Application of pueraria flower flavone extract in preparation of drugs, health care products and functional food for relieving acute ethanol poisoning

Abstract

The invention discloses application of pueraria flower flavone extract in preparation of drugs, health care products and functional food for relieving acute ethanol poisoning, and belongs to the technical field of biology and new medicine. The invention discloses a pueraria flower flavone extract for relieving rat ataxia caused by acute ethanol. The pueraria flower flavone extract promotes the acute ethanol poisoning mice to recover sober; the pueraria flower flavone extract can improve the short-term spatial memory disorder of rats suffering from subacute ethanol mania. The pueraria flower flavone extract can be selected from medicines, health-care products and functional foods for relieving acute ethanol poisoning.

Description

Technical Field

[0001] The invention belongs to the field of biology and new medicine technology, and specifically relates to the application of pueraria lobata flavonoids extract in the preparation of medicines, health products and functional foods for alleviating acute ethanol poisoning. Background Art

[0002] Ethanol is one of the most abused drugs in the world, and its consumption trend is increasing day by day. The per capita ethanol consumption level is expected to increase from 6.5 liters in 2017 to 7.6 liters in 2030. Ethanol abuse includes acute poisoning, dependence, alcoholism, etc. Among them, acute poisoning is the most common symptom for medical treatment due to ethanol abuse. It often occurs after a large amount of ethanol is consumed in a short period of time, and the nervous system is first excited and then inhibited. In severe cases, it is life-threatening.

[0003] With the rapid development of China's economy, the growth rate of ethanol consumption has exceeded that of other parts of the world. Correspondingly, the health and public safety problems caused by ethanol abuse have also increased. At present, there is a lack of effective data on the epidemiological research of acute ethanol poisoning nationwide. A survey of a large hospital in Putuo District, Shanghai showed that from 2014 to 2018, the proportion of patients with acute ethanol poisoning increased from 0.62‰ to 3.07‰, with male patients accounting for 71.4%, and the age range was concentrated between 20 and 39 years old; in addition, acute ethanol poisoning also occurs in adolescents. In 2018, the incidence rate of the student population in the hospital increased by 566.7% compared with 2014. In summary, the situation of acute ethanol poisoning is becoming increasingly serious.

[0004] Naloxone is an opioid receptor antagonist. In 1978, it was clinically reported that it could reverse coma caused by ethanol poisoning. However, in a foreign double-blind clinical trial, there was no significant difference in consciousness level between the naloxone and placebo groups. Currently, naloxone is widely used in the awakening treatment of acute ethanol poisoning in clinical practice in my country. It is considered to be safe and can be used in combination with Chinese patent medicines. The administration method is intravenous injection, but the effect lasts for a short time.

[0005] Metadoxine is composed of pyroglutamic acid and vitamin B6, and is clinically used to treat acute ethanol poisoning and alcoholic fatty liver. Clinical double-blind studies have shown that intravenous injection of metadoxine can accelerate the elimination of ethanol from the blood and promote patient sobriety. Experimental studies have shown that metadoxine can increase the activity of two enzymes, alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) in the liver of mice with acute ethanol poisoning, and reduce blood ethanol concentration, but in vitro experiments have found that it cannot activate any subtype of ADH or ALDH. At the same time, studies have also found that metadoxine can increase the mortality rate and the rate of righting reflex loss in mice with acute ethanol poisoning.

[0006] Pueraria flower, also known as kudzu flower, is the dried flower of Pueraria lobata (Willd.) Ohwi. or P. thomsonii Benth. of the Leguminosae family. It was first recorded in the "Famous Doctors' Records" and has the effects of sobering up, invigorating the spleen, and stopping bleeding. It is mainly used to treat alcohol-induced fever, thirst, headache, dizziness, abdominal distension, vomiting, acid reflux, etc. Pueraria flower has been recorded as a special medicine for sobering up in the herbal medicine of all dynasties. Modern research shows that Pueraria flower has a therapeutic effect on acute and chronic ethanol poisoning, myocardial injury, diabetic complications, and central nervous system diseases.

[0007] Up to now, acute ethanol poisoning can be treated with ethanol clearance and awakening-promoting drugs such as metadoxine or naloxone, but they have limitations such as short duration of action and many contraindications. Therefore, it is urgent to develop a safer substance to alleviate the above problems. Summary of the invention

[0008] In order to solve the above problems existing in the prior art, the present invention finds through in-depth research that flavonoid extract from Puerariae Flos (PFF) can improve acute ethanol poisoning.

[0009] The invention provides application of PFF in preparing medicines, health products or functional foods for alleviating acute ethanol poisoning.

[0010] Furthermore, the PFF is a PFF with a mass fraction of flavonoids greater than or equal to 60.29%.

