Application of a Solanum torvum extract in the preparation of a medicine for preventing and treating non-alcoholic fatty liver and indications thereof

Abstract

The application discloses application of a Solanum torvum extract in preparation of a medicine for preventing and treating non-alcoholic fatty liver and indications thereof, and the Solanum torvum extract comprises one or both of a Solanum torvum water extract and a Solanum torvum alcohol extract; the application is proved by experiments that the Solanum torvum alcohol extract can obviously reduce the weight of a non-alcoholic fatty liver NAFLD mouse; the Solanum torvum water extract can obviously reduce the inguinal white fat; the Solanum torvum extract can obviously reduce a liver weight / body weight index, a liver triglyceride (TG) content, liver lipid deposition, and liver function damage indexes, i.e., glutamic-oxalacetic transaminase (AST) and glutamic-pyruvic transaminase (ALT). The Solanum torvum extract can also reduce serum triglyceride (TG) and cholesterol (TC) levels of the mouse, improve lipid levels, and therefore has an important role in treating non-alcoholic fatty liver.

Description

TECHNICAL FIELD

[0001] The application relates to the technical field of medicines, functional foods, functional drinks and supplements thereof, and particularly relates to application of a Solanum torvum extract in preparation of medicines for preventing and treating non-alcoholic fatty liver and indications thereof. BACKGROUND

[0002] Solanum torvum Swartz. is a shrub of Solanaceae in dicotyledon, which is widely distributed in tropical regions, including Yunnan, Guangdong and Guangxi in China. The fruit of Solanum torvum can improve eyesight, and the leaf can treat sore, and the root is used as medicine, which is called 'gold button' and is a common herbal medicine in folk, has the effects of activating blood, removing blood stasis and relieving pain, can treat contusion and blood stasis, hemoptysis, stomachache, lumbar muscle strain, sore and swelling, and is one of main components of 'Shaxi herbal tea', a traditional medicine of Guangdong Yihetang, which is a Chinese old brand. The tender fruit of Solanum torvum can be cooked and used as a vegetable, and is a wild vegetable commonly eaten by ethnic groups in Yunnan (Dai, Jingpo and other ethnic groups in Xishuangbanna and Dehong) and is called'small bitter fruit'.

[0003] As a wild vegetable of Solanum torvum, the Solanum torvum has the advantages of no pollution, high nutritional value and being edible. In addition to various nutritional medicinal substances contained in the Solanum torvum, the Solanum torvum has strong absorption and enrichment of trace element selenium, is a natural selenium-rich plant, and further increases the economic value of the Solanum torvum.

[0004] The Solanum torvum has a unique flavor and is used as a wild vegetable in many ethnic groups in China due to its medicinal and edible effects. For example, the Wa people in Yunnan use bamboo pestles to grind the Solanum torvum into paste, and then add salt and chili to make a dish. The Jingpo people in Ruili, Yunnan, cook the Solanum torvum with sour bamboo shoots and seasonings, or mix the cooked fruit with sour bamboo shoot silk. The Zhuang people in Guangxi not only drink the root of the Solanum torvum, but also fry the tender leaves. In Thailand, fried catfish with Solanum torvum curry is a famous dish and is popular among international consumers.

[0005] Non-alcoholic fatty liver disease (NAFLD) is a clinical and pathological syndrome characterized by hepatocyte steatosis and lipid accumulation without excessive alcohol consumption. Its pathological changes show simple fatty liver, fatty hepatitis, fatty liver fibrosis and cirrhosis as the disease progresses. The common causes of NAFLD are obesity, type II diabetes, hyperlipidemia, hypertension, etc. In recent years, with the improvement of living standards, the incidence of NAFLD has increased significantly, and tends to be younger. Although there have been many studies on the pathogenesis of NAFLD, its exact mechanism is still unclear. At present, the drugs (rosiglitazone, obeticholic acid, metformin hydrochloride and statins) used to relieve NAFLD on the market have poor effect and large side effects, so the US Food and Drug Administration (FAD) and the American Association for the Study of Liver Diseases (AASLD) have not approved any specific drug for non-alcoholic fatty liver disease. Therefore, the development of new drugs is an important research direction for the future treatment of non-alcoholic fatty liver disease. SUMMARY

[0006] The application provides an application of a Solanum torvum extract in preparation of a medicine for preventing and treating non-alcoholic fatty liver disease and indications thereof.

