Traditional Chinese medicine composition for treating autism spectrum disorder and application thereof

Abstract

The invention discloses a traditional Chinese medicine composition for treating autism spectrum disorder and application thereof, and belongs to the technical field of traditional Chinese medicine compositions. The traditional Chinese medicine composition is prepared from the following components in parts by mass: 4 to 10 parts of cortex cinnamomi, 4 to 10 parts of radix morindae officinalis, 4 to 10 parts of radix polygalae, 4 to 10 parts of rhizoma acori graminei, 5 to 10 parts of radix rehmanniae praeparata, 4 to 10 parts of radix angelicae sinensis, 8 to 20 parts of tortoise shell and 5 to 10 parts of herba dendrobii. From the perspective of overall compatibility, the cinnamon and the morinda officinalis in the composition can warm and tonify kidney yang, the prepared rehmannia root, the tortoise shell and the dendrobe nourish yin and tonify kidney to prevent excessive warm tonification, the angelica sinensis can enrich blood and activate blood and give consideration to spleen and stomach, the polygala tenuifolia can traffic heart and kidney, soothe the nerves and promote intelligence, and the rhizoma acori graminei can reduce phlegm, induce resuscitation and induce resuscitation. The medicines are matched with one another to jointly achieve the effects of tonifying spleen and kidney, replenishing essence and benefiting marrow, and inducing resuscitation and promoting intelligence, and the symptoms of autism are comprehensively improved from the fundamental pathogenesis of spleen-kidney deficiency and by considering pathological changes such as phlegm clogging the heart and the like.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicines, and in particular relates to a traditional Chinese medicine composition for treating autism spectrum disorder and an application thereof. Background Art

[0002] Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder that begins in early development and is characterized by social interaction disorders, communication disorders, narrow interests and stereotyped behaviors, most of which are accompanied by intellectual development disorders. The global prevalence of ASD has increased year by year, but its etiology and pathogenesis have not yet been clarified. It is generally believed that ASD is a type of neurodevelopmental disorder caused by the combined effects of genetic and environmental factors. Treatment for ASD is mainly divided into two aspects: non-drug treatment and drug treatment. Non-drug treatment is currently mainly based on behavioral intervention, which changes core symptoms through rehabilitation training and educational training to improve social adaptability and quality of life. At the same time, psychotherapy is also used to improve the condition in a targeted manner. However, most ASDs affect the physical and mental health of children, have a poor prognosis, and are accompanied for life. In addition, the cost of rehabilitation training is expensive, which brings heavy mental pressure and great economic burden to families and society. There is currently no clear and effective drug for drug treatment. Usually, if there are obvious mental and neurological symptoms, drugs are needed for treatment. For example, haloperidol and benzhexol hydrochloride can be taken to control stereotyped movements, excessive activity and impulsive behavior; sulpiride can be taken to improve loneliness, calm emotions and improve speech level. However, these are also drugs to relieve mental symptoms, and long-term use has obvious side effects.

[0003] It can be seen that there is currently a great lack of drugs targeting the core symptoms of ASD. Therefore, exploring a safe and effective treatment method to improve the quality of life of patients is a hot issue that needs to be urgently addressed in the clinic. Chinese medicine compositions also have certain research value and potential in the treatment of ASD, and there are some related reports. For example, the invention patent "A Chinese medicine composition for autism spectrum disorder and its preparation method" with publication number CN 113663048 A discloses a composition made of six ingredients (tortoise shell, turtle shell, chicken gizzard lining, black plum, hawthorn and ginger). The drug combination is mainly aimed at nourishing yin and suppressing yang, strengthening the spleen and digesting food. It constructs an autistic mouse model by intraperitoneal injection of sodium valproate in pregnant female mice. The behavioral phenotype of this model is affected by multiple factors such as drug dosage, treatment time, and individual differences in animals, which may lead to its autistic behavioral phenotype being unstable.

[0004] Spleen and kidney yang deficiency and insufficient kidney essence refer to the pathological state manifested by spleen and kidney yang deficiency and insufficient kidney essence. The spleen is responsible for transporting and transforming the essence of water and grain, and the kidney is the innate foundation, storing true yin and containing primordial yang, promoting growth and development. Insufficient spleen and kidney yang cannot warm the internal organs and meridians, transform essence and qi, leading to a series of symptoms of delayed growth and development. Patients with autism spectrum disorders generally have congenital deficiencies, such as older parents, weak mothers during pregnancy, or long-term medication, which may lead to insufficient fetal endowment and affect spleen and kidney function. At the same time, there may also be problems with the acquired conditioning of patients with autism spectrum disorders, such as improper diet, excessive fatigue, etc., which may damage the spleen and kidney yang, leading to spleen and kidney yang deficiency and the pathogenesis of marrow sea insufficiency. However, the pathogenesis of spleen and kidney yang deficiency in autism spectrum disorders has not been paid attention to in the existing technology, and it is a direction that urgently needs to be broken through. Summary of the invention

[0005] In order to overcome the above-mentioned shortcomings of the prior art and to address the pathogenesis of spleen and kidney yang deficiency in autism, the present invention aims to provide a Chinese medicine composition for treating autism spectrum disorder and its application.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions: The invention discloses a traditional Chinese medicine composition for treating autism spectrum disorder. The composition is as follows by mass: 4-10 parts of cinnamon bark, 4-10 parts of Morinda officinalis, 4-10 parts of Polygala tenuifolia, 4-10 parts of Acorus calamus, 5-10 parts of Rehmannia root, 4-10 parts of Angelica sinensis, 8-20 parts of Tortoise shell and 5-10 parts of Dendrobium officinale.

