NL2031296B1

NL2031296B1 - Application of aronia melanocarpa leaf polyphenol extract in preparing antidepressants

Info

Publication number: NL2031296B1
Application number: NL2031296A
Authority: NL
Inventors: Sheng Guihua; Zhou Quancheng; Ji Wei; Nan Xijun; Li Jiayi; Wang Honglei
Original assignee: Univ Shandong Technology
Priority date: 2021-12-30
Filing date: 2022-03-15
Publication date: 2023-07-05

Abstract

U I T T R E K S E L Application of aronia melanocarpa leaf polyphenol extract in preparing medicines, functional foods or supplements for preventing, regulating or treating depression is disclosed. The aronia melanocarpa leaf polyphenol extract can obviously improve 5 the Brain—Derived Neurotrophic Factor (BDNF) level in the hippocampus of depressed mice by adjusting the richness and diversity of intestinal flora of the depressed mice to normal level, thus having a remarkable improvement effect on the depression symptoms of the mice. The aronia melanocarpa leaf lO polyphenol extract belongs to a non—toxic and harmless natural component, which does not cause side effects, so that a novel material and method may be provided for preventing, regulating or treating depression.

Description

P1124 /NL

APPLICATION OF ARONIA MELANOCARPA LEAF POLYPHENOL EXTRACT IN

PREPARING ANTIDEPRESSANTS

TECHNICAL FIELD

The disclosure discloses application of aronia melanocarpa leaf polyphenol extract in the field of prevention, regulation or treatment of depression, and belongs to the field of related prod- ucts such as medicines, functional foods and supplements.

BACKGROUND ART

Depression is an affective disorder characterized by a marked and persistent low mood, and loss of interest in daily pleasant activities. The incidence of depression is increasing year by year with increasing pressure in life of people. According to statis- tics of the World Health Organization, depression is to exceed cardiovascular diseases to become the second major disease threat- ening human health by 2020. According to study published by the team of Peking University in The Lancet magazine in 2019, it is estimated that there are 350 million people suffering from depres- sion worldwide, with more than 95 million in China. On September 11, 2020, the National Health Comnission released the Work Plan on

Exploring Featured Services for Depression Prevention and Control, which requires that “depression screening” may become a regular item of physical examination. All this shows that depression has a great impact on pecple's physical and mental health, life and work.

The pathogenesis of depression is very complicated. At pre- sent, monoamine theory has been widely accepted. According to mon- oamine theory, depression is caused by the low levels of neuro- transmitters such as serotonin and norepinephrine. Based on this, antidepressants are developed to increase the levels of both. How- ever, these medicines only benefit some patients, and adverse ef- fects such as drug resistance may occur in the patients.

Microbes in human intestinal tracts are large in quantity and rich in variety, participate in many physiological activities of human bodies, and affect the health of hosts. In recent years, it has been found that the structure and diversity of intestinal mi- crobes are closely related to depression, and intestinal microbes and depression interact with each other through the “brain- intestines-microbes” axis. Meanwhile, an animal model of depres- sion and the patients with depression have similar microbial structures. Studies have proved that the change of intestinal flo- ra can cause the change of Brain-Derived Neurotrophic Factor {BDNF) in the hippocampus of the brain. BDNF is an important regu- lator of nerve growth. The decrease of BDNF level and the increase of nerve apoptosis may cause depression symptoms. The increase of

BDNF and other neurotrophic contents in brain can reduce the apop- tosis of hippocampal neurons, promote the growth of neurons and improve cognition and emotion. Studies have shown that lactic acid bacteria and bifidobacterium can relieve depression by changing intestinal composition and diversity of intestinal flora, reducing inflammatory factors and regulating central nervous system.

Therefore, it has become the focus of people's attention to regulate depression through the intervention of nutrients in foods and to develop a depression regulation nutrition system which is efficient and has no toxic or side effects on human bodies. In re- cent years, it has been found that many foods, such as radix platycodonis, day-lily buds and semen myristicae, have the func- tion of relieving depression. Aronia melanocarpa (referred to as “black chokeberry”) is an edible berry rich in polyphenol sub- stances, which has multiple efficacies including oxidation re- sistance, immunoregulation, inflammation resistance and the like.

Aronia melanocarpa leaf polyphenol extract mainly includes procy- anidine, anthocyanin, phenolic acid and the like, and the total amount of polyphenols contained in aronia melanocarpa is higher than that of many other kinds of berries. Polyphenols have an ame- liorating effect on mouse depression caused by chronic unpredicta- ble stress, and its mechanism may be related to the inhibition of oxidative stress in brain tissue. In addition, polyphenols can in- crease the bioavailability of polyphenol metabolites by intestinal bacteria in the human body, and these polyphenol metabolites can promote psychological conditions.

