Plant extracts for alleviating pulmonary fibrosis, their preparation methods and applications

Abstract

To provide a plant extract for alleviating pulmonary fibrosis, and to provide preparation method and application thereof.SOLUTION: A method for preparing a plant extract includes the steps of: (1) drying and crushing Aronia melanocarpa fruit, mixing with ethanol, soaking, extracting, and subsequently filtering to obtain an Aronia melanocarpa extract; (2) drying and crushing Cynara scolymus flower buds, mixing with ethanol, extracting, and filtering to obtain a Cynara scolymus extract; (3) drying and crushing Brassica oleracea var. alboglabra roots, mixing with ethanol, and ultrasonically extracting to obtain a Brassica oleracea var. alboglabra extract; and (4) mixing the Aronia melanocarpa extract, Cynara scolymus extract, and Brassica oleracea var. alboglabra extract to obtain a plant extract for alleviating pulmonary fibrosis.SELECTED DRAWING: Figure 2

Description

[Technical Field]

[0001] The present invention relates to the technical field of plant extracts, and in particular to a plant extract that alleviates pulmonary fibrosis, its preparation method and application. [Background technology]

[0002] Pulmonary fibrosis is a terminal change in a large group of lung diseases characterized by fibroblast proliferation and massive accumulation of extracellular matrix, accompanied by inflammatory damage and destruction of tissue structure. Its main symptoms are dry cough and progressive dyspnea, which becomes more pronounced after exercise. Patients in the final stages may also experience symptoms of hypoxia, such as dizziness, purple lips and nails, and limited mobility. In recent years, the incidence and mortality rates of pulmonary fibrosis have been increasing. The main treatments for pulmonary fibrosis are glucocorticoids, immunosuppressants, and cytotoxic drugs. However, long-term use of these drugs has many side effects and limited therapeutic efficacy. Clinical studies have shown no significant change in the survival rate of pulmonary fibrosis patients treated with glucocorticoids. Therefore, finding drugs that can effectively alleviate pulmonary fibrosis is an urgent issue.

[0003] Black chokeberry (Aronia melanocarpa (Michx.) Elliott) is a deciduous shrub of the Aronia genus in the Rosaceae family. Its fruit is rich in substances such as flavonoids, anthocyanins, and polyphenols, and its extract is useful for treating diseases such as cancer, diabetes, and cardiovascular and cerebrovascular diseases. Artichoke (Cynara scolymus L.) is a perennial herbaceous plant of the Asteraceae family and Artichoke genus. Artichoke contains substances such as cynarin, asparagine, and flavonoids, and regular consumption of artichoke has the effects of protecting the liver and kidneys, strengthening liver detoxification, promoting digestion, improving blood circulation, preventing arteriosclerosis, and protecting the cardiovascular system. Chinese kailan (Brassica oleracea var. albiflora Kuntze) is an annual herbaceous plant of the Brassicaceae family, and its roots, stems, and leaves are used medicinally to detoxify, soothe the throat, smooth phlegm, relieve asthma, lower cholesterol, soften blood vessels, and protect the heart. However, there have been no reports to date that the extract of this plant can alleviate pulmonary fibrosis. Summary of the Invention [Problem to be solved by the invention]

[0004] The object of the present invention is to provide a plant extract that alleviates pulmonary fibrosis, its preparation method, and its application. The plant extract obtained by the preparation method of the present invention has the effect of alleviating pulmonary fibrosis, and the plant-derived components used have few side effects and are highly effective. [Means for solving the problem]

[0005] To achieve the above objectives, the present invention provides the following technical solutions:

[0006] The present invention provides a method for preparing a plant extract that alleviates pulmonary fibrosis, (1) drying and crushing black chokeberry fruit, mixing with ethanol, soaking, extracting, and filtering to obtain a black chokeberry extract; (2) drying and crushing the artichoke flower buds, mixing with ethanol to extract, and filtering to obtain an artichoke extract; (3) drying and crushing the Chinese kai root, mixing it with ethanol and ultrasonically extracting it to obtain a Chinese kai extract; (4) mixing the black chokeberry extract, the artichoke extract, and the Chinese kale extract to obtain a plant extract that alleviates pulmonary fibrosis.

