A traditional Chinese medicine composition for resisting liver ischemia-reperfusion injury and application thereof

Abstract

The application provides a traditional Chinese medicine composition resisting liver ischemia-reperfusion injury and application thereof, and belongs to the technical field of traditional Chinese medicine compositions; the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 1 part of liquorice, 1 part of salvia miltiorrhiza and 1 part of white peony root. In the application, the salvia miltiorrhiza has the effects of activating blood circulation to dissipate blood stasis, cooling blood to relieve pain and resisting oxidation; the white peony root can nourish blood and soothe liver (tonifying blood and nourishing liver) and inhibit inflammatory reaction; the liquorice has the functions of detoxification and regulating drug properties; the salvia miltiorrhiza, the white peony root and the liquorice have the effects of cooling blood to dissipate blood stasis, clearing heat to detoxify and tonifying blood to nourish liver in cooperation, and can effectively resist liver ischemia-reperfusion injury.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of traditional Chinese medicine compositions, and particularly relates to a traditional Chinese medicine composition for resisting liver ischemia-reperfusion injury and application thereof. BACKGROUND

[0002] Liver ischemia-reperfusion injury is a common secondary injury in pathological processes such as liver surgery (e.g., liver transplantation, liver resection), shock, etc., and can lead to liver failure and even multiple organ dysfunction. At present, there is a lack of specific therapeutic drugs in clinical practice, and the symptoms are mainly relieved by using antioxidants (e.g., N-acetylcysteine) or anti-inflammatory drugs, but the efficacy is limited. SUMMARY

[0003] The application aims to provide a traditional Chinese medicine composition for resisting liver ischemia-reperfusion injury and application thereof, and the traditional Chinese medicine composition has a good protective effect on liver ischemia-reperfusion injury.

[0004] The application provides a traditional Chinese medicine composition for resisting liver ischemia-reperfusion injury, which comprises the following raw materials in parts by weight: 1 part of liquorice, 1 part of salvia miltiorrhiza and 1 part of white peony root.

[0005] Preferably, the traditional Chinese medicine composition is composed of the following raw materials in parts by weight: 1 part of liquorice, 1 part of salvia miltiorrhiza and 1 part of white peony root; the salvia miltiorrhiza comprises salvia miltiorrhiza decoction pieces.

[0006] The application further provides a preparation method of the traditional Chinese medicine composition described in the above scheme, comprising the following steps:

[0007] The liquorice, salvia miltiorrhiza and white peony root are respectively subjected to water extraction to obtain liquorice water extract, salvia miltiorrhiza water extract and white peony root water extract; and the liquorice water extract, salvia miltiorrhiza water extract and white peony root water extract are mixed to obtain the traditional Chinese medicine composition.

[0008] Preferably, the water extraction of the liquorice, salvia miltiorrhiza and white peony root respectively comprises:

[0009] The liquorice, salvia miltiorrhiza and white peony root are respectively mixed with water, and are sequentially subjected to soaking and first water extraction to collect solid components and first water extract liquid respectively; the solid components are mixed with water, and are subjected to second water extraction to collect second water extract liquid; the first water extract liquid and second water extract liquid are mixed to obtain total water extract liquid; and the total water extract liquid is sequentially subjected to concentration and drying to obtain liquorice water extract, salvia miltiorrhiza water extract and white peony root water extract respectively.

[0010] Preferably, the temperature of the soaking is 20-30℃; the time of the soaking is 28-32 min; the temperature of the first water extraction and second water extraction is 95-100℃ respectively; the time of the first water extraction is 1-2 h; and the time of the second water extraction is 0.8-1.2 h.

[0011] Preferably, the drying temperature is 48~52℃; and the drying time is greater than or equal to 12 h.

[0012] The present invention also provides the application of the traditional Chinese medicine composition described in the above-described scheme or the traditional Chinese medicine composition prepared by the above-described preparation method in the preparation of drugs for treating hepatic ischemia-reperfusion injury.

[0013] Preferably, the liver ischemia-reperfusion injury includes liver transplant ischemia-reperfusion injury.

[0014] Preferably, the causes of liver ischemia include hepatic artery occlusion, portal vein occlusion, and bile duct atresia.

