同仁堂牛黄清心丸的成人给药剂量为1-20g/日。
  4. 根据权利要求3所述的应用，其特征在于，所述同仁堂牛黄清心丸的成人给药剂量为3-15g/日。
  5. 根据权利要求4所述的应用，其特征在于，所述同仁堂牛黄清心丸的成人给药剂量为6-12g/日。
  6. 根据权利要求5所述的应用，其特征在于，所述同仁堂牛黄清心丸的使用方法为口服。
  7. 根据权利要求1所述的应用，其特征在于，所述同仁堂牛黄清心丸在小鼠试验中，使用剂量为20mg/kg，显著促进海马神经的发生。
  8. 根据权利要求1所述的应用，其特征在于，所述牛黄清心丸为同仁堂牛黄清心丸的天然方产品。
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CN103099866A (zh) * 2013-01-18 2013-05-15 北京中医药大学东方医院 一种治疗焦虑、失眠的中药组合物及其制备方法

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103099866A (zh) * 2013-01-18 2013-05-15 北京中医药大学东方医院 一种治疗焦虑、失眠的中药组合物及其制备方法

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
LI, TAO: "Stagnant-fire Based Discussion on the Depressive Disorder with Insomnia", WORLD JOURNAL OF SLEEP MEDICINE, vol. 3, no. 5, 31 October 2016 (2016-10-31) *
NI, SHENGLOU: "Non-official translation: TCM Treatment for Adjusting Hardness and Softness", TCM HEALTHY LIFE-NURTURING, no. 2,, 28 February 2017 (2017-02-28) *

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