[0007] Preferably, the ratio of dried black chokeberry fruit to ethanol in step (1) is 1 g:5-10 mL, and the volume percentage concentration of the ethanol is 70-80%.

[0008] Preferably, the immersion time in step (1) is 10 to 20 hours, The extraction temperature is 50 to 60°C, the extraction is performed 2 to 4 times, and each extraction time is 1 to 3 hours.

[0009] Preferably, the ratio of dried artichoke flower buds to ethanol in step (2) is 1 g:8 to 15 mL, and the volume percentage concentration of the ethanol is 60 to 70%.

[0010] Preferably, the extraction temperature in step (2) is 50 to 60°C, the extraction is performed 1 to 2 times, and each extraction time is 2 to 3 hours.

[0011] Preferably, the ratio of dried Chinese kale root to ethanol in step (3) is 1 g:5-10 mL, and the volume percentage concentration of the ethanol is 60-80%.

[0012] Preferably, the temperature of the ultrasonic extraction in step (3) is 40 to 50°C, the time of the ultrasonic extraction is 0.5 to 2 hours, the power of the ultrasonic extraction is 50 to 100 W, and the frequency of the ultrasonic extraction is 40 to 60 kHz.

[0013] Preferably, the concentrations of the black chokeberry extract, artichoke extract, and Chinese kailan extract in step (4) are prepared so that 0.5 to 1 g of dry ingredients per mL of extract, and the volume ratio of the black chokeberry extract, artichoke extract, and Chinese kailan extract is 1:1 to 3:1 to 2.

[0014] The present invention further provides a plant extract obtained by the above-described preparation method.

[0015] The present invention further provides the application of said plant extract in the preparation of a product for alleviating pulmonary fibrosis.

[0016] Compared with the prior art, the beneficial effects of the present invention are as follows: 1. The method for preparing the plant extract of the present invention is simple, and the prepared plant extract can significantly increase the body weight of pulmonary fibrosis model rats and significantly reduce the lung index and hydroxyproline content in the lung tissue of the model rats, indicating that the plant extract of the present invention has a good effect on alleviating pulmonary fibrosis. 2. The black chokeberry, artichoke and Chinese kale used in the present invention are obtained from a wide range of sources, are low in cost, have little toxicity or side effects, and have wide application value. [Brief explanation of the drawings]

[0017] [Figure 1] 1 shows the effect of the plant extract of the present invention on the body weight of a rat model. [Figure 2] 1 shows the effect of the plant extract of the present invention on the pulmonary index of a rat model. [Figure 3] 1 shows the effect of the plant extract of the present invention on the hydroxyproline content in lung tissue of a model rat. DETAILED DESCRIPTION OF THE INVENTION

[0018] The present invention provides a method for preparing a plant extract that alleviates pulmonary fibrosis, (1) drying and crushing black chokeberry fruit, mixing with ethanol, soaking, extracting, and filtering to obtain a black chokeberry extract; (2) drying and crushing the artichoke flower buds, mixing with ethanol to extract, and filtering to obtain an artichoke extract; (3) drying and crushing the Chinese kai root, mixing it with ethanol and ultrasonically extracting it to obtain a Chinese kai extract; (4) mixing the black chokeberry extract, the artichoke extract, and the Chinese kale extract to obtain a plant extract that alleviates pulmonary fibrosis.

[0019] In the present invention, black chokeberry fruit is dried and crushed, mixed with ethanol, immersed, extracted, and then filtered to obtain a black chokeberry extract. The method for drying and crushing the black chokeberry fruit may be a method commonly used in this field.

