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 aspect of overall compatibility, the cinnamon and the morinda officinalis in the composition are used as monarch drugs for warming kidney and supporting yang to replenish essence; prepared rehmannia root and tortoise shell are used as ministerial drugs and assist monarch drugs to nourish kidney yin, replenish essence and benefit marrow; the polygala tenuifolia and the rhizoma acori graminei have the effects of reducing phlegm and inducing resuscitation, the angelica sinensis has the effects of replenishing blood and promoting blood circulation and regulating qi and blood, and the dendrobe has the effects of nourishing yin and clearing heat and restricting the warm dryness property of the monarch drugs. All the medicines are compatible to jointly achieve the effects of tonifying the kidney, warming yang, replenishing essence and benefiting marrow, kidney essence can be sufficient, and brain marrow can be nourished, so that the normal physiological function of a human body is maintained, and ASD symptoms are comprehensively improved.

Description

Technical Field

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

[0002] Autism spectrum disorder (ASD) is a neurodevelopmental disorder that begins early in development and is characterized by impairments in social interaction and communication, restricted interests, and stereotyped behaviors, often accompanied by intellectual disability. The global prevalence of ASD is increasing annually, but its etiology and pathogenesis remain unclear. It is generally believed that ASD is a neurodevelopmental disorder caused by a combination of genetic and environmental factors. Treatment for ASD primarily involves non-pharmacological and pharmacological approaches. Non-pharmacological approaches currently focus on behavioral interventions, using rehabilitation and educational training to modify core symptoms to improve social adaptability and quality of life. Psychological therapy is also used to improve specific symptoms. However, most ASDs affect the physical and mental health of children, resulting in a poor prognosis and lifelong complications. Furthermore, rehabilitation training is expensive, placing significant emotional stress and financial burden on families and society. Currently, there are no proven effective medications for this condition, and pharmacological treatment is typically indicated when significant psychiatric and neurological symptoms are present. For example, haloperidol and trihexyphenidyl hydrochloride can be used to control stereotyped movements, excessive activity, and impulsive behavior; sulpiride can be used to improve withdrawal, calm emotions, and improve speech. However, these drugs are also used to relieve mental symptoms, and long-term use can have significant side effects.

[0003] It can be seen that there is currently a severe 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 urgently needs to be addressed in the clinic. Chinese medicine compositions also have certain research value and potential in the treatment of ASD. There are also some related reports, such as the invention patent with publication number CN 113663048 A, "A Chinese medicine composition for autism spectrum disorder and its preparation method", which 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 promoting digestion. It constructs an autistic mouse model by intraperitoneal injection of sodium valproate during pregnancy in 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] "The kidneys store essence, which generates marrow that flows to the brain." Innate essence, inherited from parents and stored in the kidneys, serves as the foundation of the body's yin and yang. It then forms the brain marrow, transforming into spirit, which resides in the brain. With the spirit's residence in the brain, it exerts its influence throughout the body. Kidney essence is the original substance from which brain marrow is generated. Driven by kidney qi, kidney essence ascends via the Du meridian to the brain, where it transforms into marrow, filling the brain's orifices and forming the "kidney-marrow-brain" pathway. Therefore, congenital kidney essence deficiency is the root cause, leading to a lack of fullness in the sea of ​​marrow, resulting in brain hypoplasia, growth retardation, and intellectual disability. Children with ASD often develop due to maternal illness or emotional distress during pregnancy, or due to weak parents, resulting in inadequate fetal endowment. Alternatively, due to unexpected circumstances such as intrauterine hypoxia during childbirth, the seat of the spirit is damaged, leading to congenital kidney essence deficiency and a lack of fullness in the sea of ​​marrow. This can manifest as developmental delays in intelligence, movement, and language expression, as well as a lack of eye contact with others. Considering the pathogenesis of autism spectrum disorder, which is due to insufficient kidney essence and depletion of the sea of ​​marrow, there is still a lack of clear treatment options, which is an area that urgently needs a breakthrough. Summary of the Invention

[0005] In order to overcome the shortcomings of the above-mentioned prior art and to address the pathogenesis of autism, which is kidney essence deficiency and marrow insufficiency, 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:

[0007] The invention discloses a traditional Chinese medicine composition for treating autism spectrum disorder. The composition comprises the following components in parts 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 candidum.

