CN118634285A - A compound camellia pill for soothing the liver and strengthening the spleen and its preparation method and application - Google Patents
A compound camellia pill for soothing the liver and strengthening the spleen and its preparation method and application Download PDFInfo
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Abstract
本发明公开了一种疏肝健脾复方山茶丸及其制备方法和应用,属于中药技术领域。本发明提供的疏肝健脾复方山茶丸,按重量份计,包括以下原料:山茶花10‑15份、栀子叶6‑10份、牛舌草6‑10份、黄精叶3‑6份、橘皮3‑6份、猪胆粉1‑3份和甘草1‑3份。本发明提供疏肝健脾复方山茶丸通过合理配伍,符合“君臣佐使”配伍治则,从“扶正祛邪”两方面发挥疏肝健脾的功效;且经试验验证,本发明制备的疏肝健脾复方山茶丸可明显改善小鼠的肝脾损伤,无毒副作用,可为防治肝脾损伤提供一种新思路。
The invention discloses a compound camellia pill for soothing the liver and strengthening the spleen and its preparation method and application, and belongs to the technical field of traditional Chinese medicine. The compound camellia pill for soothing the liver and strengthening the spleen provided by the present invention includes the following raw materials by weight: 10-15 parts of camellia, 6-10 parts of gardenia leaves, 6-10 parts of ox tongue grass, 3-6 parts of polygonatum leaves, 3-6 parts of orange peel, 1-3 parts of pig gall powder and 1-3 parts of liquorice. The present invention provides a compound camellia pill for soothing the liver and strengthening the spleen, which conforms to the compatibility treatment principle of "monarch, minister, assistant and messenger" through reasonable compatibility, and exerts the effect of soothing the liver and strengthening the spleen from the two aspects of "strengthening the body and eliminating evil"; and it is verified by experiments that the compound camellia pill for soothing the liver and strengthening the spleen prepared by the present invention can significantly improve the liver and spleen damage of mice, without toxic side effects, and can provide a new idea for preventing and treating liver and spleen damage.
Description
技术领域Technical Field
本发明涉及中药技术领域,特别是涉及一种疏肝健脾复方山茶丸及其制备方法和应用。The invention relates to the technical field of traditional Chinese medicines, and in particular to a liver-soothing and spleen-strengthening compound camellia pill and a preparation method and application thereof.
背景技术Background Art
肝脏是一种重要的脏器,承担着代谢调节和储存作为三大营养素的糖、蛋白质和脂质以及分解和解毒机体不需要的物质等各种功能。酒精的过度摄取、病毒感染、不均衡的饮食习惯、压力和吸烟等会引起这些功能受到急性或慢性损伤,并且当该损伤发展时,会出现例如急性肝炎、慢性肝炎、肝硬化、酒精性脂肪肝、B型病毒肝炎、肝癌等疾病。肝脏出现疾病会累及脾脏,导致脾功能的异常。《金匮要略》“见肝之病,知肝传脾,当先实脾”。肝属木,脾属土,肝藏血而主疏泄,脾统血而主运化,为气血生化之源、气机升降之枢。二者生理上相互协调、促进,按照五行生克的规律,即土需木疏,木赖土荣。肝木疏土,土得木而达之,肝之疏泄功能正常,气机条畅,则可促进脾胃之受纳、运化,使脾升胃降协调平衡,从而使脾运健旺,生化有源,统摄有权。《血证论》云:“木之性,主于疏泄,食气入胃,全赖肝木之气以疏泄之,而水谷乃化”。土为万物之母,仓廪之本,肝木必得脾土所化生之水谷精微濡养,方遂其条达之性,而不会刚强太过,肝木才得以正常发挥。由此可见,肝脾两脏在生理和病理上相互影响,当一方太过或不及,以致打破这种平衡,即出现病变,且相互影响而表现出一系列肝脾两脏的病证。The liver is an important organ that undertakes various functions such as metabolic regulation and storage of sugar, protein and lipid as the three major nutrients, as well as decomposition and detoxification of substances not needed by the body. Excessive alcohol intake, viral infection, unbalanced eating habits, stress and smoking can cause acute or chronic damage to these functions, and when the damage develops, diseases such as acute hepatitis, chronic hepatitis, cirrhosis, alcoholic fatty liver, hepatitis B virus, liver cancer, etc. will occur. Liver diseases will affect the spleen, leading to abnormal spleen function. "Golden Chamber" says, "When you see a liver disease, you know that the liver will pass it on to the spleen, so you should first strengthen the spleen." The liver belongs to wood, and the spleen belongs to earth. The liver stores blood and is responsible for dredging, while the spleen regulates blood and is responsible for transportation and transformation. It is the source of qi and blood biochemical transformation and the pivot of qi movement. The two coordinate and promote each other physiologically, according to the law of the five