[0007] Preferably, the composition by mass is as follows: 4-6 parts of cinnamon bark, 4-6 parts of Morinda officinalis, 4-6 parts of Polygala tenuifolia, 4-6 parts of Acorus calamus, 5-8 parts of Rehmannia glutinosa, 4-6 parts of Angelica sinensis, 8-15 parts of Tortoise Shell, and 5-8 parts of Dendrobium officinale.

[0008] Preferably, 5 parts of cinnamon bark, 6 parts of Morinda officinalis, 4 parts of Polygala tenuifolia, 5 parts of Acorus calamus, 8 parts of Rehmannia glutinosa, 5 parts of Angelica sinensis, 10 parts of Carapax tortoise shell, and 8 parts of Dendrobium candidum.

[0009] Preferably, 5 parts of cinnamon bark, 5 parts of Morinda officinalis, 5 parts of Polygala tenuifolia, 5 parts of Acorus calamus, 6 parts of Rehmannia glutinosa, 5 parts of Angelica sinensis, 10 parts of Carapax tortoise shell, and 6 parts of Dendrobium candidum.

[0010] Preferably, 5 parts of cinnamon bark, 5 parts of Morinda officinalis, 5 parts of Polygala tenuifolia, 8 parts of Acorus calamus, 8 parts of Rehmannia glutinosa, 5 parts of Angelica sinensis, 8 parts of Carapax tortoise shell, and 6 parts of Dendrobium candidum.

[0011] The cinnamon bark in the Chinese medicine composition of the present invention has the effect of replenishing fire and supporting yang, and can be used to treat symptoms such as chills, cold limbs, frequent urination and enuresis caused by insufficient kidney yang and spleen and kidney yang deficiency. In addition, cinnamon bark also has the effects of sedation and analgesia, antibacterial, anti-inflammatory, anti-radiation, anti-tumor, improving immune function, relieving asthma, and killing insects.

[0012] The "Morinda officinalis" in the Chinese medicine composition of the present invention is also known as Morinda officinalis, which is a vine plant with a slightly warm nature, a pungent and sweet taste, and enters the kidney and liver meridians. It has the effects of tonifying kidney yang, strengthening muscles and bones, and removing rheumatism. It is mainly used to treat kidney deficiency, soreness of waist and knees, and weakness of muscles and bones.

[0013] The "Polygala" in the Chinese medicine composition of the present invention is a perennial herb, which is mainly produced in Northeast China, North China, Northwest China, Central China and Sichuan. Polygala is bitter, pungent and warm in nature and flavor, and enters the heart, kidney and lung meridians. Polygala can improve the situation of heart-kidney disharmony, and has a significant alleviating effect on various symptoms caused by heart-kidney disharmony, such as palpitations, soreness of waist and knees, etc. At the same time, the active ingredients in Polygala help regulate the nervous system, have a significant improvement effect on symptoms such as insomnia, dreaminess, palpitations, etc., help improve sleep quality, and enhance memory.

[0014] The "Acorus calamus" in the Chinese medicine composition of the present invention is a perennial herb. Acorus calamus tastes pungent and bitter, is warm in nature, and enters the heart and stomach meridians. Acorus calamus is good at resolving dampness and phlegm, removing filth, opening the orifices and refreshing the mind. It is often used to treat symptoms such as confusion, coma and delirium caused by phlegm, filth and evil obstructing the orifices.

[0015] The "prepared rehmannia root" in the Chinese medicine composition of the present invention is a processed product of raw rehmannia root, which is sweet and slightly warm. It enters the liver and kidney meridians. Preparing rehmannia root helps to nourish the spleen and stomach, promote appetite and digestion, improve physical strength, and is often used to treat weakness symptoms such as weakness, fatigue and indigestion; Preparing rehmannia root helps to regulate the nervous system, relieve anxiety, insomnia and emotional instability, improve mental state, and can also nourish the liver and kidney, strengthen the tendons and bones, and has a certain improvement effect on symptoms such as liver and kidney yin deficiency, soreness of waist and knees, etc.

[0016] The "Danggui" in the Chinese medicine composition of the present invention is a perennial herb with a warm nature, sweet and pungent taste, and enters the liver meridian, heart meridian and spleen meridian. Danggui can replenish blood and promote blood circulation, and can be used to treat symptoms such as blood deficiency, sallow complexion, dizziness and palpitations, and can also improve the body's resistance and enhance the immune system function.

[0017] The "tortoise shell" in the Chinese medicine composition of the present invention refers to the carapace and plastron of the turtle of the family Testudinidae, which is regarded as an important Chinese medicinal material, having multiple effects such as nourishing yin and suppressing yang, strengthening the kidney and strengthening bones, nourishing blood and nourishing the heart, consolidating menstruation and stopping metrorrhagia, etc. It is often used to treat symptoms such as yin deficiency hot flashes, bone steaming and night sweats, yin deficiency and yang hyperactivity, dizziness, internal wind movement, weakness of muscles and bones, heart deficiency and forgetfulness, metrorrhagia and heavy menstruation, etc.

