Application of solanum photeinocarpum extract in preparation of medicine or health food for treating constipation

Abstract

The invention discloses an application of a solanum photeinocarpum extract in preparation of a medicine or health food for treating constipation, which comprises the following steps: preparing solanum photeinocarpum aqueous extract dry powder from solanum photeinocarpum coarse powder by using a hot water extraction method, adding water and ethanol into the aqueous extract dry powder, centrifuging, carrying out rotary evaporation on supernate, and drying to obtain the solanum photeinocarpum extract. Freeze-drying to obtain a solanum photeinocarpum water-extract alcohol-soluble component, and freeze-drying the centrifuged precipitate part to obtain an alcohol-precipitated component. Compared with a solanum photeinocarpum alcohol precipitation component, the solanum photeinocarpum water extraction alcohol-soluble component can obviously improve the constipation symptom of mice induced by using loperamide hydrochloride, promote intestinal tract movement, improve the growth environment of intestinal flora and more effectively improve constipation.

Description

Technical Field

[0001] The invention relates to the field of laxative medicines or foods, and in particular to application of Solanum nigrum extract in preparing medicines or health foods for treating constipation. Background Art

[0002] Solanum nigrum (Solanum nigrumL) belongs to the Solanum genus of the Solanaceae family. It is a perennial herb with a bitter, slightly sweet taste, cold nature, and slight toxicity. It enters the lung, stomach, and bladder meridians. The fruit of Solanum nigrum is edible and tastes sweet. The berries are spherical, about 8mm in diameter, and black when ripe. The flowering period is from July to September, and the fruiting period is from August to October. There are five different types in my country, which can be divided into yellow fruit Solanum nigrum, purple stem Solanum nigrum, green stem Solanum nigrum, green vein few flower Solanum nigrum and brown vein few flower Solanum nigrum. According to the grade classification of esterase and peroxidase isozymes, it can be divided into two major categories, the first three are one category, and the last two are another category; according to the morphology, geographical distribution, and chromosome multiples, Solanum nigrum can be divided into two categories, namely Solanum nigrum and few flower Solanum nigrum.

[0003] Solanum nigrum (Solanum umbellatum Nakai et Odash.) is a delicate herbaceous plant of the Solanaceae family and the genus Solanum. The plant is up to 1 meter tall, with an umbel-shaped inflorescence and small flowers. The calyx is green, the corolla is white, the berries are spherical, and the young are green. The leaves are thin, and the leaves are ovate to ovate-oblong, with sparse soft hairs on both sides. The petiole is slender, the seeds are nearly ovate, and the two sides are flattened. It blooms and bears fruit almost all year round. In China, it is mainly distributed in southern Yunnan, Guangxi, Guangdong, Jiangxi, Hunan, Taiwan and other places. It mainly grows by streams, in the shade and wet places of dense forests, or in wastelands near forests.

[0004] In addition to differences in morphology, geographical distribution, and chromosome multiples, Solanum nigrum and Solanum nigrum pauciflorum also have slight differences in medicinal use: the whole plant of Solanum nigrum can be used as medicine to dissipate blood stasis and reduce swelling, clear away heat and detoxify. The leaves of Solanum nigrum pauciflorum can be eaten as vegetables, have the function of cooling and dissipating heat, and can also treat sore throats.

[0005] The phytochemical composition of Solanum nigrum is relatively complex, mainly alkaloids and polysaccharides. Many scholars at home and abroad have conducted in-depth research on its chemical composition, especially in the anti-cancer and anti-tumor aspects. However, the pharmacological effects of some chemical components are still unclear. It is necessary to conduct more in-depth research on Solanum nigrum and apply it to more fields. At the same time, paying attention to the toxicological experiments of Solanum nigrum has an extremely important and critical role in preventing and treating major diseases that threaten human health.