elements, that is, earth needs wood to dredge, and wood depends on earth to flourish. The liver and wood disperse the earth, and the earth is able to reach it with the help of wood. If the liver's function of dispersing and dispersing is normal and the qi is smooth, it can promote the spleen and stomach's reception and transportation, so that the spleen's rise and stomach's descent are coordinated and balanced, thus making the spleen's movement strong, biochemical active, and controlling power. "Blood Symptoms" says: "The nature of wood is to disperse and disperse. When food qi enters the stomach, it depends entirely on the liver and wood's qi to disperse and disperse it, and then the water and grain are transformed." The earth is the mother of all things and the foundation of the granary. The liver and wood must be nourished by the water and grain essence produced by the spleen and earth, so that it can be smooth and unobstructed, and it will not be too strong, so that the liver and wood can function normally. It can be seen that the liver and spleen affect each other in physiology and pathology. When one side is too much or too little, it breaks this balance, that is, pathological changes occur, and they affect each other and show a series of diseases and symptoms of the liver and spleen.
目前,市面上的疏肝和健脾等保健产品琳琅满目,但兼有二者功效的产品却很少,此外即使有,该产品的功效单一、疗效不佳,甚至添加了激素等可能损害机体的成分。为此,本发明特地提出一种疏肝健脾复方山茶丸来保护肝脏脾脏。At present, there are a variety of health care products on the market that can soothe the liver and strengthen the spleen, but there are few products that have both effects. In addition, even if there are, the effect of the product is single, the curative effect is poor, and even hormones and other ingredients that may damage the body are added. For this reason, the present invention specifically proposes a compound tea pill for soothing the liver and strengthening the spleen to protect the liver and spleen.
发明内容Summary of the invention
本发明的目的是提供一种疏肝健脾复方山茶丸及其制备方法和应用,以解决上述现有技术存在的问题。本发明制备的疏肝健脾复方山茶丸可有效缓解小鼠的肝脾损伤,无毒副作用,可为防治肝脾损伤提供一种新思路。The purpose of the present invention is to provide a compound camellia pill for soothing the liver and strengthening the spleen and a preparation method and application thereof to solve the problems existing in the above-mentioned prior art. The compound camellia pill for soothing the liver and strengthening the spleen prepared by the present invention can effectively alleviate liver and spleen damage in mice, has no toxic side effects, and can provide a new idea for preventing and treating liver and spleen damage.
为实现上述目的,本发明提供了如下方案:To achieve the above object, the present invention provides the following solutions:
本发明提供一种疏肝健脾复方山茶丸,包括以下重量份原料:山茶花10-15份、栀子叶6-10份、牛舌草6-10份、黄精叶3-6份、橘皮3-6份、猪胆粉1-3份和甘草1-3份。The invention provides a compound camellia pill for soothing the liver and strengthening the spleen. The compound camellia pill comprises the following raw materials in parts by weight: 10-15 parts of camellia, 6-10 parts of gardenia leaves, 6-10 parts of oxalis, 3-6 parts of polygonatum leaves, 3-6 parts of tangerine peel, 1-3 parts of pig gall powder and 1-3 parts of liquorice.
本发明还提供一种上述的疏肝健脾复方山茶丸的制备方法,包括以下步骤:The present invention also provides a method for preparing the above-mentioned compound camellia pills for soothing the liver and strengthening the spleen, comprising the following steps:
将山茶花、栀子叶、牛舌草、黄精叶、橘皮和甘草混匀,粉碎成超微粉;冷凝回流得滤液,过滤,取上清浓缩成膏状,得浸膏;Mix camellia, gardenia leaves, oxalis, polygonatum leaves, orange peel and liquorice, and grind them into ultrafine powder; condense and reflux to obtain a filtrate, filter, and take the supernatant and concentrate it into a paste to obtain an extract;
向所述浸膏中加入猪胆粉搅拌均匀,得到混浸膏;Add pig bile powder to the extract and stir evenly to obtain a mixed extract;
所述混浸膏与辅料混合,制成丸剂。The mixed extract is mixed with auxiliary materials to prepare pills.
