Application of gardenia-cinnamon composition in preparation of medicine for improving learning and memory impairment

Abstract

The invention belongs to the technical field of medicines, and particularly discloses application of a fructus gardeniae-cortex cinnamomi composition in preparation of a medicine for improving learning and memory impairment. A rat insomnia model is induced by adopting p-chlorophenylalanine, interference is given to diazepam and a fructus gardeniae-cinnamon medicine pair, an open field experiment and a Morris water maze experiment are carried out, and the medicine effect of the fructus gardeniae-cinnamon medicine pair on improving insomnia is investigated; experimental results show that the cape jasmine-cinnamon composition can improve the learning and memory impairment of an insomnia rat model caused by p-chlorophenylalanine, has an obvious effect of improving the learning and memory impairment, and can be used for preparing the medicine for improving the learning and memory impairment. The invention provides a new drug source for improving learning and memory ability impairment.

Description

Technical Field

[0001] The invention belongs to the technical field of medicine, and particularly relates to an application of a gardenia-cinnamon bark composition in preparing a medicine for improving learning and memory impairment. Background Art

[0002] Learning and memory are fundamental to cognition in higher animals and humans, and are essential for the survival of lower animals. They are closely related to human production and daily life. Learning is the acquisition of new experiences through the nervous system in response to changes in the new environment; memory is the reflection of past experiences, including the recognition of things learned, experienced, and done in the past. Learning and memory are two interconnected and inseparable processes, representing the adaptation of brain circuits to environmental changes. Learning and memory are neural activities formed by the interaction between neuronal networks and the interconnection between synapses, and are among the higher-level brain activities. Brain functional impairment and central nervous system damage are the main causes of impaired learning and memory. For example, insomnia can impair the formation and consolidation of learning and memory. Studies have shown that insomnia affects hippocampal structure and neurotransmitter levels, thereby affecting hippocampal neuroplasticity and causing learning and memory impairment. Insomnia can also indirectly impair learning and memory by inducing oxidative stress, inhibiting the production of antioxidant enzymes in the hippocampus, causing excessive increases in reactive oxygen species, lipid peroxidation, and DNA modification, leading to neuronal damage.

[0003] Among patients with learning and memory impairment, Western medicine, such as donepezil, galantamine, huperzine A, memantine hydrochloride, etc., is the most widely used in terms of number of people taking it. Although these Western medicines are very popular in China, most of them are expensive, have low therapeutic efficiency, and have many toxic and side effects. Long-term use will cause certain harm to the human body. Traditional Chinese medicine emphasizes syndrome differentiation and treatment, and overall regulation of the internal and external environmental balance of the body. It has certain advantages in improving cognitive dysfunction and has become a research hotspot in recent years. In terms of traditional Chinese medicine for improving learning and memory impairment, relevant literature such as the Chinese patent with application number 202510209692.9 was retrieved, which discloses a pharmaceutical composition for preventing and treating learning and memory impairment and its application. The pharmaceutical composition uses Gastrodia elata, Polygonatum sibiricum and Poria cocos in a specific ratio as raw materials, which can effectively prevent and treat learning and memory impairment by improving anti-inflammatory and anti-oxidative stress levels and regulating the function of the cholinergic system. However, the main defects of existing formulas include: first, there are many types of medicinal flavors, the components are relatively complex, and the cost is high; second, the Chinese medicinal raw materials often need to be pretreated (for example, soaked in rice wine), and the processing technology is cumbersome.

[0004] Gardenia jasminoides is a traditional Chinese medicinal material commonly used in my country. It is also known as yellow gardenia, mountain gardenia, etc. It is a plant of the Rubiaceae family. Gardenia jasminoidesThe dried mature fruit of Ellis was first recorded in the "Shennong Bencao Jing" of the Eastern Han Dynasty, and was also recorded in "Shanghan Lun", "Mindong Bencao", "Puji Fang" and "Shengji Zonglu". It is listed as a medium-grade wood product. It is bitter and cold, and enters the heart, lung and triple burner meridians. It has the functions of purging fire and relieving restlessness, clearing heat and dampness, cooling blood and detoxifying; external use can reduce swelling and relieve pain. Modern pharmacological studies have shown that the main active ingredients in Gardenia, iridoid glycosides, have anti-inflammatory, antioxidant, lipid-lowering, and improve insomnia, anxiety, depression and other pharmacological effects. Cinnamon is a plant of the Lauraceae family, the genus Cinnamomum. Cinnamomum cassia The dried bark of Pres. The 2020 edition of the Chinese Pharmacopoeia states that cinnamon is pungent, sweet, and highly caloric; it enters the kidney, spleen, heart, and liver meridians; it has the effects of tonifying fire and supporting yang, guiding fire back to its source, dispersing cold and relieving pain, and warming and unblocking the meridians. It is used to treat impotence, cold uterus, cold pain in the waist and knees, asthma due to kidney deficiency, rising yang deficiency, dizziness and red eyes, cold pain in the heart and abdomen, vomiting and diarrhea due to deficiency-cold, abdominal pain due to cold hernia, dysmenorrhea, and amenorrhea. Modern pharmacological research has shown that cinnamon has antibacterial, antioxidant, anti-inflammatory, anticancer, and hypoglycemic activities. However, there are currently no reports on the combination of gardenia and cinnamon to improve learning and memory impairment. Summary of the Invention

