Traditional Chinese medicine preparation for improving or treating cognitive impairment

Abstract

The application provides a traditional Chinese medicine preparation for improving or treating cognitive impairment, and relates to the technical field of traditional Chinese medicine preparation. The traditional Chinese medicine preparation is prepared from Radix Morindae Officinalis, Herba Hyperici Japonici, Platycladi Seed, Fructus Evodiae, Rhizoma Atractylodis Macrocephalae, Valeriana, Curcuma Longa, Rhizoma Acori Tatarinowii, Hirudo, Rhizoma Anemarrhenae, Dendrobium officinale, Gigeriae Cacumen, and Radix Pini. The traditional Chinese medicine preparation in the application shows significant neuroprotective and cognitive improvement effects at the cell and animal levels: at the cell level, the preparation can effectively resist A 1‑42 induced cell damage, and plays a protective role through multiple pathways such as improving cell viability, enhancing antioxidant capacity and inhibiting neuroinflammation; in animal experiments, the preparation can comprehensively improve the cognitive dysfunction of APP / PS1 model mice, including relieving behavioral abnormalities, improving spatial learning and memory ability, enhancing new object recognition ability, and effectively reducing Aβ deposition in hippocampal tissue at the molecular level. The application provides a new idea for improving or treating cognitive impairment.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine pharmaceutical technology, specifically to a traditional Chinese medicine preparation for improving or treating cognitive impairment. Background Technology

[0002] Cognitive impairment is a group of neurodegenerative diseases characterized by a decline in cognitive functions such as memory, executive function, and language. Common types include Alzheimer's disease (AD), vascular dementia (VaD), mild cognitive impairment (MCI), and post-stroke cognitive impairment (PSCI). With the increasing aging of my country's population, the prevalence of cognitive impairment is rising year by year, becoming one of the main causes of disability and long-term care requirements in the elderly. Currently, the treatment of cognitive impairment mainly relies on cholinesterase inhibitors and NMDA receptor antagonists. However, these drugs can only alleviate symptoms to a certain extent and cannot reverse the neurodegenerative process. They are ineffective for non-dementia vascular cognitive impairment (VCIND), and long-term use may be accompanied by adverse reactions.

[0003] Traditional Chinese medicine (TCM), characterized by its multi-component, multi-target, and multi-pathway approach, demonstrates unique advantages in the prevention and treatment of cognitive impairment. Its holistic view and syndrome differentiation principles align closely with the complexity of the etiology and pathogenesis of cognitive impairment. TCM classifies cognitive impairment into categories such as "dementia," "forgetfulness," and "literacy disorder," primarily affecting the brain and closely related to dysfunctions of the heart, liver, spleen, and kidneys. The core pathogenesis is described as "deficiency of kidney essence, insufficiency of qi and blood, phlegm obstruction of the orifices, and internal obstruction of blood stasis." Based on these theories, TCM has accumulated rich clinical experience in treating cognitive impairment, often employing various treatment methods such as tonifying the kidneys and replenishing essence, resolving phlegm and opening the orifices, and promoting blood circulation and unblocking the meridians. Clinically, compound preparations are frequently used, such as Rehmannia Decoction, Tongqiao Huoxue Decoction, and Guipi Decoction, showing potential and advantages in improving patients' cognitive function, enhancing daily living abilities, and overall condition.

[0004] Although traditional Chinese medicine (TCM) has shown promising prospects in the treatment of cognitive impairment, several key issues remain: First, the formulation and mechanism of action are unclear, with traditional formulas containing numerous herbs and complex relationships between the principal, assistant, and adjuvant herbs. Second, the formulation lacks systematic modern pharmacological validation; the effective component groups of most compound formulas are unclear, and their targets are dispersed, making it difficult to establish a stable efficacy evaluation system. Given the high incidence and serious harm of cognitive impairment, as well as the limitations of existing treatments, developing a scientifically formulated, effective, and safe TCM preparation is of great significance for meeting clinical treatment needs and improving patients' quality of life. Summary of the Invention

[0005] (a) Technical problems to be solved

[0006] In view of the shortcomings of the prior art, the present invention provides a traditional Chinese medicine preparation for improving or treating cognitive impairment.

[0007] (II) Technical Solution

[0008] To achieve the above objectives, the present invention provides the following technical solution:

[0009] In a first aspect, the present invention provides a traditional Chinese medicine preparation for improving or treating cognitive impairment, the preparation being made from the following raw materials in parts by weight: Morinda officinalis 15-35 parts, Eclipta prostrata 15-35 parts, Platycladus orientalis seed 12-33 parts, Alpinia oxyphylla seed 12-33 parts, Atractylodes macrocephala 10-30 parts, Valerian 10-30 parts, Curcuma longa 10-30 parts, Acorus tatarinowii 8-28 parts, Hirudo medicinalis 8-28 parts, Anemarrhena asphodeloides 5-25 parts, Dendrobium officinale 5-25 parts, Gallus gallus domesticus gizzard lining 8-28 parts, Caesalpinia sappan 3-20 parts, and Glycyrrhiza uralensis (processed) 10-30 parts.

[0010] Specifically, the traditional Chinese medicine preparation is made from the following raw materials in parts by weight: Morinda officinalis 20-30 parts, Eclipta prostrata 20-30 parts, Platycladus orientalis seed 18-27 parts, Alpinia oxyphylla seed 18-27 parts, Atractylodes macrocephala 15-25 parts, Valerian 15-25 parts, Curcuma longa 15-25 parts, Acorus tatarinowii 13-24 parts, Hirudo medicinalis 13-24 parts, Anemarrhena asphodeloides 10-20 parts, Dendrobium officinale 10-20 parts, Gallus gallus domesticus gizzard lining 13-24 parts, Caesalpinia sappan 7-15 parts, and Glycyrrhiza uralensis (processed) 15-25 parts.