[0006] Epidemiological surveys have found that the incidence of constipation is 5.0% to 15.0% worldwide and 10.0% to 15.0% in China, among which functional constipation is the most common. Elderly people are more prone to constipation, and the course of the disease is generally longer. Long-term stubborn constipation is harmful to human health and can lead to tumors, breast diseases, and even cardiovascular and cerebrovascular diseases, leading to death. In recent years, with the change of eating habits, social pressure, and the influence of mental and psychological factors, the incidence of functional constipation has increased year by year. Slow-transmission constipation is a typical type of functional constipation, which is caused by the reduction of high-amplitude propulsive contraction activity of the colon. Clinical manifestations include anorectal diseases such as difficulty in defecation, dry stools, and reduced bowel movements.

[0007] Improving intestinal motility is the main purpose of treating functional constipation, which promotes the restoration of the normal physiological function of defecation, and ultimately can eliminate the disease, enable patients to return to normal life, and improve their quality of life. Functional constipation can be treated by changing diet, physical exercise, or by drug intervention. At present, there are many side effects of using Western medicine to treat constipation. The use of plant ingredients to treat constipation has a more ameliorative effect, brings fewer side effects, and has relatively little impact on the patient's health. Looking for natural extracts with laxative and intestinal moisturizing effects, Chinese medicines that can be used as both medicine and food, or by supplementing dietary fiber, may obtain a safer, more effective, and economical health food for improving constipation. At present, there is no literature reporting that the alcohol-soluble components of the water-extracted Solanum nigrum have the effect of improving functional constipation. Summary of the invention

[0008] In view of the shortcomings of the prior art, the present invention provides the use of Solanum sparcus-flowering extract in the preparation of a medicine or health food for treating constipation. The present invention uses Solanum sparcus-flowering crude powder as a raw material to prepare an alcohol-soluble extract; uses loperamide hydrochloride as a modeling drug to establish a mouse constipation model, and verifies whether the alcohol-soluble components of the water-extracted Solanum sparcus-flowering have the effect of improving constipation from the two aspects of intestinal peristalsis and fecal properties, in order to develop health foods and medicines using Solanum sparcus-flowering as raw materials.

[0009] In order to solve the above technical problems, the technical solution of the present invention is as follows: application of Solanum nigrum extract in the preparation of medicines or health foods for treating constipation.

[0010] Furthermore, the Solanum nigrum L. extract is a water-extracted alcohol-soluble product. In the early stage of the experiment, the present invention has found through animal experiments that the water extract of Solanum nigrum L. has a laxative effect on constipated mice. In order to clarify the defecation site, the present invention uses the water extract of Solanum nigrum L. to prepare alcohol-soluble and alcohol-precipitated components, and further explores the effective ingredients for improving constipated mice. The results show that the water-extracted alcohol-soluble product is more effective in laxative.

[0011] Furthermore, the preparation method of the water-extracted alcohol-soluble product is as follows: adding the crude powder of Solanum nigrum leaf to boiling water, stirring while boiling, filtering after boiling for several minutes, and repeating the above operation 2-3 times on the residue; combining the filtrate, cooling and centrifuging in a centrifuge, and freeze-drying the supernatant to obtain the Solanum nigrum water extract dry powder;

[0012] The obtained Solanum nigrum water extract dry powder is dissolved in water, ethanol is slowly added, and the mixture is stirred until flocs appear, and then sealed with plastic wrap, and placed in a cold storage for one day (0-4° C.), and the liquid is placed in a centrifuge for centrifugation, and the supernatant is collected after the centrifugation operation; the mixture is concentrated by rotary evaporation, and the collected liquid is placed in a freeze dryer for freeze drying to obtain Solanum nigrum water extract alcohol-soluble dry powder.

[0013] Furthermore, the preparation process of the water-extracted alcohol-soluble product is as follows: the mass volume ratio of the Solanum nigrum leaf powder to water is 1:9-15; the mass volume ratio of the Solanum nigrum water extract dry powder, water and ethanol is 1:15-25:45-60; the concentration of the ethanol is 50%-80%.

