A traditional Chinese medicine composition with improved chronic sleep deprivation injury and application thereof

Abstract

The application discloses a kind of composition and its application, the composition is suitable for such as student group etc. The crowd engaged in high-intensity brain work, can improve the cognitive function decline and nerve injury caused by "chronic sleep deprivation (CSD)". It contains traditional Chinese medicine decoction pieces (according to daily amount) are: prepared 4.5~13.5g rhizoma polygonati, mulberry 4.5~13.5g, medlar seed 4.5~13.5g, longan meat 3~9g, chrysanthemum 3~9g, dried tangerine or orange peel 1.5~4.5g. Its preparation method is: the above-mentioned natural medicine is extracted by water extraction method after being separated or mixed, and the extract is spray dried to prepare powder. The powder is mixed uniformly to prepare the concentrated product. The water extract components have synergistic effect, can significantly alleviate the insufficient sleep caused by long-term passive overtime, and then the cognitive function decline, memory loss and emotional regulation imbalance caused by neuron damage.

Description

Technical Field

[0001] This invention relates to the fields of traditional Chinese medicine and food, and in particular to a traditional Chinese medicine composition (medicinal and edible traditional Chinese medicine) that improves chronic sleep deprivation (CSD) damage and its application. Background Technology

[0002] Sleep refers to the physiological process in which the body's response to external stimuli is weakened, it is in a relatively static state, and is accompanied by reversible loss of consciousness [J Biol Rhythm. 1999;14(6):557-568;Curr Opin Pulm Med. 2012; 18 6; 541-545]. Sufficient sleep plays an important role in eliminating fatigue, restoring physical strength, protecting brain function, and optimizing energy metabolism, while insufficient sleep duration and circadian rhythm disorders may lead to significant cognitive and emotional impairments [Prog Brain Res.2010; 185:105-129;Sleep Med Rev. 2010;14 4;219-226].

[0003] With the accelerated pace of modern society and increasing pressure from studies and work, young people, represented by students and working professionals, commonly experience prolonged periods of high-intensity mental activity accompanied by compressed sleep time (i.e., staying up late). Unlike patients with pathological insomnia, the sleep disorders in this group often stem from active or passive persistent sleep deprivation, which can develop into chronic sleep deprivation (CSD).

[0004] Studies have confirmed that chronic depression (CSD) can lead to decreased learning ability, attention deficit, cognitive decline, and imbalance in mood regulation, thereby inducing mental health problems such as anxiety and depression, and accelerating the progression of potential degenerative diseases [Neurobiol Learn Mem, 2011, 96 4: 564-582; Neuropsychologia, 2015, 69: 176-182]. Epidemiological studies have also shown that CSD is significantly associated with the development of type 2 diabetes, hypertension, and cardiovascular disease [Arch Ital Biol, 2014, 1522-3: 103-110; Oman Med J, 2016, 31 6: 399-403]. Its pathological mechanisms involve multiple pathways, including autonomic nervous system dysfunction, endothelial dysfunction, exacerbation of systemic inflammation, coagulation dysfunction, enhanced oxidative stress response, and neuroendocrine disorders. Through these pathophysiological mechanisms, it promotes the evolution of various chronic diseases [Sleep, 2012, 35 1: 97-101; Neurosci Biobehav Rev, 2017, 74 Pt B: 321-329].

[0005] For students and other individuals engaged in heavy mental work, chronic sleep deprivation has become a real dilemma. The continuous sacrifice of sleep to complete academic tasks leads to daytime inattention, impaired memory, and reduced learning efficiency. This, in turn, forces them to further reduce sleep to compensate for inefficient study time, creating a vicious cycle between sleep and learning efficiency. This long-term physiological compensatory overdraft not only damages individual academic performance and quality of life but may also cause irreversible long-term health damage.

[0006] Therefore, developing safe and effective traditional Chinese medicine compound preparations for the above-mentioned population to alleviate cognitive impairment and other physiological function damage induced by CSD has important clinical application value.