进一步地,所述辅料包括稀释剂、填充剂、赋形剂、粘合剂、湿润剂、崩释剂、吸收促进剂、表面活性剂、吸附载体、润滑剂和香味剂中的一种或多种。Furthermore, the auxiliary materials include one or more of diluents, fillers, excipients, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers, lubricants and flavoring agents.
进一步地,所述辅料包括可溶性淀粉0.8份、山茶油0.1份和蔗糖0.04份。Furthermore, the auxiliary materials include 0.8 parts of soluble starch, 0.1 parts of camellia oil and 0.04 parts of sucrose.
进一步地,所述冷凝回流的时间为60min。Furthermore, the condensation reflux time is 60 minutes.
进一步地,所述过滤为真空负压过滤,过滤次数为2次。Furthermore, the filtration is vacuum negative pressure filtration, and the filtration times are 2 times.
本发明还提供一种上述的疏肝健脾复方山茶丸在制备防治肝脾损伤药物中的应用。The present invention also provides an application of the above-mentioned compound camellia pills for soothing the liver and strengthening the spleen in preparing medicines for preventing and treating liver and spleen injuries.
本发明还提供一种防治肝脾损伤的药物,包括上述的疏肝健脾复方山茶丸,和辅料。The present invention also provides a medicine for preventing and treating liver and spleen damage, comprising the above-mentioned compound camellia pills for soothing the liver and strengthening the spleen, and auxiliary materials.
进一步地,所述药物的剂型包括丸剂。Furthermore, the dosage form of the drug includes pills.
进一步地,所述药物的服用方法包括口服和非经肠胃形式服用。Furthermore, the method of taking the drug includes oral and parenteral administration.
本发明的疏肝健脾复方山茶丸中,山茶花、栀子叶、牛舌草疏理肝脾瘀滞之邪为君药,黄精叶、橘皮滋阴益气、以资肝脾之阴为臣药,猪胆粉清热解毒为佐药,甘草调和诸药为使药。In the compound camellia pill for soothing the liver and strengthening the spleen of the present invention, camellia, gardenia leaves and herba cyperi are used as the main medicines for dispelling the stagnation of liver and spleen, polygonatum sibiricum leaves and tangerine peel are used as the assistant medicines for nourishing yin and qi and strengthening the yin of liver and spleen, pig bile powder is used as the adjuvant medicine for clearing away heat and detoxifying, and liquorice is used as the guiding medicine for harmonizing various medicines.
本发明公开了以下技术效果:The present invention discloses the following technical effects:
本发明提供的一种疏肝健脾复方山茶丸,组方包括山茶花、栀子叶、牛舌草、黄精叶、橘皮、猪胆粉和甘草,通过合理配伍,全方以山茶花、栀子叶、牛舌草为君药,黄精叶、橘皮为臣药,猪胆粉和甘草为佐使药,符合“君臣佐使”配伍治则,从“扶正祛邪”两方面发挥疏肝健脾的功效。且经试验验证,本发明制备的疏肝健脾复方山茶丸可明显改善小鼠的肝脾损伤,无毒副作用,可为防治肝脾损伤提供一种新思路。The compound Shancha Pill for soothing the liver and strengthening the spleen provided by the present invention comprises camellia, gardenia leaf, ox tongue grass, polygonatum leaf, tangerine peel, pig gall powder and licorice. Through reasonable compatibility, the whole prescription uses camellia, gardenia leaf and ox tongue grass as monarch medicine, polygonatum leaf and tangerine peel as minister medicine, pig gall powder and licorice as assistant medicine, which conforms to the compatibility treatment principle of "monarch, minister, assistant and assistant", and exerts the effect of soothing the liver and strengthening the spleen from the two aspects of "strengthening the body and eliminating evil". It has been verified by experiments that the compound Shancha Pill for soothing the liver and strengthening the spleen prepared by the present invention can significantly improve the liver and spleen damage of mice, has no toxic side effects, and can provide a new idea for preventing and treating liver and spleen damage.
附图说明BRIEF DESCRIPTION OF THE DRAWINGS
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图仅仅是本发明的一些实施例,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他的附图。In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.