[0014] Furthermore, in the preparation method, the centrifugal speed is 3500-4500 r / min and the centrifugal time is 8-15 min.

[0015] Preferably, the dosage of the Solanum nigrum extract is 150-600 mg / kg.

[0016] Preferably, the constipation is functional constipation.

[0017] The present invention uses Solanum nigrum crude powder to prepare Solanum nigrum water extract dry powder by hot water extraction method, adds ultrapure water and ethanol to the water extract dry powder, and after centrifugation, rotary evaporates the supernatant, freeze-dries to obtain Solanum nigrum water extract alcohol-soluble components, and freeze-dries the precipitate after centrifugation to obtain alcohol precipitation components. The present invention uses loperamide hydrochloride to induce constipation mouse model, and uses Solanum nigrum water extract alcohol-soluble components as low, medium and high dose groups to intervene in constipated mice. Experimental results show that the first black stool time of constipated mice intervened with Solanum nigrum water extract alcohol-soluble components is earlier than that of the model group, the number of fecal particles increases, the water content of feces increases, and the ink propulsion rate of the small intestine increases, indicating that Solanum nigrum alcohol-soluble components have a good improvement effect on constipation in mice. Compared with the alcohol precipitation component of Solanum nigrum L., the alcohol-soluble component of water extract of Solanum nigrum L. is more effective. After alcohol treatment, the macromolecular substances are precipitated by centrifugation, and the small molecular substances are retained in the supernatant. The small molecular substances in the alcohol-soluble component play a key role in the study of laxative effects on mice. It was identified that the main components of the small molecular substances in the alcohol-soluble component of water extract are: nucleotides and their derivatives, aromatic compounds, phenols and their derivatives, carboxylic acids and their derivatives, amino acids and their derivatives, fatty acyl compounds, alkaloids, triazines, purine nucleosides and indoles and their derivatives; among them, the monomer compounds with relatively high contents are adenosine, ethyl p-aminobenzoate, guanosine, 2-phenylacetamide, guanine, 2-chlorophenylalanine, D-2-aminobutyric acid, 4-hydroxybenzaldehyde, inosine and deethyl atrazine.

[0018] Compared with the prior art, the present invention has the following beneficial effects: the alcohol-soluble components of the water-extract of Solanum nigrum provided by the present invention can significantly improve the constipation symptoms of mice induced by loperamide hydrochloride, promote intestinal peristalsis, improve the growth environment of intestinal flora, and effectively improve constipation. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 It is the relative content of alcohol-soluble components in Solanum nigrum water extract;

[0020] Figure 2 Example 2 Animal grouping and drug administration;

[0021] Figure 3 Effects of alcohol-soluble fractions extracted from Solanum nigrum on body weight of constipated mice;

[0022] Figure 4 Effects of alcohol-soluble fractions extracted from Solanum nigrum on water drinking and food intake in constipated mice (# indicates comparison with blank control group; * indicates comparison with model control group, the same below), A. water intake; B. food intake;

[0023] Figure 5 Effects of alcohol-soluble fractions extracted from Solanum nigrum on defecation parameters in constipated mice;

[0024] Figure 6 Example 2 Fecal characteristics of mice in different groups;

[0025] Figure 7 Effects of alcohol-soluble components of Solanum nigrum water extract on ink propulsion rate in constipation; A. Small intestine length; B. Ink propulsion rate; C. Small intestine picture;

[0026] Figure 8 Effects of water-extracted and alcohol-precipitated components of Solanum nigrum on body weight of constipated mice;

[0027] Fig. 9 Effects of water-extracted and alcohol-precipitated components of Solanum nigrum on drinking water and food intake in constipated mice;

[0028] Fig.10 Effects of the water-extracted and alcohol-precipitated fractions of Solanum nigrum on defecation parameters in constipated mice;

[0029] Fig.11 Effects of water-extracted and alcohol-precipitated components of Solanum nigrum on small intestinal motility in constipated mice; Note: Compared with the blank control group, the symbols of the model group are: ###p<0.001 extremely significant, ##p<0.01 extremely significant, #p<0.05 significant; Compared with the model control group, the symbols of each intervention group are: *p<0.05 significant, **p<0.01 extremely significant, ***p<0.001 extremely significant. DETAILED DESCRIPTION

[0030] The technical solution of the present invention is further described in detail below in conjunction with the accompanying drawings and specific embodiments, but the present invention is not limited to the following technical solution.