[0007] Traditional Chinese medicine theory indicates that sleep nourishes liver blood and preserves kidney essence. The *Huangdi Neijing* states, "When a person lies down, blood returns to the liver, and the liver, receiving blood, enables vision"; "Insufficient sleep depletes essence"; and "The kidneys store essence, produce marrow, and connect to the brain." Furthermore, *Introduction to Medicine* records, "Prolonged sitting injures the muscles, prolonged lying injures the qi, prolonged standing injures the bones, prolonged walking injures the tendons, and prolonged staring injures the blood." Therefore, prolonged sitting, prolonged staring, and long-term sleep deprivation easily lead to liver and kidney yin deficiency, heart and spleen deficiency, depletion of essence and blood, and malnourishment of the eyes, resulting in symptoms such as fatigue, forgetfulness, dizziness, dry eyes, and lower back and knee weakness. Treatment should focus on nourishing the liver and kidneys, strengthening the spleen and replenishing qi, and nourishing blood to improve vision.

[0008] Classic Chinese herbal formulas include: Gui Pi Wan (primarily tonifying qi and blood, strengthening the spleen and nourishing the heart), An Shen Bu Nao Ye (warming and tonifying, strengthening the brain and generating marrow), Sheng Mai Yin (tonifying qi and yin), and Qi Ju Di Huang Wan (nourishing the kidneys and liver), all of which are related to this treatment principle. These formulas have undergone extensive clinical verification, demonstrating definite clinical efficacy and clear indications: Gui Pi Wan is selected for qi and blood deficiency; An Shen Bu Nao Ye is selected for kidney essence deficiency with qi and blood deficiency; Sheng Mai Yin is selected for qi and yin deficiency; and Qi Ju Di Huang Wan is selected for liver and kidney yin deficiency.

[0009] With the development of the times, the severe sleep deprivation caused by the increasingly heavy academic pressure among contemporary teenagers has a complex underlying pathogenesis. Currently, there is still a lack of comprehensive treatment plans that address the dysfunction of multiple organs in this condition. This group often exhibits a complex pathogenesis of heart and liver blood deficiency, kidney essence depletion, spleen qi deficiency, and upward disturbance of deficient fire. Simple tonification or purging is insufficient to address all aspects. It is advisable to optimize the combination of traditional formulas. The following formula is based on food and medicine homology, integrating yin-nourishing and blood-tonifying, calming the mind and improving intelligence, and tonifying the liver and kidneys. Summary of the Invention

[0010] The purpose of this invention is to provide a traditional Chinese medicine composition (medicinal and edible traditional Chinese medicine) that can effectively improve learning ability decline, attention deficit, cognitive decline and emotional regulation imbalance, and improve chronic sleep deprivation (CSD) damage, as well as its application.

[0011] To achieve the above objectives, the technical solution adopted by the present invention is as follows: A traditional Chinese medicine composition for improving chronic sleep deprivation injury, the traditional Chinese medicine composition (based on daily dosage) is: 4.5~13.5g of processed Polygonatum sibiricum, 4.5~13.5g of mulberry, 4.5~13.5g of wolfberry, 3~9g of longan pulp, 3~9g of chrysanthemum, and 1.5~4.5g of dried tangerine peel.

[0012] Preferably, the traditional Chinese medicine composition (based on daily dosage) consists of the following traditional Chinese medicine decoction pieces, based on daily dosage: 4.5-9g of prepared Polygonatum sibiricum, 4.5-9g of mulberry, 4.5-9g of wolfberry, 4.5-9g of jujube seed, 3-6g of longan pulp, 3-6g of chrysanthemum, and 1.5-3g of dried tangerine peel.

[0013] The application of the aforementioned traditional Chinese medicine composition in the preparation of pharmaceutical or food formulations for improving learning impairment, attention deficit, cognitive decline, mood dysregulation, and nerve damage caused by chronic sleep deprivation (CSD).

[0014] The application of the traditional Chinese medicine composition in the preparation of drug or food preparations for cognitive decline and memory loss caused by long-term passive sleep deprivation due to insufficient sleep, which in turn leads to neuronal damage.

[0015] An formulation for improving cognitive decline and neurological damage caused by chronic sleep deprivation (CSD), wherein the composition is the active ingredient.