[0005] The present invention aims to use a gardenia-cinnamon composition in the preparation of a medicament for improving learning and memory impairment. The raw materials of the composition are simple and easily available. The compound combination of gardenia and cinnamon can effectively improve learning and memory impairment, providing a new source of medicine for improving learning and memory impairment.

[0006] The technical solution adopted in the present invention is as follows: Application of a gardenia-cinnamon bark composition in preparing a medicine for improving learning and memory impairment.

[0007] Furthermore, in the above application, the gardenia-cinnamon bark composition is used to prepare a medicine for improving learning and memory impairment caused by insomnia.

[0008] Preferably, in the gardenia-cinnamon composition, the weight ratio of gardenia:cinnamon is 6:1.

[0009] Furthermore, in the above application, the gardenia-cinnamon composition is used alone or in combination with other drugs.

[0010] Furthermore, the aforementioned gardenia-cinnamon combination is used in the preparation of a drug for improving learning and memory impairment. The drug comprises active ingredients including gardenia and cinnamon, and is prepared into a pharmaceutically acceptable dosage form using pharmaceutically acceptable carriers or excipients and processes. The dosage forms include powders, granules, oral liquid preparations, and injections.

[0011] The present invention adopts p-chlorophenylalanine ( DLA rat model of insomnia was induced by PCPA (4-chlorophenylalanine). After intervention with diazepam and the Gardenia-Cinnamon drug pair, the open field test and Morris water maze test were performed to investigate the efficacy of the Gardenia-Cinnamon drug pair in improving insomnia. The experimental results showed that compared with the model group, the total distance traveled in the open field test in the Gardenia-Cinnamon group was significantly reduced ( P <0.05), the immobility time was significantly prolonged ( P The results showed that Gardenia-Cinnamon Bark could improve learning and memory impairment in a rat model of insomnia induced by PCPA.

[0012] The beneficial effects of the present invention are: The present invention discloses the use of a gardenia-cinnamon combination in the preparation of a drug for improving learning and memory impairment, providing a new source of drugs for improving learning and memory impairment. In addition, the raw materials of the gardenia-cinnamon combination provided by the present invention are simple, easily available, and low in cost. The present invention has shown through research that the gardenia-cinnamon combination can improve learning and memory impairment in a rat model of insomnia induced by p-chlorophenylalanine, has a significant effect of improving learning and memory impairment, and can be used to prepare a drug for improving learning and memory impairment. The present invention explores the effect of gardenia-cinnamon on improving p-chlorophenylalanine-induced insomnia in rats. DL The study investigated the pharmacological effects of Gardenia jasminoides and Cinnamon bark on learning and memory in a rat model of insomnia induced by PCPA, providing a scientific basis for the pharmacological effects of Gardenia jasminoides and Cinnamon bark on the treatment of learning and memory impairment. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 Effects of Gardenia jasminoides and Cinnamon bark on the open field test in insomnia rats ; Figure 1 Middle, positive: diazepam group, granules: gardenia-cinnamon group; Figure 2 Effects of Gardenia jasminoides and Cinnamon bark on the localization and navigation abilities of insomnia rats ; Figure 2 Middle, positive: diazepam group, granules: gardenia-cinnamon group; Figure 3 Morris water maze movement trajectory diagram of rats in each group; Figure 3 Middle, positive: diazepam group, granules: gardenia-cinnamon group; Figure 4 Effects of Gardenia-Cinnamon Pair on Spatial Exploration Ability in Insomnia Rats ; Figure 4 In the middle, positive: diazepam group, granule: gardenia-cinnamon group. DETAILED DESCRIPTION Example 1

[0014] Study on the improvement of learning and memory impairment in the p-chlorophenylalanine-induced insomnia rat model by Gardenia jasminoides and cinnamon bark 1 Experimental Materials 1.1 Experimental Animals Thirty-two SPF-grade Sprague-Dawley rats of both sexes, weighing 180–200 g, were purchased from Hunan Slake Jingda Laboratory Animal Co., Ltd. (license number SCXK (Xiang) 2019-0004). The animals were housed in an environment with a humidity of 50 ± 10%, a temperature of 25 ± 2°C, and a 12-h day / night cycle. They had free access to food and water, with feed added at regular intervals and water changes daily. Bedding consisted of dry, clean corncob pellets, which were changed every three days. Animals were acclimated for seven days before the experiment began.