[0007] The application provides a scheme of the application.

[0008] The application provides an application of a Solanum torvum extract in preparation of a medicine for preventing and treating non-alcoholic fatty liver disease and indications thereof, which comprises the Solanum torvum extract, and the Solanum torvum extract comprises one or both of a Solanum torvum water extract and a Solanum torvum alcohol extract.

[0009] The Solanum torvum alcohol extract can effectively reduce the body weight of a non-alcoholic fatty liver disease patient, and the Solanum torvum extract can prevent and treat non-alcoholic fatty liver disease.

[0010] As a preferred technical scheme, the Solanum torvum water extract can effectively reduce the inguinal white fat.

[0011] As a preferred technical scheme, the Solanum torvum extract can effectively reduce the liver index.

[0012] As a preferred technical scheme, the Solanum torvum extract can effectively reduce transaminase, and the Solanum torvum extract can effectively reduce triglyceride and total cholesterol.

[0013] As a preferred technical scheme, the Solanum torvum extract can improve liver damage and liver steatosis caused by a high-fat diet.

[0014] As a preferred technical scheme, a preparation method of the Solanum torvum water extract is as follows: fresh Solanum torvum is dried, soaked in pure water overnight, then decocted, and decocted for 50 min under a micro-boiling state, repeated for three times, the water decoction is combined, and rotary water vapor at 60 DEG C is obtained to obtain an extract, that is, the Solanum torvum water extract, which is stored at 4 DEG C.

[0015] As a preferred technical solution, the preparation method of the water egg alcohol extract: take fresh water egg, mortar and pestle, soak it in 95% ethanol with a solid-liquid ratio of 1:4-6 overnight, then ultrasonic at 35℃ for 0.5h, collect the filtrate, repeat three times, combine the filtrate, rotate and evaporate 95% ethanol at 40℃, obtain the alcohol extract, then filter the alcohol extract through a macroporous resin chromatographic column, collect the filtrate and evaporate it on a rotary evaporator to obtain the extract, which is the water egg alcohol extract, and store it in a 4℃ refrigerator for standby.

[0016] As a preferred technical solution, soak it in 95% ethanol with a solid-liquid ratio of 1:5 overnight; weigh the alcohol extract, calculate the extract ratio as 0.0481, and then filter the alcohol extract through a macroporous resin chromatographic column.

[0017] Due to the adoption of the above technical solution, the water egg extract in the preparation of the application for preventing and treating non-alcoholic fatty liver and its indications, which contains water egg extract, the water egg extract includes one or both of water egg water extract and water egg alcohol extract; the water egg alcohol extract can effectively reduce the body weight of non-alcoholic fatty liver patients; and the water egg extract can prevent and treat non-alcoholic fatty liver.

[0018] As a preferred technical solution, the water egg water extract can effectively reduce the inguinal white fat.

[0019] As a preferred technical solution, the water egg extract can effectively reduce the liver index.

[0020] As a preferred technical solution, the water egg extract can effectively reduce transaminase, triglyceride and total cholesterol.

[0021] As a preferred technical solution, the water egg extract can improve liver damage and liver steatosis caused by high-fat diet.

[0022] Advantages of the present application:

[0023] The present application has been proved by experiments that the water egg alcohol extract can significantly reduce the body weight of non-alcoholic fatty liver patients; the water egg water extract can significantly reduce the inguinal white fat; the water egg extract can significantly reduce the liver weight / body weight index, liver triglyceride (TG) content, liver lipid deposition, and liver function damage indicators-glutamic-oxalacetic transaminase (AST) and glutamic-pyruvic transaminase (ALT). It can also reduce the serum triglyceride (TG) and total cholesterol (TC) levels of mice, and improve the lipid level. It can be seen that the water egg extract plays an important role in the treatment of non-alcoholic fatty liver. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1Figure of the influence of the aqueous extract and the alcohol extract of Solanum torvum in the present embodiment on the liver pathology of mice; wherein A is a blank control group, B is a model control group, C is a Solanum torvum aqueous extract group (100 mg / kg), and D is a Solanum torvum alcohol extract group (100 mg / kg);

[0025] Figure 2 Figure of the influence of the aqueous extract and the alcohol extract of Solanum torvum on the lipid deposition level in the liver of mice; wherein A is a blank control group, B is a model control group, C is a Solanum torvum aqueous extract group (100 mg / kg), and D is a Solanum torvum alcohol extract group (100 mg / kg). DETAILED DESCRIPTION

[0026] The present application provides a use of a Solanum torvum extract in the preparation of a drug for preventing and treating non-alcoholic fatty liver and indications thereof.