[0016] When the preparation is a pharmaceutical preparation, the dosage form is one or more of the following: granules, capsules, tablets, pills, oral liquid preparations, and powders.

[0017] When the preparation is a food preparation, it is in the form of one or more of the following: biscuits, tea, and powders.

[0018] The above composition is based on traditional Chinese medicine theory and is designed for syndromes of deficiency of both heart and spleen, deficiency of liver and kidney yin, and deficiency of essence and blood. The formula should focus on tonifying the spleen and replenishing qi, nourishing the liver and kidneys, and nourishing blood and improving eyesight. Therefore, this invention divides the components into two groups: 1. Spleen-tonifying, qi-replenishing, and blood-nourishing group (see Table 1); 2. Liver-kidney-nourishing and vision-improving group (see Table 2). The "properties, flavors, meridian tropisms," and "functions" of each medicinal material in Tables 1 and 2 are derived from the 2020 edition of the Chinese Pharmacopoeia.

[0019] Table 1. Spleen-tonifying, Qi-boosting, and Blood-nourishing Group

[0020] Table 2. Liver and Kidney Nourishing and Vision Improving Group

[0021] Meanwhile, the compatibility mechanism and synergistic effect of the composition of the present invention are as follows: This formula contains prepared Polygonatum sibiricum to strengthen the spleen and kidneys, Lycium barbarum to nourish the liver and kidneys, and Morus alba to nourish yin and replenish blood. These three ingredients work together to replenish essence. Longan pulp nourishes the heart and spleen, while chrysanthemum calms the liver, improves eyesight, and clears the head and eyes, making it especially suitable for those suffering from spleen deficiency due to prolonged sitting or blood deficiency due to prolonged visual strain. Tangerine peel regulates qi and harmonizes the middle jiao, aiding in spleen and stomach function. It is a balanced formula that is not stagnant or excessively nourishing, with mild medicinal properties and no harm even with long-term use. It is particularly suitable for long-term use by adolescents to improve symptoms such as memory loss, poor concentration, mood swings, and eye strain caused by sleep deprivation, and has a positive effect on improving learning efficiency and overall physical and mental health.

[0022] The similarities and differences between this formula and traditional Chinese medicine classic prescriptions are shown in Table 3: Table 3. Comparison of the formulations of this invention with classic formulations

[0023] Note: Chinese Pharmacopoeia 2020 Edition.

[0024] The daily dosage of the above-mentioned composition of the present invention is shown in Table 4.

[0025] Table 4. Daily dosage and proportion of each component in the composition

[0026] The preparation of the above-mentioned formulation containing the composition can be as follows: Method 1: Take the above-mentioned drugs and mix them in the preferred proportions. Add water and decoct twice, one hour each time. Combine the decoctions, filter and concentrate to an appropriate amount, which can be made into a decoction or further dried into powder or granules. The resulting preparation can be formulated into pharmaceutical preparations (such as granules or capsules, tablets, pills, oral liquid preparations, powders and other dosage forms) or food preparations (such as biscuits, tea, granules, etc.) as needed.

[0027] Method 2: After weighing the above-mentioned drugs according to the preferred proportions, they are pulverized, sieved, and mixed evenly, and then directly filled into pharmaceutical capsules or compressed into tablets; or appropriate excipients can be added to prepare pharmaceutical preparations (such as granules or capsules, tablets, pills, oral liquid preparations, powders and other dosage forms), or food preparations (such as biscuits, tea drinks, granules, etc.).

[0028] Method 3: According to the preferred ratio, each group of medicinal slices is extracted separately, and the effective components are obtained by water extraction. The extracts are then filtered, concentrated, and dried to make powder. The qualified medicinal powders are then mixed evenly according to the original proportions of the medicinal materials. Appropriate excipients are added according to the preparation requirements to prepare pharmaceutical preparations (such as granules or capsules, tablets, pills, oral liquid preparations, powders, and other dosage forms), or food preparations (such as biscuits, teas, granules, etc.).