[0008] 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.

[0009] 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.

[0010] 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.

[0011] 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.

[0012] The cinnamon bark in the traditional 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 kidney yang deficiency and spleen and kidney yang deficiency. Cinnamon bark also has sedative and analgesic effects, antibacterial and anti-inflammatory properties, anti-radiation and anti-tumor effects, immune function enhancement, asthma relief, and insecticide effects.

[0013] The Morinda officinalis in the Chinese medicine composition of the present invention is also known as Baji, which is a vine plant with a slightly warm nature, a pungent and sweet taste. It enters the kidney and liver meridians and has the effects of tonifying kidney yang, strengthening muscles and bones, and removing rheumatism. It is mainly used to treat kidney deficiency, soreness of the waist and knees, and weakness of muscles and bones.

[0014] The "Polygala tenuifolia" in the Chinese medicine composition of the present invention is a perennial herb primarily grown in Northeast, North, Northwest, and Central China, as well as in Sichuan. Polygala tenuifolia is bitter, pungent, and warm in nature, and enters the heart, kidney, and lung meridians. It can improve heart-kidney disharmony and significantly alleviate various symptoms caused by this condition, such as palpitations and soreness of the waist and knees. Furthermore, the active ingredients in Polygala tenuifolia help regulate the nervous system, significantly improving symptoms such as insomnia, frequent dreams, and palpitations, thereby enhancing sleep quality and memory.

[0015] 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, dispelling filth, opening the orifices and refreshing the mind. It is often used to treat symptoms such as confusion, coma and delirium caused by phlegm and filth, and evil obstructing the orifices.

[0016] The "prepared rehmannia root" in the Chinese medicine composition of this invention is a processed product of raw rehmannia root. It is sweet and slightly warm, and enters the liver and kidney meridians. Preparing rehmannia root helps nourish the spleen and stomach, promote appetite and digestion, and enhance physical strength. It is commonly used to treat debilitating symptoms such as weakness, fatigue, and indigestion. Preparing rehmannia root helps regulate the nervous system, alleviates anxiety, insomnia, and mood swings, and improves mental state. It also nourishes the liver and kidneys, strengthens the muscles and bones, and has a certain effect on treating liver and kidney yin deficiency, as well as soreness of the waist and knees.

[0017] The Chinese angelica root in the Chinese medicinal composition is a perennial herb with a warm nature, sweet and pungent flavor, and enters the liver, heart, and spleen meridians. It nourishes and invigorates blood circulation and can be used to treat symptoms such as anemia, sallow complexion, dizziness, and palpitations. It also boosts the body's resistance and strengthens the immune system.

[0018] The "tortoise shell" in the present Chinese medicine composition refers to the carapace and plastron of the tortoise, a member of the family Testudinidae. It is considered an important Chinese medicinal material, boasting multiple benefits, including nourishing yin and suppressing yang, tonifying the kidneys and strengthening bones, nourishing blood and tonifying the heart, and strengthening menstruation and arresting metrorrhagia. It is commonly used to treat symptoms such as hot flashes due to yin deficiency, night sweats caused by bone steaming, yin deficiency and yang hyperactivity, dizziness, internal wind movement, weakness of muscles and bones, heart deficiency and forgetfulness, and metrorrhagia.

[0019] The dendrobium in the Chinese medicine composition of the present invention is a plant of the orchid family with a cold nature and a sweet, light, slightly salty taste. It enters the stomach, lung, and kidney meridians. Dendrobium nourishes the stomach, produces body fluids, nourishes yin, and clears heat. It is primarily used to treat symptoms such as loss of body fluids due to febrile illness, dry mouth and thirst, insufficient stomach yin, loss of appetite, retching, persistent post-illness fever, yin deficiency and excessive fire, bone steaming and fatigue fever, blurred vision, and weakness of muscles and bones.