[0027] In order to make the technical means, creative features, purposes and effects achieved by the present application easy to understand, the present application is further described below in combination with specific embodiments.

[0028] EMBODIMENT

[0029] Improving effect of Solanum torvum aqueous extract and Solanum torvum alcohol extract on non-alcoholic fatty liver mice

[0030] 1. MATERIALS AND METHODS

[0031] 1.1 Main reagents

[0032] Detection kits for aspartate aminotransferase (AST), alanine aminotransferase (ALT), total cholesterol (TC), and triglyceride (TG) (C009-2, C010-2, A110-1, and A111-1) were purchased from Nanjing Jiancheng Bioengineering Institute; macroporous adsorption resin D101: National Pharmaceutical Group Chemical Reagent Co., Ltd., item number: 30266071; oil red O staining kit: Biyun Tian, product number: C0157S.

[0033] 1.2 Main instruments

[0034] RM2235 paraffin microtome, CM1900 freezing microtome, ASP300S dehydrator, DM2500 upright microscope + image system (Germany Leica), EYEL4 rotary evaporator, Epoch full-wavelength microplate reader.

[0035] 1.3 Preparation of Solanum torvum extract

[0036] 1.3.1 Solanum torvum aqueous extract

[0037] Take fresh water eggplant 1500g (purchased from Jinghong City, Yunnan Province), dry, soak in pure water overnight, decoct for 50 min in a boiling state, repeat three times, combine the water decoction, rotate the water vapor at 60°C to obtain the extract, store at 4°C.

[0038] 1.3.2 Water eggplant alcohol extract

[0039] Take fresh water eggplant 524.1g, crush in a mortar, soak in 95% ethanol at a ratio of 1:5 overnight, ultrasonic at 35°C for 0.5h, collect the filtrate, repeat three times, combine the filtrate, rotate 95% ethanol at 40°C to obtain the extract, weigh and calculate the extract ratio as 0.0481. The obtained alcohol extract extract was filtered through a macroporous resin chromatographic column, the filtrate was collected and rotary evaporated to dryness on a rotary evaporator, the extract was obtained and stored in a refrigerator at 4°C for standby.

[0040] 1.4 Experimental animals

[0041] C57BL / 6 female mice (8 weeks old, 18-22g), purchased from Sibeifu (Beijing) Biotechnology Co., Ltd., animal production certificate number: SCXK (Jing) 2019-0010. The mice were raised in SPF level animal room, temperature (22±1) °C, humidity 55%±5%, 12h light and dark alternation, free access to sterilized feed and drinking water, adaptive feeding for one week before experiment.

[0042] 1.5 NAFLD model establishment and water eggplant extract administration

[0043] 50 C57BL / 6J mice were adaptively fed for 1 week, and then randomly divided into blank control group, model control group, water egg water extraction group, water egg alcohol extraction group. The model control group had 14 mice, and the other groups had 12 mice each. The blank control group was fed with normal size mouse maintenance feed (feed purchased from: Sbi Bio-technology Co., Ltd. (Beijing)), and the other groups were fed with high-fat feed (high-fat feed purchased from: Changzhou Mouse Mouse Biological Technology Co., Ltd., item number: D18402002, batch number: 23022804). All mice were fed in a free diet mode. After 4 weeks of modeling, 2 mice were randomly selected from the model control group for pathological morphological observation of liver tissue: under the microscope, fatty degeneration was observed, indicating successful modeling. Subsequently, the model control group, water egg water extraction group, and water egg alcohol extraction group continued to be fed with high-fat feed, and the water egg water extraction group and the water egg alcohol extraction group were given oral administration, with a dose of 100 mg / kg. The other groups were given oral administration of the same amount of drinking water, and the administration was continued for 5 weeks. The mice were observed every other day for hair, diet, autonomous activity, and mental state. The mice's body weight was recorded once a week. After the modeling and administration were completed, the mice were fasted and deprived of water for 12 h. The mice were anesthetized with 1% sodium pentobarbital and sacrificed. The mouse liver was removed and weighed. The inguinal, perirenal, and epididymal white adipose tissue was weighed. After weighing the liver, a portion of the liver tissue was placed in 10% neutral formaldehyde solution for fixation. Another portion of the liver was subjected to frozen section staining. The remaining liver tissue was stored in a -80°C freezer for future use.