[0020] In the present invention, the ratio of dried black chokeberry fruit to ethanol is 1 g:5 to 10 mL, preferably 1 g:6 to 9 mL, and more preferably 1 g:7 to 8 mL, and the volume percentage concentration of the ethanol is 70 to 80%, preferably 72 to 78%, and more preferably 74 to 76%.

[0021] In the present invention, the immersion time is 10 to 20 hours, preferably 12 to 18 hours, and more preferably 14 to 16 hours.

[0022] In the present invention, the extraction temperature is 50 to 60°C, preferably 52 to 58°C, and more preferably 54 to 56°C, the extraction is performed 2 to 4 times, and preferably 3 times, and the extraction time for each time is 1 to 3 hours, preferably 1.5 to 2.5 hours, and more preferably 2 hours.

[0023] In the present invention, the filtration may be carried out by a filtration method generally used in this field.

[0024] In the present invention, artichoke flower buds are dried and crushed, mixed with ethanol, extracted, and then filtered to obtain an artichoke extract. The artichoke flower buds can be dried and crushed using methods commonly used in this field.

[0025] In the present invention, the ratio of dried artichoke flower buds to ethanol is 1 g:8 to 15 mL, preferably 1 g:9 to 14 mL, and more preferably 1 g:10 to 12 mL, and the volume percentage concentration of the ethanol is 60 to 70%, preferably 62 to 68%, and more preferably 64 to 66%.

[0026] In the present invention, the extraction temperature is 50 to 60°C, preferably 52 to 58°C, and more preferably 54 to 56°C, the extraction is performed 1 to 2 times, and the extraction time for each time is 2 to 3 hours, and preferably 2.5 hours.

[0027] In the present invention, the filtration may be carried out by a filtration method generally used in this field.

[0028] In the present invention, Chinese kale root is dried and crushed, mixed with ethanol, and subjected to ultrasonic extraction to obtain a Chinese kale extract. The method for drying and crushing the Chinese kale root may be a conventional method used in this field.

[0029] In the present invention, the ratio of dried Chinese kale root to ethanol is 1g:5-10mL, preferably 1g:6-9mL, and more preferably 1g:7-8mL, and the volume percentage concentration of the ethanol is 60-80%, preferably 65-75%, and more preferably 68-72%.

[0030] In the present invention, the temperature of the ultrasonic extraction is 40 to 50°C, preferably 42 to 48°C, and more preferably 44 to 46°C, the time of the ultrasonic extraction is 0.5 to 2 hours, and preferably 1 to 1.5 hours, the output of the ultrasonic extraction is 50 to 100 W, preferably 60 to 90 W, and more preferably 70 to 80 W, and the frequency of the ultrasonic extraction is 40 to 60 kHz, preferably 45 to 55 kHz, and more preferably 48 to 52 kHz.

[0031] In the present invention, the black chokeberry extract, artichoke extract, and Chinese kailan extract are mixed to obtain a plant extract that alleviates pulmonary fibrosis, and the concentrations of the black chokeberry extract, artichoke extract, and Chinese kailan extract are prepared so that the dry ingredients are 0.5 to 1 g per mL of extract, preferably 0.6 to 0.9 g per mL of extract, and more preferably 0.7 to 0.8 g per mL of extract, and the volume ratio of the black chokeberry extract, artichoke extract, and Chinese kailan extract is preferably 1:1 to 3:1 to 2.

[0032] The present invention further provides a plant extract obtained by the above-described preparation method.

[0033] The present invention further provides the application of said plant extract in the preparation of a product for alleviating pulmonary fibrosis.

[0034] The following describes in detail the technical solutions provided by the present invention in conjunction with examples, which should not be construed as limiting the protection scope of the present invention.