[0018] The "dendrobe" in the Chinese medicine composition of the present invention is a plant of the orchid family, which is cold in nature, sweet, light, slightly salty, and enters the stomach, lung, and kidney meridians. Dendrobium can benefit the stomach and produce body fluid, nourish yin and clear away heat, and is mainly used to treat symptoms such as damage to body fluid due to febrile diseases, dry mouth and thirst, insufficient stomach yin, poor appetite and dry retching, persistent deficiency heat after illness, yin deficiency and excessive fire, bone steaming and fatigue fever, blurred vision, and weakness of tendons and bones.

[0019] The present invention also discloses the use of the above-mentioned traditional Chinese medicine composition for treating autism spectrum disorders in preparing medicine for treating autism spectrum disorders.

[0020] Preferably, the drug is a drug that improves the user's social skills and reduces stereotyped behaviors.

[0021] Preferably, the drug is added with pharmaceutically acceptable adjuvants or excipients to prepare an oral preparation.

[0022] More preferably, the oral preparation is a tablet, powder, pill, capsule, granule or decoction.

[0023] More preferably, the auxiliary materials or excipients include one or more of stabilizers, solubilizers, lubricants and disintegrants.

[0024] Compared with the prior art, the present invention has the following beneficial effects: The present invention uses traditional Chinese medicine theory, and successfully prepares a Chinese medicine composition containing cinnamon, Morinda officinalis, Polygala tenuifolia, Acorus calamus, Rehmannia glutinosa, Angelica sinensis, Tortoise shell and Dendrobium officinale based on the theory and practice of traditional Chinese medicine on autism spectrum disorder. The composition has simple and reasonable compatibility, significant efficacy, convenient material preparation, and easy promotion. The selection of each medicine in the composition has been carefully studied, and the final composition emphasizes the reasonable compatibility between the drugs to achieve the purpose of synergistic enhancement. Specifically, the cinnamon in the drug composition has the effects of warming yang and dispersing cold, replenishing fire and assisting yang, which can improve the symptoms of cold limbs and enhance body immunity; Morinda officinalis has the effects of warming and replenishing kidney yang and strengthening muscles and bones, and is suitable for symptoms such as five delays, five softness, and developmental delay caused by insufficient kidney yang, and can promote body development; Polygala tenuifolia has the effects of calming the mind and improving intelligence, and connecting the heart and kidney, and has a good effect on insomnia, dreaminess, poor memory, abnormal emotions, and inattention caused by disharmony between the heart and kidney; Acorus calamus has the effects of opening the mind and resolving phlegm, refreshing the mind and improving intelligence, and is suitable for phlegm-dampness and sedation. It can relieve symptoms such as unclear consciousness, poor memory, and lack of concentration caused by blocking the clear orifices; Rehmannia root has the functions of nourishing yin and blood, replenishing essence and filling marrow, providing a material basis for growth and development; Angelica sinensis has the functions of replenishing blood and activating blood circulation, which can improve the circulation of qi and blood and nourish the internal organs; Tortoise shell has the functions of nourishing yin and suppressing yang, replenishing the kidney and strengthening bones, restraining the hyperactivity of yang in cinnamon and Morinda officinalis, harmonizing yin and yang, and at the same time helping bone development; Dendrobium has the functions of nourishing the stomach and promoting fluid, nourishing yin and clearing away heat, and has a good effect on improving symptoms such as dry mouth and tongue, internal heat due to yin deficiency, and irritability caused by insufficient stomach yin and kidney yin. From the perspective of overall compatibility, the composition of the present application focuses on the problem of both spleen and kidney deficiency, and selects cinnamon and Morinda officinalis to warm and nourish the spleen and kidney yang, Rehmannia glutinosa, tortoise shell, and Dendrobium to nourish yin and nourish the kidney to prevent excessive warming and nourishing, Angelica sinensis to nourish blood and promote blood circulation while taking into account the spleen and stomach, Polygala tenuifolia to connect the heart and kidney, calm the mind and improve intelligence, and Acorus calamus to resolve phlegm, open the orifices and refresh the mind. These medicines cooperate with each other to achieve the effects of nourishing the spleen and kidney, replenishing essence and marrow, opening the orifices and improving intelligence. Starting from the fundamental pathogenesis of both spleen and kidney deficiency, taking into account pathological changes such as phlegm covering the brain orifices, the symptoms of autism are comprehensively improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is a design flow chart of the whole animal experiment of the present invention; Figure 2 This is the total distance result of the open field test in behavioral experiments; Figure 3 The results of central area movement time in the behavioral open field test; Figure 4 The number of buried beads for behavioral experiments; Figure 5 Grooming time results for behavioral experiments; Figure 6 For behavioral experiments, the results of social ability sniffing time (s) were obtained; Figure 7The result of behavioral experiment social ability index (S1-E) / (S1+E); Figure 8 For behavioral experiments, the results of social preference sniffing time (s); Fig. 9 The social preference index of behavioral experiments is (S2-S1) / (S1+S2). Fig.10 The results of spontaneous alternation rate (%) in the behavioral experiment Y maze; Fig.11 It is the test result of MDA content (nmol / mg).

