Compound traditional Chinese medicine essential oil and application thereof in improvement of Parkinson disease

Abstract

The invention relates to compound traditional Chinese medicine essential oil and application thereof to improvement of Parkinson disease, the compound traditional Chinese medicine essential oil comprises rosemary essential oil, lavender essential oil, lemon essential oil, peppermint essential oil, magnolia flower essential oil, sharpleaf galangal fruit essential oil and acorus gramineus essential oil, and further comprises 36% of rosemary essential oil, 12% of lavender essential oil, 24% of lemon essential oil and 10% of peppermint essential oil. , 4% of magnolia flower essential oil, 10% of fructus alpiniae oxyphyllae essential oil and 4% of rhizoma acori Experiments prove that the compound traditional Chinese medicine essential oil disclosed by the invention can be used for remarkably improving Parkinson-like dyskinesia of model animals.

Description

Technical Field

[0001] The invention discloses a Chinese medicinal essential oil, and particularly relates to a compound Chinese medicinal essential oil and application thereof in improving Parkinson's disease. Background Art

[0002] Parkinson's disease, also known as Parkinson's paralysis, is the second most common chronic neurodegenerative disease after Alzheimer's disease. Its main pathological features are the loss of dopamine (DA) neurons in the substantia nigra compacta of the midbrain and the appearance of Lewy bodies (mainly composed of α-synuclein) in neurons. In clinical practice, behavioral abnormalities are often used as indicators for diagnosing Parkinson's disease, mainly including resting tremor, bradykinesia, muscle rigidity, postural instability and other Parkinson-like symptoms (Parkinsonism). As the disease develops and worsens, a series of non-motor symptoms may appear in the late stage, such as depression, anxiety, cognitive impairment, sleep disorders, olfactory disorders, constipation, and impairment of spatial learning and memory functions.

[0003] Parkinson's disease is a major problem in modern medicine. The cause is not fully understood and there is a lack of standardized treatment methods. At present, the most widely used is the levodopa preparation Madopar, which has been used for more than 40 years since it was launched in 1973. However, only 5% to 10% of levodopa can actually pass through the blood-brain barrier, and the rest is dispersed in the body, causing a variety of side effects including movement disorders and orthostatic hypotension. Monoamine oxidase B inhibitors such as selegiline combined with Madopar can relatively reduce the dosage of levodopa, but after the honeymoon period, there will be a decrease in efficacy, fluctuations in symptoms, and obvious side effects. Therefore, the existing Western medicine treatments cannot effectively prevent the progression of the disease, and patients need to take medication for life to improve their symptoms. The use of traditional Chinese medicine with less toxic side effects has become a preferred or auxiliary treatment method.

[0004] In traditional Chinese medicine, Parkinson's disease belongs to the category of "tremor syndrome", and its symptoms were described as early as the Yellow Emperor's Classic of Internal Medicine. Sun Yikui, a doctor in the Ming Dynasty, named Parkinson's disease "tremor syndrome" for the first time in "Red Water Mysterious Pearl". Scholars of all generations have made corresponding discussions and accumulated experience. At present, it is believed that the pathogenesis of this disease is generally a combination of deficiency of the root and excess of the symptoms. The deficiency of the root is liver and kidney deficiency, and blood deficiency, while the excess of the symptoms is wind, fire, phlegm, blood stasis, poison, and cold. The Third Academic Symposium on Geriatric Encephalopathy of the All-China Association of Traditional Chinese Medicine has formulated the syndrome of "tremor syndrome" as phlegm-heat causing wind, blood stasis causing wind, deficiency of both qi and blood, deficiency of liver and kidney, and deficiency of both yin and yang.