[0020] The present invention also discloses the use of the above-mentioned traditional Chinese medicine composition for treating autism spectrum disorder in preparing medicine for treating autism spectrum disorder.

[0021] Preferably, the drug is a drug that improves the user's social skills and reduces stereotyped behaviors.

[0022] Preferably, the drug is added with pharmaceutically acceptable excipients or excipients to prepare an oral preparation.

[0023] More preferably, the oral preparation is a tablet, powder, pill, capsule, granule or decoction.

[0024] More preferably, the auxiliary materials or excipients include one or more of a stabilizer, a solubilizer, a lubricant and a disintegrant.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] The present invention uses traditional Chinese medicine theory, and successfully prepares a Chinese medicine composition containing cinnamon, Morinda officinalis, Polygala tenuifolia, Acorus gramineus, Rehmannia glutinosa, Chinese angelica, tortoise shell and Dendrobium candidum based on the theory and practice of traditional Chinese medicine on autism spectrum disorder. The composition has a simple and reasonable compatibility, significant efficacy, convenient material preparation, and is easy to promote. The selection of each medicine in the composition has been carefully considered, and the final composition emphasizes the reasonable compatibility between the medicines to achieve the purpose of synergistic enhancement. Specifically, the cinnamon in the pharmaceutical 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 the body's immunity; Morinda officinalis has the effects of warming and replenishing kidney yang, strengthening muscles and bones, and is suitable for symptoms such as five delays, five softness, and developmental delay caused by kidney yang deficiency, 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 gramineus has the effects of opening the mind and resolving phlegm, refreshing the mind and improving intelligence, and is suitable for phlegm-dampness obstruction It can treat symptoms such as unclear consciousness, poor memory, and lack of concentration caused by blocking the clear orifices; Rehmannia root has the effects of nourishing yin and blood, replenishing essence and filling marrow, providing a material basis for growth and development; Angelica sinensis has the effects 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 effects of nourishing yin and suppressing yang, replenishing the kidneys 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 effects 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 insufficient kidney essence and insufficient marrow. Cinnamon bark and Morinda officinalis are selected as the main drugs to warm the kidney and assist yang to help replenish essence; Rehmannia root and tortoise shell are selected as the ministers to assist the main drugs in nourishing kidney yin and nourishing the brain; Polygala root, Acorus calamus, Angelica sinensis, and Dendrobium are selected as the adjuvants. Polygala root and Acorus calamus resolve phlegm and open the orifices, Angelica sinensis replenishes blood and activates blood circulation, regulates qi and blood, and Dendrobium nourishes yin and clears heat, restraining the warming and dry nature of the main drugs. The combination of all the drugs together plays the role of tonifying the kidney and warming yang, replenishing essence and benefiting marrow, starting from the fundamental pathogenesis of insufficient kidney essence, and comprehensively improving the symptoms of autism. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is a design flow chart for the whole animal experiment of the present invention;

[0028] Figure 2 This is the total distance result of the open field test in behavioral experiments;

[0029] Figure 3 The results of central area movement time in the open field test of behavioral experiments;

[0030] Figure 4 The number of buried beads for behavioral experiments;

[0031] Figure 5 Grooming time results for behavioral experiments;

[0032] Figure 6 For behavioral experiments, the results of social ability sniffing time (s) were obtained;

[0033] Figure 7 The result of behavioral experiment social ability index (S1-E) / (S1+E);

[0034] Figure 8 The results of social preference sniffing time (s) in behavioral experiments;

[0035] Figure 9 The social preference index (S2-S1) / (S1+S2) of the behavioral experiment;

[0036] Figure 10 The results of spontaneous alternation rate (%) in the behavioral experiment Y maze;

[0037] Figure 11 This is the test result of MDA content (nmol / mg).