[0015] The present invention also provides a drug for treating hepatic ischemia-reperfusion injury, comprising a traditional Chinese medicine composition and pharmaceutically acceptable excipients; the traditional Chinese medicine composition comprises the traditional Chinese medicine composition described in the above scheme or the traditional Chinese medicine composition prepared by the above preparation method.

[0016] This invention provides a traditional Chinese medicine composition for combating hepatic ischemia-reperfusion injury, comprising the following raw materials in parts by weight: 1 part licorice root, 1 part salvia miltiorrhiza root, and 1 part white peony root. In this invention, salvia miltiorrhiza root has the effects of promoting blood circulation, removing blood stasis, cooling blood, relieving pain, and antioxidation; white peony root can nourish blood and soften the liver (tonify blood and nourish the liver) and inhibit inflammatory responses; licorice root has both detoxification and harmonizing medicinal properties; salvia miltiorrhiza root, white peony root, and licorice root work synergistically to have the effects of cooling blood, removing blood stasis, clearing heat and detoxifying, tonifying blood and nourishing the liver, effectively combating hepatic ischemia-reperfusion injury. Attached Figure Description

[0017] 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.

[0018] Figure 1 The figure shows the results of the combined use of Danshen, Licorice, and White Peony to protect the liver from ischemia-reperfusion in mice; where A represents serum ALT activity and B represents serum AST activity. One-way ANOVA and Tukey post-test analysis were used to compare statistical differences between the groups. Compared with the HIRI model group... * P <0.05, ** P <0.01, *** P <0.001;

[0019] Figure 2 The results of HE staining of liver sections. Detailed Implementation

[0020] The application provides a traditional Chinese medicine composition for resisting liver ischemia-reperfusion injury, which comprises the following raw materials in parts by weight: 1 part of licorice, 1 part of salvia miltiorrhiza and 1 part of white peony root.

[0021] As an implementation form, the traditional Chinese medicine composition is composed of the following raw materials in parts by weight: 1 part of licorice, 1 part of salvia miltiorrhiza and 1 part of white peony root; the salvia miltiorrhiza comprises salvia miltiorrhiza decoction pieces; the licorice comprises licorice decoction pieces; and the white peony root comprises white peony root decoction pieces.

[0022] In the application, the salvia miltiorrhiza has the effects of activating blood circulation to dissipate blood stasis, cooling blood to relieve pain and resisting oxidation; the white peony root can nourish blood and soothe liver (tonifying blood and nourishing liver) and inhibit inflammatory reaction; the licorice has the functions of detoxification and regulating drug properties; the salvia miltiorrhiza, the white peony root and the licorice have the effects of cooling blood to dissipate blood stasis, clearing heat to detoxify and tonifying blood to nourish liver in cooperation, and can effectively resist liver ischemia-reperfusion injury.

[0023] The application further provides a preparation method of the traditional Chinese medicine composition in the above-mentioned scheme, comprising the following steps:

[0024] The licorice, the salvia miltiorrhiza and the white peony root are respectively subjected to water extraction to obtain licorice water extract, salvia miltiorrhiza water extract and white peony root water extract respectively;

[0025] The licorice water extract, the salvia miltiorrhiza water extract and the white peony root water extract are mixed to obtain the traditional Chinese medicine composition.

[0026] As an implementation form, the water extraction of the licorice, the salvia miltiorrhiza and the white peony root respectively comprises:

[0027] The licorice, the salvia miltiorrhiza and the white peony root are respectively mixed with water, and are sequentially subjected to soaking and first water extraction to collect solid components and first water extract respectively; the solid components are mixed with water to perform second water extraction to collect second water extract; and the first water extract and the second water extract are mixed to obtain total water extract; and the total water extract is sequentially subjected to concentration and drying to obtain licorice water extract, salvia miltiorrhiza water extract and white peony root water extract respectively.

[0028] The licorice, the salvia miltiorrhiza and the white peony root are respectively mixed with water, and are sequentially subjected to soaking and first water extraction to collect solid components and first water extract respectively.

[0029] As an implementation form, the mass of the licorice, the salvia miltiorrhiza or the white peony root and the volume of water are in a ratio of 1 g:10 mL; and the water comprises distilled water.

[0030] After the solid components are collected, the solid components are mixed with water to perform second water extraction to collect second water extract.

[0031] After the first water extract and the second water extract are obtained, the first water extract and the second water extract are mixed to obtain total water extract.

[0032] The present application extracts twice and then mixes, which can avoid saturation of extracted components in one time and improve yield accuracy.