At present, there is little research on the aronia melanocar-

pa leaf polyphenol extract, and meanwhile, there is no report on application of the aronia melanocarpa leaf polyphenol extract in the aspect of preventing, regulating or treating depression.

Therefore, it is of great significance to develop application of the aronia melanocarpa leaf polyphenol extract in the aspect of antidepressants, functional foods or supplements.

SUMMARY

The disclosure provides an aronia melanocarpa leaf polyphenol extract and a preparation method and application of the aronia melanocarpa leaf polyphenol extract. The aronia melanocarpa leaf polyphenol extract has the effect of preventing, regulating or treating depression and can be applied to the fields of medicines, functional foods, supplements and foods with homology of medicine and food.

The aronia melanocarpa leaf polyphenol extract can be ex- tracted from aronia melanocarpa, its extraction method is also re- ported in a lot of existing documents, and aronia melanocarpa can be obtained through commercial purchase.

Preferably, the application dose of the aronia melanocarpa leaf polyphenol extract is 15-25 mL/kg body weight.

Preferably, an appropriate dose of aronia melanocarpa leaf polyphenol extract has a positive effect on reducing the diversity of intestinal flora in depressed mice.

Preferably, the aronia melanocarpa leaf polyphenol extract has a certain inhibition effect on harmful bacteroidetes, and is beneficial to the proliferation of beneficial firmicutes.

Preferably, the aronia melanocarpa leaf polyphenol extract can restore the richness and diversity of intestinal flora of the depressed mice to normal levels, which is beneficial to release of

BDNF and improvement of depression symptoms.

Preferably, application of the aronia melanocarpa leaf poly- phenol extract in preparing functional foods and supplements for preventing, regulating or treating depression is disclosed.

Preferably, application of the aronia melanocarpa leaf poly- phenol extract in preparing medicines for preventing regulating or treating depression is disclosed.

Furthermore, appropriate amount of carriers, auxiliary mate-

rials or auxiliary components can be added into the medicines, functional foods or supplements according to needs.

The disclosure has the following beneficial effects. 1. The disclosure verifies that the aronia melanocarpa leaf polyphenol extract can reduce relative abundance of harmful flora in intestinal tracts of mice, inhibit proliferation of bacteroide- tes and prevent inflammation. Meanwhile, the aronia melanocarpa leaf polyphenol extract is beneficial to proliferation of firmicu- tes, so that the energy acquisition of intestinal epithelial cells is guaranteed, the entry of pathogenic bacteria is prevented, and the secretion of 5-HT is promoted. The aronia melanocarpa leaf polyphenol extract can reduce adverse effects caused by intestinal microbial disorder and regulate the richness and diversity of in- testinal microbial flora to normal levels. 2. The aronia melanocarpa leaf polyphenol extract has the ef- fects of regulating the intestinal microenvironment, improving the intestinal flora structure and improving the BDNF expression level in the hippocampus of the brain. 3. The disclosure provides the application of the aronia mel- anocarpa leaf polyphenol extract in antidepressants, functional foods or supplements, which belongs to development of novel prod- ucts of aronia melanocarpa. The aronia melanocarpa leaf polyphenol extract can be added into the medicines, functional foods or sup- plements as an additive to improve its value. The aronia melano- carpa leaf polyphenol extract can effectively improve the intesti- nal flora, and is used for preventing, regulating or treating de- pression, and toxic and side effects caused by traditional medi- cines are avoided.

DETAILED DESCRIPTION OF THE EMBODIMENTS

The disclosure will be described in detail below with refer- ence to embodiments.

Embodiment 1

Aronia melanocarpa polyphenol extract and reagents: aronia melanocarpa is taken from Tieshan Forest Farm in Zibo

City. Preparation method of aronia melanocarpa leaf polyphenol ex- tract: fresh aronia melanocarpa leaves are washed up and pulped. 5 g of pulp is weighed, an appropriate amount of 50% ethanol is add-

ed according to the material-liquid ratio of 1:5, ultrasonic treatment is carried out for 40 min, suction filtration is carried out, the filtrate is subjected to reduced pressure at 50 °C to re- move ethanol, and after freeze drying, freeze storage is carried 5 out for later use.