[0029] Compared with the prior art, the present invention has the following advantages: This invention is the first to propose a treatment strategy and formulation approach for treating syndromes of deficiency of both heart and spleen, deficiency of liver and kidney yin, and deficiency of essence and blood, by tonifying the spleen and replenishing qi, nourishing the liver and kidneys, and nourishing blood and improving eyesight. All the medicinal materials used are of the "medicine and food homology" type, ensuring high safety. They can be added to food for long-term consumption without significant toxic side effects, making them particularly suitable for improving the learning ability decline caused by chronic sleep deprivation in adolescents. Attached Figure Description

[0030] Figure 1 The water maze test and the content of inflammatory factors in the serum of mice in each group provided in the embodiments of the present invention (n=6, Based on the prepared Chinese medicinal materials of Example 1, the following groups were formed: 1. Complete formula group (i.e., the complete composition formulated according to the original formula ratio); 2. Formula group without processed Polygonatum; 3. Formula group without Mulberry; 4. Formula group without Goji berries; 5. Formula group without Longan pulp; 6. Formula group without Chrysanthemum; 7. Formula group without Tangerine peel; 8. Formula group with Ziziphus jujuba seed. The proportion of Ziziphus jujuba seed added was 3 parts, consistent with the proportion of the original processed Polygonatum. Group 9 was the model group, and group 10 was the blank control group. Detailed Implementation

[0031] The following examples further illustrate specific embodiments of the present invention. It should be noted that the specific embodiments described herein are merely for illustration and explanation and are not intended to limit the scope of the present invention.

[0032] Example 1. Effects of the composition on spatial learning and memory in chronically sleep-deprived mice Experimental animals: Clean-grade, healthy 4-week-old female C57BL / 6J mice, with free access to food, housed in a 12-hour light / dark cycle environment, with the room temperature maintained at 22±2℃. Mice were acclimatized for one week before the experiment, and then randomly divided into a control group, a model group, a low-dose composition group, and a high-dose composition group, with 6 mice in each group.

[0033] Preparation of the composition: Take each Chinese herbal medicine slices, including 3 parts of Polygonatum sibiricum, 3 parts of mulberry, 3 parts of wolfberry, 2 parts of longan pulp, 2 parts of chrysanthemum, and 1 part of dried tangerine peel. Add water and decoct twice, 1 hour each time. Combine the decoctions, filter, and concentrate to 0.5g of solids per mL for later use (approximately 2.5g of original medicinal materials).

[0034] During the experiment, except for the control group which was fed normally, all other groups were subjected to 10 hours of sleep deprivation per day using a sleep deprivation device (XR-XS 107, Shanghai Xinsoft Co., Ltd.). Sleep deprivation was achieved through tactile stimulation using a parallel movement of a metal rod at the bottom of the device, employing a "gentle touch" motion. The metal rod moved for 60 seconds followed by 15 seconds of stillness, repeating this cycle. The deprivation period was from 8:00 AM to 6:00 PM. Sleep deprivation continued for 8 weeks. After the 4th week, the low-dose group and the high-dose group were administered the composition of this invention by gavage at 1.5 g / kg and 3 g / kg daily, respectively. The control group and the model group were administered an equal volume of physiological saline by gavage daily. After 4 weeks of continuous administration, a water maze test was performed to assess the spatial learning and memory abilities of the mice.

[0035] Morris Water Maze (MWM) Test: A circular pool is filled with opaque water mixed with white dye, making the platform beneath the surface invisible. The pool is divided into four quadrants, with the hidden platform located in one quadrant. High-contrast visual markers are placed on the walls as spatial cues to help the mice locate the platform. A camera tracking system is positioned above the pool to record the mouse's path, swimming speed, and time from entry into the water to finding the platform.

[0036] The first phase was location training. Mice were placed in the water from different entry points and were required to find a hidden platform within 60 seconds. If they failed to find it within the time limit, they were guided to the platform and kept there for 30 seconds. This training was conducted for 5 consecutive days, 4 times a day, with 30-minute intervals between each session. The time it took for the mice to find the platform from entering the water was recorded (escape latency).