[0026] In the figure: # P<0.05, ## P<0.01, ### P < 0.001, #### P<0.0001, compared with the blank control group; *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, compared with the model group) DETAILED DESCRIPTION In order to enable those skilled in the art to better understand the scheme of the present invention, the technical scheme in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0027] It should be noted that the terms "first", "second", etc. in the specification and claims of the present invention and the above-mentioned drawings are used to distinguish similar objects, and are not necessarily used to describe a specific order or sequence. It should be understood that the data used in this way can be interchanged where appropriate, so that the embodiments of the present invention described herein can be implemented in an order other than those illustrated or described herein. In addition, the terms "including" and "having" and any variations thereof are intended to cover non-exclusive inclusions, for example, a process, method, system, product or device that includes a series of steps or units is not necessarily limited to those steps or units that are clearly listed, but may include other steps or units that are not clearly listed or inherent to these processes, methods, products or devices.

[0028] The present invention is further described in detail below in conjunction with the accompanying drawings: 1. Examples of Chinese medicine compositions Example 1 A traditional Chinese medicine composition for treating autism spectrum disorder, comprising: 5g of cinnamon bark, 6g of Morinda officinalis, 4g of Polygala tenuifolia, 5g of Acorus calamus, 8g of Rehmannia glutinosa, 5g of Angelica sinensis, 10g of Tortoise Shell, and 8g of Dendrobium officinale.

[0029] Example 2 A traditional Chinese medicine composition for treating autism spectrum disorder, comprising: 5g of cinnamon bark, 5g of Morinda officinalis, 5g of Polygala tenuifolia, 5g of Acorus calamus, 6g of Rehmannia glutinosa, 5g of Angelica sinensis, 10g of Tortoise Shell, and 6g of Dendrobium candidum.

[0030] Example 3 A traditional Chinese medicine composition for treating autism spectrum disorder, comprising: 5g of cinnamon bark, 5g of Morinda officinalis, 5g of Polygala tenuifolia, 8g of Acorus calamus, 8g of Rehmannia glutinosa, 5g of Angelica sinensis, 8g of Tortoise Shell, and 6g of Dendrobium officinale.

[0031] The above embodiments of the present invention can be prepared into common medicaments for use when they are used specifically. The following is an explanation using granules as an example. According to the characteristics of the raw medicinal materials in the pharmaceutical composition, the corresponding granulation operation is performed. Taking the composition of Example 2 as an example, 5g of cinnamon bark, 5g of Morinda officinalis, 5g of Polygala tenuifolia, 5g of Acorus calamus, 6g of Rehmannia glutinosa, 5g of Angelica sinensis, 10g of Carapax tortoise shell, and 6g of Dendrobium officinale are prepared.

[0032] 1) Cinnamon: Take 5g of cinnamon slices, add 8 times water and boil for 60 minutes, extract volatile oil at the same time, and filter; add 6 times water and boil for 45 minutes, extract volatile oil at the same time. Filter with a 150-mesh filter and combine the filtrate. Concentrate the filtrate to a clear paste with a relative density of 1.05-1.10 (the dry extract yield is 4%~8%). Volatile oil and β-cyclodextrin are mixed at a weight ratio of 1:6~10 in a colloid mill at a constant temperature for 2 hours. Spray dry (liquid supply temperature 40℃~60℃, air inlet temperature 150℃~170℃). Add an appropriate amount of auxiliary material dextrin, mix well, and granulate.

[0033] 2) Morinda officinalis: Take 5g of Morinda officinalis slices, add 10 times water, decoct for 60 minutes, filter; add 8 times water, decoct for 45 minutes, filter. Filter with a 150-mesh filter and combine the filtrate. Concentrate the filtrate to a clear paste with a relative density of 1.05-1.10 (the dry extract yield is 36%~45%), add an appropriate amount of auxiliary material dextrin, spray dry (liquid supply temperature 50℃~65℃, air inlet temperature 150℃~170℃). Add an appropriate amount of auxiliary material dextrin, mix well, and dry granulate.

[0034] 3) Polygala: Take 5g of Polygala slices, add 10 times water, decoct for 60 minutes, filter; add 8 times water, decoct for 45 minutes, filter. Filter with a 150-mesh filter and combine the filtrate. Concentrate the filtrate to a clear paste with a relative density of 1.05-1.10 (the dry extract yield is 21.5%~40.5%), add an appropriate amount of auxiliary material dextrin, spray dry (liquid supply temperature 50℃~60℃, air inlet temperature 150℃~170℃). Add an appropriate amount of auxiliary material dextrin, mix well, and dry granulate.

[0035] 4) Acorus calamus: Take 5g of Acorus calamus slices, add 8 times water and boil for 60 minutes, extract volatile oil at the same time, and filter; add 6 times water and boil for 45 minutes, extract volatile oil at the same time. Filter with a 150-mesh filter and combine the filtrate. Concentrate the filtrate to a clear paste with a relative density of 1.05-1.10 (the dry extract yield is 8%~13%). Volatile oil and β-cyclodextrin are mixed at a weight ratio of 1:8~10 in a colloid mill at a constant temperature for 2 hours. Spray dry (liquid supply temperature 40℃~60℃, air inlet temperature 150℃~170℃). Add an appropriate amount of auxiliary material dextrin, mix well, and granulate.