[0011] Specifically, the traditional Chinese medicine preparation is made from the following raw materials in parts by weight: Morinda officinalis 25 parts, Eclipta prostrata 25 parts, Platycladus orientalis seed 22 parts, Alpinia oxyphylla seed 22 parts, Atractylodes macrocephala 20 parts, Valerian 20 parts, Curcuma longa 20 parts, Acorus tatarinowii 18 parts, Hirudo medicinalis 18 parts, Anemarrhena asphodeloides 15 parts, Dendrobium officinale 15 parts, Gallus gallus domesticus gizzard lining 18 parts, Caesalpinia sappan 12 parts, and Glycyrrhiza uralensis (processed) 20 parts.

[0012] Secondly, the present invention provides a method for preparing a traditional Chinese medicine preparation for improving or treating cognitive impairment. The traditional Chinese medicine preparation is prepared into an oral dosage form by conventional pharmaceutical methods and with medically acceptable excipients, including any one of pills, powders, granules, oral liquids, decoctions, tablets, capsules, and ointments.

[0013] Thirdly, the present invention provides the use of the aforementioned traditional Chinese medicine preparation in the preparation of a medicament for improving and / or treating cognitive impairment.

[0014] The pharmacological effects of each traditional Chinese medicine in the herbal preparation of this invention are as follows:

[0015] Morinda officinalis: The dried root of Morinda officinalis, a plant in the Rubiaceae family. It is sweet and pungent in taste, slightly warm in nature, and enters the kidney and liver meridians. It has the effects of tonifying kidney yang, strengthening tendons and bones, and dispelling wind and dampness. It is used for impotence and seminal emission, infertility due to cold uterus, irregular menstruation, lower abdominal pain due to cold, rheumatic pain, and weakness of tendons and bones.

[0016] Eclipta prostrata: The dried aerial parts of Eclipta prostrata, a plant of the Asteraceae family. It is sweet and sour in taste, cold in nature, and enters the kidney and liver meridians. It has the effects of nourishing the liver and kidneys, cooling the blood and stopping bleeding. It is used for liver and kidney yin deficiency, loose teeth, premature graying of hair, dizziness and tinnitus, soreness and weakness of the waist and knees, vomiting blood, epistaxis, hematuria, bloody dysentery, metrorrhagia, and external bleeding.

[0017] Arborvitae seed kernel: The dried, mature seed kernel of Arborvitae, a plant of the Cupressaceae family. It is sweet in taste and neutral in nature, and enters the heart, kidney, and large intestine meridians. It has the effects of nourishing the heart and calming the mind, moistening the intestines and relieving constipation, and stopping sweating. It is used for yin and blood deficiency, vexation and insomnia, palpitations, constipation due to intestinal dryness, and night sweats due to yin deficiency.

[0018] Alpinia oxyphylla: The fruit of Alpinia oxyphylla, a plant in the ginger family. It is pungent and warm in nature, and enters the spleen and kidney meridians. It has the effects of warming the spleen to stop diarrhea and reduce salivation; warming the kidneys to reduce urination and consolidate essence. It is used for spleen and stomach deficiency and cold, vomiting, diarrhea, cold in the abdomen, excessive salivation, kidney deficiency and enuresis, frequent urination, seminal emission, and leukorrhea.

[0019] Atractylodes macrocephala: The dried rhizome of Atractylodes macrocephala, a plant of the Asteraceae family. It is bitter and sweet in taste, warm in nature, and enters the spleen and stomach meridians. It has the effects of strengthening the spleen and replenishing qi, drying dampness and promoting diuresis, stopping sweating, and calming the fetus. It is used for spleen deficiency with poor appetite, abdominal distension and diarrhea, phlegm retention with dizziness and palpitations, edema, spontaneous sweating, and threatened abortion.

[0020] Valerian: The root and rhizome of Valeriana and Valeriana purpurea, both belonging to the Valerianaceae family. It has a pungent and sweet taste, is warm in nature, and enters the heart and liver meridians. It has the effects of calming the mind, regulating qi, and relieving pain. It is used for neurasthenia, insomnia, hysteria, epilepsy, abdominal distension and pain, lower back and leg pain, and traumatic injuries.

[0021] Turmeric: The dried rhizome of the turmeric plant (Zingiberaceae family). It has a pungent and bitter taste, is warm in nature, and enters the spleen and liver meridians. It has the effects of promoting blood circulation, regulating qi, and relieving pain. It is used for chest and rib pain, chest pain, dysmenorrhea, amenorrhea, abdominal masses, rheumatic shoulder and arm pain, and swelling and pain from falls.

[0022] Acorus calamus: The dried rhizome of Acorus calamus, a plant of the Araceae family. It is pungent and bitter in taste, warm in nature, and enters the heart and stomach meridians. It has the effects of opening the orifices and resolving phlegm, refreshing the mind and improving intelligence, and resolving dampness and stimulating the appetite. It is used for coma and epilepsy, forgetfulness and insomnia, tinnitus and deafness, epigastric fullness and lack of appetite, and aphthous stomatitis and diarrhea.

[0023] Leech: The dried whole body of the leech (Hirudo medicinalis), the leech (Hirudo spp.), or the willow leech (Hirudo spp.). It is salty and bitter in taste and neutral in nature. It enters the liver meridian. It has the effects of breaking up blood stasis and promoting menstruation, removing blood stasis and eliminating masses. It is used for amenorrhea due to blood stasis, abdominal masses, hemiplegia due to stroke, and injuries from falls.

[0024] Anemarrhena asphodeloides: The dried rhizome of Anemarrhena asphodeloides, a plant of the Liliaceae family. It is bitter and sweet in taste, cold in nature, and enters the lung, stomach, and kidney meridians. It has the effects of clearing heat and purging fire, nourishing yin and moistening dryness. It is used for exogenous febrile diseases, high fever and thirst, dry cough due to lung heat, bone steaming fever, internal heat and thirst, and constipation due to intestinal dryness.