[0038] In the figure (except Figure 6 outside):( # P<0.05, ## P < 0.01, ### P < 0.001, compared with the blank control group; *P < 0.05, **P < 0.01, ***P < 0.001, compared with the model group); Figure 6 Middle: (***P<0.001, S1 compared with E). DETAILED DESCRIPTION

[0039] In order to enable those skilled in the art to better understand the solutions of the present invention, the technical solutions 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 embodiments described 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 making creative efforts should fall within the scope of protection of the present invention.

[0040] It should be noted that the terms "first", "second", etc. in the description 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 numbers 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 clearly listed, but may include other steps or units that are not clearly listed or inherent to these processes, methods, products or devices.

[0041] The present invention is described in further detail below with reference to the accompanying drawings:

[0042] 1. Examples of Chinese medicine compositions

[0043] Example 1

[0044] 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 candidum.

[0045] Example 2

[0046] 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.

[0047] Example 3

[0048] 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 candidum.

[0049] The above embodiments of the present invention can be prepared into common medicaments for use. The following is an example of a granule. The corresponding granulation operation is performed according to the characteristics of the raw medicinal materials in the pharmaceutical composition. 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 root, 5g of Angelica sinensis, 10g of Carapax tortoise shell, and 6g of Dendrobium candidum are prepared.

[0050] 1) Cinnamon: Take 5g of cinnamon slices and add 8 times the amount of water to the cinnamon bark. Boil for 60 minutes while extracting the volatile oil, then filter. Add 6 times the amount of water and boil for another 45 minutes while extracting the volatile oil. Filter through a 150-mesh filter and combine the filtrates. Concentrate the filtrates to a clear paste with a relative density of 1.05-1.10 (dry extract yield is 4%-8%). Combine the volatile oil and β-cyclodextrin in a colloid mill at a weight ratio of 1:6-10 for 2 hours at a constant temperature. Spray dry (liquid temperature 40°C-60°C, air inlet temperature 150°C-170°C). Add an appropriate amount of dextrin as an auxiliary, mix thoroughly, and granulate.

[0051] 2) Morinda officinalis: Take 5g of Morinda officinalis slices, add 10 times the amount of water, and boil for 60 minutes. Filter. Add 8 times the amount of water and boil for 45 minutes. Filter. Filter through a 150-mesh filter and combine the filtrates. Concentrate the filtrates to a clear paste with a relative density of 1.05-1.10 (dry extract yield of 36%-45%). Add an appropriate amount of dextrin as an auxiliary material, and spray dry (liquid supply temperature 50°C-65°C, air inlet temperature 150°C-170°C). Add an appropriate amount of dextrin as an auxiliary material, mix thoroughly, and dry granulate.

[0052] 3) Polygala: Take 5g of Polygala root slices, add 10 times the amount of water, and boil for 60 minutes. Filter. Add 8 times the amount of water and boil for 45 minutes. Filter. Filter through a 150-mesh filter and combine the filtrates. Concentrate the filtrates to a clear paste with a relative density of 1.05-1.10 (dry extract yield is 21.5%-40.5%). Add an appropriate amount of dextrin as an auxiliary material, and spray dry (liquid temperature 50°C-60°C, air inlet temperature 150°C-170°C). Add an appropriate amount of dextrin as an auxiliary material, mix thoroughly, and dry granulate.

[0053] 4) Acorus calamus: Take 5g of Acorus calamus slices and add 8 times the amount of water to boil for 60 minutes while extracting the volatile oil. Filter. Add 6 times the amount of water again and boil for 45 minutes while extracting the volatile oil. Filter through a 150-mesh filter and combine the filtrates. Concentrate the filtrates to a clear paste with a relative density of 1.05-1.10 (dry extract yield is 8%-13%). Combine the volatile oil and β-cyclodextrin in a colloid mill at a weight ratio of 1:8-10 for 2 hours at a constant temperature. Spray dry (liquid temperature 40°C-60°C, air inlet temperature 150°C-170°C). Add an appropriate amount of dextrin as an auxiliary, mix thoroughly, and granulate.