[0015] 1.2 Experimental drugs and reagents

[0016] 1.3 Experimental equipment

[0017] 2 Experimental methods 2.1 Preparation of Gardenia-Cinnamon Drug Solution The Gardenia jasminoides and Cinnamon bark Chinese medicine granules were mixed in a crude drug ratio of 6:1 and dissolved in pure water to obtain a Gardenia jasminoides-Cinnamon bark drug pair solution with a crude drug concentration of 0.735 g / mL, which was stored in a -4°C refrigerator.

[0018] 2.2 Establishment of PCPA insomnia model 2.2.1 Preparation of PCPA suspension Prepare 0.5% sodium carboxymethylcellulose solution with normal saline and add it to PCPA powder in small amounts and multiple times while stirring or vortexing continuously to prepare a suspension with a mass concentration of 40 mg / mL.

[0019] 2.2.2 Preparation of PCPA-induced insomnia rat model After one week of animal adaptation, 48 SD rats were intraperitoneally injected with PCPA suspension (400 mg / kg) between 8:00 and 9:30 daily for four consecutive days. Another 16 rats served as a blank group.

[0020] 2.2.3 Determination of insomnia model General behavioral observation: After intraperitoneal injection of PCPA suspension on the first day, the behavior of the two groups of rats was observed. Compared with the blank group, the rats in the modeling group were irritable, aggressive, sensitive to external stimuli, had dull hair, and soft and gray feces, indicating that the modeling was successful.

[0021] Body weight test: Before and after modeling, rats in each group were weighed at the same time in the morning and their body weights were recorded. Compared with the blank group, the body weights of the modeling groups were reduced, indicating that the modeling was successful.

[0022] 2.3 Grouping and Dosing After modeling, rats were randomly divided into three groups based on body weight: a model group, a diazepam (2 mg / kg) group, and a Gardenia-Cinnamon (2.5 g / kg, equivalent to 10 times the adult clinical dose based on crude drug dosage). Each group consisted of 16 rats, 8 males and 8 females. Drugs were administered by gavage at the same time every morning. Rats were weighed before gavage. Each treatment group received the corresponding drug (10 mL / kg) intragastrically. The blank and model groups received an equal volume of normal saline intragastrically once daily for 7 consecutive days before specimen collection.

[0023] 2.4 Animal behavior evaluation 2.4.1 Open field test On the third day of dosing, an open field test was conducted. Thirty minutes after dosing, rats were placed in the center of the mine box to acclimate for 1 minute. The rats were then removed and returned to the center of the mine box. The total distance traveled and the time spent stationary within 5 minutes were recorded for each group. Once the experiment began, all experimenters left the laboratory and the doors and windows were closed to prevent their movements from affecting the rats' condition and thus the evaluation data. After the previous rat was observed, feces was cleaned with a paper towel. The interior and bottom of the mine box were then thoroughly sprayed with 75% alcohol and wiped clean with a towel to prevent information left by the previous rat from influencing the observation results of the next rat. This procedure was repeated between experiments.

[0024] 2.4.2 Morris water maze test On the fourth day of drug administration, the Morris water maze test was performed. The experiment was conducted in a circular pool with a black inner wall. The pool was surrounded by curtains to prevent the movement of the experimenter from startling the rats and affecting their judgment. The pool was divided into four equal quadrants in a rectangular coordinate system. Four pieces of hard cardboard of different colors and shapes were affixed to the inner wall of each quadrant. Quadrant IV was designated as the target quadrant. The circular target platform for the rats to climb onto and rest was located in quadrant IV, 1 cm below the water surface. The water temperature was maintained at a constant 22±2°C, and the light was constant, with no influence from daylight fluctuations. The experimental process was recorded using a video tracking and analysis system.