[0035] Example 1 This example provides a method for preparing a plant extract that alleviates pulmonary fibrosis, (1) Dry and crush black chokeberry fruit, weigh out 100 g of black chokeberry fruit powder, mix with 500 mL of 80% by volume ethanol, soak at room temperature for 10 hours, then heat to 50°C and extract for 3 hours, and filter the residue and extract it with 500 mL of 80% by volume ethanol at 50°C for 3 hours. The two filtrates are combined and concentrated until the concentration is adjusted to 0.5 g of dried black chokeberry fruit per mL of filtrate, thereby obtaining a black chokeberry extract; (2) Dry and crush artichoke flower buds, weigh 100g of artichoke flower bud powder, mix with 800mL of 70% ethanol by volume, heat to 50℃ and extract for 3 hours, and then extract the residue after filtration with 800mL of 70% ethanol by volume at 50℃ for 3 hours. Combine the two filtrates and concentrate the filtrate until the concentration of 0.5g of artichoke flower buds per mL of filtrate is adjusted to obtain an artichoke extract; (3) Dry and crush the Chinese kale root, weigh 100g of the Chinese kale root powder, mix with 500mL of ethanol, and ultrasonically extract the powder. The ultrasonic extraction parameters are 40℃, 2h, 50W, and 60kHz. The concentration of the filtrate after filtration is adjusted to 0.5g of dried Chinese kale root per 1mL of filtrate, to obtain a Chinese kale extract. (4) mixing the black chokeberry extract, artichoke extract, and cabbage extract in a volume ratio of 1:3:2 to obtain a plant extract that alleviates pulmonary fibrosis.

[0036] Example 2 This example provides a method for preparing a plant extract that alleviates pulmonary fibrosis, (1) drying and crushing black chokeberry fruit, weighing 100 g of black chokeberry fruit powder, mixing with 1000 mL of 70% by volume ethanol, soaking at room temperature for 20 hours, and then heating to 60°C and extracting for 1 hour, and extracting the residue after filtration two more times, combining the three filtrates, and concentrating the filtrate until the concentration is adjusted to 0.8 g of dried black chokeberry fruit per mL of filtrate, thereby obtaining a black chokeberry extract; (2) Dry and crush artichoke flower buds, weigh 100g of artichoke flower bud powder, mix with 1500mL of 60% ethanol by volume, heat to 60℃ and extract for 2 hours, and then extract the residue after filtration with 1500mL of 60% ethanol by volume at 60℃ for 2 hours. Combine the two filtrates and concentrate the filtrate until the concentration of 0.8g of artichoke flower buds per mL of filtrate is adjusted to obtain an artichoke extract; (3) Dry and crush the Chinese kale root, weigh 100g of the Chinese kale root powder, mix with 1000mL of ethanol, and ultrasonically extract the powder. The ultrasonic extraction parameters are temperature 50℃, time 0.5h, power 100W, and frequency 40kHz. The concentration of the filtrate after filtration is adjusted to 0.8g of dried Chinese kale root per mL of filtrate, to obtain a Chinese kale extract. (4) mixing the black chokeberry extract, artichoke extract, and cabbage extract in a volume ratio of 1:2:2 to obtain a plant extract that alleviates pulmonary fibrosis.

[0037] Example 3 This example provides a method for preparing a plant extract that alleviates pulmonary fibrosis, (1) Dry and crush black chokeberry fruit, weigh out 100 g of black chokeberry fruit powder, mix with 800 mL of 75% vol ethanol, soak at room temperature for 15 hours, then heat to 55°C and extract for 2 hours, and extract the residue after filtration with 800 mL of 75% vol ethanol at 55°C for 2 hours. The two filtrates are combined and concentrated until the concentration is adjusted to 1 g of dried black chokeberry fruit per mL of filtrate, thereby obtaining a black chokeberry extract; (2) Dry and crush artichoke flower buds, weigh 100g of artichoke flower bud powder, mix with 1000mL of 65% ethanol by volume, heat to 55℃ and extract for 2.5 hours, and extract the residue after filtration with 1000mL of 65% ethanol by volume at 55℃ for 2.5 hours. Combine the two filtrates and concentrate the filtrate until the concentration is adjusted to 1g of artichoke flower buds per 1mL of filtrate to obtain an artichoke extract; (3) Dry and crush the Chinese kale root, weigh 100g of the Chinese kale root powder, mix with 800mL of ethanol, and ultrasonically extract the powder. The ultrasonic extraction parameters are 45℃, 1h, 80W, and 50kHz. The concentration of the filtrate after filtration is 1g of dried Chinese kale root per 1mL of filtrate to obtain a Chinese kale extract. (4) mixing the black chokeberry extract, artichoke extract, and cabbage extract in a volume ratio of 1:2:1 to obtain a plant extract that alleviates pulmonary fibrosis.