[0054] 5) Rehmannia glutinosa: Take 6g of Rehmannia glutinosa slices, add 10 times the amount of water, and boil for 90 minutes. Filter. Add 8 times the amount of water and boil for 75 minutes. Filter. Filter through a 150-mesh filter and combine the filtrates. Concentrate the filtrate to a clear paste with a relative density of 1.05-1.10 (dry extract yield is 44%-76.5%). Add an appropriate amount of dextrin as an auxiliary material, and spray dry (liquid temperature 50°C-60°C, air inlet temperature 150°C-170°C). Add an appropriate amount of dextrin as an auxiliary material, mix thoroughly, and dry granulate.

[0055] 6) Angelica: Take 5g of Angelica slices and add 8 times the amount of water, boil for 75 minutes, and extract the volatile oil. Filter. Add 6 times the amount of water and boil for 60 minutes, extracting the volatile oil. Filter through a 150-mesh filter and combine the filtrates. Concentrate the filtrates to a clear paste with a relative density of 1.05-1.10 (dry extract yield is 35%-56%). Combine the volatile oil and β-cyclodextrin in a colloid mill at a weight ratio of 1:6-10 for 2 hours at a constant temperature. Spray dry (liquid temperature 40°C-60°C, air inlet temperature 150°C-170°C). Add an appropriate amount of dextrin as an auxiliary, mix thoroughly, and granulate.

[0056] 7) Turtle Shell: Take 10g of tortoise shell slices, add 10 times the amount of water, and boil for 90 minutes. Filter. Add 8 times the amount of water and boil for 75 minutes. Filter. Filter through a 150-mesh filter and combine the filtrates. Concentrate the filtrate to a clear paste with a relative density of 1.05-1.10 (dry extract yield of 3%-7%). Add an appropriate amount of dextrin as an auxiliary material, and spray dry (liquid supply temperature 50°C-60°C, air inlet temperature 150°C-170°C). Add an appropriate amount of dextrin as an auxiliary material, mix thoroughly, and dry granulate.

[0057] 8) Dendrobium: Take 6g of Dendrobium officinale (Dendrobium tasseliensis) slices, add 10 times the amount of water, and boil for 80 minutes. Filter. Add 8 times the amount of water, and boil for 60 minutes. Filter. Filter through a 150-mesh filter, and combine the filtrates. Concentrate the filtrates to a clear paste with a relative density of 1.05-1.10 (dry extract yield is 9%-14.5%). Add an appropriate amount of dextrin as an auxiliary material, and spray dry (liquid temperature 50°C-60°C, air inlet temperature 150°C-170°C). Add an appropriate amount of dextrin as an auxiliary material, mix thoroughly, and dry granulate.

[0058] Note that in the actual application development process, the Chinese medicine composition of the present invention can also be prepared into oral dosage forms such as tablets, powders, pills, capsules, etc. according to conventional processes in the pharmaceutical field as needed.

[0059] 2. In vivo animal pharmacology experiments

[0060] See also Figure 1 , which is a design flow chart of the whole animal experiment of the present invention.

[0061] 1. Experimental Materials

[0062] 1.1 Experimental animals

[0063] 12 male C57BL / 6J mice at 4 - 5 weeks of age were used as the blank control group, provided by Beijing Vital River Laboratory Animal Technology Co., Ltd. (License number: SCXK(Beijing)2021 - 0011). 36 male SPF - grade BTBR mice at 4 - 5 weeks of age were provided by the Clinical Research Institute of China - Japan Friendship Hospital (License number: SYXK(Beijing)2023 - 0001). They were raised in a 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 approval number: ZRDWLL240005.