[0025] Dendrobium officinale: The dried stem of Dendrobium officinale, a plant of the Orchidaceae family. It is sweet in taste and slightly cold in nature, and enters the stomach and kidney meridians. It has the effects of nourishing the stomach and promoting the production of body fluids, nourishing yin and clearing heat. It is used for febrile diseases with depletion of body fluids, dry mouth and thirst, insufficient stomach yin, poor appetite and dry retching, persistent low fever after illness, yin deficiency and fire excess, bone steaming fever, blurred vision, and weakness of muscles and bones.

[0026] Chicken gizzard lining: The dried inner wall of the gizzard of the domestic chicken (Pheasantidae). It is sweet in taste and neutral in nature, and enters the spleen, stomach, small intestine, and bladder meridians. It has the effects of strengthening the stomach and promoting digestion, astringing essence and stopping seminal emission, and clearing urinary stones and dissolving stones. It is used for indigestion, vomiting and diarrhea, infantile malnutrition, enuresis, seminal emission, painful urination with stones, and gallbladder distension and hypochondriac pain.

[0027] Sappanwood: The dried heartwood of the legume Sappanwood, sweet and salty in taste, neutral in nature, and enters the heart, liver, and spleen meridians; it has the effects of promoting blood circulation and removing blood stasis, reducing swelling and relieving pain; it is used for traumatic injuries, fractures and sprains, swelling and pain due to blood stasis, amenorrhea and dysmenorrhea, postpartum blood stasis, chest and abdominal pain, and carbuncles and boils.

[0028] Prepared licorice root: A processed product of the dried roots and rhizomes of licorice, licorice inflatum, or licorice glabra, all belonging to the legume family. It is sweet in taste and neutral in nature, and enters the heart, lung, spleen, and stomach meridians. It has the effects of tonifying the spleen and stomach, replenishing qi and restoring pulse. It is used for spleen and stomach weakness, fatigue, palpitations, and irregular pulse.

[0029] (III) Beneficial Effects

[0030] This invention provides a traditional Chinese medicine preparation for improving or treating cognitive impairment. The preparation is made from Morinda officinalis, Eclipta prostrata, Platycladus orientalis seed, Alpinia oxyphylla, Atractylodes macrocephala, Valeriana, Curcuma longa, Acorus tatarinowii, Hirudo medicinalis, Anemarrhena asphodeloides, Dendrobium officinale, Gallus gallus domesticus gizzard lining, Caesalpinia sappan, and Glycyrrhiza uralensis (processed). The kidney is the foundation of innate essence; sufficient kidney essence leads to abundant marrow, which is the basis for the overall improvement or treatment of cognitive impairment. This formula uses Morinda officinalis and Eclipta prostrata as the principal herbs. Morinda officinalis excels at warming the kidney and strengthening yang, replenishing essence and marrow, and strengthening tendons and bones; Eclipta prostrata nourishes yin and kidney, cools the blood and stops bleeding, and replenishes essence and marrow. The two herbs, one warming and one cooling, one tonifying yang and one nourishing yin, together serve as the principal herbs, forming a core of simultaneously tonifying kidney yin and yang and nourishing essence and marrow, directly addressing the fundamental pathogenesis of insufficient marrow. The formula consists of cypress seed, alpinia oxyphylla seed, atractylodes macrocephala, valerian, and turmeric. Cypress seed nourishes the heart and calms the mind, benefits blood and brain, and moistens the intestines to relieve constipation; alpinia oxyphylla warms the kidneys and consolidates essence, benefits intelligence and calms the mind, and warms the spleen to stop diarrhea; atractylodes macrocephala invigorates qi and strengthens the spleen, dries dampness and promotes diuresis, and generates qi and blood; valerian calms the mind and soothes the nerves, regulates qi and relieves pain, and calms the heart and relieves palpitations; turmeric breaks up blood stasis and promotes qi circulation, and relieves pain. The five herbs work together to strengthen the spleen and benefit intelligence, calm the mind and stabilize the will, and unblock the meridians and assist the marrow, thereby enhancing the kidney-tonifying and marrow-replenishing power of the principal herb, forming a synergistic brain-nourishing mechanism of kidney-tonifying, spleen-generating, and heart-calming. The formula is supplemented with Acorus tatarinowii, leech, Anemarrhena asphodeloides, Dendrobium officinale, chicken gizzard lining, and Sappanwood as adjuvant herbs. Acorus tatarinowii dispels dampness, opens the orifices, refreshes the mind, improves intelligence, and calms the nerves; leech breaks up blood stasis and clears the brain meridians; Sappanwood invigorates blood circulation, clears the meridians, disperses blood stasis, and reduces swelling; Dendrobium officinale benefits the stomach, generates fluids, nourishes yin and the brain, and clears heat and moistens dryness; chicken gizzard lining strengthens the spleen, promotes digestion, and resolves stagnation; Anemarrhena asphodeloides clears heat, nourishes yin, moistens dryness, and purges fire. The six adjuvant herbs work together to address both the symptoms of cognitive impairment caused by phlegm and blood stasis obstructing the orifices and to balance the bias of the principal and assistant herbs, avoiding excessive heat and dryness that could damage yin or excessive purgation. Prepared licorice root is used as the guiding herb. Prepared licorice root is warm in nature and enters the heart, lung, spleen, and stomach meridians. It harmonizes the properties of the other herbs and moderates the strong effects of drastic herbs such as leech and Acorus tatarinowii, ensuring a well-balanced and moderate formula.

[0031] The traditional Chinese medicine preparation of this invention exhibits significant neuroprotective and cognitive-improving effects at both the cellular and animal levels. At the cellular level, this preparation can effectively resist Aβ. 1-42 The induced SH-SY5Y cell damage is protected through multiple pathways, including increasing cell viability, enhancing antioxidant capacity (manifested as increased SOD activity and decreased MDA content), and inhibiting neuroinflammation (such as reducing the release of pro-inflammatory factors like TNF-α, IL-1β, and IL-18). In animal experiments, this formulation comprehensively improved cognitive dysfunction in APP / PS1 model mice, including alleviating behavioral abnormalities, improving spatial learning and memory abilities, enhancing the ability to recognize new objects, and effectively reducing Aβ deposition in the hippocampus at the molecular level. This traditional Chinese medicine formulation, through the synergistic effects of multiple components and multiple targets, demonstrates comprehensive potential for improving or treating cognitive impairment, from cell protection to overall behavioral improvement, and has promising application prospects. Attached Figure Description

[0032] Figure 1Comparison of MDA content and SOD activity in cell supernatants of each group; Note: Compared with the blank control group, # P<0.05, ## P<0.01; compared with the model control group, * P<0.05, ** P<0.01.