[0044] 1.6 Liver wet weight, inguinal, perirenal, and epididymal white adipose tissue weight determination

[0045] At the end of the experiment, the mice were sacrificed immediately after intraperitoneal injection of an overdose of anesthetic. The inguinal, perirenal, and epididymal adipose tissue was removed and weighed. The liver tissue was rinsed with pre-cooled physiological saline and weighed.

[0046] 1.7 Liver alanine transaminase (ALT), aspartate transaminase (AST), and triglyceride (TG), cholesterol (TC), low-density lipoprotein (LDL) content determination

[0047] The liver of each group of mice was homogenized with PBS, and the AST, ALT, TG, TC, and LDL contents in the mouse liver were determined according to the Nanjing Jiancheng kit instructions.

[0048] 1.8 HE staining to observe liver damage and lipid deposition changes

[0049] After the liver tissue was fixed with 10% neutral formaldehyde solution, it was dehydrated with different concentrations of ethanol, embedded in paraffin, sectioned, stained according to the HE staining kit, and observed under a upright microscope and image was collected. 1.9 Oil red O staining to observe liver lipid deposition

[0050] Fresh liver tissue was taken for OCT embedding, immediately placed into a constant temperature cold box microtome for frozen section, and oil red O staining was performed according to the kit instructions. The degree of lipid deposition in the liver tissue of each group of mice was observed under a microscope.

[0051] 2. Results

[0052] 2.1 Effect of Solanum torvum extract on body weight of NAFLD mice

[0053] Compared with the normal control group, the body weight of the model control group mice increased significantly (P<0.01 or P<0.001); after 2 and 3 weeks of administration, the body weight of the Solanum torvum alcohol extract group decreased significantly (P<0.01), and the body weight of the mice was lighter than that of the model group at the fourth and fifth weeks, but there was no significant statistical difference, and Solanum torvum water extract had no significant effect on the body weight of mice, as shown in Table 1.

[0054] Table 1 Effect of Solanum torvum extract on body weight of NAFLD mice (x±s, n=12)

[0055]

[0056] Compared with the model control group *P<0.05, **P<0.01, ***P<0.001 (same below).

[0057] 2.2 Effect of Solanum torvum extract on liver weight of NAFLD mice

[0058] Compared with the normal control group, the liver weight of NAFLD mice increased significantly (P<0.01); after intervention with Solanum torvum water extract and alcohol extract, the liver weight of NAFLD mice decreased significantly (P<0.01 or P<0.001), as shown in Table 2.

[0059] Table 2 Effect of Solanum torvum extract on liver weight of mice (x±s, n=12)

[0060] Group Dose (mg / kg) Liver weight (g) Normal control group 1.33±0.13 Model control group 1.58 ± 0.20 ** ]] Water caltrop water extraction group 100 1.42 ± 0.12 * ]] Water caltrop alcohol extraction group 100 1.28 ± 0.13 *** ]]

[0061] 2.3 Effect of Solanum torvum extract on the weight of inguinal, perirenal and epididymal white adipose tissue

[0062] Compared with the normal control group, the weight of inguinal, perirenal and epididymal white adipose tissue of NAFLD mice increased significantly (P<0.01, P<0.001); compared with the model group, the weight of inguinal and perirenal white adipose tissue of mice in the Solanum torvum water extract and alcohol extract groups decreased significantly (P<0.05), but there was no significant effect on epididymal white adipose tissue, as shown in Table 3.

