Use of Iris lactea Pall. var. chinensis (Fisch.) Koidz. extract

Abstract

The present invention provides the use of an Iris fruit extract in preparing a product for preventing and / or treating nerve cell damage. The present invention establishes an AD model, and after treatment with the Iris fruit extract, the activities of SOD and GSH can be significantly increased, and the increase in MDA levels can be inhibited, and the AchE content can also be inhibited. This indicates that the Iris fruit extract can effectively improve the oxidative stress response in the pathological process of AD and improve the cholinergic nerve damage of AD.

Description

Technical Field

[0001] The present invention relates to the field of medical applications of plant extracts, in particular to applications of an Iris striata fruit extract. Background Art

[0002] Iris lactea Pall.var. Chinensis (Fisch.) Koidz is a perennial herbaceous perennial plant of the genus Iris, family Iridaceae, in the phylum Angiospermae. So far, a variety of chemical components have been isolated from the seeds of Iris, including flavonoids, benzoquinones, oligostilbenes, etc. Modern pharmacology has studied the main active ingredients in Iris and found that Iris has a variety of pharmacological activities, such as radiosensitization, anti-radiation, immunity enhancement, anti-fertility, and anti-implantation effects. However, in recent years, research on the chemical components and pharmacological effects of Iris has been limited to the radiosensitization effect of irisin, while research on other chemical components and active effects has not been in-depth. Therefore, it is necessary to conduct more in-depth and detailed research on other chemical components and pharmacological effects of Iris and explore the active mechanisms based on existing research.

[0003] Neurological diseases are harming people's health, and long-term use of traditional therapeutic drugs will have side effects on the patient's body. Therefore, there is an urgent need to develop safer and more effective drugs from natural plant sources. Summary of the Invention

[0004] The present invention provides use of an Iris fruit extract in preparing a product for preventing and / or treating nerve cell damage; the Iris fruit extract is an alcohol extract.

[0005] In the technical solution of the present invention, the alcohol extract of Iris seeds refers to the extract of Iris seeds kernel extracted with ethanol, and the extraction method includes but is not limited to conventional methods such as leaching, hot reflux, and ultrasonic extraction.

[0006] In a specific embodiment of the present invention, the active substance is extracted by an immersion method, and the extraction conditions are: 70-90% ethanol, 3-5 extraction times, each time 1-5 hours, temperature 55°C-65°C, and after extraction, reduced pressure concentration is obtained to obtain a seed kernel alcohol extract.

[0007] In a specific embodiment of the present invention, the product is a product for preventing and / or treating cholinergic nerve cell damage.

[0008] Cholinergic nerves refer to nerve fibers that release acetylcholine as a chemical transmitter from their endings. Acetylcholine is a common neurotransmitter that acts as a "postman" and is responsible for moving between nerve cells to transmit information.

[0009] In a particular embodiment of the invention, the product is a product that increases ChAT activity.

[0010] In a specific embodiment of the present invention, the product is an AchE inhibitor.

[0011] The activities of choline acetyltransferase (ChAT) and acetylcholinesterase (AchE) are both related to cholinergic neurons. For example, in the development of Alzheimer's disease (AD), cholinergic neurons are severely lost, ChAT activity decreases, and AchE activity increases. Acetylcholine is hydrolyzed while the acetylcholine content decreases, nerve conduction is terminated, and ultimately leads to clinical manifestations mainly characterized by learning and memory impairment and cognitive impairment, resulting in dementia symptoms.

[0012] The present invention provides use of an Iris fruit extract in preparing a product for preventing and / or treating cerebral lipid peroxidation.

[0013] The peroxidative environment in the brain is an important factor leading to neuronal cell damage; when the oxidative product ROS is excessive and cannot be completely consumed, an oxidative stress state will occur; under the oxidative stress state, oxygen free radicals attack various unsaturated fatty acids in the biological membrane, causing lipid peroxidation.

[0014] In a specific embodiment of the present invention, the product is a product that increases the activity of antioxidant enzymes; further, the antioxidant enzymes include SOD, SRX-PRX, GSH-Px and CAT.

[0015] In a specific embodiment of the present invention, the antioxidant enzymes are SOD and GSH-Px.

[0016] In a particular embodiment of the invention, the product is a product that reduces the concentration of MDA.

[0017] In the present invention, the nerve cell damage is damage caused by metal ions.

[0018] In a specific embodiment of the present invention, the nerve cell damage is Al 3+ Caused damage.

[0019] The present invention provides use of an Iris fruit extract in preparing a product for preventing and / or treating any one of movement disorders, memory loss, cognitive impairment, consciousness disorders, muscle atrophy, dementia, and brain dysfunction.

[0020] In some specific embodiments, the product is at least one of the drugs for preventing and / or treating Alzheimer's disease, vascular dementia, amyotrophic lateral sclerosis, Parkinson's disease, dementia with Lewy bodies, and Sheehan syndrome-related diseases.

[0021] In the present invention, the product is a medicine, a health product or a food.