[0036] 5) Rehmannia glutinosa: Take 6 g of Rehmannia glutinosa slices, add 10 times the amount of water, boil for 90 minutes, and filter; add 8 times the amount of water, boil for 75 minutes, and filter. Filter through a 150-mesh filter and combine the filtrate. Concentrate the filtrate to a clear paste with a relative density of 1.05-1.10 (the dry extract yield is 44%~76.5%), add an appropriate amount of auxiliary material dextrin, and spray dry (liquid supply temperature 50℃~60℃, air inlet temperature 150℃~170℃). Add an appropriate amount of auxiliary material dextrin, mix well, and dry granulate.

[0037] 6) Angelica: Take 5g of Angelica slices, add 8 times water and boil for 75 minutes, extract volatile oil at the same time, filter; add 6 times water and boil for 60 minutes, extract volatile oil at the same time. Filter with a 150-mesh filter and combine the filtrate. Concentrate the filtrate to a clear paste with a relative density of 1.05-1.10 (the dry extract yield is 35%~56%). Volatile oil and β-cyclodextrin are mixed at a weight ratio of 1:6~10 in a colloid mill at a constant temperature for 2 hours. Spray dry (liquid supply temperature 40℃~60℃, air inlet temperature 150℃~170℃). Add appropriate amount of auxiliary material dextrin, mix well, and granulate.

[0038] 7) Turtle shell: Take 10g of tortoise shell slices, add 10 times water, boil for 90 minutes, filter; add 8 times water, boil for 75 minutes, filter. Filter with a 150-mesh filter and combine the filtrate. Concentrate the filtrate to a clear paste with a relative density of 1.05-1.10 (the dry extract yield is 3%~7%), add an appropriate amount of auxiliary material dextrin, spray dry (liquid supply temperature 50℃~60℃, air inlet temperature 150℃~170℃). Add an appropriate amount of auxiliary material dextrin, mix well, and dry granulate.

[0039] 8) Dendrobium officinale: Take 6 g of Dendrobium officinale (Dendrobium fimbriatum) decoction pieces, add 10 times the amount of water and boil for 80 minutes, then filter; add 8 times the amount of water and boil for 60 minutes, then filter. Filter through a 150-mesh sieve and combine the filtrates. Concentrate the filtrates to a clear paste with a relative density of 1.05 - 1.10 (the extraction rate of dry extract is 9% - 14.5%), add an appropriate amount of auxiliary material dextrin, and spray dry (the supply liquid temperature is 50°C - 60°C, and the inlet air temperature is 150°C - 170°C). Then add an appropriate amount of auxiliary material dextrin again, mix evenly, and granulate by dry granulation method.

[0040] Note that during the actual application and development process, according to the needs, the traditional Chinese medicine composition of the present invention can also be prepared into oral dosage forms such as tablets, powders, pills, and capsules according to the conventional processes in the pharmaceutical field.

[0041] II. Pharmacological experiments in animals See Figure 1 , which is the design flow chart of the whole animal experiment of the present invention.

[0042] 1. Experimental materials 1.1 Experimental animals 12 male C57BL / 6J mice at 4 - 5 weeks old were used as the blank control group; provided by Beijing Vital River Laboratory Animal Technology Co., Ltd. (License number: SCXK(Jing)2021 - 0011). 36 male SPF - grade BTBR mice at 4 - 5 weeks old were provided by the Clinical Research Institute of China - Japan Friendship Hospital (License number: SYXK(Jing)2023 - 0001), and were raised in an SPF - grade barrier animal room. After 1 - week of adaptive feeding, they were randomly divided into a model group, a low - dose Bushen Qizhi formula group, and a high - dose Bushen Qizhi formula group according to body weight stratification. Ethical batch number: ZRDWLL240005.

[0043] 1.2 Experimental instruments Open - field experiment instrument, three - chamber social experiment instrument, Y - maze experiment instrument, Tracking Master V5.0 behavioral tracking software (Beijing Zhongshi Dichuang Technology Development Co., Ltd.); 4% paraformaldehyde (Beijing Sawell Co., Ltd.). Malondialdehyde test kit (MDA Kit, Beijing Regbio Technology Development Co., Ltd. & China, batch number: 20250810.40003, product number: RGB - 40003).

[0044] 1.3 Experimental drugs For the convenience of experimental investigation, the experimental drugs were selected as the traditional Chinese medicine granules prepared from the composition in Example 2 above, and were administered by gavage after being dissolved in pure water according to the body weight of the mice.

[0045] 2. Experimental methods 2.1 Experimental grouping and administration methods Blank control group: After one week of adaptive feeding, mice were gavaged with a corresponding volume of pure water daily, 0.2 mL for every 10 g of mouse body weight, for 8 weeks.

[0046] Model group: After one week of adaptive feeding, mice were gavaged with a corresponding volume of pure water daily, 0.2 mL for every 10 g of mouse body weight, for 8 weeks.

[0047] The Chinese medicine granules prepared from the Chinese medicine composition of the present invention are: after one week of adaptive feeding, the corresponding volumes of Chinese medicine granules of different concentrations are administered orally daily, with 0.2 mL administered for every 10 g of mouse body weight, for 8 weeks.

[0048] The small-dose and large-dose groups of the Chinese medicine composition for autism spectrum disorders in children of the present invention are treated by daily oral gavage of a small-dose (12 g / kg) or large-dose (24 g / kg) Chinese medicine granules of corresponding volumes after birth, with 0.2 mL being administered for every 10 g of mouse body weight for 8 weeks.