图1为各组小鼠血清AST水平的统计图;*表示P<0.05,差异具有统计学意义;Figure 1 is a statistical graph of serum AST levels in mice in each group; * indicates P < 0.05, and the difference is statistically significant;
图2为各组小鼠血清ALT水平的统计图;*表示P<0.05,差异具有统计学意义;Figure 2 is a statistical chart of serum ALT levels in mice in each group; * indicates P < 0.05, and the difference is statistically significant;
图3为各组小鼠肝脏HE染色结果图;A:空白组,B:肝损伤组,C:实例1组,D:实例2组,E:实例3组;Figure 3 is the HE staining results of the liver of mice in each group; A: blank group, B: liver injury group, C: Example 1 group, D: Example 2 group, E: Example 3 group;
图4为各组小鼠脾脏HE染色结果图;A:空白组,B:脾损伤组,C:实例1组,D:实例2组,E:实例3组;Figure 4 is a diagram showing HE staining results of spleen of mice in each group; A: blank group, B: spleen injury group, C: Example 1 group, D: Example 2 group, E: Example 3 group;
图5为各组小鼠血清中IgG(A)和IgM(B)含量统计图;*表示P<0.05,差异具有统计学意义。FIG5 is a statistical diagram of the IgG (A) and IgM (B) levels in the serum of mice in each group; * indicates P < 0.05, and the difference is statistically significant.
具体实施方式DETAILED DESCRIPTION
现详细说明本发明的多种示例性实施方式,该详细说明不应认为是对本发明的限制,而应理解为是对本发明的某些方面、特性和实施方案的更详细的描述。Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.
应理解本发明中所述的术语仅仅是为描述特别的实施方式,并非用于限制本发明。另外,对于本发明中的数值范围,应理解为还具体公开了该范围的上限和下限之间的每个中间值。在任何陈述值或陈述范围内的中间值,以及任何其他陈述值或在所述范围内的中间值之间的每个较小的范围也包括在本发明内。这些较小范围的上限和下限可独立地包括或排除在范围内。It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.
除非另有说明,否则本文使用的所有技术和科学术语具有本发明所述领域的常规技术人员通常理解的相同含义。虽然本发明仅描述了优选的方法和材料,但是在本发明的实施或测试中也可以使用与本文所述相似或等同的任何方法和材料。本说明书中提到的所有文献通过引用并入,用以公开和描述与所述文献相关的方法和/或材料。在与任何并入的文献冲突时,以本说明书的内容为准。Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and/or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.
在不背离本发明的范围或精神的情况下,可对本发明说明书的具体实施方式做多种改进和变化,这对本领域技术人员而言是显而易见的。由本发明的说明书得到的其他实施方式对技术人员而言是显而易见得的。本发明说明书和实施例仅是示例性的。It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.
关于本文中所使用的“包含”、“包括”、“具有”、“含有”等等,均为开放性的用语,即意指包含但不限于。The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.
下述实施例中的实验方法,如无特殊说明,均为常规方法,按照本领域内的文献所描述的技术或条件或者按照产品说明书进行。下述实施例中所用的材料、试剂等,如无特殊说明,均可从商业途径得到。The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.
雌性ICR小鼠,购自河南斯克贝斯生物科技股份有限公司,体重18±2g,所有试验动物温度保持在22-26℃之间,相对湿度在40%-70%之间,所有小鼠都被关在SPF笼子中,昼夜周期为12h光照和12h黑暗(每天8:00至20:00)。每只试验动物被允许随意喂食标准的小鼠食物和水。适应性喂养一周后再进行试验。本发明的具体研究如下:Female ICR mice were purchased from Henan Sikebes Biotechnology Co., Ltd., weighing 18±2g. The temperature of all experimental animals was maintained between 22-26°C, and the relative humidity was between 40%-70%. All mice were kept in SPF cages with a day and night cycle of 12h light and 12h dark (8:00 to 20:00 every day). Each experimental animal was allowed to feed standard mouse food and water at will. The experiment was carried out again after one week of adaptive feeding. The specific research of the present invention is as follows:
实施例1Example 1
一种疏肝健脾复方山茶丸,包括以下重量份原料:山茶花12份,栀子叶8份,牛舌草9份,黄精叶4份,橘皮3份,猪胆粉1份和甘草1份。A compound camellia pill for soothing the liver and strengthening the spleen comprises the following raw materials in parts by weight: 12 parts of camellia, 8 parts of gardenia leaves, 9 parts of oxalis, 4 parts of polygonatum leaves, 3 parts of tangerine peel, 1 part of pig gall powder and 1 part of liquorice.