[0031] The following examples were carried out at the Engineering Center for the Development and Utilization of Food and Drug Resources of Yunnan Agricultural University in 2022. The sources of the reagents and raw materials used are as follows: Solanum nigrum dry powder was purchased from Simao District, Pu'er City. Loperamide hydrochloride was purchased from Sigma. Phenolphthalein was purchased from Shandong Renhetang Pharmaceutical Co., Ltd. Ethanol was purchased from Chengdu Chemical Reagent Co., Ltd. Gum arabic and activated carbon powder were purchased from Aladdin.

[0032] Example 1

[0033] Preparation of Solanum nigrum water extract

[0034] Use an electronic balance to accurately weigh 100g of Solanum nigrum crude powder and add it to 900mL of boiling ultrapure water, with a solid-liquid ratio of 1:9 (g / mL). Stir with a glass rod, boil for three minutes, and filter with 7 layers of nylon cloth. After filtering, take the filter residue and add it to 900mL of boiling ultrapure water again. After boiling for three minutes, filter and repeat the above operation once. Finally, mix the three filtrates and let them cool, then put them into a large high-speed centrifuge for centrifugation (4000r / min, 10min), take the supernatant and put it into a freeze-drying plate for freeze-drying, and finally obtain the dry powder of Solanum nigrum water extract with a yield of 31.6%.

[0035] Preparation of alcohol solution extracted from Solanum nigrum

[0036] 20g of dry powder of water extract of Solanum nigrum was accurately weighed with an electronic balance, added to 480mL ultrapure water, with a solid-liquid ratio of 1:24, stirred with a glass rod to completely dissolve the dry powder of water extract in water, and then slowly added 70% ethanol, with a solid-liquid ratio of 1:56 (g / mL). In order to ensure that the concentration gradient of the solution remains stable during the addition of ethanol, a glass rod should be used to stir during the process; Stir continuously until completely mixed, seal with plastic wrap after floccules appear, put in a cold storage for one day, centrifuge the liquid in a large high-speed centrifuge (4000r / min, 10min), collect the supernatant after centrifugation; Use a rotary evaporator to perform rotary evaporation at a temperature of 55-65℃, the purpose of which is to remove alcohol and water, so that the liquid is concentrated, collect the liquid and put it into a freeze dryer for freeze drying to obtain water extract alcohol-soluble dry powder of Solanum nigrum. The precipitated part after centrifugation is freeze-dried to obtain the alcohol precipitation component of water extract of Solanum nigrum. The yield ratio of alcohol-soluble component to alcohol precipitation component is about 10:3. The dosage in the later experimental design was set according to the 10:3 ratio.

[0037] Based on the detection of extensive targeted metabolomics technology, 846 compounds were obtained from the analysis of the alcohol-soluble dry powder of Solanum nigrum water extract. The top 10 substances were classified as follows: nucleotides and their derivatives, aromatic compounds, phenols and their derivatives, carboxylic acids and their derivatives, amino acids and their derivatives, fatty acyl compounds, alkaloids, triazines, purine nucleosides, indoles and their derivatives; the top 10 monomer compounds in terms of relative content were adenosine, ethyl p-aminobenzoate, guanosine, 2-phenylacetamide, guanine, 2-chlorophenylalanine, D-2-aminobutyric acid, 4-hydroxybenzaldehyde, inosine, and deethylatrazine. The relative contents were as follows: Figure 1 shown.