[0037] The second stage was a spatial exploration test. The hidden platform was removed from the water, and the mice were placed in the pool from the original opposite quadrant. The number of times the mice crossed the original platform position within 60 seconds and the percentage of time spent in the target quadrant were recorded.

[0038] The escape latency of mice in each group during the first stage is shown in Table 5. With increasing training days, the escape latency of all groups showed a shortening trend, indicating a gradual improvement in their spatial learning ability. By day 2, the low-dose group showed a significant difference from the model group (P=0.037), and the high-dose group showed an even greater difference in latency compared to the model group (P=0.011). This trend continued until day 5.

[0039] Table 5. Changes in escape latency during navigation training in different groups of mice (unit: seconds) (n=6, )

[0040] The results of the second phase of spatial exploration testing are shown in Table 6. There was no significant difference in swimming speed among the groups (P>0.05), indicating that the motor abilities of the groups were basically the same. The number of times the low-dose group and the high-dose group crossed the original platform position was significantly more than that of the model group, at 2.54±0.8 times and 3.01±0.25 times, respectively. The proportion of time spent in the target quadrant was 36.56±4.2% and 40.37±4.5%, respectively, which were significantly higher than those of the model group (2.12±0.31 times and 31.21±3.2%).

[0041] Table 6. Number of times mice crossed the original platform location and percentage of time spent in the target quadrant during the spatial exploration test (n=6, )

[0042] The above results indicate that spatial learning and memory abilities were significantly improved in both the low-dose and high-dose groups after drug intervention, showing a dose-dependent trend. This suggests that the composition has a significant protective effect against cognitive impairment caused by chronic sleep deprivation.

[0043] Example 2. Improvement of anxiety and irritability in chronically sleep-deprived mice by the composition. Experimental animals: Referring to Example 1, the experiment was divided into 5 groups (10 animals in each group), namely blank control group, model group, positive drug control group 1, positive drug control group 2 and composition group.

[0044] Preparation of the composition: Refer to Example 1.

[0045] Sleep deprivation modeling: Refer to Example 1.

[0046] Testing of anxiety-like behavior in mice: Dosage regimen: Sleep deprivation was continued for 8 weeks. After week 4, the composition group, positive control group 1 and 2 were administered the composition of the present invention 3g / kg (based on raw materials), fluoxetine 20mg / kg and liver-soothing and depression-relieving capsules 0.25g / kg (National Drug Approval Number Z20080580) by gavage daily. The control group (without sleep deprivation) and the model group were administered the same volume of physiological saline by gavage daily for 4 weeks until the chronic sleep deprivation ended.

[0047] After 4 weeks of continuous administration, the elevated plus-maze (EPM) test was performed.

[0048] Test method: Mice were placed in the center of an elevated cross maze (EPM) with their heads facing the open arms. The time spent in the open and closed arms (OT and CT) and the number of entries (OE and CE) were recorded over 5 minutes. The completion of the test was marked by the mouse's foot leaving a certain arm. The percentage of total entries into the open arm (OE) to total entries into the arm (CE) (OE%), and the percentage of time spent in the open arm (OT) to total time spent in the arm (CT) (OT%) were used as indicators of the anti-anxiety effect.

[0049] The improvement of anxiety-like behavior in mice after chronic sleep deprivation by the composition is shown in Table 7.

[0050] Table 7. Effects of the composition of the present invention on the behavior of mice in the elevated cruciate maze (n=10, )

[0051] As shown in Table 7, the composition of this invention significantly increased the dwell time and number of entries in the open arm of sleep-deprivation model mice, with both OT% and OE% being significantly higher than those in the model group (P<0.001), indicating that the composition has a significant effect on improving anxiety behavior induced by chronic sleep deprivation. Simultaneously, the OT% and OE% of the composition group were also significantly better than those of the positive control group 2 (P=0.019 & P=0.015), suggesting that its anti-anxiety effect is stronger than that of Shugan Jieyu capsules.

[0052] Testing of aggressive behavior in mice: The Resident-intruder test (RIT) was used to assess the aggressive behavior of mice in each group, in order to evaluate the aggressive emotional response of mice after chronic sleep deprivation. During the test, unfamiliar foreign mice (intruders) were placed in the cages of the experimental mice (residents), and the latency, number of attacks, and total attack time of aggressive behavior were recorded within 10 minutes. The results are shown in Table 8.