[0064] 1.2 Experimental instruments

[0065] Open - field test instrument, three - chamber social interaction test instrument, Y - maze test instrument, Tracking Master V5.0 behavioral tracking software (Beijing Zhongshi Dichuang Technology Development Co., Ltd.); 4% paraformaldehyde (Beijing Saierwei Co., Ltd.). Malondialdehyde test kit (MDAKit, Beijing Regbio Technology Development Co., Ltd. & China, batch number: 20250810.40003, product number: RGB - 40003).

[0066] 1.3 Experimental drugs

[0067] For the convenience of experimental investigation, the experimental drug was the traditional Chinese medicine granule prepared from the composition of Example 上述实施例2. It was dissolved in pure water according to the body weight of the mice and administered by gavage when used.

[0068] 2. Experimental methods

[0069] 2.1 Experimental grouping and administration methods

[0070] Blank control group: After 1 week of adaptive feeding, pure water of the corresponding volume was administered by gavage every day. For every 10 g of the mouse body weight, 0.2 mL was administered by gavage, lasting for 8 weeks.

[0071] Model group: After 1 week of adaptive feeding, pure water of the corresponding volume was administered by gavage every day. For every 10 g of the mouse body weight, 0.2 mL was administered by gavage, lasting for 8 weeks.

[0072] Traditional Chinese medicine granule prepared from the traditional Chinese medicine composition of the present invention: After 1 week of adaptive feeding, different - concentration traditional Chinese medicine granule infusions of the corresponding volume were administered by gavage for treatment every day. For every 10 g of the mouse body weight, 0.2 mL was administered by gavage, lasting for 8 weeks.

[0073] Low - and high - dose groups of the traditional Chinese medicine composition of the present invention for children with autism spectrum disorder: After 4 - 5 weeks of birth, low - dose (12 g / kg) and high - dose (24 g / kg) traditional Chinese medicine granule infusions of the corresponding volume were administered by gavage for treatment every day. For every 10 g of the mouse body weight, 0.2 mL was administered by gavage, lasting for 8 weeks.

[0074] 2.2 Observation of mouse behavior

[0075] 2.2.1 Open field test

[0076] The experiment used an open-field behavioral chamber to assess spontaneous locomotion, exploratory behavior, and anxiety in mice. The chamber was an open-top cube measuring 40 cm × 40 cm × 40 cm. The mouse was placed in the center of the chamber with its back facing the experimenter and allowed to explore freely for 5 minutes. The mouse's free movement within the chamber was recorded over the 5-minute period, and the chamber was then cleaned with 70% ethanol. The total distance traveled and the time spent in the central zone were recorded.

[0077] 2.2.2 Buried bead test

[0078] During the experiment, fresh, unscented bedding was added to a 5cm depth in a brand new cage and compacted flat. Standard black glass marbles (approximately 1cm in diameter and weighing approximately 1g) were gently placed on the bedding in five rows of four marbles spaced evenly apart. A camera was used to record the mice's movements for 20 minutes. Between each experiment, the marbles were washed with a mild detergent and allowed to dry before use. The number of marbles buried within 20 minutes was counted, with a bead buried two-thirds of the way inside being counted as one.

[0079] 2.2.3Y-maze spontaneous alternation experiment

[0080] 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 recorded as a correct alternation response. The mouse was placed in the device for 5 minutes for the adaptation phase, and then continued to observe for another 5 minutes, stop the video recording and save the file. 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.

[0081] 2.2.4 Self-grooming experiment

[0082] During the experiment, mice were placed in a brand new cage. A camera was turned on and the mice's movements were recorded for 20 minutes. Between each experiment, the cage was cleaned with 75% ethanol. The time the mice spent grooming was recorded for 20 minutes.

[0083] 2.2.5 Three-Box Social Experiment

[0084] (1) Acclimation phase: The experimental mouse was placed in the central chamber of the behavior box with its back facing the experimenter. It was allowed to move freely for 10 minutes to fully familiarize itself with the environment. After the test, the instrument was wiped with 75% ethanol. The second phase was carried out simultaneously in both channels.