制备方法:将山茶花、栀子叶、牛舌草、黄精叶、橘皮和甘草混匀,破壁机粉碎成超微粉;向超微粉中加入15倍体积的水,冷凝回流提取60min,得提取液,真空负压过滤,收集滤液,再次过滤取上清,浓缩成膏状,得浸膏;向浸膏中加入猪胆粉搅拌均匀,得到混浸膏,按照重量份计,取混浸膏1份与辅料(可溶性淀粉0.8份、山茶油0.1份和蔗糖0.04份),混合均匀,制成丸剂,密封保存。即得疏肝健脾复方山茶丸。Preparation method: Mix camellia, gardenia leaves, oxalis, polygonatum leaves, orange peel and licorice, and crush them into ultrafine powder by a wall-breaking machine; add 15 times the volume of water to the ultrafine powder, condense and reflux for 60 minutes to obtain an extract, vacuum negative pressure filter, collect the filtrate, filter again to obtain the supernatant, concentrate into a paste, and obtain an extract; add pig gall powder to the extract and stir evenly to obtain a mixed extract, take 1 part of the mixed extract and auxiliary materials (0.8 parts of soluble starch, 0.1 parts of camellia oil and 0.04 parts of sucrose) by weight, mix evenly, make pills, and seal for storage. Soothing the liver and strengthening the spleen compound camellia pills are obtained.
实施例2Example 2
一种疏肝健脾复方山茶丸,包括以下重量份原料:山茶花10份,栀子叶9份,牛舌草6份,黄精叶6份,橘皮4份,猪胆粉2份和甘草3份。A compound camellia pill for soothing the liver and strengthening the spleen comprises the following raw materials in parts by weight: 10 parts of camellia, 9 parts of gardenia leaves, 6 parts of ox tongue grass, 6 parts of polygonatum sibiricum leaves, 4 parts of tangerine peel, 2 parts of pig gall powder and 3 parts of liquorice.
制备方法同实施例1。The preparation method is the same as that of Example 1.
实施例3Example 3
一种疏肝健脾复方山茶丸,包括以下重量份原料:山茶花15份,栀子叶6份,牛舌草8份,黄精叶3份,橘皮6份,猪胆粉3份和甘草2份。A compound camellia pill for soothing the liver and strengthening the spleen comprises the following raw materials in parts by weight: 15 parts of camellia, 6 parts of gardenia leaves, 8 parts of oxalis, 3 parts of polygonatum leaves, 6 parts of tangerine peel, 3 parts of pig gall powder and 2 parts of liquorice.
制备方法同实施例1。The preparation method is the same as that of Example 1.
效果例Effect example
1.疏肝健脾复方山茶丸对肝损伤小鼠的治疗作用1. Therapeutic effect of Soothing Liver and Strengthening Spleen Compound Shancha Pill on mice with liver injury
取25只健康雌性ICR小鼠,随机均分为五组:空白组、肝损伤组、实例1组、实例2组、实例3组。肝损伤组、实例1组、实例2组、实例3组每天按照0.01mL/g的剂量腹腔注射四氯化碳(2%),连续3天建立肝损伤模型。模型建立后,实例1组、实例2组、实例3组分别使用实施例1、实施例2、实施例3制备的疏肝健脾复方山茶丸进行灌胃治疗(0.5g/kg体重剂量),连续治疗3d,治疗结束后处死小鼠,采集血液用于分离血清,采集肝脏于4%多聚甲醛保存。进行以下实验。25 healthy female ICR mice were randomly divided into five groups: blank group, liver injury group, Example 1 group, Example 2 group, and Example 3 group. The liver injury group, Example 1 group, Example 2 group, and Example 3 group were intraperitoneally injected with carbon tetrachloride (2%) at a dose of 0.01 mL/g every day for 3 consecutive days to establish a liver injury model. After the model was established, Example 1 group, Example 2 group, and Example 3 group were treated with oral gavage (0.5 g/kg body weight dose) of the Soothing Liver and Strengthening Spleen Compound Camellia Pills prepared in Example 1, Example 2, and Example 3, respectively, for 3 consecutive days. After the treatment, the mice were killed, blood was collected for serum separation, and the liver was collected and stored in 4% paraformaldehyde. The following experiments were performed.