[0053] Table 8. Effects of the compositions of the present invention on aggressive behavior in mice (n=10, )

[0054] [1] Comparison of P-values ​​between the composition group and the modeling group.

[0055] [2] Compare the P value between the composition group and the positive control group 2.

[0056] As shown in Table 8, the mice in the composition group showed significantly better performance in all aggressive behavior indicators (except for the time of the first bite) than the model group (P<0.001), and were also better than the positive control group 2 in terms of the number of attacks, the number of bites, and the number of upright attacks (P=0.006, P=0.007, P<0.001), suggesting that the composition of the present invention has a significant intervention effect in alleviating aggressive emotional responses caused by chronic sleep deprivation.

[0057] As can be seen from the results shown in Table 8, in addition to significantly improving cognitive function impairment caused by sleep deprivation, the composition of the present invention can also effectively regulate emotional disorders caused by sleep deprivation, especially in the inhibition of anxiety and aggressive behavior.

[0058] Example 3. Effects of the composition on oxidative stress and neuroinflammation in chronically sleep-deprived mice In Example 1, after completing the water maze test, mice in each group were completely anesthetized, their eyeballs were removed and blood was collected for later use. After blood collection, the chest was opened and the right atrial appendage was cut open. Pre-cooled 0.9% saline was injected into the aortic arch and rapidly perfused until the liver and limbs turned pale. The brain was completely removed and the hippocampus was dissected on ice and frozen at -80°C. Then the liver was removed and frozen at -80°C.

[0059] Detection of serum TNF-α, IL-6, and IL-1β levels The concentrations of inflammatory factors in serum were calculated and the levels of inflammatory factors were determined using the ELISA method (TNF-α / IL-6 / IL-1β ELISA kit, Xinbosheng Biotechnology Co., Ltd.) based on the instructions and CurveExpert 1.4 software.

[0060] Measurement of SOD and GSH activity in hippocampus and liver tissue The SOD and GSH kits were developed using products from Nanjing Jiancheng Bioengineering Institute. Hippocampal and liver tissues were homogenized and diluted to appropriate concentrations. The kits were then operated according to the instructions. The absorbance values ​​were measured using a spectrophotometer, and the SOD activity and GSH content were calculated.

[0061] The relative expression levels of TNF-α, P53, BAX, and BCL-2 proteins in hippocampus and liver tissues were determined using Western blot. Total protein was extracted from hippocampus and liver tissues, quantified using the BCA method, and loaded onto a plate in equal volumes. The samples were separated by SDS-PAGE electrophoresis and transferred to a membrane for blocking. Primary antibodies against rabbit anti-TNF-α / P53 (Boster Biologics Inc.), rabbit anti-BAX / BCL-2 (Wanlei Biotechnology Co., Ltd.), and mouse anti-GAPDH / β-Actin (Beijing Zhongshan Jinqiao Biotechnology Co., Ltd.) were added and incubated overnight at 4°C. The next day, after washing with TBST, the corresponding HRP-labeled secondary antibodies were added and incubated. ECL staining was performed, and the gray values ​​of the protein bands were analyzed using Image Lab 5.2 software to calculate the relative protein expression levels.

[0062] The levels of inflammatory factors in the serum of mice in each group are shown in Table 9. The levels of TNF-α, IL-6, and IL-1β in the model group were significantly higher than those in the control group (P<0.01), indicating that chronic sleep deprivation can induce a significant systemic inflammatory response. After drug intervention, the levels of the above-mentioned inflammatory factors decreased in both the low-dose and high-dose groups. The low-dose group did not show a statistically significant difference (P>0.05), while the high-dose group showed a significant decrease (P<0.05), suggesting that the high-dose composition can effectively inhibit the inflammatory response caused by chronic sleep deprivation.