[0038] The polysaccharide content of the alcohol precipitation extracted from Solanum nigrum water-extracted by phenol-sulfuric acid spectrophotometry was 22.91%, and the protein content was 1.85%.

[0039] According to the above experimental process, a scaled-up experiment was carried out to obtain multiple groups of alcohol-soluble components and alcohol-precipitated components.

[0040] 250 g of water extract dry powder gave about 180 g of water extract alcohol soluble components and 54 g (there is an error) of water extract alcohol precipitate components.

[0041] 500 g of water extract dry powder obtained about 360 g of water-extract alcohol-soluble components and 108 g (there is an error) of water-extract alcohol-precipitated components.

[0042] 750 g of water extract dry powder obtained about 540 g of water extract alcohol soluble components and 162 g (there is an error) of water extract alcohol precipitate components.

[0043] The corresponding three groups of water-extracted alcohol-soluble components were used in Example 2 to carry out a laxative effect test; the three groups of water-extracted alcohol-precipitated components were used in Example 3 to carry out a laxative effect test.

[0044] Example 2: Laxative effect test of water-extracted alcohol-soluble substance

[0045] The preparation of loperamide hydrochloride suspension, ink and phenolphthalein suspension was prepared by referring to the existing method.

[0046] Animal grouping and dosing A total of 72 Kunming mice (specific pathogen-free animals) weighing about 20-25 g were fed in the animal laboratory of the Engineering Research Center for the Development and Utilization of Food and Drug Resources of the Ministry of Education, Yunnan Agricultural University. Male Kunming mice (20-25 g) aged 6 weeks were purchased from Hunan Slake Jingda Experimental Animal Co., Ltd. All mice were housed under specific pathogen-free barrier conditions (24±1°C, 30-50% humidity, 12-hour daylight cycle, lights off at 20:00), with a normal diet (10.8% fat, 68.7% carbohydrates and 20.5% protein) and free drinking water. After 7 days of adaptive feeding, the mice were divided into six groups according to body weight, with 12 mice in each group. Three mice were housed in the same cage, and the weight, food intake and water intake of the mice were recorded every day. Loperamide hydrochloride (8 mg / kg·bw, purchased from Sigma) was used to establish the STC mouse model.

[0047] Group settings: Figure 2As shown, the control group (gavage sterile water), the model control group (gavage loperamide hydrochloride, sterile water as solvent), the positive control group: (gavage 200mg / kg·bw phenolphthalein tablet suspension, produced by Shandong Renhetang Pharmaceutical Co., Ltd.), the low-dose group (gavage 8mg / kg·bw loperamide and 200mg / kg·bw alcohol-soluble fraction of Solanum nigrum water extract), the medium-dose group (gavage 8mg / kg·bw loperamide and 400mg / kg·bw alcohol-soluble fraction of Solanum nigrum water extract) and the high-dose group (gavage 8mg / kg·bw loperamide and 600mg / kg·bw alcohol-soluble fraction of Solanum nigrum water extract). The STC model mice were induced by administering LOP for 8 days, during which the mice were given an oral dose of 0.1mL / 10g.bw per day by gavage. The required doses of phenolphthalein tablets and alcohol-soluble fractions of Solanum nigrum water extract were dissolved in sterile water. All mice were deprived of food but not water 10 h before the defecation test and gastrointestinal transit test. All procedures were approved by the Animal Ethics Committee of Yunnan Agricultural University.