[0085] (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 was kept empty (empty). The mouse was allowed to move freely for 10 minutes and the activity was recorded. After the test, the instrument was wiped with 75% alcohol. After the wiping, the third stage was carried out.

[0086] (3) Third stage social preference test: Place the mouse back in the central chamber, remove stranger 1 and place it back in the restraint cage on one side, then place another mouse of the same species and similar age (stranger 2) in the restraint cage on the other side. Allow the mouse to move freely for 10 minutes and record the behavior. At the end of the experiment, place the mouse back in the home cage and clean the behavior box. Statistical experimental data:

[0087] ① Sociability test: Count the total time, S1, the mouse's nose spent near stranger 1's restraint cage (time starts when the mouse's nose touches the experimental mouse and stops when it leaves); and the total time, E, the mouse spent in the empty cage (time starts when the nose touches the experimental mouse and stops when it leaves). Sociability index = (S1-E) / (S1+E).

[0088] ② Social novelty test: Count the total time the mouse's nose tip spent near stranger 1's restraint cage (S1) (time starts when the mouse's nose tip sniffs the experimental mouse and stops when it leaves); and the total time the mouse spent in stranger 2's chamber (S2) (time starts when the mouse's nose tip sniffs and stops when it leaves). Social preference index = (S2 - S1) / (S2 + S1).

[0089] 2.3 Biochemical detection of MDA

[0090] The kit was used to detect the MDA content in mouse hippocampal tissue and the operation was carried out according to the corresponding method.

[0091] 2.4 Sample preparation

[0092] Samples were collected the day after behavioral testing. After anesthesia, mice were quickly decapitated and brains removed. Brain tissue was quickly dissected on an ice box and rinsed with pre-chilled RNase-free water. The right hemisphere was fixed in 4% paraformaldehyde. The hippocampus and prefrontal lobe of the left brain were quickly frozen in liquid nitrogen in cryovials and stored at -80°C until use.

[0093] 2.5 Statistical analysis

[0094] The measurement data are expressed as mean ± standard deviation Normality was tested using the Shapiro-Wilk method, and homogeneity of variance was analyzed using the Levene test. Comparisons between two groups were performed with the independent sample t-test, and multiple groups were compared with one-way analysis of variance (ANOVA). Multiple comparisons were performed with the Bonferroni test, and when the data were nonnormal, the Mann-Whitney U test was used. Statistical analysis was performed with GraphPad Prism 10.0. P < 0.05 indicated statistical significance.

[0095] 3. Results

[0096] 3.1 Effect of the Chinese herbal composition of the present invention on the motor ability of BTBR mice

[0097] 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 increasing trend, but the difference was not statistically significant. The results of 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.

[0098] 3.2 The Chinese herbal composition of the present invention reduces stereotypic behavior in BTBR mice

[0099] The buried bead test is used to detect repetitive stereotyped behaviors in mice. Figure 4 Compared 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 the 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 low-dose group and the high-dose group have similar efficacy.

[0100] Grooming time is used to test repetitive stereotyped behaviors in mice. Figure 5Compared with the blank control group, the model group spent more time grooming, and the difference was statistically significant (P < 0.001). Compared with the model group, the low-dose and high-dose groups of the Chinese herbal composition of the present invention spent less time grooming, 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 was suggested that the low-dose and high-dose groups of the Chinese herbal composition of the present invention can reduce the repetitive stereotyped behavior of BTBR mice. There was no statistically significant difference between the low-dose and high-dose groups of the Chinese herbal composition of the present invention, indicating that the low-dose and high-dose groups have equivalent efficacy.

