Application of Pinellia ternata root extract in the preparation of drugs for treating and / or preventing liver injury

Abstract

This invention relates to the field of traditional Chinese medicine extracts, and particularly to the application of *Pinellia ternata* root extract in the preparation of drugs for treating and / or preventing liver injury. The *Pinellia ternata* root extract is an aqueous extract of *Pinellia ternata* root, and the extraction methods include decoction extraction or reflux extraction. This invention has found that the aqueous extract of *Pinellia ternata* root can inhibit carbon tetrachloride-induced increases in AST and ALT, and reduce carbon tetrachloride-induced production of TNF-α, IL-β, and IL-6, exhibiting good protective effects against carbon tetrachloride-induced chemical liver injury and alcoholic liver injury.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine extract technology, and in particular to the application of Pinellia ternata root extract in the preparation of drugs for treating and / or preventing liver damage. Background Technology

[0002] The liver is one of the most important core organs in the human body, primarily responsible for metabolic functions, and plays an irreplaceable role in maintaining homeostasis and vital activities. Therefore, research on the mechanisms of liver protection and the development of related drugs has significant clinical value and practical implications. Liver injury is a group of diseases caused by various pathogenic factors such as drugs, alcohol, chemical toxins, and viral infections, resulting in abnormal liver function and structural damage. If it continues to progress, it can develop into liver fibrosis, cirrhosis, and even liver cancer, seriously threatening human health.

[0003] In the classic model of chemical liver injury, carbon tetrachloride is metabolized by hepatic cytochrome P450 hydroxylase to generate highly reactive trichloromethyl radicals (·CCl3), which in turn trigger a chain of lipid peroxidation reactions, disrupting cell membrane permeability and fluidity, leading to hepatocyte necrosis and apoptosis. The main pathological mechanism of alcoholic liver injury is as follows: ethanol is oxidized in the liver through three pathways: alcohol dehydrogenase, the hepatic microsomal ethanol oxidation system, and catalase, to produce acetaldehyde. Acetaldehyde can directly damage intracellular enzyme systems and organelle functions, while simultaneously inducing oxidative stress and immune inflammatory responses, ultimately leading to liver damage.

[0004] *Elaeagnus multiflora* Thunb., a deciduous shrub belonging to the genus *Elaeagnus* in the family Elaeagnaceae, is rich in chemical components, mainly including alkaloids, fatty acids, volatile oils, flavonoids, aromatic phenolic acids, terpenes, and steroidal compounds. Traditional Chinese medicine theory holds that different medicinal parts of *Elaeagnus multiflora* have different effects: the root bark has the effects of promoting qi and blood circulation, stopping diarrhea, and astringing sores; the leaves can relieve asthma and promote blood circulation. However, to date, there are few reports, both domestically and internationally, on systematic modern pharmacological studies using *Elaeagnus multiflora* root extract as a single active ingredient to target chemical or alcoholic liver injury. Therefore, developing *Elaeagnus multiflora* root extract and applying it to the preparation of drugs for the treatment and prevention of chemical and alcoholic liver injury has significant theoretical importance and broad application prospects. Summary of the Invention

[0005] The purpose of this invention is to provide the application of Pinellia ternata root extract in the preparation of drugs for treating and / or preventing liver damage. The Pinellia ternata root extract prepared by this invention can reduce indicators of carbon tetrachloride-induced or alcoholic liver damage and repair pathological damage to liver tissue.

[0006] To achieve the above objectives, the present invention provides the application of Pinellia ternata root extract in the preparation of drugs for treating and / or preventing liver injury, wherein the Pinellia ternata root extract is a water extract of Pinellia ternata root, and the water extraction method includes decoction extraction or reflux extraction.

[0007] Preferably, the concentration of the aqueous extract of Pinellia ternata root is 1.5-2.5 mg / L.

[0008] In an optional embodiment of the present invention, the concentration of the aqueous extract of Pinellia ternata root is 2 mg / L.

[0009] Preferably, when the water extraction method is decoction extraction, it includes the following steps: placing the root of Pinellia ternata in water, decoct and extract, concentrating the extract to obtain water extract of Pinellia ternata root.

[0010] Preferably, during the decoction extraction, the root of *Pinellia ternata* is in the form of flakes, segments, or powder; the mass ratio of the root of *Pinellia ternata* to water is 1:6-20.

[0011] In an optional embodiment of the present invention, during the decoction extraction, the mass ratio of the root of Pinellia ternata to water is 1:10.

[0012] Preferably, the decoction extraction is performed 1-4 times.