[0033] Figure 2 Results of the Morris water maze experiment; a: Escape latency of mice in each group during the navigation experiment; b: Number of times mice in each group traversed platforms during the spatial exploration experiment; c: Time spent in the target quadrant by mice in each group during the spatial exploration experiment; Note: Compared with the blank control group, # P<0.05, ## P<0.01; compared with the model control group, * P<0.05, ** P<0.01.

[0034] Figure 3 Results of the new object recognition experiment; Note: Compared with the blank control group, # P<0.05, ## P<0.01; compared with the model control group, * P<0.05, ** P<0.01. Detailed Implementation

[0035] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0036] Example 1

[0037] A traditional Chinese medicine preparation for improving or treating cognitive impairment, the preparation being made from the following raw materials in parts by weight: Morinda officinalis 25 parts, Eclipta prostrata 25 parts, Platycladus orientalis seed 22 parts, Alpinia oxyphylla seed 22 parts, Atractylodes macrocephala 20 parts, Valerian 20 parts, Curcuma longa 20 parts, Acorus tatarinowii 18 parts, Hirudo medicinalis 18 parts, Anemarrhena asphodeloides 15 parts, Dendrobium officinale 15 parts, Gallus gallus domesticus gizzard lining 18 parts, Sappanwood 12 parts, and Glycyrrhiza uralensis (processed) 20 parts.

[0038] Example 2

[0039] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine preparation is made from the following raw materials in parts by weight: 15 parts Morinda officinalis, 15 parts Eclipta prostrata, 12 parts Platycladus orientalis seed, 12 parts Alpinia oxyphylla seed, 10 parts Atractylodes macrocephala, 10 parts Valerian, 10 parts Curcuma longa, 8 parts Acorus tatarinowii, 8 parts Hirudo medicinalis, 5 parts Anemarrhena asphodeloides, 5 parts Dendrobium officinale, 8 parts Gallus gallus domesticus gizzard lining, 3 parts Sappanwood, and 10 parts Glycyrrhiza uralensis (processed).

[0040] Example 3

[0041] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine preparation is made from the following raw materials in parts by weight: Morinda officinalis 30 parts, Eclipta prostrata 30 parts, Platycladus orientalis seed 27 parts, Alpinia oxyphylla seed 27 parts, Atractylodes macrocephala 23 parts, Valerian 23 parts, Curcuma longa 23 parts, Acorus tatarinowii 21 parts, Hirudo medicinalis 21 parts, Anemarrhena asphodeloides 20 parts, Dendrobium officinale 20 parts, Gallus gallus domesticus gizzard lining 24 parts, Caesalpinia sappan 17 parts, and Glycyrrhiza uralensis (processed) 25 parts.

[0042] Example 4

[0043] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine preparation is made from the following raw materials in parts by weight: Morinda officinalis 35 parts, Eclipta prostrata 35 parts, Platycladus orientalis seed 33 parts, Alpinia oxyphylla seed 33 parts, Atractylodes macrocephala 30 parts, Valerian 30 parts, Curcuma longa 30 parts, Acorus tatarinowii 28 parts, Hirudo medicinalis 28 parts, Anemarrhena asphodeloides 25 parts, Dendrobium officinale 25 parts, Gallus gallus domesticus gizzard lining 28 parts, Caesalpinia sappan 20 parts, and Glycyrrhiza uralensis (processed) 30 parts.

[0044] Example 5

[0045] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine preparation is made from the following raw materials in parts by weight: Morinda officinalis 28 parts, Eclipta prostrata 28 parts, Platycladus orientalis seed 25 parts, Alpinia oxyphylla seed 24 parts, Atractylodes macrocephala 21 parts, Valerian 22 parts, Curcuma longa 20 parts, Acorus tatarinowii 20 parts, Hirudo medicinalis 20 parts, Anemarrhena asphodeloides 18 parts, Dendrobium officinale 18 parts, Gallus gallus domesticus gizzard lining 21 parts, Caesalpinia sappan 15 parts, and Glycyrrhiza uralensis (processed) 22 parts.

[0046] Example 6

[0047] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine preparation is made from the following raw materials in parts by weight: Morinda officinalis 18 parts, Eclipta prostrata 18 parts, Platycladus orientalis seed 15 parts, Alpinia oxyphylla seed 15 parts, Atractylodes macrocephala 13 parts, Valerian 12 parts, Curcuma longa 14 parts, Acorus tatarinowii 10 parts, Hirudo medicinalis 10 parts, Anemarrhena asphodeloides 7 parts, Dendrobium officinale 7 parts, Gallus gallus domesticus gizzard lining 10 parts, Caesalpinia sappan 5 parts, and Glycyrrhiza uralensis (processed) 12 parts.

[0048] Example 7

[0049] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine preparation is made from the following raw materials in parts by weight: 20 parts Morinda officinalis, 20 parts Eclipta prostrata, 18 parts Platycladus orientalis seed, 17 parts Alpinia oxyphylla, 15 parts Atractylodes macrocephala, 15 parts Valerian, 15 parts Curcuma longa, 12 parts Acorus tatarinowii, 12 parts Hirudo medicinalis, 10 parts Anemarrhena asphodeloides, 10 parts Dendrobium officinale, 12 parts Gallus gallus domesticus gizzard lining, 6 parts Sappanwood, and 15 parts Glycyrrhiza uralensis (processed).