1.1小鼠肝功能试验1.1 Mouse liver function test
按照南京建成生物研究所提供谷草转氨酶(AST)和谷丙转氨酶(ALT)试剂盒说明书的要求检测小鼠的肝功能。The liver function of mice was detected according to the instructions of the aspartate aminotransferase (AST) and alanine aminotransferase (ALT) kits provided by Nanjing Jiancheng Biological Research Institute.
试验结果如图1-2所示,与空白组相比,肝损伤组小鼠血清AST和ALT水平显著升高(P<0.05),与肝损伤组相比,实例1组、实例2组、实例3组小鼠血清AST和ALT水平显著降低(P<0.05)。表明本发明成功建立肝损伤模型,并且,本发明实施例1-3制备的疏肝健脾复方山茶丸均可有效改善小鼠的肝损伤。The test results are shown in Figures 1-2. Compared with the blank group, the serum AST and ALT levels of mice in the liver injury group were significantly increased (P<0.05), and compared with the liver injury group, the serum AST and ALT levels of mice in Example 1, Example 2, and Example 3 were significantly reduced (P<0.05). This shows that the liver injury model was successfully established by the present invention, and the compound Shancha pills for soothing the liver and strengthening the spleen prepared in Examples 1-3 of the present invention can effectively improve the liver injury of mice.
1.2肝脏病理组织切片1.2 Liver pathological tissue sections
将于4%多聚甲醛溶液保存的肝脏组织取出,用手术刀片修成0.5mm3的组织块,经常规组织包埋技术进行石蜡包埋,组织切片机切片、脱蜡后,按照HE染色方法制片,并在数码摄像显微镜下拍照,观察肝脏的病理组织学变化。The liver tissue preserved in 4% paraformaldehyde solution was removed and cut into 0.5 mm3 tissue blocks with a surgical blade. The tissue blocks were embedded in paraffin using conventional tissue embedding techniques. After slicing and dewaxing with a tissue slicer, the tissue sections were prepared according to the HE staining method and photographed under a digital camera microscope to observe the pathological histological changes of the liver.
HE染色结果如图3所示,与空白组相比,肝损伤组小鼠肝脏细胞出现大量的坏死,肝脏细胞排列紊乱,与肝损伤组相比,实例1组、实例2组、实例3组肝脏坏死细胞显著减少,肝细胞排列趋于正常。试验结果同样表明本发明实施例1-3制备的疏肝健脾复方山茶丸均可有效改善小鼠的肝损伤。The HE staining results are shown in Figure 3. Compared with the blank group, the liver cells of the mice in the liver injury group showed a large amount of necrosis, and the liver cells were arranged in disorder. Compared with the liver injury group, the liver necrotic cells in the Example 1 group, the Example 2 group, and the Example 3 group were significantly reduced, and the liver cell arrangement tended to be normal. The test results also show that the compound Shancha pills for soothing the liver and strengthening the spleen prepared in Examples 1-3 of the present invention can effectively improve the liver injury of mice.
2.疏肝健脾复方山茶丸对脾损伤小鼠的治疗作用2. The therapeutic effect of Shancha Pills for soothing the liver and strengthening the spleen on mice with spleen injury
取25只健康雌性ICR小鼠,随机均分为五组:空白组、脾损伤组、实例1组、实例2组、实例3组。实例1组、实例2组、实例3组分别使用实施例1、实施例2、实施例3制备的疏肝健脾复方山茶丸进行灌胃(0.1g/kg体重剂量),连续灌胃7d,在灌胃的第5-7d,脾损伤组、实例1组、实例2组、实例3组每天按照5mg/kg体重剂量的环磷酰胺进行腹腔注射,连续3天建立脾损伤模型。治疗结束后处死小鼠,采集血液用于分离血清,采集脾脏于4%多聚甲醛保存。进行以下实验。25 healthy female ICR mice were randomly divided into five groups: blank group, spleen injury group, Example 1 group, Example 2 group, and Example 3 group. Example 1 group, Example 2 group, and Example 3 group were gavaged with the compound Shancha pills for soothing the liver and strengthening the spleen prepared in Example 1, Example 2, and Example 3 (0.1 g/kg body weight dose) for 7 consecutive days. On the 5th to 7th day of gavage, the spleen injury group, Example 1 group, Example 2 group, and Example 3 group were intraperitoneally injected with cyclophosphamide at a dose of 5 mg/kg body weight every day for 3 consecutive days to establish a spleen injury model. After the treatment, the mice were killed, blood was collected for serum separation, and the spleen was collected and stored in 4% paraformaldehyde. The following experiments were performed.