[0025] (1) Positioning and navigation experiment: The farthest position from the target platform (quadrant II) was selected as a fixed location. 30 minutes after drug administration, rats were placed in the water in quadrant II facing the inner wall. All rats were trained twice a day for two consecutive days. The time taken by each group of rats from entering the water to climbing onto the target platform and staying there for 15 seconds during the fourth training session was recorded, which was the rat's escape latency. During the training period, if a rat did not find the target platform within 90 seconds, it was guided and stayed on the platform for 15 seconds.

[0026] (2) Spatial exploration experiment: The platform was removed and rats from each group were placed in the water at the same position facing the inner wall of the pool. The movement trajectory, average swimming speed, and the number of times, time, and distance of the rats crossing the target quadrant within 60 seconds were recorded.

[0027] 2.5 Data Processing Data The data were processed using Graphpad Prism 8.0.2 software.

[0028] 3 Results 3.1 General behavioral observations of rats After modeling, the rats in each group showed significant differences in their condition. The blank group had shiny, smooth fur, normal weight gain, black, hard feces, and dry cages. Their mental state was normal, with a clear circadian rhythm. The model group had matted, matted fur, lost weight, soft, grayish-white feces, increased urine and feces, and unusually damp cages. The rats also showed lethargy, low energy, irritability, aggressive behavior, and sensitivity to external stimuli. Following treatment, the rats in each group gradually returned to normal. In summary, the PCPA insomnia model was successfully established.

[0029] After the model was successfully established, drug treatment was given. It was observed that except for the blank group, the hair of the rats in the other drug-treated groups gradually became smooth, their weight gradually increased, the color of their stool gradually turned black, their mental state became good, and the mouse cage gradually became dry, indicating that the drug has a therapeutic effect on insomnia rats.

[0030] 3.2 Weight change results After modeling, the body weight of rats in the model group decreased significantly, while the body weight of rats in the blank group continued to increase steadily.

[0031] After drug treatment, the body weight of the rats in the blank group continued to increase steadily, while the body weight of the rats in the other groups showed a trend of first decreasing and then increasing. The results are shown in Table 3.

[0032]

[0033] 3.3 Open field test results This experiment used the open field test to detect changes in the autonomous activities of the four groups of rats after drug intervention. The open field test is a classic method for evaluating the autonomous and exploratory behaviors of experimental animals in a new environment. The specific detection indicators are as follows: (1) Total distance of movement - positively correlated with the activity level of the experimental animals. (2) Still time - reflects the anxiety level of the experimental animals. Figure 1 As shown in the results, compared with the blank group, the total distance traveled by rats in the model group was significantly increased, and the immobility time was significantly decreased (P < 0.001); compared with the model group, the total distance traveled by rats in each drug-treated group was significantly decreased, and the immobility time was significantly increased (P < 0.01, P < 0.05). This indicates that each drug-treated group can inhibit the abnormal excitement state of insomnia rats, alleviate anxiety, and have a sedative and hypnotic effect. Figure 1 Effects of the Gardenia-Cinnamon herbal pair on the open field test in insomnia rats ,In the figure, Note: Positive: diazepam group; Granule: Gardenia-Cinnamon group; Compared with the blank group, * P< 0.05,** P< 0.01,*** P< 0.001, **** P< 0.0001; compared with the model group, # P< 0.05, ## P< 0.01, ### P< 0.001, ### P< 0.0001; the same applies to the following figures.

[0034] 3.4 Morris water maze behavioral experimental results 3.4.1 Results of positioning navigation experiment The escape latency was used as an indicator to evaluate the exploration ability of insomnia rats in space. Figure 2 Compared with the blank group, the escape latency of rats in the model group was extremely significantly increased (P < 0.0001); compared with the model group, the escape latency of rats in the diazepam group and the Gardenia-Cinnamon group was significantly decreased (P < 0.001, 0.01).

[0035] 3.4.2 Space Exploration Experiment Results The movement trajectory of rats in each group in the pool was recorded using the rat water maze tester tracking imaging system. Figure 3The quadrant where the original platform was located was quadrant IV. After entering the water, the rats in the blank group were able to autonomously search for the platform in quadrant IV, and their tracks were mainly concentrated in quadrant IV. The tracks of the rats in the model group were disorganized, and they moved in the pool almost aimlessly and irregularly. The rats in the diazepam group paraded around the wall after entering the water, as if searching for the closest hard cardboard to the target platform, and their activity tracks were mostly concentrated in quadrant IV. The rats in the gardenia-cinnamon group also paraded around the wall after entering the water, but their tracks mostly wandered in quadrants I and IV, with less activity in quadrant II. This suggests that after administration of the gardenia-cinnamon drug pair, the rats' spatial exploration ability was enhanced, and they had certain autonomous and exploratory behaviors.