[0033] As an embodiment, the temperature of the soaking is 20-30℃; the time of the soaking is 28-32 min, further 30 min; the temperature of the first and second water extraction is 95-100℃ respectively; the time of the first water extraction is 1-2 h, further 1.5 h; the time of the second water extraction is 0.8-1.2 h, further 1 h.

[0034] After obtaining the total water extract, the present application concentrates and dries the total water extract in turn to obtain the water extract of licorice, the water extract of salvia miltiorrhiza and the water extract of radix paeoniae alba respectively.

[0035] As an embodiment, the drying includes vacuum drying; the temperature of the drying is 48-52℃, further 50℃; the time of the drying is greater than or equal to 12 h.

[0036] The present application also provides the application of the traditional Chinese medicine composition in the above-mentioned scheme or the traditional Chinese medicine composition in the preparation of a medicine for resisting liver ischemia-reperfusion injury.

[0037] As an embodiment, the resisting liver ischemia-reperfusion injury includes preventing, improving and / or protecting liver ischemia-reperfusion injury.

[0038] As an embodiment, the resisting liver ischemia-reperfusion injury includes in vivo resisting liver ischemia-reperfusion injury; ALT mainly exists in the cytoplasm of liver cells, when the liver cells are damaged, ALT is released into the blood, resulting in the increase of ALT level in the blood; when the liver cells are severely damaged, AST in the mitochondria is released into the blood, resulting in the increase of AST level in the blood; the in vivo resisting liver ischemia-reperfusion injury includes reducing the activity of serum alanine aminotransferase (ALT) and / or aspartate aminotransferase (AST).

[0039] As an embodiment, the liver ischemia-reperfusion injury includes liver transplantation ischemia-reperfusion injury.

[0040] As an embodiment, the inducement of liver ischemia includes hepatic artery occlusion, portal vein occlusion and bile duct atresia; the time of the liver ischemia is 45 min.

[0041] As an embodiment, the dosage form of the medicine includes oral dosage form.

[0042] As an embodiment, taking mice as an example, the unit preparation of the medicine is prepared to facilitate the administration dose of 4-4.1 g / kg.

[0043] The application further provides a medicine for resisting liver ischemia-reperfusion injury, which comprises a traditional Chinese medicine composition and pharmaceutically acceptable adjuvants; the traditional Chinese medicine composition comprises the traditional Chinese medicine composition described in the above scheme or prepared by the preparation method described above.

[0044] In order to further illustrate the application, the application of the traditional Chinese medicine composition for resisting liver ischemia-reperfusion injury and the application thereof are described in detail below in combination with the drawings and examples, but they should not be understood as limiting the protection scope of the application.

[0045] Example 1

[0046] The mass ratio of Danshen, Gancao and Baishao is 1:1:1.

[0047] The water extract of Danshen, Gancao and Baishao is extracted by the following steps:

[0048] Take 500 g of Danshen decoction pieces, and add distilled water according to the amount of 1 g of medicinal materials: 10 mL of water. First, soak in water at room temperature for 30 min, then heat to 100℃ for 1.5 h. Then extract the Danshen decoction pieces in 100℃ water again for 1 h. Combine the water extracts of the two extractions, and transfer to a rotary evaporator to obtain a concentrated liquid. Finally, place the concentrated liquid in a vacuum drying oven at 50℃ and dry for more than 12 h. After drying, take out and obtain 218 g of Danshen water extract.

[0049] The water extracts of Gancao and Baishao are extracted by the same method. 500 g of Gancao obtains 159 g of Gancao water extract. 500 g of Baishao obtains 112.5 g of Baishao water extract.

[0050] Test Example 1

[0051] The steps of animal modeling are as follows:

[0052] After the male C57BL / 6J mice are adapted to the environment for 7 days, they are randomly divided into 12 groups according to the body weight, which are sham operation group (sham, n = 6), liver ischemia-reperfusion model group (HIRI, n = 6), Gancao administration group (n = 6), Danshen administration group (n = 6), Baishao administration group (n = 6), Gancao: Danshen = 1:1 (mass ratio, the same below) administration group (n = 6), Danshen: Baishao = 1:1 administration group (n = 6), Gancao: Baishao = 1:1 administration group (n = 6), Gancao: Danshen: Baishao = 1:1:1 administration group (n = 6), Gancao: Danshen: Baishao = 1:1:2 administration group (n = 6), Gancao: Danshen: Baishao = 2:1:1 administration group (n = 6), Gancao: Danshen: Baishao = 1:2:1 administration group (n = 6).