[0011] The present invention provides a pharmaceutical composition, comprising the PFF and pharmaceutically acceptable excipients.

[0012] The invention provides application of the pharmaceutical composition in preparing a medicine for alleviating acute ethanol poisoning.

[0013] The present invention provides a health product or functional food composition, comprising the PFF and acceptable auxiliary materials.

[0014] The present invention provides application of the health care product or functional food composition in preparing the health care product or functional food for alleviating acute ethanol poisoning.

[0015] Furthermore, the dosage form of the drug is an oral dosage form.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] The PFF (mass fraction of flavonoids greater than or equal to 60.29%) in the present invention can alleviate acute ethanol poisoning. The present invention has confirmed through animal experiments that: PFF can improve ataxia caused by acute ethanol; PFF can promote acute ethanol poisoning mice to recover sobriety; PFF can improve short-term spatial memory impairment in subacute ethanol binge drinking rats. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The quantitative analysis of PFF in Example 1 of the present invention is as follows: Figure 1 (A) is a high performance liquid chromatography diagram of the mixed standard; Figure 1 (B) is a high performance liquid chromatography diagram of PFF; Figure 1 (C) is the standard curve of rutin;

[0019] Figure 2 This is a diagram showing that PFF improves acute ethanol-induced ataxia in rats in Example 2 of the present invention, wherein Figure 2 (A) Graph showing the increase in rotarod residence time in rats with acute ethanol-induced ataxia due to PFF; Figure 2 (B) Graph showing the increase in PFF between the front paws of rats with acute ethanol-induced ataxia;

[0020] Figure 3 This is a diagram showing that PFF promotes the recovery of acute ethanol poisoning mice in Example 3 of the present invention, wherein Figure 3 (A) Graph showing the latency of righting reflex loss in mice with acute ethanol intoxication after PFF increase; Figure 3 (B) Schematic diagram showing that PFF shortens the duration of loss of righting reflex in mice with acute ethanol intoxication;

[0021] Figure 4 This is a diagram showing that PFF improves short-term spatial memory impairment in subacute ethanol binge drinking rats in Example 4 of the present invention, wherein Figure 4 (A) Graph showing the displacement object discrimination index of subacute ethanol binge drinking rats with increased PFF; Figure 4 (B) Graph showing the percentage of time spent in the novel arm increased by PFF in subacute ethanol binge drinking rats; DETAILED DESCRIPTION

[0022] Example 1: Preparation and quantitative analysis of PFF

[0023] Weigh 1500g of dried kudzu flower, place in a round-bottom flask, add 10L of petroleum ether, heat and reflux for 2h, filter and discard the yellow-green filtrate after cooling, repeat this operation 6 times, and the last petroleum ether filtrate is light yellow. Add 10L of 70% ethanol to the kudzu flower residue, heat and reflux for 2h, filter and collect the filtrate after cooling, repeat this operation 5 times, combine the extracts and distill under reduced pressure to obtain 443.4g of kudzu flower ethanol extract (yield: 29.56%). Use 2L of distilled water to suspend the above extract, add 3L of water-saturated n-butanol for extraction, repeat this operation 5 times, combine the n-butanol extracts, and distill under reduced pressure to obtain 358g of kudzu flower n-butanol extract (yield: 25.67%). The extract was separated by macroporous adsorption resin and gradient eluted with water, 30% ethanol, 60% ethanol and 90% ethanol respectively. Finally, the 60% and 90% ethanol elution products were combined (yield: 11%) and freeze-dried to finally obtain light brown powder PFF.

[0024] The above ethanol concentrations are all volume concentrations.

[0025] PFF was dissolved in methanol solution for high performance liquid chromatography analysis. The analysis process used mobile phase gradient treatment, the injection volume was 10 μL, and the chromatographic conditions were as follows:

[0026] Chromatographic column YMC-Pack, 5μm, C18, 250×4.6mm

[0027] DetectorOD265 UV detector

[0028] Mobile phase 1‰Formic acid:Acetonitrile

[0029] Flow rate 0.8mL / min

[0030] The mobile phase conditions were as follows:

[0031] 0-20 minutes 1‰ Formic acid: acetonitrile = 86%: 14%

[0032] 30 minutes 1‰ Formic acid: acetonitrile = 75%: 25%

[0033] 40 minutes 1‰ Formic acid: acetonitrile = 72%: 28%

[0034] 50 minutes 1‰ Formic acid: acetonitrile = 70%: 30%

[0035] 60 minutes 1‰ Formic acid: acetonitrile = 60%: 40%

[0036] 80 minutes 1‰ Formic acid: acetonitrile = 0%: 100%

[0037] The above 1‰ formic acid is a volume concentration, and the ratio of 1‰ formic acid to acetonitrile is a volume ratio.