[0063] Table 9. Serum inflammatory factor levels (pg / ml) in each group of mice (n=6, )

[0064] Table 10 shows the SOD and GSH activities in the hippocampus and liver tissues of mice in each group. The SOD activity and GSH content in the model group's hippocampus and liver tissues were significantly lower than those in the control group (P<0.01), suggesting that chronic sleep deprivation can lead to a decrease in antioxidant capacity. After drug intervention, both the low-dose and high-dose groups showed a significant increase in SOD activity and GSH content in the hippocampus compared to the model group (P<0.05), exhibiting a dose-dependent increasing trend. However, no significant improvement was observed in the liver tissue in either the low-dose or high-dose groups (P>0.05), suggesting that the composition has a certain tissue selectivity in regulating oxidative stress in the hippocampus.

[0065] Table 10. SOD activity (U / mg prot) and GSH content (μmol / g prot) in hippocampus and liver tissues of mice in each group (n=6, )

[0066] Since sleep deprivation induces systemic oxidative stress (Table 10), the expression levels of apoptosis activator P53, pro-apoptotic protein BAX, and anti-apoptotic protein BCL-2 in hippocampal and liver tissues were further examined. The results are shown in Table 11. In the model group, the expression of P53 and BAX proteins was significantly upregulated in both hippocampal and liver tissues, while the expression of BCL-2 was significantly downregulated (P<0.01), indicating activation of the apoptosis pathway. After drug intervention, the high-dose group showed significant regulatory effects on both the hippocampus and liver, manifested as a significant decrease in the expression levels of inflammatory factors, apoptosis activator factors, and pro-apoptotic proteins, and a significant increase in the expression levels of anti-apoptotic proteins (P<0.05).

[0067] In the low-dose group, besides a significant upregulation of BCL-2 in liver tissue (P=0.012), there was no significant effect on the expression of inflammatory factors TNF-α, P53, and BAX (P>0.05). However, it showed a significant improvement in hippocampal tissue (P<0.05). This further suggests that the composition's inhibitory effect on sleep deprivation-induced hippocampal cell apoptosis is tissue-specific, and its protective effect is more likely to target neural tissue rather than act broadly on peripheral organs.

[0068] Table 11. Relative expression levels of TNF-α, p53, BAX, and BCL-2 proteins in the hippocampus and liver tissues of mice in each group (n=6, )

[0069] The hippocampus is a key structure for learning and memory, and the number of hippocampal neurons is closely related to cognitive functions such as learning and memory. Studies have confirmed that sleep deprivation impairs cognitive function by increasing hippocampal oxidative stress, altering synaptic structure and function, and reducing neuronal plasticity, leading to a decline in spatial learning and memory abilities [Science, 2017, 355(6324):511-515.]. The results of this embodiment, combined with behavioral improvement, further corroborate that its protective effect against cognitive impairment caused by sleep deprivation stems from the composition's protective effect on hippocampal neurons. In particular, it exhibits significant anti-inflammatory and anti-apoptotic activity in the hippocampus, providing strong support for the subsequent expansion of the composition's functions.

[0070] Example 3. Analysis of the contribution of each component of the composition of the present invention to the overall efficacy. The composition was prepared based on the prepared Chinese herbal medicine pieces of Example 1, divided into 10 groups: 1. Complete formula group (i.e., the complete composition formulated according to the original formula ratio); 2. Formula group without processed Polygonatum; 3. Formula group without Mulberry; 4. Formula group without Lycium; 5. Formula group without Longan pulp; 6. Formula group without Chrysanthemum; 7. Formula group without Tangerine Peel; 8. Formula group with Ziziphus jujuba seed added. The proportion of Ziziphus jujuba seed added was 3 parts, consistent with the proportion of the original processed Polygonatum. Group 9 was the model group, and group 10 was the blank control group. Each group with added or removed ingredients prepared corresponding samples according to the same process. The spatial learning and memory abilities of mice in each group were evaluated using the same behavioral experimental protocol as in Example 1, and the levels of various inflammatory factors in the mouse serum were measured. The results are as follows: Figure 1 As shown.