[0022] The beneficial effects of the present invention are:

[0023] (1) The present invention studies the activity of the alcohol extract of Iris seed kernel, uses AlCl3 to induce zebrafish to establish an AD model, and after treatment with the alcohol extract of Iris seed kernel, the SOD and GSH activities in the zebrafish induced by AlCl3 can be significantly increased, and the increase in MDA levels can be inhibited, and the AchE content can also be inhibited; this shows that the alcohol extract of Iris seed kernel can effectively improve the oxidative stress response in the pathological process of AD and improve the cholinergic nerve damage of AD.

[0024] (2) The present invention provides a new use for the alcohol extract of Iris seeds.

[0025] The abbreviations in the present invention have the following meanings:

[0026] ChAT indicates choline acetyltransferase;

[0027] AchE denotes acetylcholinesterase;

[0028] SOD stands for superoxide dismutase;

[0029] SRX-PRX indicates thioredoxin peroxidase;

[0030] GSH-Px indicates glutathione peroxidase;

[0031] GSH denotes glutathione;

[0032] CAT indicates catalase;

[0033] MDA stands for malondialdehyde;

[0034] AD indicates Alzheimer's disease;

[0035] Dpz indicates donepezil;

[0036] HA stands for huperzine A. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 The diagram shows the movement trajectory of zebrafish in each drug group;

[0038] Figure 2 is the total distance moved in each drug group;

[0039] Figure 3 is the average speed of movement in each drug group;

[0040] Figure 4 The effect of Iris odorata extract on acetylcholinesterase;

[0041] Figure 5 The effect of Iris odorata extract on acetylcholine transferase;

[0042] Figure 6 The effect of Iris odorata extract on SOD;

[0043] Figure 7 The effect of Iris odorata extract on GSH;

[0044] Figure 8 This is the effect of Iris odorata extract on MDA. DETAILED DESCRIPTION

[0045] The following is a clear and complete description of the technical solution of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0046] Example 1 Preparation of Iris Fruit Extract

[0047] Dried Iris seeds were processed in a grinder for 3-5 seconds to obtain the seed coat and kernel, respectively. The kernel was extracted with 85% ethanol at 60°C three times for 3 hours each time. The three extracts were combined and concentrated under reduced pressure to obtain the kernel ethanol extract, which is the Iris seed extract (C) of the present invention.

[0048] Example 2 Effect of Iris Fruit Extract on Nerve Damage

[0049] 1 Experimental Materials

[0050] 1.1 Experimental Animals

[0051] Adult wild-type zebrafish (AB strain) aged 6 to 8 months used in this invention were acclimated to the culture system for at least one week prior to the experiments. The temperature was 25 ± 2°C, the pH was 7.4 ± 0.2, the photoperiod was a 14-hour light / 10-hour dark cycle, and brine shrimp were fed twice daily. Juveniles were obtained by natural mating. Zebrafish juveniles 3 days post fertilization (dpf) were used in this invention. All experiments followed standard ethical guidelines and were conducted under the supervision of the Animal Ethics Committee of the Northwest Plateau Institute of Biology.

[0052] 1.2 Instruments and reagents

[0053] Analytical grade crystalline aluminum chloride hexahydrate (AlCl3·6H2O) was purchased from Sigma Reagent Company;

[0054] zebrafish culture system (Beijing Aisheng Technology Development Co., Ltd.);

[0055] Biochemical index (AChE, ChAT, SOD, MDA, GSH) detection kits were purchased from Nanjing Jiancheng Bioengineering Institute;

[0056] The automatic computer video tracking system and VisuTrack-AI software were purchased from Shanghai Xinruan Information Technology Co., Ltd.

[0057] Iris fruit extract: obtained by separation and extraction in Example 1.

[0058] 2 Experimental methods

[0059] 2.1 Experimental Methods

[0060] The same batch of juveniles (3 dpf) from the same sire and dam were selected and randomly divided into 6 groups, with 90 juveniles in each group. Zebrafish juveniles were co-treated with 5 μg / mL and 10 μg / mL of Iris irritans extract and 80 μM AlCl3 from 3 to 6 dpf. AChE inhibitors (donepezil and huperzine A) were used as positive drugs. In the positive group, juveniles were co-treated with 80 μM AlCl3 and 4 μM donepezil (Dpz) / huperzine A (HA) from 3 to 6 dpf. After treatment, 12 larvae were randomly selected from each group for image acquisition. The culture medium was replenished regularly every day. The light cycle and water temperature remained unchanged during the modeling period, and attention was paid to the cleanliness of the culture dishes to prevent interference with the experimental results.

[0061] 2.2 Behavioral Analysis

[0062] 12 larvae from each group were randomly collected and cleaned using embryo culture medium (1mM MgSO4, 0.5mM KCl, 15mM NaCl, 0.05mM (NH4)3PO4, 0.15mM KH2PO4, 0.7mM NaHCO3 and 1mM CaCl2). They were then placed in a 96-well plate. After a 10-minute adaptation period, the movement behavior of each larva was recorded using an automatic computer video tracking system (VisuTrack-AI, Shanghai Xinruan Information Technology Co., Ltd.). The behavioral test consisted of three 30-minute alternating light and dark cycles (5 minutes of illumination and 5 minutes of darkness). VisuTrack-AI software was used to record and analyze the movement distance and speed changes of zebrafish.