[0038] Comparative Example 1 Compared with Example 1, this comparative example does not contain Chinese kale extract, and other preparation methods and parameters are the same as those of Example 1.

[0039] Comparative Example 2 Compared with Example 1, the volume ratio of black chokeberry extract, artichoke extract and Chinese kale extract in this comparative example is 1:1:1, and other preparation methods and parameters are the same as those in Example 1.

[0040] Experimental Example 1 In this experiment, the effects of the plant extracts prepared in Examples 1 to 3 on the body weight and lung index of rats were investigated, and the method was as follows.

[0041] 1. Construction of a rat pulmonary fibrosis model and grouping Eighty healthy SD rats, weighing 230g ± 10g, were selected, half male and half female, and divided into a blank group, a model group, Example 1 group, Example 2 group, Example 3 group, Comparative Example 1 group, Comparative Example 2 group, and a positive control group. Each group contained 10 rats. After one week of conventional care, each rat was intraperitoneally injected with 10mL / kg of 4% chloral hydrate. Anesthetized rats were placed in a supine position, their necks were cleaned, disinfected with iodine alcohol, and a longitudinal incision was made to expose the trachea. For each group (except the blank group), a syringe was inserted 1cm into the trachea, passing through the gap between the two tracheal cartilage rings toward the heart. After the syringe was withdrawn without resistance, bleomycin was injected intratracheally at 5mg / kg. The blank group was injected with an equal volume of saline. Immediately after injection, the rats were held upright and rotated for 5 minutes to ensure complete drug distribution within the lungs. The tissues were sutured and partially disinfected with alcohol. 24 hours after model construction, the blank and model groups received daily intragastric administration of 5 mL / kg saline. The positive control group received daily intragastric administration of 5 mg / kg prednisone acetate. Example groups 1-3 and Comparative groups 1-2 received daily intragastric administration of the corresponding plant extract at 5 mL / kg. During this period, the rats in each group were housed in the usual manner.

[0042] 2.Detection indicators (1) Body weight: The rats of each group are weighed on the day of intragastric administration, and then every 7 days. (2) Lung index: Five rats from each group were sacrificed on days 14 and 28, and the lung tissues were removed and weighed to calculate the lung index. Lung index = wet lung weight (mg) / body weight (mg) × 100%.

[0043] 3. Experimental Results (1) Weight measurement results The results are shown in Table 1 and Figure 1. Table 1 and Figure 1 show that compared with the blank group, the body weight of the rats in the model group was significantly reduced, and the difference was statistically significant (P<0.05). Compared with the rats in the model group, the body weight of the rats in Examples 1 to 3 was significantly increased (P<0.05), indicating that the plant extracts prepared in Examples 1 to 3 can improve the constitution of pulmonary fibrosis model rats and significantly alleviate the effects of pulmonary fibrosis on the rats.

[0044] [Table 1]

[0045] (2) Lung index measurement results The results are shown in Table 2 and Figure 2. Table 2 and Figure 2 show that the lung index of the rats in the model group was significantly increased compared to the blank group, and the difference was statistically significant (P<0.05). After 14 and 28 days of administration, the lung index of the rats in Examples 1 to 3 was significantly decreased compared to the rats in the model group (P<0.05), indicating that the plant extracts prepared in Examples 1 to 3 can improve bleomycin-induced pulmonary fibrosis in mice and significantly alleviate the progression of pulmonary fibrosis in model mice.