[0071] Depend on Figure 1 As is known, in the water maze test, the target quadrant dwell time and number of crossings in the deflavored group and the model group were generally lower than those in the whole formula group, especially in the groups without processed Polygonatum and wolfberry. Interestingly, the addition of Ziziphus jujuba seed did not significantly improve spatial learning and memory ability; instead, it reduced the target quadrant dwell time and number of crossings compared to the whole formula group, suggesting that Ziziphus jujuba seed may not have played a synergistic role in this combination. Further detection of serum inflammatory factors revealed that the levels of IL-6, TNF-α, and IL-1β in the deflavored group were generally higher than those in the whole formula group, which is consistent with the behavioral trend. However, the serum levels of the above inflammatory factors in the flavored group were not significantly lower than those in the whole formula group, and even IL-6 and IL-1β were slightly increased, indicating that the addition of Ziziphus jujuba seed did not enhance the anti-inflammatory efficacy of the composition, but may have interfered with the balance of the original formula. Combining behavioral and molecular indicators, the whole formula group showed the best performance in spatial learning and memory ability and anti-inflammatory effect, suggesting a synergistic effect among the components.

[0072] Example 4. Preparation of the composition of the present invention According to the formulation of the composition in Example 1, take the medicinal materials of each slice, wash them, add water with 8 times the total mass of each medicinal material and decoct twice, 1 hour each time. Filter the dregs, combine the filtrates, and concentrate under reduced pressure to 0.3g of solids per 1mL for later use.

[0073] The concentrate is mixed with appropriate excipients, including maltodextrin, β-cyclodextrin and lactose (at a dry weight ratio of 1:3 to the solids of the extract), and granulated with 80% ethanol as a wetting agent. The granules are dried at 65°C for 1 hour and then granulated to obtain the final product.

[0074] Alternatively, the concentrate can be spray-dried into dry extract powder. Add 15 kg of maltodextrin, 1 kg of micronized silica gel, and 5 kg of β-cyclodextrin to 100 kg of concentrate (30% solid content); with an inlet air temperature of 160℃, an outlet air temperature of 80℃, and an atomization pressure of 2.0 MPa. After obtaining the dry extract powder, package it according to conventional processes to obtain the finished product.

Claims

1. A traditional Chinese medicine composition for improving chronic sleep deprivation injury, characterized in that: The traditional Chinese medicine composition (based on daily dosage) is as follows: 4.5-13.5g of processed Polygonatum sibiricum, 4.5-13.5g of mulberry, 4.5-13.5g of wolfberry, 3-9g of longan pulp, 3-9g of chrysanthemum, and 1.5-4.5g of dried tangerine peel.

2. The traditional Chinese medicine composition for improving chronic sleep deprivation injury according to claim 1, characterized in that: The traditional Chinese medicine composition (based on daily dosage) consists of the following traditional Chinese medicine decoction pieces, based on daily dosage: 4.5-9g of prepared Polygonatum sibiricum, 4.5-9g of mulberry, 4.5-9g of wolfberry, 4.5-9g of jujube seed, 3-6g of longan pulp, 3-6g of chrysanthemum, and 1.5-3g of dried tangerine peel.

3. The application of the traditional Chinese medicine composition according to claim 1, characterized in that: The application of the traditional Chinese medicine composition in the preparation of pharmaceutical or food formulations that improve learning impairment, attention deficit, cognitive decline, mood dysregulation, and nerve damage caused by chronic sleep deprivation (CSD).

4. The application of the traditional Chinese medicine composition according to claim 3, characterized in that: The application of the traditional Chinese medicine composition in the preparation of drug or food preparations for cognitive decline and memory loss caused by long-term passive sleep deprivation due to insufficient sleep, which in turn leads to neuronal damage.

5. An agent for improving cognitive decline and neurological damage caused by chronic sleep deprivation (CSD), characterized in that: The composition according to claim 1 is the active ingredient.

6. The formulation according to claim 5, characterized in that: When the preparation is a pharmaceutical preparation, the dosage form is one or more of the following: granules, capsules, tablets, pills, oral liquid preparations, and powders.

7. The formulation according to claim 5, characterized in that: When the preparation is a food preparation, it is in the form of one or more of the following: biscuits, tea, and powders.

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