Corticosterone is provided by Jiangsu Jiaerke Pharmaceutical

Group Co., Ltd.; propylene glycol is provided by Jinan Huaying

Chemical Co., Ltd.; fluoxetine is provided by Shandong Xiya Chemi- cal Industry Co., Ltd.; gallic acid standard and other commonly used reagents are purchased from Shanghai Aichun Biotechnology

Co., Ltd.

Embodiment 2

A mouse model of depression is established. 100 male BALB/C mice are injected subcutaneously with 0.2 mL of corticosterone to establish the mouse model of depression. Nor- mal mice are used to establish a normal group. The successfully modeled depressed mice are divided into a model group, a solvent group (0.2 mL of propylene glycol), a medicine group (0.2 mL of fluoxetine), and a low-dose group, a medium-dose group and a high- dose group of aronia melanocarpa leaf polyphenol extract. 10 mice in each cage are subjected to a gavage experiment for 4 weeks. The normal group and model group are given the same volume of normal saline. After the feeding experiment is finished, intestinal feces of the mice are taken and stored in sterilized EP tubes in liquid nitrogen. Determination of the diversity of the intestinal flora, flora community, PCA and other analysis are carried out.

Embodiment 3

After the feeding experiment of the mice is finished, intes- tinal feces of the mice are taken for carrying out intestinal flo- ra dilution curve analysis. Compared with a blank group, the Sobs value of the model group is significantly higher than that of the blank group, indicating that depression can improve the diversity of the intestinal flora of mice. Compared with the model group, the diversity of intestinal flora in the medium-dose group of the aronia melanocarpa leaf polyphenol extract is significantly re- duced, indicating that an appropriate dose of aronia melanocarpa leaf polyphenol extract has a positive effect on reducing the di-

versity of intestinal flora of depressed mice.

Embodiment 4

And after the feeding experiment of the mice is finished, in- testinal feces of the mice are taken for carrying out Alpha diver- sity analysis of intestinal flora (see Table 1). Compared with a blank group, the Chao value and the Ace value of the model group decrease significantly, indicating that depression can reduce the richness of the intestinal flora of mice. However, the richness of the intestinal flora of the medicine group decreases significant- ly, indicating that medicines do not have a positive effect on the improvement of the richness of the intestinal flora of the mice.

Compared with the model group, the richness of the intestinal flo- ra of the low-dose group of the aronia melanocarpa leaf polyphenol extract obviously increases, indicating that appropriate dose of aronia melanocarpa leaf polyphenol extract has a positive effect on improving the richness of the intestinal flora of the depressed mice. The richness and diversity of the intestinal flora of the mice in the high-dose group of the aronia melanocarpa leaf poly- phenol extract are similar to those in the blank group, indicating that high dose of aronia melanocarpa leaf polyphenol extract can maintain the intestinal flora of the mice at a normal level.

Table 1 ALPHA diversity analysis of the effect of the aronia melanocarpa leaf polyphenol extract on intestinal flora of depressed mice

Richness Dwesty

Group “Ace Chao Shannon(107) Simpson(107) Sobs

D 38157:002° 383.97:006° 38.0%0.02° 05:0.01° 326.00:034°

M 380.11+0.37° 383.29+0.23° 38.4%0.04° 0.610.01° 354.00+0.38"

Y 350.29+0.343 349.60:0.07 37.00.045 0.140.071’ 316.00+0.34°

HGd 445.70+0.11° 451.28+0.01° 37.7+0.03' 0.6+0.01° 404.00+0.24°

HGz 415.63+0.03° 431.00:0.33° 41.640.02° 0.3+0.01° 378.00+0.24°

HGg 338.33+0.12° 358,78+0.20° 36.5:0.04" 0.640.01° 288.00£0.29"

HYd 426.7640.165 431.89+0.41 45.4+0.05° 0.3+0.01° 401.00£0.29"

HYz 273.69+0.15' 274.87+0.04% 29.7+0.03' 1.140.01° 203.00+0.18'

HYg 363.04+0.41 370.13:0.01" 38.6+0.04° 0.8+0.01° 322.00+0.35"

Note: compared with the blank group, *p<0.05. D: blank group;

M: model group; Y: medicine group; and HYd, HYz, HYg: the low-dose group, medium-dose group and high-dose group of the aronia melano- carpa leaf polyphenol extract.