[0046] [Table 2]

[0047] Experimental Example 2 In this experiment, the effect of the plant extracts prepared in Examples 1 to 3 on the hydroxyproline content in lung tissue was examined.

[0048] The construction and grouping of the rat pulmonary fibrosis model are shown in Experimental Example 1. The rats in each group were intragastrically administered for 28 days and then sacrificed. The hydroxyproline content in the lung tissue was measured using a hydroxyproline content kit (purchased from Shanghai Enzyme Biotechnology Co., Ltd.). The results are shown in Table 3 and Figure 3.

[0049] Table 3 and Figure 3 show that, compared with the blank group, the hydroxyproline content in the lung tissue of the rats in the model group was significantly increased, and the difference was statistically significant (P<0.05). Compared with the rats in the model group, the hydroxyproline content in the lung tissue of the rats in Examples 1 to 3 was significantly decreased (P<0.05). The hydroxyproline content in lung tissue is one of the important indicators for determining the degree of pulmonary fibrosis. The above results indicate that the plant extracts prepared in Examples 1 to 3 can significantly alleviate the progression of pulmonary fibrosis in model mice.

[0050] [Table 3]

[0051] As can be seen from the above examples, the present invention provides a plant extract for alleviating pulmonary fibrosis, its preparation method and application, and the plant extract obtained by the preparation method of the present invention has the effect of alleviating pulmonary fibrosis, and the plant-derived ingredients used have few side effects and significant effects.

[0052] The above description is merely a preferred embodiment of the present invention, and those skilled in the art may make some further improvements and modifications without departing from the principles of the present invention, and these improvements and modifications should also be considered as within the protection scope of the present invention.

Claims

[Claim 1] (1) drying and crushing black chokeberry fruit, mixing with ethanol, macerating, extracting, and filtering to obtain a black chokeberry extract; (2) drying and crushing the artichoke flower buds, mixing with ethanol to extract, and filtering to obtain an artichoke extract; (3) drying and crushing the Chinese kale root, mixing with ethanol and ultrasonically extracting to obtain a Chinese kale extract; (4) A method for preparing a plant extract for alleviating pulmonary fibrosis, the method comprising: mixing the black chokeberry extract, the artichoke extract, and the Chinese kale extract to obtain a plant extract for alleviating pulmonary fibrosis, In step (1), the ratio of dried black chokeberry fruit to ethanol is 1 g:5-10 mL, and the volume percentage concentration of the ethanol is 70-80%; The immersion time in step (1) is 10 to 20 hours; In step (1), the extraction temperature is 50 to 60°C, the extraction is performed 2 to 4 times, and the extraction time for each time is 1 to 3 hours; In step (2), the ratio of dried artichoke flower buds to ethanol is 1 g:8-15 mL, and the volume percentage concentration of the ethanol is 60-70%; In step (2), the temperature of the extraction is 50 to 60°C, the number of extractions is 1 to 2, and the extraction time for each extraction is 2 to 3 hours; In step (3), the ratio of dried Chinese kale root to ethanol is 1 g:5-10 mL, and the volume percentage concentration of the ethanol is 60-80%; In step (3), the temperature of the ultrasonic extraction is 40-50°C, the time of the ultrasonic extraction is 0.5-2 hours, the power of the ultrasonic extraction is 50-100W, and the frequency of the ultrasonic extraction is 40-60kHz; a concentration of the black chokeberry extract, the artichoke extract, and the Chinese kale extract in step (4) being 0.5-1 g of dry ingredients per mL of extract, and a volume ratio of the black chokeberry extract, the artichoke extract, and the Chinese kale extract being 1:1-3:1-2.

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