[0049] 2.2 Behavioral observation of mice 2.2.1 Open field test The experiment used an open field behavior box to detect the spontaneous activity, exploratory behavior and anxiety of mice. The behavior box was an open cube of 40cm×40cm×40cm. During the experiment, the mouse was placed in the center of the behavior box with its back facing the experimenter and allowed to explore freely for 5 minutes. The free activity data of the mouse in the behavior box was recorded within 5 minutes, and the behavior box was cleaned with 70% ethanol. The total distance traveled by the mouse and the central area movement time were counted.

[0050] 2.2.2 Buried bead test During the experiment, fresh, odorless bedding was added to a brand new cage to a depth of 5 cm, and the bedding was flattened and compacted; standard black glass marbles (about 1 cm in diameter and 1 g in weight) were gently placed on the bedding in 5 rows of 4 marbles at equal distances; the camera was turned on and the movement of the mice was recorded within 20 minutes. Between each experiment, the marbles were washed with a mild detergent and dried before use. The number of beads buried by the mice in 20 minutes was counted, and a bead buried 2 / 3 was counted as 1.

[0051] 2.2.3 Y-maze spontaneous alternation test The mouse was gently placed in the center of the maze (starting arm) and allowed to explore freely. The order and total number of times the mouse entered each arm were recorded. When the mouse entered different arms three times in a row, it was counted as a correct alternation response. The mouse was placed in the device for 5 minutes as the adaptation phase, and then observed for another 5 minutes, the video was stopped and the file was saved. The number of correct alternation responses and the total number of arm entries for each mouse within 5 minutes were counted, and the spontaneous alternation rate was calculated.

[0052] 2.2.4 Self-grooming experiment During the experiment, the mice were placed in a brand new cage, the camera was turned on, and the movement of the mice was recorded for 20 minutes. Between each experiment, the cage was cleaned with 75% ethanol. The grooming time of the mice was counted for 20 minutes.

[0053] 2.2.5 Three-Box Social Experiment (1) Adaptation phase: The experimental mouse is placed in the central chamber of the behavior box with its back facing the experimenter, and allowed to move freely for 10 minutes to fully familiarize itself with the environment. After the test, the instrument is wiped with 75% ethanol, and the second phase is carried out after the wiping is completed. Both channels are carried out simultaneously.

[0054] (2) Second stage of social ability test: A mouse of the same species and similar age (stranger 1) was placed in one side of the restraint in the left and right chambers, and the other one remained empty (empty). The mouse was allowed to move freely for 10 min and the activity was recorded. After the test, the instrument was wiped with 75% alcohol. After the wiping, the third stage was carried out.

[0055] (3) The third stage of social preference detection: put the mouse back into the central chamber, take out stranger 1 and put it back into the restraint cage on one side, then put another mouse of the same species and similar age (stranger 2) into the restraint cage on the other side, open the small doors on the left and right baffles to let it move freely for 10 minutes and record. At the end of the experiment, put the mouse back into the cage and clean the behavior box. Statistics of experimental data: ①Social ability test: Count the total time S1 that the mouse's nose touches the stranger 1's restraint cage (start timing when the mouse's nose sniffs the experimental mouse and stop timing when it leaves); and the total time E that the mouse stays in the empty cage (start timing when the nose sniffs and stop timing when it leaves). Sociability index = (S1-E) / (S1+E).

[0056] ② Social novelty test: Count the total time S1 that the mouse's nose tip touches the cage of stranger 1 (start timing when the mouse's nose tip sniffs the experimental mouse and stop timing when it leaves); and the total time S2 that the mouse spends in stranger 2 (start timing when the nose tip sniffs and stop timing when it leaves). Social preference index = (S2-S1) / (S2+S1).

[0057] 2.3 Biochemical detection of MDA The kit was used to detect the MDA content in mouse hippocampal tissue and the operation was performed according to the corresponding method.

[0058] 2.4 Sample preparation Samples were collected the day after the behavioral study. After the mice were anesthetized, blood was collected from the eyeballs and serum was prepared for later use. After the blood was collected, the mice were quickly decapitated and the brains were removed. The brain tissues of the mice were quickly separated on an ice box and rinsed with precooled RNase-free water. The right hemisphere was fixed in 4% paraformaldehyde solution. The hippocampus and prefrontal lobe of the left brain tissue were placed in a cryovial and quickly frozen with liquid nitrogen and stored in a -80°C refrigerator for later use.

[0059] 2.5 Statistical analysis The measurement data are expressed as mean ± standard deviation ( ) The normality test was performed with the Shapiro-Wilk method, and the variance homogeneity analysis was performed with the Levene test. The independent sample t test was used for comparison between two groups, and one-way analysis of variance (ANOVA) was used for comparison between multiple groups. The Bonferroni method was used for multiple comparisons. When the overall distribution of the data was non-normal, the Mann-Whitney U test was used. GraphPad Prism 10.0 was used for statistical analysis, and P < 0.05 indicated that the difference was statistically significant.