[0053] The sham operation group and the liver ischemia-reperfusion model group are given Milli-Q ultrapure water by gavage every day;

[0054] The drug administration group was given the water extract of the medicine prepared with Milli-Q ultrapure water by gavage every day, and the administration volume was 0.2 mL / 10 g. The administration dose of the water extract of the medicine was converted according to the crude drug amount.

[0055] The liquorice administration group was given 1.30 g / kg of the water extract of liquorice by gavage every day;

[0056] The Danshen administration group was given 1.79 g / kg of the water extract of Danshen by gavage every day;

[0057] The Baishao administration group was given 0.92 g / kg of the water extract of Baishao by gavage every day;

[0058] The liquorice: Danshen = 1:1 administration group was given 1.3 g / kg of the water extract of liquorice and 1.79 g / kg of the water extract of Danshen by gavage every day;

[0059] The Danshen: Baishao = 1:1 administration group was given 1.79 g / kg of the water extract of Danshen and 0.92 g / kg of the water extract of Baishao by gavage every day;

[0060] The liquorice: Baishao = 1:1 administration group was given 1.3 g / kg of the water extract of liquorice and 0.92 g / kg of the water extract of Baishao by gavage every day;

[0061] The liquorice: Danshen: Baishao = 1:1:1 administration group was given 1.79 g / kg of the water extract of Danshen, 1.3 g / kg of the water extract of liquorice and 0.92 g / kg of the water extract of Baishao by gavage every day;

[0062] The liquorice: Danshen: Baishao = 1:1:2 administration group was given 1.79 g / kg of the water extract of Danshen, 1.30 g / kg of the water extract of liquorice and 1.84 g / kg of the water extract of Baishao by gavage every day;

[0063] The liquorice: Danshen: Baishao = 2:1:1 administration group was given 1.79 g / kg of the water extract of Danshen, 2.6 g / kg of the water extract of liquorice and 0.92 g / kg of the water extract of Baishao by gavage every day;

[0064] The liquorice: Danshen: Baishao = 1:2:1 administration group was given 3.58 g / kg of the water extract of Danshen, 1.3 g / kg of the water extract of liquorice and 0.92 g / kg of the water extract of Baishao by gavage every day.

[0065] After the continuous gavage for 7 days, each group was fasted for 12 h but could drink water freely, and then the liver ischemia-reperfusion modeling was performed.

[0066] The liver ischemia-reperfusion modeling scheme is as follows: intraperitoneal injection of 0.1% sodium pentobarbital anesthetize mice, use a constant temperature heating pad to maintain body temperature at 37°C during operation. Fix the mouse, wipe the abdomen with alcohol cotton, then make an incision along the midline of the abdomen to expose the hepatic portal area. Use a non-invasive vascular clamp to clamp the hepatic portal triad structure (hepatic artery, portal vein and bile duct), and confirm the success of ischemia by the pale color of the left and middle lobes of the liver. After 45 min of ischemia, remove the vascular clamp to restore blood flow, and suture the muscle layer and skin layer with sutures. The sham operation group exposes the hepatic portal structure but does not block the blood flow.

[0067] After 6 h of reperfusion, anesthetize the mouse, and collect the whole blood sample of the mouse through the inferior vena cava. After the mouse blood sample is left at room temperature for 1.5 h, centrifuge at 4°C and 4000 rpm for 15 min, take the upper serum and store it in a -80°C ultra-low temperature refrigerator for subsequent serum biochemical detection. Use a full-automatic biochemical analyzer (BS-240Vet) to detect the activity of ALT and AST in the serum of the mouse.