[0101] 3.3 The Chinese herbal composition of the present invention improves the sociability of BTBR mice

[0102] 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 unfamiliar mice than the empty cage, and the difference was statistically significant (P < 0.001), 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 unfamiliar mice, indicating that the model group mice had abnormal social ability. The sniffing time of the unfamiliar 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 < 0.001, P < 0.001), indicating that the social ability of BTBR mice was improved after medication. Figure 7 Compared with the blank control group, the model group had a lower sociability index, with a statistically significant difference (P < 0.05). Compared with the model group, the sociability index of the low-dose and high-dose groups of the Chinese herbal composition of the present invention was significantly improved (P < 0.05), indicating that the medication improved the sociability of BTBR mice. In the second phase, the results of social time and sociality index showed no statistically significant difference between the low-dose and high-dose groups of the Chinese herbal composition of the present invention, indicating that the low-dose and high-dose groups were equally effective.

[0103] 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 spent more time sniffing unfamiliar mice than familiar mice, but the difference was not statistically significant, which is 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 the 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), suggesting that the social preference of the model group was abnormal; compared with the model group, the social preference index of the low-dose group and the high-dose group of the Chinese medicine composition of the present invention had no statistically significant difference, 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 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 was equivalent. See the results for details. Figure 9 .

[0104] 3.4 The Chinese herbal composition of the present invention improves the memory ability of BTBR mice

[0105] The Y-maze spontaneous alternation test was used to assess the spatial working memory ability of mice. Figure 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 rate of the low-dose group and the high-dose group of the Chinese medicine composition of the present invention increased, and the difference between the two groups was 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 low-dose group and the high-dose group of the Chinese medicine composition of the present invention have equivalent efficacy.

[0106] 3.5 The Chinese herbal composition of the present invention reduces MDA levels

[0107] 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 is one of the most useful markers for monitoring oxidative damage. Figure 11 Compared with the blank control group, the MDA content in the model group increased, with a statistically significant difference (P < 0.001), indicating that the model group had lipid peroxidation. Compared with the model group, the MDA content in the low-dose and high-dose groups of the Bushen Qizhi Recipe decreased, with statistically significant differences between the two groups (P < 0.01, P < 0.01), indicating that the Chinese herbal composition of the present invention can reverse oxidative damage. There was no statistically significant difference between the low-dose and high-dose groups of the Chinese herbal composition of the present invention, indicating that the low-dose and high-dose groups of the Chinese herbal composition of the present invention have comparable efficacy.

[0108] In summary, the present invention, based on the stable genetic background of BTBR mice and their relatively stable behavioral phenotype, selects BTBR T+Itpr3tf / J (BTBR) autism model mice to better simulate the stereotyped behaviors, social deficits, and cognitive impairments seen in clinical ASD. Furthermore, the present invention combines the pathogenesis of autism: insufficient kidney essence and insufficient marrow, and uses the principle of tonifying the kidney and warming yang, replenishing essence and nourishing marrow as a treatment approach. Innovative considerations are used to propose the present invention's traditional Chinese medicine composition, which, combined with multiple behavioral studies to validate the efficacy of the drug, can provide a more rigorous and comprehensive scientific basis. Experimental data show that the use of the present invention's traditional Chinese medicine composition improves the social and cognitive abilities of BTBR mice and reduces repetitive stereotyped behaviors. Its mechanism of action may be related to reducing MDA levels and alleviating lipid peroxidation. The low-dose group of the present invention's traditional Chinese medicine composition exhibited comparable effects to the high-dose group. Furthermore, clinical studies have shown that the use of the present invention's traditional Chinese medicine composition can effectively improve eye contact in children with ASD, increase their interest in external things, and enhance the effectiveness of behavioral intervention training.

[0109] The above content is 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 root, 4-10 parts of Angelica sinensis, 8-20 parts of Tortoise shell, and 5-10 parts of Dendrobium candidum.

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 root, 4-6 parts of Angelica sinensis, 8-15 parts of Tortoise shell, and 5-8 parts of Dendrobium.

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 Carapax tortoise shell, and 8 parts of Dendrobium candidum.

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 Carapax tortoise shell, and 6 parts of Dendrobium candidum.

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 traditional 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.

Images