[0060] 3. Results 3.1 Effect of the Chinese medicine composition of the present invention on the motor ability of BTBR mice The open field test was used to detect whether the pharmaceutical composition of the present invention changes the motor ability and exploration ability of BTBR mice. Figure 2 and Figure 3 , the total distance reflects the activity ability of mice, and the increase in central area movement time is closely related to the exploratory behavior of mice. Compared with the blank control group, the total distance of the model group was longer, and the difference was statistically significant (P < 0.05); compared with the model group, the total distance of the low-dose group and the high-dose group of the Chinese medicine composition of the present invention showed an upward trend, but the difference was not statistically significant. The results of the central area time analysis showed that there was no statistically significant difference between the model group and the blank control group; compared with the model group, the time of the low-dose group and the high-dose group of the Chinese medicine composition of the present invention showed an increasing trend, but the difference was not statistically significant. The total distance and central area time showed that there was no statistically significant difference between the low-dose group and the high-dose group of the Chinese medicine composition of the present invention, indicating that the efficacy of the low-dose group and the high-dose group is equivalent.

[0061] 3.2 The Chinese herbal composition of the present invention reduces stereotyped behavior in BTBR mice The buried bead test detects repetitive stereotyped behaviors in mice. Figure 4Compared with the blank control group, the number of buried beads in the model group increased, and the difference was statistically significant (P < 0.05); compared with the model group, the number of buried beads in the low-dose group and high-dose group of the Chinese medicine composition of the present invention decreased, and the difference was statistically significant (P < 0.001, P < 0.001). There was no statistically significant difference between the low-dose group and the high-dose group of the Chinese medicine composition of the present invention, indicating that the efficacy of the low-dose group and the high-dose group is equivalent.

[0062] Grooming time is used to measure repetitive stereotyped behaviors in mice. Figure 5 Compared with the blank control group, the model group had a longer grooming time, and the difference was statistically significant (P < 0.0001); compared with the model group, the grooming time of the low-dose group and the high-dose group of the Chinese medicine composition of the present invention was reduced, and the difference was statistically significant (P < 0.001, P < 0.01). Combined with the results of the bead burying experiment and the grooming experiment, it is suggested that the low-dose group and the high-dose group of the Chinese medicine composition of the present invention can reduce the repetitive stereotyped behavior of BTBR mice. There was no statistically significant difference between the low-dose group and the high-dose group of the Chinese medicine composition of the present invention, indicating that the efficacy of the low-dose group and the high-dose group is equivalent.

[0063] 3.3 The Chinese medicine composition of the present invention improves the social ability of BTBR mice The three-box social experiment was used to assess the mice's sociability and their preference for new mice. Figure 6 The blank control group spent more time sniffing the strange mice than the empty cage, and the difference was statistically significant (P < 0.0001), indicating that the blank control group had normal social ability; while the model group had no statistically significant difference in the sniffing time between the empty cage and the strange mice, indicating that the model group mice had abnormal social ability. The sniffing time of the strange mice in the low-dose group and the high-dose group of the Chinese medicine composition of the present invention was longer than the sniffing time in the empty cage, and the differences were statistically significant (P < 0.0001, P < 0.001), indicating that the social ability of BTBR mice was improved after medication. Figure 7 Compared with the blank control group, the social ability index of the model group was low, and the difference was statistically significant (P < 0.05); compared with the model group, the social index of the low-dose and high-dose groups of the Chinese medicine composition of the present invention was improved, and the difference was statistically significant (P < 0.05), indicating that the medication has an improvement effect on the social ability of BTBR mice. In the second stage, the results of social time and social index showed that there was no statistically significant difference between the low-dose group and the high-dose group of the Chinese medicine composition of the present invention, indicating that the efficacy of the low-dose group and the high-dose group was equivalent.

[0064] In the third stage, the sniffing time of mice towards familiar mice (S1) and unfamiliar mice (S2) was counted. Figure 8, the blank control group sniffed the unfamiliar mice more than the familiar mice, but the difference was not statistically significant, which was considered to be related to the specific behavioral operating environment and the small number of mice; while the model group, the low-dose group of the Chinese medicine composition of the present invention, and the high-dose group had differences in the sniffing time of S1 and S2, but the differences were not statistically significant. In each group, S1>S2, indicating that the social preference of the mice in the model group was abnormal, and the medication did not improve the social preference of BTBR mice. However, the social preference index of the model group was lower than that of the blank control group, and the difference was statistically significant (P<0.05), indicating that the social preference of the model group was abnormal; compared with the model group, there was no statistically significant difference in the social preference index between the low-dose group and the high-dose group of the Chinese medicine composition of the present invention, indicating that the Chinese medicine composition of the present invention had no significant improvement in social preference ability. In the third stage, the results of social preference sniffing time and social preference index indicated that there was no statistically significant difference between the low-dose group of the Chinese medicine composition of the present invention and the high-dose group, indicating that the efficacy of the low-dose group of the Chinese medicine composition of the present invention was equivalent to that of the high-dose group. For results, see Fig. 9 . 3.4 The Chinese herbal composition of the present invention improves the memory ability of BTBR mice The Y-maze spontaneous alternation test was used to assess the spatial working memory ability of mice. Fig.10 Compared with the blank control group, the spontaneous alternation rate of the model group decreased, and the difference was statistically significant (P < 0.05); compared with the model group, the spontaneous alternation rates of the low-dose group and the high-dose group of the Chinese medicine composition of the present invention increased, and the differences between the two groups were statistically significant (P < 0.05, P < 0.05), indicating that the Chinese medicine composition of the present invention can improve the memory ability of mice. There was no statistically significant difference between the low-dose group and the high-dose group of the Chinese medicine composition of the present invention, indicating that the efficacy of the low-dose group and the high-dose group of the Chinese medicine composition of the present invention is equivalent.