[0038] like Figure 1 As shown in (AB), the peaks marked 1-4 are tectoriusin-7-oxo-xylosyl glucoside, genistin, tectoriusin, and tectoriusin, respectively, and the four compound standards were effectively separated within 80 minutes. The PFF also contains the above four compounds; Figure 1 As shown in (C), the rutin standard curve has good linearity, and the flavonoid content in PFF was measured to be 60.29%.

[0039] Example 2: PFF improves acute ethanol-induced ataxia in rats

[0040] Rats were randomly divided into 7 groups, blank control group, ethanol group, ethanol + PFF (25, 50, 100, 200, 400 mg / kg) group. Rats in the blank control group and ethanol group were intragastrically administered with 0.5% CMC-Na for 3 consecutive days, and rats in the ethanol + PFF groups were intragastrically administered with PFF 25, 50, 100, 200, 400 mg / kg for 3 consecutive days. One hour after the last administration, rats in the ethanol group and ethanol + PFF groups were intraperitoneally injected with 20% (w / v) ethanol 2g / kg, and rats in the blank control group were intraperitoneally injected with an equal volume of normal saline. Rotarod and gait behavior experiments were performed 60 minutes later.

[0041] like Figure 2 As shown in (AB), compared with the blank control group, 60 minutes after ethanol injection, the rotarod residence time and the distance between the front paws of the rats in the ethanol group were significantly reduced; compared with the ethanol group, the rotarod residence time and the distance between the front paws of the rats in the PFF (25 mg / kg) and PFF (400 mg / kg) groups were significantly increased. This suggests that PFF (25, 400 mg / kg) can significantly improve acute ethanol-induced ataxia in rats.

[0042] Example 3: PFF promotes recovery of acute ethanol intoxication mice

[0043] Mice were randomly divided into 6 groups, ethanol group, ethanol + PFF (25, 50, 100, 200, 400 mg / kg) group. Mice in the ethanol group were given 0.5% CMC-Na by gavage for 3 consecutive days, and mice in the ethanol + PFF group were given PFF 25, 50, 100, 200, 400 mg / kg by gavage for 3 consecutive days. One hour after the last administration, mice in each group that had fasted for 12 hours were given 30% (w / v) ethanol 6 g / kg by gavage, and then behavioral tests were performed.

[0044] like Figure 3 As shown in (A), PFF increased the latency of loss of righting reflex in mice with acute ethanol intoxication; Figure 3(B) As shown, PFF reduces the duration of loss of righting reflex in mice with acute ethanol intoxication;

[0045] Example 4: PFF improves short-term spatial memory impairment in rats with subacute ethanol binge drinking

[0046] The rats were randomly divided into 7 groups: blank control group, ethanol group, and ethanol+PFF (25, 50, 100, 200, 400 mg / kg) group. Rats in the blank control group and ethanol group were intragastrically administered with 0.5% CMC-Na for 14 consecutive days. Rats in the ethanol+PFF group were intragastrically administered with PFF 25, 50, 100, 200, and 400 mg / kg for 14 consecutive days. One hour after the last administration, the rats in the ethanol group were intragastrically administered with ethanol ansu solution (25% w / v, 50% v / v) once every 8 hours, 3 times a day, for 4 days. The first dose of ethanol was 5 g / kg (25% w / v), and the subsequent dose of ethanol was determined according to the behavioral scores of the rats, 0 points -5 g / kg, 1 point -4 g / kg, 2 points -3 g / kg, 3 points -2 g / kg, 4 points -1 g / kg, 5 points -0 g / kg. Rats in the ethanol+PFF groups were intragastrically administered with the average dose of ethanol in the ethanol group, 3 times a day, for 4 days. The blank control group was given isocaloric dextrin ansu solution (35% w / v, 50% v / v). The object displacement memory experiment was conducted on the 13th day, and the spatial novelty preference experiment was conducted on the 14th day.

[0047] like Figure 4 As shown in (A), PFF increased the displacement object discrimination index in rats with subacute ethanol binge drinking; Figure 4 (B) As shown, PFF increased the percentage of time spent in the novel arm in rats subjected to subacute ethanol binge drinking.

Claims

1. Application of kudzu flower flavonoids extract in the preparation of drugs, health products and functional foods for relieving acute ethanol poisoning.

2. The use according to claim 1, characterized in that: The flavonoids content in the kudzu flower flavonoids extract is greater than or equal to 60.29% by mass.

3. A pharmaceutical and / or food composition, characterized in that The invention comprises pueraria flower flavonoids extract and auxiliary materials acceptable in medicine and / or food.

4. Use of the medicine and / or food composition according to claim 3 in the preparation of medicines, health products or functional foods for alleviating acute ethanol poisoning.

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