[0063] Table 3 Effect of Solanum torvum extract on the weight of inguinal, perirenal and epididymal white adipose tissue (x±s, n=12)

[0064]

[0065] 2.4 Effects of Solanum torvum Extracts on Liver Function and Lipid Levels in NAFLD Mice

[0066] Compared with the normal control group, the AST and ALT enzyme activities in the livers of NAFLD mice were significantly increased (P < 0.05), and the TG and TC contents were also significantly increased (P < 0.001). Compared with the model control group, the AST and ALT enzyme activities in the water extract group were reduced (P < 0.05), and the TG and TC contents in the water extract and alcohol extract groups were significantly reduced (P < 0.01, P < 0.001). The water extract group also significantly reduced the low-density lipoprotein (LDL) content (P < 0.05), as shown in Table 4.

[0067] Table 4 Effects of Solanum torvum Extracts on Liver Function and Lipid Levels in NAFLD Mice (x ± s, n = 8)

[0068]

[0069]

[0070] 2.5 Effects of Solanum torvum Water Extract and Alcohol Extract on Liver Pathology in Mice

[0071] HE staining results showed that the liver tissue structure was normal in the normal control group, with regular liver cell arrangement and clear structure. In the model control group, the liver cells were arranged in disorder, with a large number of fat vacuoles of varying sizes and ballooning degeneration, and inflammatory cell infiltration. In the Solanum torvum water extract and alcohol extract groups, the liver cell fat vacuoles were significantly reduced, the cell arrangement was regular, and the structure was clearer than that in the model control group, indicating that Solanum torvum extracts can significantly improve the damage and fatty degeneration of the liver in high-fat diet-induced mice. Figure 1 .

[0072] 2.6 Effects of Solanum torvum Water Extract and Alcohol Extract on Lipid Deposition in the Liver of Mice

[0073] Oil red O staining showed that a small amount of orange-red lipid droplets was observed in the liver of the normal control group, and the nucleus was clearly stained blue. In the model control group, a large number of orange-red fat deposits were observed in the liver of the mice, with a large number of lipid droplets in the cytoplasm, and the nucleus was not obvious. In the Solanum torvum water extract and alcohol extract groups, the orange-red lipid droplets were significantly reduced, and the nucleus was relatively clear, indicating that Solanum torvum extracts can significantly improve the lipid deposition in the liver of non-alcoholic fatty liver mice. Figure 2 .

[0074] 3. Conclusion

[0075] After the intervention of the water extract and ethanol extract of Solanum torvum for 5 weeks, the liver tissue weight, pathological morphology changes and lipid deposition of the mice in each group were detected, combined with the detection of liver function and liver lipid deposition index. The results showed that the water extract (100 mg / kg) and ethanol extract (100 mg / kg) of Solanum torvum could improve the liver injury and lipid deposition of the mice with NAFLD to different degrees.

[0076] The above shows and describes the basic principles, main features and advantages of the present application. Those skilled in the art should understand that the present application is not limited to the above examples, and the above examples and descriptions in the specification are only to illustrate the principles of the present application. Without departing from the spirit and scope of the present application, various changes and improvements can be made to the present application, and these changes and improvements all fall within the scope of the claimed present application. The scope of protection of the present application is defined by the appended claims and their equivalents.

Claims

1. The use of a water extract of Solanum torvum in the preparation of a medicament for preventing and treating non-alcoholic fatty liver, characterized in that: The water extract of Solanum torvum is prepared by water extraction of Solanum torvum. ​ 2. Use according to claim 1, characterized in that: The medicine is used for reducing liver index.

3. Use according to claim 1, characterized in that: The medicine is used for reducing transaminase.

4. The use according to claim 1, characterized in that: The medicine is used for reducing triglyceride and total cholesterol.

5. The use according to claim 1, characterized in that: The medicine is used for improving liver injury and liver steatosis caused by high-fat diet.

6. The use according to claim 1, wherein The preparation method of the water extract of Solanum torvum is as follows: fresh Solanum torvum is dried, soaked in pure water overnight, then decocted, and the decoction is repeated for three times under the condition of micro-boiling, the decoction is combined, and water vapor is volatilized at 60 DEG C to obtain extract, namely the water extract of Solanum torvum, which is stored at 4 DEG C.

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