[0065] 3.5 The Chinese medicine composition of the present invention reduces MDA levels Biomarkers of oxidative stress can help assess the level of oxidative stress in the body. Malondialdehyde (MDA) is a common research product of lipid peroxidation and one of the most useful markers for monitoring oxidative damage. Fig.11 Compared with the blank control group, the MDA content in the model group increased, and the difference was statistically significant (P < 0.001), indicating that the model group had lipid peroxidation; compared with the model group, the MDA content in the low-dose group and the high-dose group of the Bushen Qizhi Recipe decreased, and the difference between the two groups was statistically significant (P < 0.01, P < 0.01), indicating that the Chinese medicine composition of the present invention can reverse oxidative damage. There was no statistically significant difference between the low-dose group and the high-dose group of the Chinese medicine composition of the present invention, indicating that the efficacy of the low-dose group and the high-dose group of the Chinese medicine composition of the present invention is equivalent.

[0066] In summary, the present invention is based on the fact that BTBR mice have a stable genetic background, so the behavioral phenotype is relatively stable. The BTBR T+Itpr3tf / J (BTBR for short) autism model mice can better simulate the stereotyped behavior, social defects, cognitive impairment and other problems of ASD in clinical practice. At the same time, the present invention combines the pathogenesis of autism: spleen and kidney yang deficiency, marrow sea insufficiency, warming the spleen and kidney, replenishing essence and marrow, and resolving phlegm and opening the orifices as the treatment principle, and innovatively considers the proposal of the Chinese medicine composition of the present invention, and combines the efficacy of various behavioral verification drugs to provide a more rigorous and comprehensive scientific basis. The experimental results show that the use of the Chinese medicine composition of the present invention improves the social ability and cognitive ability of BTBR mice, reduces repetitive stereotyped behavior, and its mechanism of action may be related to reducing MDA levels and alleviating lipid peroxidation. The low-dose group of the Chinese medicine composition of the present invention plays a comparable role compared with the high-dose group. At the same time, the present invention has found through clinical studies that the use of the Chinese medicine composition of the present invention can effectively improve the eye contact of children with ASD, increase interest in external things, and improve the effect of behavioral intervention training.

[0067] The above contents are only for explaining the technical idea of ​​the present invention and cannot be used to limit the protection scope of the present invention. Any changes made on the basis of the technical solution in accordance with the technical idea proposed by the present invention shall fall within the protection scope of the claims of the present invention.

Claims

1. A Chinese medicine composition for treating autism spectrum disorder, characterized in that: The composition by mass is as follows: 4-10 parts of cinnamon bark, 4-10 parts of Morinda officinalis, 4-10 parts of Polygala tenuifolia, 4-10 parts of Acorus calamus, 5-10 parts of Rehmannia glutinosa, 4-10 parts of Angelica sinensis, 8-20 parts of Tortoise Shell, and 5-10 parts of Dendrobium officinale.

2. The Chinese medicine composition for treating autism spectrum disorder according to claim 1, characterized in that: The composition by mass is as follows: 4-6 parts of cinnamon bark, 4-6 parts of Morinda officinalis, 4-6 parts of Polygala tenuifolia, 4-6 parts of Acorus calamus, 5-8 parts of Rehmannia glutinosa, 4-6 parts of Angelica sinensis, 8-15 parts of Tortoise Shell, and 5-8 parts of Dendrobium officinale.

3. The Chinese medicine composition for treating autism spectrum disorder according to claim 1, characterized in that: The composition by mass is as follows: 5 parts of cinnamon bark, 6 parts of Morinda officinalis, 4 parts of Polygala tenuifolia, 5 parts of Acorus calamus, 8 parts of Rehmannia glutinosa, 5 parts of Angelica sinensis, 10 parts of Carapace of Tortoise Shell, and 8 parts of Dendrobium officinale.

4. The Chinese medicine composition for treating autism spectrum disorder according to claim 1, characterized in that: The composition by mass is as follows: 5 parts of cinnamon bark, 5 parts of Morinda officinalis, 5 parts of Polygala tenuifolia, 5 parts of Acorus calamus, 6 parts of Rehmannia glutinosa, 5 parts of Angelica sinensis, 10 parts of Carapace of Tortoise Shell, and 6 parts of Dendrobium officinale.

5. The Chinese medicine composition for treating autism spectrum disorder according to claim 1, characterized in that: The composition by mass is as follows: 5 parts of cinnamon bark, 5 parts of Morinda officinalis, 5 parts of Polygala tenuifolia, 8 parts of Acorus calamus, 8 parts of Rehmannia glutinosa, 5 parts of Angelica sinensis, 8 parts of Carapax tortoise shell, and 6 parts of Dendrobium candidum.

6. Use of the Chinese medicine composition for treating autism spectrum disorder according to any one of claims 1 to 5 in the preparation of a medicament for treating autism spectrum disorder.

7. The use according to claim 6, characterized in that The medicine is a medicine for improving the user's social ability and reducing stereotyped behavior.

8. The use according to claim 6, characterized in that The drug is added with pharmaceutically acceptable adjuvants or excipients to prepare an oral preparation.

9. The use according to claim 8, characterized in that The oral preparation is in the form of tablets, powders, pills, capsules, granules or decoctions.

10. The use according to claim 8, characterized in that The auxiliary materials or excipients include one or more of stabilizers, solubilizers, lubricants and disintegrants.

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