[0048] Defecation experiment

[0049] The mouse defecation test was carried out according to China's "Guidelines for the Functional Evaluation of Health Foods". On the evening of the sixth day of continuous gavage, the food of the mice was removed but the water was not removed. The empty tubes for feces were prepared, and the empty tubes were weighed and the data was recorded. On the morning of the seventh day, a clean cage was prepared in advance, and a slightly damp sterilized gauze was laid in the cage, and the corresponding label and tubes for feces were placed on the cage; after the mice were gavaged with the test substance, waited for 30 minutes, and the mice were filled with ink and placed in the cages prepared in advance. When the last mouse in each group was placed in the cage, the timing started immediately. Carefully observe the defecation of each mouse, record the time and number of feces particles of each defecation, and put the feces into the empty tube. When the first black feces appeared, the feces were put into a new empty tube, and the time at this time was marked. After two hours of defecation observation, food was added to the mice. After six hours, the defecation experiment was over. Arrange the tubes for feces of each mouse, weigh the wet feces of the mice and record the data. After the above operation, the feces were placed in an oven and dried at 50°C to a constant weight, and the dry feces were weighed and the data were recorded. The feces moisture content of each group can be calculated by the formula feces moisture content (%) = [feces wet weight (g) - feces dry weight (g)] / feces wet weight (g) × 100.

[0050] Small Intestinal Propulsion Test

[0051] The mouse small intestine propulsion test was carried out according to China's "Guidelines for the Functional Evaluation of Health Foods". At 10 pm on the 7th day of continuous intervention, the mice were deprived of food, but the water bottle was retained. On the morning of the 8th day, after the test substance was instilled into the mice, wait for 30 minutes, and then instill ink into the mice as an indicator. After waiting for 20 minutes, the mice were induced to coma with carbon dioxide, the abdominal cavity was cut open to separate the mesentery, and the intestinal tissue from the pylorus to the ileocecal region was completely removed with scissors, and the excess fat tissue was removed. The intestinal tissue was carefully stretched and flattened, and the total length of the small intestine was measured with a ruler. The length from the pylorus to the front of the ink was measured, which was the ink propulsion distance, and the data was recorded. According to the formula, the ink propulsion rate (%) = ink propulsion distance (cm) / total length of the small intestine (cm) × 100, the ink propulsion rate of each group was calculated.

[0052] Data analysis

[0053] Graphpadprism 8.0 software was used for statistical analysis of the test data. According to the Chinese "Guidelines for the Evaluation of Health Food Functions", independent T-test (two-tailed method) was used to analyze the weight, water intake, time to first black stool, ink propulsion rate, small intestine length, number of feces discharged by mice within 6 hours, total weight of feces discharged by mice, and water content of feces of mice, etc., and the differences between the blank control group and the model control group, and between the model control group and each intervention group were significant. If one of the results of the feces weight and the number of feces discharged by mice is positive (p<0.05), at the same time, one of the results of the small intestine propulsion test and the time to the first black stool of mice is positive (p<0.05), it can be determined that the test substance helps to relieve constipation.

[0054] Effects of alcohol-soluble fractions extracted from Solanum nigrum on body weight and water and food intake in constipated mice

[0055] like Figure 3 As shown, the body weight of each group of mice was recorded while gavage was continued for seven consecutive days. From the recorded data, it can be seen that although there were small changes in the weight of the mice during the experiment, it did not exceed the normal weight range and remained steadily increasing overall, indicating that after treatment with various test substances, the weight of the mice still increased normally.

[0056] like Figure 4As shown, the model group constipated mice drank the least amount of water, the positive group and low-dose group mice drank similar amounts of water, the medium-dose group and high-dose group mice drank similar amounts of water, but all were lower than the blank group; data analysis showed that there was no significant difference between the groups of mice (P>0.05), indicating that the alcohol-soluble components of the water extract of Solanum nigrum had no significant effect on the water intake of constipated mice. The blank group mice ate the most, the low-dose group mice ate relatively less, and the other four groups ate similar amounts of food. Data analysis showed that there was no significant difference in food intake between the groups of mice (P>0.05), indicating that the alcohol-soluble components of the water extract of Solanum nigrum had no significant effect on the food intake of the mice in the intervention groups.