[0036] The number of times rats entered the target quadrant in the pool, average speed, distance traveled to the target quadrant, and time spent in the target quadrant were used as indicators for analysis. Figure 4 As shown in the results, compared with the blank group, the number of times, average speed, distance and time of the rats in the model group entering the IV quadrant after the platform was removed were significantly reduced (P < 0.01, 0.05); compared with the model group, the number of times, average speed, distance and time of the rats in each drug group entering the target quadrant were significantly prolonged (P < 0.05), indicating that the Gardenia-Cinnamon drug pair can improve the unstable psychological state such as anxiety or depression caused by insomnia in rats, and has a certain sedative effect.

[0037] 4 Discussion In this study, a PCPA rat model of insomnia was established and behavioral tests such as open field and Morris water maze were used to evaluate the effect of Gardenia-Cinnamon on the insomnia response to para-chlorophenylalanine ( DL The study investigated the pharmacological effects of Gardenia-Cinnamon combination on learning and memory in a rat model of insomnia induced by PCPA, providing a scientific basis for the pharmacological effects of Gardenia-Cinnamon combination on the treatment of learning and memory impairment.

[0038] The PCPA-induced insomnia model is the most widely used pharmacological model for evaluating the sedative and hypnotic effects of drugs. Intraperitoneal injection of PCPA inhibits tryptophan hydroxylation and blocks 5-HT synthesis, resulting in a significant decrease in 5-HT levels in the peripheral and central nervous systems, abrogation of circadian rhythms, and insomnia. Animal behavioral tests can effectively reflect the mood and spontaneous activity of rats. The open field test and water maze test are the most commonly used behavioral tests for assessing anxiety and depression in animals. The open field test is a classic behavioral test that assesses an animal's exploratory behavior, cognitive ability, and arousal level in a novel environment. The total distance traveled and the time spent inactive in the mine box are used to assess the emotional state of insomniac rats in a novel environment. Total distance traveled reflects the rat's ability to explore horizontally, while the time spent inactive reflects its avoidance, anxiety or depression, and autonomous activity. The water maze is a commonly used behavioral test for assessing rat cognitive function and emotional state. The escape latency of rats can, to some extent, reflect their anxiety. Track maps clearly show the path of the rats, which can be used to assess their psychological processes and exploratory abilities. Normal rats exhibit exploratory behavior in novel environments, are emotionally stable, have relatively short escape latencies, and increase the number of target quadrant entries, average speed, distance traveled, and time spent in the target quadrant. Conversely, anxious rats exhibit increased escape latency and decreased number of target quadrant entries, average speed, distance traveled, and time spent in the target quadrant. This study found that the gardenia-cinnamon herb combination significantly reduced the total distance traveled in the open field, prolonged immobility time, and reduced the escape latency of the water maze parameters in insomnia rats. It is suggested that gardenia-cinnamon can stimulate spontaneous activity and learning and memory in insomnia rats.

[0039] 5 Conclusion The Gardenia-Cinnamon bark combination has the effect of improving learning and memory impairment in the PCPA-induced insomnia rat model.

[0040] Example 2 A gardenia-cinnamon combination is used in the preparation of a medicament for improving learning and memory impairment. The gardenia-cinnamon combination comprises a weight ratio of gardenia to cinnamon of 6:1. The medicament comprises active ingredients including gardenia and cinnamon, and is prepared into a pharmaceutically acceptable dosage form using pharmaceutically acceptable carriers or excipients and processes. Furthermore, the gardenia-cinnamon combination is used in the preparation of a medicament for improving learning and memory impairment caused by insomnia.

Claims

1. Use of a gardenia-cinnamon bark combination in the preparation of a medicament for improving learning and memory impairment.

2. The use according to claim 1, characterized in that The gardenia-cinnamon bark composition is used to prepare a medicine for improving learning and memory impairment caused by insomnia.

3. The use according to claim 1 or 2, characterized in that In the gardenia-cinnamon composition, the weight ratio of gardenia:cinnamon is 6:

1.

4. The use according to claim 1 or 2, characterized in that The gardenia-cinnamon bark composition is used alone or in combination with other drugs.

5. The use according to claim 1 or 2, characterized in that: The active ingredients of the drug include Gardenia jasminoides and Cinnamon bark, and are prepared into pharmaceutically acceptable dosage forms using pharmaceutically acceptable carriers or excipients and processes. The dosage forms include powders, granules, oral liquid preparations, or injections.

Images