[0005] Aromatherapy refers to the process of making aromatic plants into different dosage forms, and adjusting the physiological or psychological state of a person through smearing, sniffing, massage, and taking, so as to relieve pain and restore health. As an auxiliary therapy, aromatherapy is often used to relieve neuropsychiatric diseases and improve physical and mental health. Aromatherapy has a history of thousands of years. At first, the ancient Egyptians used the technique of immersion to extract essential oils with aromatic smells from plants for massage. In ancient my country, aromatic medicines were used to make sachets and aromatherapy to prevent and treat diseases, avoid filth and disinfect. The Compendium of Materia Medica also recorded the efficacy of a variety of aromatic plants, and summarized and innovated their applications. It was not until 1936 that French chemist Gattefossé first proposed aromatherapy, which attracted more and more attention and promoted the rapid development of aromatherapy around the world. In recent years, my country's aromatherapy industry has been in a stage of rapid development. Due to the continuous improvement of standardization and specialization, it has been favored by more and more people. At present, the opportunities for application in daily and clinical care are gradually increasing. As a natural and organic alternative therapy, it has great development potential.

[0006] As the part that is most closely connected to the brain, many literatures have shown that aromatic Chinese medicine can regulate the brain center and limbic system through the olfactory pathway, thereby regulating brain function. However, there are no reports on the research on the nasal administration of compound Chinese herbal aromatic essential oils in the treatment of Parkinson's disease. Summary of the invention

[0007] In view of this, the technical problem to be solved by the present invention is to provide a compound Chinese herbal essential oil, which can improve Parkinson's disease by nasal inhalation.

[0008] In the present invention, improving Parkinson's disease includes protecting against Parkinson-like dyskinesia.

[0009] The present invention uses behavioral data as the best indicator for the improvement of Parkinson's-like symptoms, and proves that nasal inhalation of composite Chinese herbal essential oil has a significant protective effect on Parkinson's disease through pole climbing experiment, rotating rod experiment and CatWalk gait analysis.

[0010] In the present invention, the protection of Parkinson's-like movement disorders includes improving motor coordination ability, improving agility, improving balance maintenance ability, improving limb use and movement state and gait stability. The pathogenic mechanism of Parkinson's disease is diverse. In the present invention, the accumulation of α-synuclein caused by the A53T point mutation of the SNCA gene causes Parkinson's disease.

[0011] The concentration of the compound Chinese medicinal essential oil for improving Parkinson's disease by sniffing is 5%.

[0012] In the present invention, the compound Chinese medicine essential oil comprises: rosemary essential oil, lavender essential oil, lemon essential oil, peppermint essential oil, magnolia essential oil, oxyphylla essential oil and calamus essential oil.

[0013] Furthermore, the compound Chinese medicine essential oil used in the present invention comprises: 30-40% rosemary essential oil, 10-15% lavender essential oil, 20-30% lemon essential oil, 8-12% peppermint essential oil, 2-6% magnolia essential oil, 5-15% oxyphylla essential oil and 2-6% calamus essential oil.

[0014] In some specific embodiments, the compound Chinese medicine essential oil used in the present invention includes: rosemary essential oil accounting for 36%, lavender essential oil 12%, lemon essential oil 24%, peppermint essential oil 10%, magnolia essential oil 4%, Alpinia oxyphylla essential oil 10%, and Acorus calamus essential oil 4%.

[0015] The present invention uses the Parkinson's disease animal model (SNCA-AAV (A53T) brain localized injection model) as the research object to study the protective effect of nasal administration of compound Chinese herbal essential oil on Parkinson's-like movement disorder. The results show that the compound Chinese herbal essential oil can significantly improve the Parkinson's-like movement disorder of the model animal. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 This is a curve chart of weight changes of experimental mice in each group for 3 weeks;

[0017] Figure 2 It is a bar graph of the time of mice climbing the pole in each drug group;

[0018] Figure 3 The bar graph is the rotarod residence time of mice in each drug group;

[0019] Figure 4 It is a bar graph of the average speed of mice in the CatWalk gait analysis experiment for each drug group;

[0020] Figure 5 It is a bar graph of the maximum speed change rate in the CatWalk gait analysis experiment of mice for each group of drugs;

[0021] Figure 6 It is a bar graph of the step frequency in the CatWalk gait analysis experiment of mice in each drug group;

[0022] Figure 7 It is a bar graph of the number of steps in the CatWalk gait analysis experiment of mice for each group of drugs. DETAILED DESCRIPTION

[0023] The present invention provides the use of nasal inhalation of composite Chinese herbal essential oil to improve PD. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The methods and applications of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications of this article without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0024] The test materials used in the present invention are all common commercial products and can be purchased in the market.