[0013] In an optional embodiment of the present invention, when the decoction extraction is performed twice, the preparation process includes: (1) placing the root of *Pinellia ternata* in water, decocting and extracting, filtering for the first time to obtain filtrate A and residue A; (2) placing residue A in water, decocting and extracting, filtering for the second time to obtain filtrate B and residue B; (3) mixing filtrate A and filtrate B to obtain an extract, concentrating the extract to obtain an aqueous extract of *Pinellia ternata* root. The mass ratio of residue A to water in step (2) is 1:6-20.

[0014] In an optional embodiment of the present invention, the mass ratio of filter residue A to water in step (2) is 1:8.

[0015] Preferably, the single decoction extraction process includes sequential boiling over high heat and simmering over low heat, with each simmering session lasting 0.5-4 hours.

[0016] In one optional embodiment of the present invention, the simmering time for a single simmering is 1-1.5 hours.

[0017] Preferably, when the water extraction method is reflux extraction, it includes the following steps: placing the root of Pinellia ternata in water, reflux extraction, concentrating the extract, and obtaining the water extract of Pinellia ternata root.

[0018] Preferably, during the decoction extraction, the root of *Pinellia ternata* is in the form of flakes, segments, or powder; the mass ratio of the root of *Pinellia ternata* to water is 1:6-20.

[0019] In an optional embodiment of the present invention, during the reflux extraction, the mass ratio of the root of Pinellia ternata to water is 1:10.

[0020] Preferably, the reflux extraction is performed 1-4 times.

[0021] In an optional embodiment of the present invention, when the reflux extraction is performed twice, the preparation process includes: S1, placing the root of *Pinellia ternata* in water, reflux extraction, and initial filtration to obtain filtrate 1 and residue 1; S2, placing residue 1 in water, reflux extraction, and secondary filtration to obtain filtrate 2 and residue 2; S3, mixing filtrate 1 and filtrate 2 to obtain an extract, concentrating the extract to obtain an aqueous extract of *Pinellia ternata* root. The mass ratio of residue 1 to water in S2 is 1:6-20.

[0022] In an optional embodiment of the present invention, the mass ratio of filter residue 1 to water in S2 is 1:8.

[0023] Preferably, the temperature for a single reflux extraction is 50-100℃, and the extraction time is 0.5-4h.

[0024] In one optional embodiment of the present invention, the temperature of a single reflux extraction is 50°C, and the time of a single reflux extraction is 1-1.5 h.

[0025] Preferably, the liver injury is alcoholic liver injury or carbon tetrachloride-induced liver injury.

[0026] The present invention has the following beneficial effects: This invention provides the application of *Pinellia ternata* root extract in the preparation of drugs for treating and / or preventing liver injury. The *Pinellia ternata* root extract is an aqueous extract of *Pinellia ternata* root, and the aqueous extraction method includes decoction extraction or reflux extraction. This invention uses only *Pinellia ternata* root as raw material, without the addition of other medicinal materials, simplifying the selection of raw materials. Furthermore, the aqueous extraction method is simple.

[0027] Aqueous extracts of Pinellia ternata root can inhibit the increase of AST and ALT induced by carbon tetrachloride, and reduce the production of TNF-α, IL-β and IL-6 induced by carbon tetrachloride. It has a good protective effect against carbon tetrachloride-induced liver injury and alcoholic liver injury.

[0028] This invention breaks through the traditional medicinal limitations of water extract of Pinellia ternata root in promoting qi circulation, activating blood circulation, and stopping diarrhea, and opens up a new pharmaceutical application for it in the preparation of drugs for anti-chemical liver damage and alcoholic liver damage, providing a new source of natural raw materials for liver-protecting drugs. Attached Figure Description

[0029] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0030] Figure 1 This is a bar chart showing the effect of the aqueous extract of Pinellia ternata root prepared in Example 1 of the present invention on serum ALT and AST in rats with carbon tetrachloride-induced liver injury. in, Figure 1 In the figure, 'a' represents the bar chart showing the effect of water extract from Pinellia ternata root on ALT. Figure 1 In the figure, b is a bar chart showing the effect of water extract of Pinellia ternata root on AST; Figure 2 Histopathological section of liver tissue in a rat model of acute liver injury induced by carbon tetrachloride; in, Figure 2 A in the figure represents the blank control group. Figure 2 B in the model group is the model group. Figure 2 C in the sample represents the high-level group of Pinellia ternata root extract. Figure 2 D in the figure represents the level group in the root extract of Pinellia ternata. Figure 2 E in the sample represents the low-level group of Pinellia ternata root extract. Figure 2 The F in the text represents the liver-protecting tablet group; Figure 3 The bar chart shows the effect of the aqueous extract of Pinellia ternata root prepared in Example 1 on inflammatory factors of carbon tetrachloride-induced liver injury. in, Figure 3 In the figure, 'a' represents the bar chart showing the effect of water extract of Pinellia ternata root on TNF-α. Figure 3 In the figure, b is a bar chart showing the effect of water extract of Pinellia ternata root on IL-β. Figure 3 In the figure, c represents the bar chart showing the effect of water extract of Pinellia ternata root on IL-6. Detailed Implementation