[0063] 2.3 Analysis of neurobiochemical indicators

[0064] After co-treatment with AlCl₃ and each drug, zebrafish were sacrificed at 6 dpf using the anesthetic diazepam. Ninety zebrafish from each group were placed in a 1.5 mL EP tube and filled with pre-chilled saline at a ratio of 1:9 (mass:volume) without additional water. The tissues were then mechanically homogenized using an ultrasonic homogenizer in an ice bath to completely disrupt the tissue. The homogenates from each group were centrifuged at 12,000 rpm for 15 minutes, and the supernatants were used for analysis of the biochemical markers AChE, SOD, MDA, and GSH. All measurements were repeated three times, and the average value was calculated.

[0065] 3 Experimental results

[0066] 3.1 Behavioral Experimental Results

[0067] like Figure 1 As shown in the figure, the total distance moved by zebrafish in the AD model group was significantly shorter than that in the untreated group, and their average speed was also significantly weakened after the light and dark stimulation changed; these results indicate that AlCl3 reduced the movement ability of zebrafish, indicating that the zebrafish AD model was successfully established.

[0068] like Figure 2 and Figure 3 As shown in the figure, after treatment with Iris irritans extract C, the travel distance and average speed of zebrafish increased compared with those in the AD group, and the 5μg / mL and 10μg / mL treatment groups were both significant;

[0069] This indicates that Iris extract C improves AlCl3-induced movement disorders in zebrafish.

[0070] 3.2 Effects of acetylcholinesterase

[0071] Acetylcholine is a common neurotransmitter that acts like a "postman," responsible for moving between nerve cells to transmit information. During the development of AD, cholinergic neurons are severely lost, ChAT activity decreases, AchE activity increases, and while hydrolyzing acetylcholine, the acetylcholine content decreases, nerve conduction is terminated, ultimately leading to clinical manifestations characterized by learning and memory impairment and cognitive impairment, resulting in dementia symptoms. Therefore, improving the function of the cholinergic nervous system has become one of the important methods for the prevention and treatment of AD.

[0072] like Figure 4 and Figure 5 As shown in the results, after AlCl3 induction, AchE levels in the model zebrafish were significantly increased and ChAT levels were significantly decreased. The positive drugs donepezil and huperzine A also significantly decreased AchE levels and increased ChAT levels. Iris extract C (5 μg / mL and 10 μg / mL) also significantly decreased AchE levels while significantly increasing ChAT levels (5 μg / mL).

[0073] 3.3 Effect of redox level

[0074] MDA is the final product of lipid peroxidation, and its cytotoxic effects can induce neuronal degeneration and necrosis in AD. MDA content can serve as an important indicator of the degree of lipid peroxidation. SOD is an oxygen free radical scavenging enzyme that, through dismutation reactions, can reduce or eliminate the damage caused by superoxide anion radicals. The body's ability to scavenge free radicals is assessed by measuring SOD activity in vivo or in tissues. Furthermore, glutathione (GSH) is the most abundant antioxidant in the brain and plays a key role in ROS detoxification. GSH levels are reduced in the brains of AD patients, and changes in GSH are crucial in the pathogenesis of AD.

[0075] like Figures 6 to 8 As shown in the results, after AlCl3 induction in the model group, the MDA content increased, while the SOD and GSH levels decreased significantly. These results suggest that the degree of peroxidation in AD zebrafish is increased, causing damage to hippocampal neurons, which is closely related to learning and memory dysfunction. Treatment with Iris iris extract significantly reduced the MDA content while increasing the SOD and GSH levels.

[0076] In summary, the AD model was established by inducing zebrafish with AlCl3, and treatment with Iris irritans extract can significantly increase the AlCl3-induced increase in SOD and GSH activities in zebrafish, inhibit the increase in MDA levels, inhibit AchE content and increase ChAT levels; this shows that Iris irritans extract can effectively improve the oxidative stress response in the pathological process of AD and improve the cholinergic nerve damage of AD.

[0077] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. Use of an Iris fruit extract in preparing a product for preventing and / or treating Alzheimer's disease; the Iris fruit extract is an ethanol extract of Iris fruit kernels obtained by ethanol extraction and concentration; the ethanol concentration is 85%.

2. The use according to claim 1, characterized in that The product can improve cholinergic nerve cell damage.

3. The use according to claim 1, characterized in that The product can increase the level of ChAT.

4. The use according to claim 2, characterized in that The product is an AchE inhibitor.

5. The use according to claim 1, characterized in that The product can reduce the level of lipid peroxidation in the brain.

6. The use according to claim 5, characterized in that The product can increase antioxidant enzyme activity.

7. The use according to claim 5, characterized in that The product can reduce MDA concentration.

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