[0068] Experimental results:

[0069] The results of the activity of ALT and AST in the serum of the mouse are shown in Table 1. Figure 1 Compared with the liver ischemia-reperfusion model group, the activity of ALT and AST in the serum of the mouse in the licorice: salvia miltiorrhiza: white peony root = 1:1:1 administration group is significantly decreased, indicating that the licorice: salvia miltiorrhiza: white peony root = 1:1:1 combined administration has a protective effect on the liver ischemia-reperfusion of the mouse. In addition, although the level of ALT and AST in the serum of the mouse in the salvia miltiorrhiza administration group, the white peony root administration group, the licorice: salvia miltiorrhiza = 1:1 administration group, the salvia miltiorrhiza: white peony root = 1:1 administration group, the licorice: white peony root = 1:1 administration group, the licorice: salvia miltiorrhiza: white peony root = 1:1:2 administration group and the licorice: salvia miltiorrhiza: white peony root = 1:2:1 administration group is slightly decreased compared with the liver ischemia-reperfusion model group, and the level of ALT in the serum of the mouse in the licorice administration group is slightly decreased, but the effect is slightly worse than that of the licorice: salvia miltiorrhiza: white peony root = 1:1:1 administration group, indicating that the protective effect of the licorice: salvia miltiorrhiza: white peony root = 1:1:1 combined administration on the liver ischemia-reperfusion of the mouse is better than that of other administration schemes. The level of ALT and AST in the serum of the mouse in the licorice: salvia miltiorrhiza: white peony root = 2:1:1 administration group does not decrease significantly compared with the liver ischemia-reperfusion model group, indicating that these combinations have no effect or insignificant effect on improving the liver ischemia-reperfusion injury.

[0070] The results of HE staining of liver sections are shown in Table 2. Figure 2As shown, the liver showed obvious necrosis after ischemia-reperfusion injury, while the liver of the mice in the combined administration of glycyrrhiza: salvia: white peony root = 1: 1: 1 group showed no obvious necrosis characteristics, which was basically consistent with the results of the control group, indicating that the combined administration of glycyrrhiza: salvia: white peony root = 1: 1: 1 can reduce the damage caused by liver ischemia-reperfusion, which is consistent with the conclusion of serum ALT and AST. Other drug combinations have a protective effect on liver ischemia-reperfusion injury, but the effect is not as good as that of the combined administration of glycyrrhiza: salvia: white peony root = 1: 1: 1.

[0071] Although the above embodiment describes the present application in detail, it is only a part of the embodiments of the present application, not all the embodiments, and other embodiments can be obtained under the premise of no creativity according to the present embodiment, which belongs to the protection scope of the present application.

Claims

1. Use of a traditional Chinese medicine composition in the preparation of a medicament for resisting liver ischemia-reperfusion injury; the traditional Chinese medicine composition is composed of the following raw materials in parts by weight: Glycyrrhiza 1 part, Salvia miltiorrhiza 1 part and Radix Paeoniae Alba 1 part; ​ The preparation method of the traditional Chinese medicine composition comprises the following steps: Glycyrrhiza, Salvia miltiorrhiza and Radix Paeoniae Alba are mixed with water respectively, and are sequentially subjected to soaking and first water extraction, and the solid components and the first water extract are collected respectively; The solid components and water are mixed, and second water extraction is carried out, and the second water extract is collected; The first water extract and the second water extract are mixed to obtain a total water extract; The total water extract is sequentially subjected to concentration and drying to obtain Glycyrrhiza water extract, Salvia miltiorrhiza water extract and Radix Paeoniae Alba water extract respectively; The Glycyrrhiza water extract, the Salvia miltiorrhiza water extract and the Radix Paeoniae Alba water extract are mixed to obtain a traditional Chinese medicine composition; The mass of the Glycyrrhiza, the Salvia miltiorrhiza or the Radix Paeoniae Alba and the volume of water are 1 g:10 mL; The temperature of the soaking is 20-30℃, the time of the soaking is 28-32 min, the temperature of the first water extraction and the second water extraction is 95-100℃ respectively, the time of the first water extraction is 1-2 h, and the time of the second water extraction is 0.8-1.2 h.

2. Use according to claim 1, characterized in that, The Salvia miltiorrhiza comprises Salvia miltiorrhiza decoction pieces.

3. Use according to claim 1, characterized in that, The temperature of the drying is 48-52℃, and the time of the drying is greater than or equal to 12 h.

4. Use according to claim 1, characterized in that, The liver ischemia-reperfusion injury comprises liver transplantation ischemia-reperfusion injury.

5. Use according to claim 1 or 4, characterized in that, The inducement of the liver ischemia comprises hepatic artery occlusion, portal vein occlusion and bile duct atresia.

6. Use according to claim 1, characterized in that, The medicine comprises the traditional Chinese medicine composition and a pharmaceutically acceptable adjuvant.

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