[0050] Example 8

[0051] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine preparation is made from the following raw materials in parts by weight: Morinda officinalis 32 parts, Eclipta prostrata 32 parts, Platycladus orientalis seed 31 parts, Alpinia oxyphylla 30 parts, Atractylodes macrocephala 26 parts, Valerian 25 parts, Curcuma longa 25 parts, Acorus tatarinowii 24 parts, Hirudo medicinalis 25 parts, Anemarrhena asphodeloides 23 parts, Dendrobium officinale 22 parts, Gallus gallus domesticus gizzard lining 26 parts, Caesalpinia sappan 18 parts, and Glycyrrhiza uralensis (processed) 27 parts.

[0052] Experimental Example 1

[0053] 1. Materials and Methods

[0054] 1.1 Investigational Drug

[0055] Experimental Group 1: Morinda officinalis 25 g, Eclipta prostrata 25 g, Platycladus orientalis seed 22 g, Alpinia oxyphylla seed 22 g, Atractylodes macrocephala 20 g, Valerian 20 g, Curcuma longa 20 g, Acorus tatarinowii 18 g, Hirudo medicinalis 18 g, Anemarrhena asphodeloides 15 g, Dendrobium officinale 15 g, Gallus gallus domesticus gizzard lining 18 g, Caesalpinia sappan 12 g, Glycyrrhiza uralensis (processed) 20 g.

[0056] Experimental Group 2: Eclipta prostrata 25 g, Platycladus orientalis seed 22 g, Alpinia oxyphylla seed 22 g, Atractylodes macrocephala 20 g, Valerian 20 g, Curcuma longa 20 g, Acorus tatarinowii 18 g, Dendrobium officinale 15 g, Gallus gallus domesticus gizzard lining 18 g, Caesalpinia sappan 12 g, Glycyrrhiza uralensis (processed) 20 g.

[0057] Experimental Group 3: Morinda officinalis 25 g, Platycladus orientalis seed 22 g, Alpinia oxyphylla seed 22 g, Atractylodes macrocephala 20 g, Valerian 20 g, Curcuma longa 20 g, Acorus tatarinowii 18 g, Hirudo medicinalis 18 g, Anemarrhena asphodeloides 15 g, Caesalpinia sappan 12 g, Glycyrrhiza uralensis (processed) 20 g.

[0058] Experimental Group 4: Morinda officinalis 25 g, Eclipta prostrata 25 g, Atractylodes macrocephala 20 g, Valerian 20 g, Curcuma longa 20 g, Hirudo medicinalis 18 g, Anemarrhena asphodeloides 15 g, Dendrobium officinale 15 g, Gallus gallus domesticus gizzard lining 18 g, Glycyrrhiza uralensis (processed) 20 g.

[0059] Experimental Group 5: Morinda officinalis 25 g, Eclipta prostrata 25 g, Platycladus orientalis seed 22 g, Alpinia oxyphylla 22 g, Acorus tatarinowii 18 g, Anemarrhena asphodeloides 15 g, Dendrobium officinale 15 g, Gallus gallus domesticus gizzard lining 18 g, Caesalpinia sappan 12 g, Glycyrrhiza uralensis (processed) 20 g.

[0060] Weigh each raw material according to the above weight ratio, wash them, add 3 times the total weight of distilled water and soak for 30 minutes, bring to a boil over high heat and then simmer over low heat for 40 minutes. Repeat the simmering process twice, filter, combine the filtrates and concentrate under reduced pressure until the raw drug content is 0.1 g / mL to obtain the original Chinese medicine liquid of test groups 1-5.

[0061] 1.2 Cell Culture, Drug Pretreatment and Modeling

[0062] The experiment included a blank control group, a model control group, and experimental groups 1-5, with three replicates per group. SH-SY5Y cells were resuscitated in DMEM complete medium containing 10% FBS and 1% penicillin antibiotics and cultured at 37°C in a 5% CO2 incubator, with the medium changed every 2-3 days. When cell confluence reached 80%-90%, cells were digested with 0.25% trypsin and passaged. Logarithmic growth phase cells were collected and prepared into 2×10⁶ cells / wells using complete medium. 5 Cell suspensions of 100 μL / mL were seeded into 96-well plates and cultured for 24 h to allow cell adhesion. The original culture medium was then discarded, and 100 μL of complete culture medium containing 300 μg / mL of the corresponding drug was added to groups 1-5. The blank control group and model group received an equal volume of complete culture medium and were incubated for 2 h. Subsequently, except for the blank control group, all other groups received 20 μM Aβ. 1-42 The cells were treated, and an equal volume of complete culture medium was added to the blank control group. After culturing for another 48 h, the cells and supernatant were collected for various tests.

[0063] 1.3 Cell viability assay

[0064] Discard the original culture medium, add 20 μL of CCK-8 and 180 μL of complete culture medium to each well, and incubate at 37℃ and 5% CO2 for 2 h. Collect the supernatant and measure the absorbance at 450 nm using a microplate reader. Set up blank wells (containing only 200 µL of complete culture medium). Calculate cell viability according to the following formula:

[0065] Cell viability (%) = (OD) 实验组 -OD 空白孔 ) / (OD 空白对照组 -OD 空白孔 ) × 100%

[0066] 1.4 Detection of Oxidative Stress Indicators

[0067] Discard the original culture medium, add 100 μL of PBS to each well, scrape cells with a cell scraper, collect the cell suspension, centrifuge at 10,000 rpm for 5 min, discard the supernatant, and retain the cell pellet. Following the kit instructions, add lysis buffer to the cell pellet to fully lyse the cells, centrifuge at 12,000 rpm for 10 min, collect the supernatant, and use the xanthine oxidase method to detect superoxide dismutase (SOD) activity and the barbituric acid sulfate method to detect malondialdehyde (MDA) content, respectively.

[0068] 1.5 Detection of inflammatory factor levels

[0069] Collect the cell supernatant, centrifuge at 1000 rpm for 10 min, take the supernatant, and detect the levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-18 (IL-18) in the cell supernatant according to the ELISA kit instructions.