[0025] In the present invention, the compound Chinese medicine essential oil comprises: rosemary essential oil, lavender essential oil, lemon essential oil, peppermint essential oil, magnolia essential oil, oxyphylla essential oil and calamus essential oil.

[0026] Furthermore, in the compound Chinese medicinal essential oil adopted by the present invention, rosemary essential oil accounts for 30-40%, lavender essential oil accounts for 10-15%, lemon essential oil accounts for 20-30%, peppermint essential oil accounts for 8-12%, magnolia essential oil accounts for 2-6%, alkyl oxyphylla essential oil accounts for 5-15%, and calamus essential oil accounts for 2-6%.

[0027] As a preferred embodiment, in the compound Chinese medicine essential oil adopted by the present invention, rosemary essential oil accounts for 36%, lavender essential oil 12%, lemon essential oil 24%, peppermint essential oil 10%, magnolia essential oil 4%, oxyphylla essential oil 10%, and calamus essential oil 4%. The preparation of the compound Chinese medicine essential oil of the present invention includes 36% rosemary essential oil, 12% lavender essential oil, 24% lemon essential oil, 10% peppermint essential oil, 4% magnolia essential oil, 10% oxyphylla essential oil, and 4% calamus essential oil.

[0028] The present invention is further described below:

[0029] 1. Experimental Materials and Instruments

[0030] 1. Experimental Animals

[0031] C57BL / 6J mice, male, 6 weeks old, weighing 20-22 g, were purchased from the Experimental Animal Management Center of Southern Medical University (License: SCXK (Guangdong) 2021-0041). The animals were raised in the Medical Experimental Animal Center of Jinan University at a temperature of 23±2°C, a humidity of 50±10%, and standard mouse feed. The experimental animals in each group were allowed to eat and drink freely.

[0032] 2. Main experimental reagents

[0033] Rosemary essential oil (Guangzhou Wuyuan Essential Oil Co., Ltd.), lavender essential oil (Guangzhou Wuyuan Essential Oil Co., Ltd.), lemon essential oil (Guangzhou Wuyuan Essential Oil Co., Ltd.), peppermint essential oil (Guangzhou Wuyuan Essential Oil Co., Ltd.), magnolia essential oil (Jiangxi Juncai Biotechnology Co., Ltd.), Alpinia oxyphylla essential oil (Jiangxi Juncai Biotechnology Co., Ltd.), Acorus calamus essential oil (Jiangxi Juncai Biotechnology Co., Ltd.), SNCA-AAV tool: AAV2 / 9-SNCA (A53T) (Shanghai Taiertu Biotechnology Co., Ltd., S1024-9), levodopa (Sigma, D9628), benserazide (Sigma, B7283). Madopar: levodopa and benserazide were mixed in a ratio of 4:1.

[0034] 3. Main experimental instruments

[0035] Mouse stereotaxic instrument (Shenzhen Ruiwode Life Science Co., Ltd.), mouse rotarod instrument (UgoBasile, Italy), high-speed low-temperature centrifuge (Sartorius, Germany, A-14C), chemiluminescence developer (Tanon-5200), electronic balance (Sartorius, Germany, BS210), and smart aromatherapy machine.

[0036] 2. Experimental Methods and Results

[0037] 1. PD Model Preparation

[0038] First, SNCA-AAV was used to construct a PD model: After seven days of adaptive feeding, the mice were weighed and deeply anesthetized with 1% sodium pentobarbital. The head hair was shaved with a razor and the mice were fixed in a brain stereotaxic instrument. The left and right ear bars were adjusted to make the fixation more stable. The skin of the mouse head was cut along the midline to expose the mouse skull, and hydrogen peroxide was used to wipe to fully expose the bregma and lambda. With the bregma coordinate origin as the reference of the Mouse Brain Stereotaxic Atlas, the injection coordinate site was AP: +0.3; L: +2.3, DV: -2.9. A small hole was carefully drilled with a skull drill (to avoid damaging brain tissue), and 1.5 μL of 5 μg / μL SNCA-AAV was slowly injected at a speed of 0.1 μL / min. After the injection, the needle was retained for 5 minutes, and the needle was slowly returned to perform scalp suture to complete the brain localization injection.