Embodiment 5

After the feeding experiment of the mice is finished, intes- tinal feces of the mice are taken to determine and analyze the composition of intestinal flora community (see Table 2). The pro- portion of bacteroides in intestinal flora of the model mice is significantly higher than that of the blank group, and the propor- tions of bacteroides in the aronia melanocarpa leaf polyphenol ex- tract groups are all lower than that of the model group, indicat- ing that the aronia melanocarpa leaf polyphenol extract has a cer- tain inhibition effect on bacteroides. The proportion of firmicu- tes in intestinal flora of the model group is significantly lower than that of the blank group, and the proportions of firmicutes in the aronia melanocarpa leaf polyphenol extract groups are all higher than that in the model group, indicating that polyphenol compounds in aronia melanocarpa are beneficial to proliferation of firmicutes.

Table 2 Composition of horizontal community of intestinal flora of mice

D ossw0s o03wmo03

M 0.76£0.02° 0.17+0.01’

Y 0.69+0.02" 0.11+0.02

HGd 0.64+0.05 0.26+0.01"

HGz 0.72+0.03" 0.25+0.03°

HGg 0.71+0.02" 0.23+0.01°

Note: compared with the blank group, *p<0.05. D: blank group;

Embodiment 6

After the feeding experiment of the mice is finished, intes- tinal feces of the mice are taken for carrying out intestinal flo- ra PCA analysis. The difference between the blank group and other groups is mainly in the direction of PC2, which can explain the difference of 23.83%. The similarity among the blank group, the medicine group and the aronia melanocarpa leaf polyphenol extract groups is extremely high. These results indicate that the intesti- nal flora of the mice in the model group is out of order, the aro- nia melanccarpa leaf polyphenol extract can regulate and improve the intestinal flora of the depressed mice, and the high-dose group has the optimal regulation effect.

Embodiment 7

After the feeding experiment of the mice is finished, BDNF assay is carried out. After the mice are denecked and lethal, the skull is cut off, the brain of the mice is cut open with a dissec- tion blade, the hippocampus tissue located at the bottom of the cerebral cortex is taken out, a proper amount of concentrated SDS-

PAGE protein buffer solution is added into the treated sample for protein separation, and the voltage is 120 V. Membrane conversion is performed with a PVDF membrane, then washing is carried out twice, and sealing is carried out for overnight standing at 4 °C.

Incubating is carried out in the original diluent for 1h, washing is carried out for 5 min and 3 times, the second time of antibody incubating adopts the steps as above, and finally, exposing and developing are carried out.

The result shows that compared with the blank group, the BDNF level of the model group obviously decreases. The intestinal flora of the mice is out of order due to depression, and the change of the flora induces mental disorder, so that the BDNF expression in the brain of the mice decreases. Compared with the model group, the BDNF levels in the low-dose group and the high-dose group of the aronia melanocarpa leaf polyphenol extract significantly in- crease, indicating that appropriate dose of aronia melanocarpa leaf polyphenol extract is beneficial to release of BDNF.

Claims

CONCLUSIONS

The use of aronia melanocarpa leaf polyphenol extract in the preparation of medicines, functional foods or supplements for the prevention, regulation or treatment of depression, wherein the aronia melanocarpa leaf polyphenol extract consists of rutin, chlorogenic acid and isomer, 3-O-caffeoylquinic acid, durcetin and derivative quercetin 3-0-0-L arabinopyranosyl-{(1-+6}) β-D-glucopyranoside, caffeic acid and derivative hyperoside.

Use according to claim 1, wherein the administration dose of the polyphenol extract of aronia melanocarpa leaf is 15 to 25 ml/kg body weight.

The use according to claim 1, wherein the polyphenol extract of aronia melanocarpa leaf can effectively reduce the relative abundance of some harmful bacteria in intestinal microbes, regulate the intestinal microenvironment, and maintain the diversity and richness of the intestinal flora at a normal level.

The use according to claim 1, wherein the polyphenol extract of aronia melanocarpa leaf can regulate the flora structure of the intestinal microenvironment and significantly improve the BDNF expression level in the hippocampus of the brain of depressed mice.

Use of the polyphenol extract of aronia melanocarpa leaf according to claim 1, wherein the polyphenol extract of aronia melanocarpa leaf is used in antidepressants.

Use of the polyphenol extract of aronia melanocarpa leaf according to claim 1, wherein the polyphenol extract of aronia melanocarpa leaf is used in functional foods and supplements against depression.

Use according to any one of claims 5 to 6, wherein an acceptable carrier is added to the medicines, functional foods or supplements.

Use according to any one of claims 5 to 6, wherein an acceptable adjuvant material or adjuvant ingredient is added to the drugs, functional foods or supplements for depression.

Use according to any one of claims 5 to 6, wherein the dosage form of the drugs, functional foods or supplements for depression can be any pharmaceutical dosage form.