[0070] 1.6 Statistical Methods

[0071] All data were analyzed using SPSS 26.0 software and are expressed as mean ± standard deviation. One-way ANOVA was used for comparisons between groups, with P < 0.05 considered statistically significant.

[0072] 2 Results

[0073] 2.1 Cell viability

[0074] As shown in Table 1, the cell viability of the model control group was significantly lower than that of the blank control group (P<0.01), indicating that Aβ 1-42 A model of induced SH-SY5Y cell damage was successfully established; the cell viability of groups 1-5 in the experiment was significantly higher than that of the model control group (P<0.01), indicating that the traditional Chinese medicine preparation of this invention can inhibit Aβ. 1-42 Induced SH-SY5Y cell damage and improved cell survival.

[0075] 2.2 Oxidative stress

[0076] Depend on Figure 1 It was found that, compared with the blank control group, the model control group showed significantly decreased SOD activity and significantly increased MDA content (P<0.01), indicating that Aβ... 1-42 The study successfully induced oxidative stress damage in cells. Compared with the model control group, the SOD activity in experimental groups 1-5 showed an increasing trend, while the MDA content decreased. Among them, experimental group 1 showed the most significant improvement (P<0.01), indicating that the traditional Chinese medicine preparation of this invention can effectively reduce Aβ by enhancing the antioxidant capacity of cells. 1-42 Induced oxidative stress damage.

[0077] 2.3 Inflammatory Factors

[0078] Table 1 shows that the levels of TNF-α, IL-1β, and IL-18 in the model control group were significantly higher than those in the blank control group (P<0.01), indicating that Aβ... 1-42 The inflammatory response of SH-SY5Y cells was successfully induced. The levels of the aforementioned inflammatory factors in groups 1-5 were significantly lower than those in the model control group (P<0.05 or P<0.01), with group 1 showing the most significant effect. This indicates that the traditional Chinese medicine preparation of this invention can inhibit Aβ. 1-42 Induced release of inflammatory factors from SH-SY5Y cells, thereby alleviating inflammatory damage.

[0079] Table 1 Comparison of cell viability and serum inflammatory factor levels in each group

[0080] Grouping Cell viability (%) TNF-α (pg / mL) IL-1β (pg / mL) IL-18 (pg / mL) Blank control group 100±0.00 24.66±4.07 19.81±3.23 46.92±3.77 Model control group <![CDATA[52.59±3.73 ## ]]> <![CDATA[132.51±6.37 ## ]]> <![CDATA[47.76±2.12 ## ]]> <![CDATA[129.68±5.70 ## ]]> Experiment 1 group <![CDATA[100.57±2.39 ** ]]> <![CDATA[38.39±4.09 ** ]]> <![CDATA[24.05±1.64 ** ]]> <![CDATA[75.19±3.54 ** ]]> Experimental Group 2 <![CDATA[99.85±2.98 ** ]]> <![CDATA[56.74±3.96 ** ]]> <![CDATA[31.00±1.85 ** ]]> <![CDATA[92.58±4.77 ** ]]> Experiment 3 groups <![CDATA[97.50±2.89 ** ]]> <![CDATA[77.48±4.64 ** ]]> <![CDATA[36.14±1.57 ** ]]> <![CDATA[81.77±4.53 ** ]]> Experiment 4 groups <![CDATA[102.30±3.20 ** ]]> <![CDATA[117.87±4.28 * ]]> <![CDATA[42.03±1.61 * ]]> <![CDATA[116.34±4.53 * ]]> Experiment 5 groups <![CDATA[96.08±2.32 ** ]]> <![CDATA[99.52±5.61 ** ]]> <![CDATA[40.49±2.14 ** ]]> <![CDATA[105.91±4.52 ** ]]>

[0081] Note: Compared with the blank control group # P<0.05, ## P<0.01; compared with the model control group, * P<0.05, ** P<0.01.

[0082] 3. Conclusion

[0083] The above experimental results show that the traditional Chinese medicine preparation of this invention has an effect on Aβ. 1-42 The induced SH-SY5Y cell damage exhibits significant protective effects. Specifically, it effectively enhances cell viability and maintains cell morphology; enhances cellular antioxidant capacity and reduces oxidative damage by increasing SOD activity and decreasing MDA content; and reduces the release of pro-inflammatory factors such as TNF-α, IL-1β, and IL-18, thereby alleviating neuroinflammatory responses. Among these, the traditional Chinese medicine preparation of this invention (experimental group 1) showed the best improvement effect in all the above indicators. The effects of experimental groups 2-5 were weakened to varying degrees due to the lack of certain herbs in the formula. This phenomenon verifies at the molecular level the multi-component, multi-target, and synergistic effects of the traditional Chinese medicine preparation of this invention—the various herbs in the formula exert synergistic effects through the compatibility of the principal, assistant, adjuvant, and guide herbs, jointly constructing a multi-dimensional protection against neuronal damage.

[0084] Experimental Example 2

[0085] 1.1 Laboratory Animals

[0086] Thirty-five 6-8 week old APP / PS1 double transgenic male mice and five age-matched wild-type C57BL / 6J male mice, weighing 25±3 g, were used. All mice were housed in an SPF-grade animal facility under environmental conditions of (24±1)℃ and (50±5)% relative humidity, with a 12h light / 12h dark circadian rhythm. After one week of acclimatization, the mice were allowed free access to food and water during the experiment.

[0087] 1.2 Experimental Drugs

[0088] The traditional Chinese medicine preparation of this invention consists of: Morinda officinalis 25 g, Eclipta prostrata 25 g, Platycladus orientalis seed 22 g, Alpinia oxyphylla seed 22 g, Atractylodes macrocephala 20 g, Valerian 20 g, Curcuma longa 20 g, Acorus tatarinowii 18 g, Hirudo medicinalis 18 g, Anemarrhena asphodeloides 15 g, Dendrobium officinale 15 g, Gallus gallus domesticus gizzard lining 18 g, Caesalpinia sappan 12 g, and Glycyrrhiza uralensis (processed) 20 g. Each ingredient is weighed according to the above weight ratio, washed, and soaked in distilled water (3 times the total weight) for 30 minutes. The mixture is brought to a boil over high heat, then simmered over low heat for 40 minutes. This simmering process is repeated twice. The mixture is filtered, and the filtrates are combined and concentrated under reduced pressure to a crude drug content of 0.1 g / mL. It is then stored at 4℃ for later use. Before use, it is diluted with physiological saline to the corresponding concentration according to the required dosage.