2.1脾脏病理组织切片2.1 Spleen pathological tissue sections
将于4%多聚甲醛溶液保存的脾脏组织取出,用手术刀片修成0.5mm3的组织块,经常规组织包埋技术进行石蜡包埋,组织切片机切片、脱蜡后,按照HE染色方法制片,并在数码摄像显微镜下拍照,观察脾脏的病理组织学变化。The spleen tissue preserved in 4% paraformaldehyde solution was removed and cut into 0.5 mm3 tissue blocks with a surgical blade. The blocks were embedded in paraffin using conventional tissue embedding techniques. After sectioning and dewaxing with a tissue slicer, the blocks were prepared according to the HE staining method and photographed under a digital camera microscope to observe the pathological histological changes of the spleen.
HE染色结果如图4所示,结果显示空白组红髓与白髓结构清晰,分界明显,厚度适中,未发现明显病变;脾损伤组红髓与白髓边界模糊,淋巴细胞排列稀疏且水肿,局部明显病变;实例1组、实例2组、实例3组白髓与红髓界限清晰,脾索与脾窦形态规则,淋巴细胞水肿减少,局部病变明显改善。表明本发明实施例1-3制备的疏肝健脾复方山茶丸均可有效改善小鼠的脾损伤。The HE staining results are shown in Figure 4, and the results show that the red pulp and white pulp of the blank group have clear structures, obvious boundaries, moderate thickness, and no obvious lesions; the red pulp and white pulp boundaries of the spleen injury group are blurred, the lymphocytes are sparsely arranged and edematous, and local lesions are obvious; the boundaries of the white pulp and red pulp of Example 1, Example 2, and Example 3 are clear, the splenic cords and splenic sinuses have regular morphology, the lymphocyte edema is reduced, and the local lesions are significantly improved. This shows that the Soothing Liver and Strengthening Spleen Compound Shancha Pills prepared in Examples 1-3 of the present invention can effectively improve the spleen injury of mice.
2.2血清免疫因子水平测定2.2 Determination of serum immune factor levels
按照武汉Elabscience小鼠IgG和IgM ELISA说明书的要求检测小鼠血清中IgG和IgM的含量。The levels of IgG and IgM in mouse serum were detected according to the instructions of Wuhan Elabscience Mouse IgG and IgM ELISA.
检测结果如图5所示,相较于空白组,脾损伤组小鼠血清中IgG和IgM含量显著降低(P<0.05),与脾损伤组相比,实例1组、实例2组、实例3组血清中IgG和IgM含量显著增加(P<0.05)且与空白组相比无显著差异。表明脾损伤组小鼠免疫能力下降,本发明实施例1-3制备的疏肝健脾复方山茶丸均可显著增强小鼠的免疫能力,发挥保护脾功能的作用。The test results are shown in Figure 5. Compared with the blank group, the IgG and IgM levels in the serum of mice in the spleen injury group were significantly reduced (P<0.05). Compared with the spleen injury group, the IgG and IgM levels in the serum of Example 1, Example 2, and Example 3 groups were significantly increased (P<0.05) and there was no significant difference compared with the blank group. This shows that the immune ability of mice in the spleen injury group is reduced, and the liver-soothing and spleen-strengthening compound Shancha pills prepared in Examples 1-3 of the present invention can significantly enhance the immune ability of mice and play a role in protecting spleen function.
综上所述,本发明制备的疏肝健脾复方山茶丸可明显改善小鼠的肝脾损伤,无毒副作用,可为防治肝脾损伤提供一种新思路。In summary, the compound Shancha pills for soothing the liver and strengthening the spleen prepared by the present invention can significantly improve the liver and spleen damage in mice, have no toxic side effects, and can provide a new idea for preventing and treating liver and spleen damage.
以上所述的实施例仅是对本发明的优选方式进行描述,并非对本发明的范围进行限定,在不脱离本发明设计精神的前提下,本领域普通技术人员对本发明的技术方案做出的各种变形和改进,均应落入本发明权利要求书确定的保护范围内。The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.
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