[0039] 2. Experimental Grouping and Weight Monitoring

[0040] The experimental mice were divided into 4 groups: 8 mice without treatment were set as the blank group (Sham). The mice with successful PD modeling were randomly divided into 3 groups, 8 in each group, namely: SNCA-AAV group, SNCA-AAV+compound Chinese medicine essential oil group (administered after modeling, fumigation inhalation 5% concentration, 2 weeks) and SNCA-AAV+Medopar group (administered after modeling, 1 mg / kg, gavage for 2 weeks).

[0041] Weight monitoring: Figure 1 As shown, the body weight data for 3 consecutive weeks showed that the modeling and drug administration methods used in this experiment had no significant effect on the body weight of mice.

[0042] 3. Pole climbing experiment methods and results

[0043] Method description: The pole climbing experiment is a behavioral test experiment that records the difference in the time mice spend on the pole to detect the coordination of the mouse's limb movements. The entire experiment was completed in a quiet and dark environment. The climbing pole is composed of an iron frame with a diameter of 1 cm and a height of 60 cm and a small ball with a diameter of 1.5 cm fixed on the top. Before the experiment, the surface of the climbing pole was wrapped with gauze to prevent the mouse from slipping. Before modeling, the mice were trained for one week in advance to adapt, and each mouse was trained five rounds each time. After modeling, the test was conducted once a week. During the experiment, the mouse was placed head up on the climbing pole, and the time required for the mouse to climb to the bottom from the beginning was recorded. If it exceeded 1 minute, it was counted as 1 minute. Each mouse was tested 5 rounds each time and the average value was taken.

[0044] Experimental results: The time required for mice to climb from the top of the pole to the bottom was used to evaluate the coordination and agility of the mice's limbs. Figure 2 As shown. Compared with the blank control, the time required for mice in the SNCA group to climb the pole was significantly prolonged (***p<0.001vs. blank group, n=8). Both the compound Chinese herbal essential oil and the positive drug Madopar can significantly shorten the climbing time of model mice (###p<0.001vs. SNCA-AAV group, n=8), and the compound Chinese herbal essential oil has a better effect.

[0045] 4. Rotarod test method and results

[0046] Method description: The rotarod test uses a mouse rotarod fatigue instrument to record the time the mouse stays on the cylindrical rotarod to detect the motor coordination ability of the mouse. The experiment was carried out in a quiet and dark environment. The mouse rotarod fatigue instrument can accommodate 5 mice for each experiment. Each mouse has a separate runway. The connected computer software records the time when the mouse falls. When the mouse passively rotates with the rotarod for more than two consecutive circles, the timing should also be stopped. Before modeling, the mice were trained for one week in advance to adapt, and each mouse was trained three times each time. After modeling, the test was conducted once a week, and the parameters were set to increase from 5r / min to 30r / min at a constant speed for 5min. The time of the mouse on the rod within 5min was recorded. If it exceeded 5min, it was counted as 5min. Each mouse was tested for 3 rounds each time, and the average value was taken. The test interval of each group of mice was at least 30min. Before changing the mice each time, the instrument was cleaned with 75% alcohol to remove the odor before the next group of experiments.

[0047] Experimental results: The motor coordination ability of mice was evaluated by the anti-fatigue time of mice on the rotarod. Figure 3 As shown. Compared with the blank control, the rod-study time of the mice in the MPTP group was significantly shortened (***p<0.001 vs. blank group, n=8). Both the compound Chinese medicine essential oil and the positive drug Madopar were able to significantly restore the rod-study time of the model mice (###p<0.001 vs. SNCA-AAV group, n=8).