[0089] Positive control drug: donepezil hydrochloride.

[0090] 1.3 Animal grouping and administration

[0091] APP / PS1 mice were randomly divided into a model group, a low-dose group, a high-dose group, and a positive control group, with 5 mice in each group; wild-type C57BL / 6J male mice served as the blank control group. All mice were administered the drug by gavage at a volume of 10 mL / kg, once daily for 4 weeks, as follows:

[0092] Blank control group and model group: were given an equal volume of physiological saline.

[0093] Low-dose group: The traditional Chinese medicine preparation of the present invention was administered at a dose of 50 mg / kg (based on crude drug).

[0094] High-dose group: The traditional Chinese medicine preparation of the present invention was administered at a dose of 100 mg / kg (based on crude drug).

[0095] Positive control group: given donepezil hydrochloride suspension at a dose of 0.65 mg / kg.

[0096] 1.4 Detection Indicators

[0097] 1.4.1 General Case

[0098] The mental state, food and water intake, spontaneous activity intensity, fur color, and urination and defecation of mice in each group were observed and recorded daily.

[0099] 1.4.2 Water Maze Experiment

[0100] The Morris water maze test was used to assess the spatial learning and memory abilities of mice. The experimental setup consisted of a circular water tank with a diameter of 90 cm, inside which was a 5.0 cm × 5.0 cm platform. An appropriate amount of titanium dioxide powder was added to the water and stirred until the water became cloudy. The experiment consisted of three stages, the specific procedures of which are as follows:

[0101] Orientation and navigation training: 5 consecutive days of training, once a day, with a 1-hour interval between each training session. Mice were randomly placed in one of the four quadrants facing the pool wall. The time required for the mouse to successfully climb onto the platform (escape latency) was recorded. If the time was >60 seconds, the mouse was manually guided to swim to the platform and remain there for 10 seconds to reinforce memory. The mouse was then dried, cared for, and returned to its cage.

[0102] Orientation and navigation test: conducted on day 6, mice were placed into the water tank from the quadrant opposite to the platform, and the escape latency of the mice was recorded.

[0103] Space exploration experiment: conducted on day 7. At this time, the platform was removed, and the quadrant opposite to the platform was used as the entry point for the mice. The mice were allowed to swim freely for 60 seconds, and the number of times they crossed the original platform position and the time spent in the target quadrant were recorded.

[0104] 1.4.3 New Object Recognition Experiment

[0105] The working memory and non-spatial memory abilities of mice were assessed using a novel object recognition experiment. The experiment was conducted over two days, with the following procedures: On the first day, mice were placed in a 40 cm × 40 cm × 30 cm test box and allowed to explore freely for 5 minutes. This process was repeated once after a 1-hour interval to familiarize them with the environment. After familiarization, two identical objects were placed diagonally opposite each other in the test box, and the mice were placed back in the test box and allowed to explore freely for 5 minutes. Their exploration behavior was recorded. On the second day, one of the two identical objects was replaced with a new object of different appearance. The test box environment and objects were cleaned and odors were removed. The mice were then allowed to explore freely for 5 minutes, and the exploration time for the new object and the old object was recorded. The Recognition Index (RI) was calculated.

[0106] RI = (New object exploration time / New object exploration time + Old object exploration time) × 100%

[0107] 1.4.4 Determination of Aβ content in hippocampal tissue

[0108] After the behavioral experiment, the mice were fasted for 12 hours. The next day, the mice were sacrificed by cervical dislocation, and the brains were removed. The hippocampus tissue was isolated, and residual blood on the surface of the tissue was washed away with pre-cooled physiological saline. Pre-cooled physiological saline was added at a ratio of 1:15 (mass / volume) to prepare a tissue homogenate. The tissue homogenate was centrifuged at 3500 r / min for 15 min at 4°C, and the supernatant was collected. The Aβ content in the hippocampus tissue was measured according to the ELISA kit instructions. 1-40 Aβ 1-42 The content of.

[0109] 1.5 Statistical Methods

[0110] SPSS 26.0 software was used for analysis, and results are expressed as mean ± standard deviation. Graphpad Prism 8.0 software was used for statistical analysis and graphing. One-way ANOVA was used to compare differences between groups, and a p-value < 0.05 was considered statistically significant.

[0111] 2 Results

[0112] 2.1 General Case

[0113] Compared with the blank control group, the model group mice showed lethargy, reduced spontaneous activity, increased sleep time, dull and lackluster fur, and decreased food and water intake. Compared with the model group, the general condition of mice in each drug administration group was improved to varying degrees, specifically, their mental state recovered, their food and water intake tended to be normal, their fur gradually regained its luster, and their spontaneous activity and sensitivity increased.

[0114] 2.2 Water Maze

[0115] Depend on Figure 2 It can be seen that, compared with the blank control group, the escape latency of the model group mice in the localization and navigation test was significantly prolonged, and the number of times they crossed the original platform and the time spent in the target phenomenon in the spatial exploration experiment were significantly reduced (P<0.01). Compared with the model group, the escape latency of mice in each drug administration group was significantly shortened, and the number of times they crossed the platform and the time spent in the target phenomenon were significantly increased (P<0.05 or P<0.01), and showed a dose-dependent improvement trend, indicating that the traditional Chinese medicine preparation of the present invention has an enhancing effect on the spatial learning ability of APP / PS1 mice.