[0031] The present invention will be further described below with reference to the accompanying drawings and embodiments. Unless otherwise defined, the technical or scientific terms used in this invention should be understood in their ordinary sense by those skilled in the art. The features mentioned above or in the specific examples mentioned in this invention can be combined arbitrarily, and these specific embodiments are only used to illustrate the invention and are not intended to limit the scope of the invention.

[0032] Experimental materials: healthy SD rats, carbon tetrachloride (Tianjin Guangfu Technology Development Co., Ltd.), peanut oil (Shandong Luhua Group Co., Ltd.), ALT and AST reagent kits (Nanjing Jiancheng Bioengineering Institute), eosin and hematoxylin (Zhuhai Beso Biotechnology Co., Ltd.).

[0033] Example 1 (1) Place 700g of segmented Pinellia root in 7kg of water, bring to a boil over high heat and then simmer over low heat for 1.5h. After initial filtration with gauze, obtain residue A and filtrate A. (2) Add water to filter residue A (the mass ratio of filter residue A to water is 1:8), bring to a boil over high heat and then simmer over low heat for 1 hour. Filter the residue twice with gauze to obtain filter residue B and filtrate B. (3) After mixing filtrate A and filtrate B, the extract is obtained. The extract is concentrated in a rotary evaporator to obtain a water extract of Pinellia ternata root with a concentration of 2 g / mL.

[0034] Example 2 S1. Place 700g of thin slices of Pinellia ternata root in 7kg of water and reflux at 50℃ for 1.5h. After initial filtration with gauze, filter residue 1 and filtrate 1 are obtained. S2. Add water to filter residue 1 (the mass ratio of filter residue 1 to water is 1:8), reflux at 50°C for 1 hour, and filter twice with gauze to obtain filter residue 2 and filtrate 2. S3. After mixing filtrate 1 and filtrate 2, the extract is obtained. The extract is concentrated in a rotary evaporator to obtain a water extract of Pinellia ternata root with a concentration of 2 g / mL.

[0035] Test example: Establishing a carbon tetrachloride-induced acute liver injury model in rats: Healthy adult male SD rats, weighing 230-250g, were acclimatized for one week, with free access to food and water, and a 12-hour day-night cycle. They were randomly divided into six groups: a blank control group (Ctrl), a model group (CCl4), groups with high (H-MB-CCl4), medium (M-MB-CCl4), and low (L-MB-CCl4) levels of the water extract of *Pinellia ternata* root prepared in Example 1 (40g / kg, 20g / kg, 10g / kg), and a liver-protecting tablet group (HGP-CCl4, 3.5g / kg, Heilongjiang Sunflower Pharmaceutical Co., Ltd.). The rats were administered the drug by gavage for 14 consecutive days. The blank control group and the model group were administered the same volume of ddH2O by gavage. One hour after the last administration, the rats in the other groups were intraperitoneally injected with a 50% carbon tetrachloride peanut oil solution at a dose of 2mL / kg, while the blank control group received an equal volume of peanut oil. 24 hours later, blood was collected from the abdominal aorta of rats in each group, and serum was separated; liver tissue was collected and preserved in tissue fixative.

[0036] Indicator detection: Following the operating procedures of the ALT and AST kits (Nanjing Jiancheng Bioengineering Institute), the activities of AST and ALT in serum were detected; TNF-α, IL-β, and IL-6 were detected using the ELISA kits (Jiangsu Enzyme Immunoassay Co., Ltd.).

[0037] Liver tissue processing: Liver tissue was dehydrated, embedded, sectioned, dewaxed, and stained to obtain pathological sections. After mounting, the pathological changes of liver tissue in each group were observed under an optical microscope.

[0038] The effects of the aqueous extract of Pinellia ternata root prepared in Example 1 of this invention on carbon tetrachloride-induced serum AST and ALT levels in rats are as follows: Figure 1 As shown. By Figure 1 It can be seen that, compared with the model group, the serum ALT and AST levels in each treatment group were significantly reduced. P <0.05, P The result was <0.01, indicating that the aqueous extract of Pinellia ternata root has a protective effect against carbon tetrachloride-induced chemical liver injury in rats.