[0048] 5. CatWalk gait analysis method and results

[0049] Method description: CatWalk gait analysis is a system for rodent gait analysis, including balance maintenance ability, limb use, movement state and gait stability analysis. The main accessories are built-in red LED light board and green fluorescent glass plate, high-frequency camera under the glass plate, image transmission system and computer hardware and software. Before the test, the camera height was adjusted to 60cm, the effective runway length and width were framed in the software, and the camera gain was set to 28.12dB. The process of each mouse passing through the glass plate smoothly was recorded at room temperature, and the maximum time was set to 10s, that is, the mouse passed the set distance within 10s as a valid data. The principle of recording is: the mouse's paw print is illuminated by the fluorescent reflection of the fluorescent lamp in the glass plate, captured by the camera below, and transmitted to the computer. The system software analyzes and counts the recorded video data. The mouse gait is qualitatively and quantitatively analyzed based on the position of the paw print, the pressure of the paw on the glass plate, the distance between different paw prints, the speed of the mouse gait, and the regularity of the gait.

[0050] Before the formal test, the mice need to be trained 3 times in advance, and then the test will start, and each mouse will be recorded at least 5 times. The experiment should be carried out in a dark and quiet environment without human interference. After each mouse walks, the runway will be cleaned and wiped with 75% alcohol to avoid interference with the next mouse. After the experiment, the data obtained were statistically analyzed using the CatWalkXT9.1 system of Noldus.

[0051] Experimental results: The gait changes caused by motor defects and other reasons were evaluated by comprehensive analysis of the movement trajectory of mice during natural walking. Figures 4 to 7 As shown in the figure, the evaluation indicators used were average speed, maximum speed change rate, cadence and number of steps. Compared with the blank control, the above evaluation indicators of the SNCA-AAV group mice were significantly impaired (***p<0.001 vs. blank group, n=8). Both the compound Chinese medicine essential oil inhalation and the positive drug Madopar were able to significantly restore the gait behavior disorder of the model mice (###p<0.001 vs. SNCA-AAV group, n=8).

[0052] The above description of the embodiments is to facilitate the understanding and application of the present invention by those skilled in the art. It is obvious that those skilled in the art can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without creative work. Therefore, the present invention is not limited to the embodiments herein, and improvements and modifications made to the present invention by those skilled in the art based on the disclosure of the present invention should be within the scope of protection of the present invention.

Claims

1. A compound Chinese medicinal essential oil, characterized in that: Including rosemary essential oil, lavender essential oil, lemon essential oil, peppermint essential oil, magnolia essential oil, oxyphylla essential oil, and calamus essential oil.

2. The compound Chinese medicinal essential oil as claimed in claim 1, characterized in that Including rosemary essential oil accounting for 30-40%, lavender essential oil 10-15%, lemon essential oil 20-30%, peppermint essential oil 8-12%, magnolia essential oil 2-6%, Alpinia oxyphylla essential oil 5-15%, and Acorus calamus essential oil 2-6%.

3. The compound Chinese medicinal essential oil as claimed in claim 2, characterized in that Including rosemary essential oil accounting for 36%, lavender essential oil 12%, lemon essential oil 24%, peppermint essential oil 10%, magnolia essential oil 4%, Alpinia oxyphylla essential oil 10%, and Acorus calamus essential oil 4%.

4. Application of the compound Chinese medicinal essential oil as claimed in claim 1, 2 or 3 in improving Parkinson's disease.

5. The use according to claim 4, characterized in that The compound Chinese medicinal essential oil is inhaled through the nose, and the inhaled concentration is 5%.

6. The use according to claim 4, characterized in that Improvement of Parkinson's disease includes protection against parkinsonian movement disorders.

7. Use of the compound Chinese medicinal essential oil as claimed in claim 1, 2 or 3 in combination with a levodopa preparation in the preparation of a medicament for treating Parkinson's disease.

8. The use according to claim 7, characterized in that The levodopa preparation is Madopar.

9. A combination drug, characterized in that The invention comprises a levodopa preparation and the compound Chinese medicinal essential oil as claimed in claim 1, 2 or 3, wherein the compound Chinese medicinal essential oil is administered nasally.

10. The combination drug according to claim 9, characterized in that The levodopa preparation is Madopar.

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