[0116] 2.3 New Object Recognition

[0117] Depend on Figure 3It can be seen that, compared with the blank control group, the RI index of the model group mice was significantly reduced (P<0.01); compared with the model group, the RI index of the low-dose group and the high-dose group of the traditional Chinese medicine preparation increased by 5.99% and 22.02% respectively (P<0.05 or P<0.01), indicating that the traditional Chinese medicine preparation of the present invention can significantly improve the working memory and non-spatial memory ability of APP / PS1 mice, and the high-dose group has a more significant effect.

[0118] 2.4 Aβ content

[0119] Table 2 shows that, compared with the blank control group, the Aβ levels in the hippocampus of the model group mice were significantly higher. 1-40 Aβ 1-42 The levels of Aβ in the hippocampus of mice in each treatment group were significantly increased (P<0.01); compared with the model group, the levels of Aβ in the hippocampus of mice in each treatment group were significantly increased. 1-40 Aβ 1-42 The content of all drugs was significantly reduced (P<0.05 or P<0.01), with the high-dose group showing a significantly greater decrease than the low-dose group, which was comparable to the improvement effect of donepezil hydrochloride as a positive control drug.

[0120] Table 2 Comparison of Aβ content in hippocampus tissue of mice in different groups

[0121] Grouping <![CDATA[Aβ 1-40 (ng / mL)]]> <![CDATA[Aβ 1-42 (ng / mL)]]> Blank control group 66.61±3.74 23.80±3.11 Model control group <![CDATA[141.07±4.94 ## ]]> <![CDATA[58.78±2.36 ## ]]> low-dose group <![CDATA[128.54±4.40 * ]]> <![CDATA[50.83±2.35 * ]]> High-dose group <![CDATA[85.36±3.40 ** ]]> <![CDATA[38.53±2.97 ** ]]> Positive control group <![CDATA[81.07±3.39 ** ]]> <![CDATA[32.18±3.13 ** ]]>

[0122] Note: Compared with the blank control group # P<0.05, ## P<0.01; compared with the model control group, * P<0.05, ** P<0.01.

[0123] 3. Conclusion

[0124] The above experimental data show that the traditional Chinese medicine preparation of this invention has a significant therapeutic effect on cognitive impairment in APP / PS1 mice, effectively improving their behavioral performance, enhancing learning, memory, and recognition abilities, and intervening in key pathological processes. The specific mechanism of action is as follows: This preparation can effectively alleviate core behavioral deficiencies in APP / PS1 mice, improve their mental state, spontaneous activity, and coat luster, and enhance their overall vitality; in terms of cognitive function, this preparation can significantly shorten the escape latency of mice, increase the number of platform crossings, and the dwell time in the target quadrant, thereby improving their spatial learning and memory abilities; simultaneously, it can significantly improve the recognition index, improving key cognitive aspects such as non-spatial recognition and working memory; at the molecular level, it can significantly reduce Aβ levels in the hippocampus of mice. 1-40 Aβ 1-42The content of Aβ is reduced, thus decreasing Aβ deposition. This invention's traditional Chinese medicine preparation improves cognitive impairment in APP / PS1 mice from multiple levels, including overall behavioral performance, cognitive function, and molecular pathological basis, providing a new approach for improving or treating cognitive impairment.

[0125] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A traditional Chinese medicine preparation for improving or treating cognitive impairment, characterized in that, The traditional Chinese medicine preparation is made from the following raw materials in parts by weight: Morinda officinalis 15-35 parts, Eclipta prostrata 15-35 parts, Platycladus orientalis seed 12-33 parts, Alpinia oxyphylla seed 12-33 parts, Atractylodes macrocephala 10-30 parts, Valerian 10-30 parts, Curcuma longa 10-30 parts, Acorus tatarinowii 8-28 parts, Hirudo medicinalis 8-28 parts, Anemarrhena asphodeloides 5-25 parts, Dendrobium officinale 5-25 parts, Gallus gallus domesticus gizzard lining 8-28 parts, Caesalpinia sappan 3-20 parts, and Glycyrrhiza uralensis (processed) 10-30 parts.

2. The traditional Chinese medicine preparation for improving or treating cognitive impairment according to claim 1, characterized in that, The traditional Chinese medicine preparation is made from the following raw materials in parts by weight: Morinda officinalis 20-30 parts, Eclipta prostrata 20-30 parts, Platycladus orientalis seed 18-27 parts, Alpinia oxyphylla seed 18-27 parts, Atractylodes macrocephala 15-25 parts, Valerian 15-25 parts, Curcuma longa 15-25 parts, Acorus tatarinowii 13-24 parts, Hirudo medicinalis 13-24 parts, Anemarrhena asphodeloides 10-20 parts, Dendrobium officinale 10-20 parts, Gallus gallus domesticus gizzard lining 13-24 parts, Caesalpinia sappan 7-15 parts, and Glycyrrhiza uralensis (processed) 15-25 parts.

3. The traditional Chinese medicine preparation for improving or treating cognitive impairment according to claim 1, characterized in that, The traditional Chinese medicine preparation is made from the following raw materials in parts by weight: Morinda officinalis 25 parts, Eclipta prostrata 25 parts, Platycladus orientalis seed 22 parts, Alpinia oxyphylla seed 22 parts, Atractylodes macrocephala 20 parts, Valerian 20 parts, Curcuma longa 20 parts, Acorus tatarinowii 18 parts, Hirudo medicinalis 18 parts, Anemarrhena asphodeloides 15 parts, Dendrobium officinale 15 parts, Gallus gallus domesticus gizzard lining 18 parts, Sappanwood 12 parts, and Glycyrrhiza uralensis (processed) 20 parts.

4. The method for preparing a traditional Chinese medicine preparation for improving or treating cognitive impairment as described in any one of claims 1 to 3, characterized in that, The traditional Chinese medicine preparation is prepared into an oral dosage form according to conventional pharmaceutical methods and with medically acceptable excipients. The oral dosage form is any one of pills, powders, granules, oral liquids, decoctions, tablets, capsules, and ointments.

5. The use of the traditional Chinese medicine preparation according to any one of claims 1 to 3 in the preparation of a medicament for improving and / or treating cognitive impairment.

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