[0039] The results of the aqueous extract of Pinellia ternata root prepared in Example 1 of this invention on the pathological changes of rat liver tissue induced by carbon tetrachloride are as follows: Figure 2 As shown. By Figure 2 It is evident that, compared with the blank control group, the liver tissue pathological damage of rats in the model group was aggravated, the liver lobule structure was severely destroyed, and vacuoles and inflammatory cells appeared. Compared with the model group, the liver tissue damage of rats in each treatment group was significantly reduced, which indicates that the water extract of Pinellia ternata root prepared in Example 1 alleviated the chemical liver tissue damage induced by carbon tetrachloride.

[0040] The effect of the aqueous extract of Pinellia ternata root prepared in Example 1 of this invention on inflammatory factors of carbon tetrachloride-induced liver injury is as follows: Figure 3 As shown. By Figure 3 It can be seen that, compared with the model group, the levels of liver injury inflammatory factors (TNF-α, IL-β, IL-6) in each treatment group were significantly reduced, indicating that the aqueous extract of Pinellia ternata root can inhibit the production of inflammatory factors TNF-α, IL-β, and IL-6. This suggests that the aqueous extract of Pinellia ternata root has a protective effect against carbon tetrachloride-induced chemical liver injury in rats.

[0041] Alcohol-induced rat liver injury model test Healthy adult male SD rats, weighing 200-220g, were acclimatized for one week with free access to food and water, and a 12-hour diurnal cycle. They were randomly divided into a model group, high, medium, and low dose groups of *Pinellia ternata* root (40g / kg, 20g / kg, and 10g / kg), a blank control group, and a liver-protecting tablet group (3.5g / kg). Administered the medication by gavage for 14 consecutive days. From day 15 onwards, they were administered 50% ethanol by gavage at a dose of 5g / kg every 12 hours for three consecutive times. The blank control group received an equal volume of ultrapure water by gavage.

[0042] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. The application of Pinellia ternata root extract in the preparation of drugs for treating and / or preventing liver injury, characterized in that, The root extract of Pinellia ternata is an aqueous extract of Pinellia ternata root, and the aqueous extraction method includes decoction extraction or reflux extraction.

2. The use of the Pinellia ternata root extract according to claim 1 in the preparation of drugs for treating and / or preventing liver injury, characterized in that, The concentration of the aqueous extract of Pinellia ternata root is 1.5-2.5 mg / L.

3. The use of the Pinellia ternata root extract according to claim 2 in the preparation of drugs for treating and / or preventing liver injury, characterized in that, When the water extraction method is decoction extraction, it includes the following steps: The root of Pinellia ternata was placed in water, decocted and extracted, and the extract was concentrated to obtain the water extract of Pinellia ternata root.

4. The use of the Pinellia ternata root extract according to claim 3 in the preparation of drugs for treating and / or preventing liver injury, characterized in that, During the decoction extraction, the root of Pinellia ternata is in the form of flakes, segments, or powder. The mass ratio of the root of Pinellia ternata to water is 1:6-20.

5. The use of the root extract of Pinellia ternata according to claim 3 in the preparation of drugs for treating and / or preventing liver injury, characterized in that, The decoction extraction is performed 1-4 times. Each decoction extraction process includes boiling over high heat and simmering over low heat in sequence, with each simmering time being 0.5-4 hours.

6. The use of the root extract of Pinellia ternata according to claim 2 in the preparation of drugs for treating and / or preventing liver injury, characterized in that, When the water extraction method is reflux extraction, it includes the following steps: The root of Pinellia ternata was placed in water and refluxed for extraction. The extract was then concentrated to obtain an aqueous extract of Pinellia ternata root.

7. The use of the root extract of Pinellia ternata according to claim 6 in the preparation of drugs for treating and / or preventing liver injury, characterized in that, During the reflux extraction, the root of Pinellia ternata is in the form of flakes, segments, or powder. The mass ratio of the root of Pinellia ternata to water is 1:6-20.

8. The use of the root extract of Pinellia ternata according to claim 6 in the preparation of drugs for treating and / or preventing liver injury, characterized in that, The reflux extraction is performed 1-4 times; The temperature for a single reflux extraction is 50-100℃, and the extraction time is 0.5-4h.

9. The use of the root extract of Pinellia ternata according to claim 1 in the preparation of drugs for treating and / or preventing liver injury, characterized in that, The liver injury referred to includes alcoholic liver injury and carbon tetrachloride-induced liver injury.

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