[0057] Effects of alcohol-soluble fractions extracted from Solanum nigrum on defecation parameters in constipated mice

[0058] The time when the mouse excretes the first black stool is one of the most important indicators of bowel movement, indicating the time when the ink is excreted from the entire mouse intestine. The earlier the ink-containing black stool is excreted, the better the digestive function of the mouse. Figure 5 It can be seen that compared with the model group, the time for the first black stool of the mice in the blank group constipated was earlier, and there was a significant difference (P<0.05). Compared with the time for the first black stool of the mice in the positive group constipated and the model group constipated, the time for the first black stool of the mice in the positive group constipated was earlier than that of the model group, and there was a significant difference between the two groups (P<0.05). The time for the first black stool of the mice in the constipated group treated with low, medium and high doses of the alcohol-soluble components of the water extract of Solanum nigrum was earlier than that of the model group, and there was a significant difference compared with the model group (P<0.05). In summary, compared with the model group, the time for the first black stool of the mice in the blank group was earlier than that of the model group, the time for the first black stool of the mice in the constipated group treated with phenolphthalein was significantly shortened, and the time for the first black stool of the mice in the constipated group treated with low, medium and high doses of the alcohol-soluble components of the water extract of Solanum nigrum was also significantly shortened.

[0059] Compared with the blank group, the number of fecal particles of mice in the model group was less than that in the blank group, and there was a very significant difference (P<0.01). Compared with the model group, the number of fecal particles of mice in the positive group was more than that in the model group, and there was a very significant difference (P<0.001). The number of fecal particles of constipated mice treated with low and medium doses of alcohol-soluble components of water extract of Solanum nigrum was more than that of constipated mice in the model group, and there was a significant difference (P<0.05). The number of fecal particles of constipated mice treated with high doses of alcohol-soluble components of water extract of Solanum nigrum was higher than that of constipated mice in the model group, but there was no significant difference (P>0.05). In summary, low and medium doses of phenolphthalein and alcohol-soluble components of water extract of Solanum nigrum can increase the number of fecal particles in constipated mice.

[0060] The total feces weight of the constipated mice in the blank group was higher than that in the model group, and the total feces weight of the constipated mice in the blank group was the largest, with significant differences between the two groups (P<0.05). The total feces weight of the constipated mice in the positive group was higher than that in the model group, with significant differences (P<0.05). The total feces weight of the constipated mice in the low, medium and high doses of the alcohol-soluble fractions of the water extract of Solanum nigrum was higher than that of the model group, but the total feces weight of the constipated mice in the medium and high dose groups was significantly different from that in the model group (P<0.05), and the total feces weight of the constipated mice in the low dose group was not significantly different from that in the model group. In summary, the medium and high doses of the alcohol-soluble fractions of the water extract of Solanum nigrum had a significant effect on the feces weight of constipated mice.

[0061] The fecal water content of constipated mice in the model group was the lowest, the fecal water content of constipated mice in the blank group was higher than that in the model group, and there was a significant difference (P<0.05), the fecal water content of constipated mice in the positive group was higher than that in the model group, and there was an extremely significant difference between the two groups (P<0.001), and the fecal water content of constipated mice in the low, medium and high dose groups of the alcohol-soluble fraction of the water extract of Solanum nigrum was higher than that in the model group, among which there was an extremely significant difference between the constipated mice in the low dose group and the model group (P<0.001), and there was a significant difference between the constipated mice in the medium and high dose groups and the model group (P<0.05). In summary, low doses of phenolphthalein and alcohol-soluble fractions of the water extract of Solanum nigrum can significantly increase the fecal water content of constipated mice, and medium and high doses of Solanum nigrum can also significantly increase the fecal water content of constipated mice.

[0062] comprehensive Figure 5 ABCD, Figure 6 The above analysis showed that the time of the first black stool of the constipated mice in the model group was longer than that of the blank group, the positive group, and the low, medium and high dose groups of the alcohol-soluble fraction of water-extracted Solanum nigrum, and the time of the first black stool of the positive group and the low, medium and high doses of the alcohol-soluble fraction of water-extracted Solanum nigrum was significantly different from that of the model group (P<0.05). The number of fecal pellets of the constipated mice in the positive group was the largest, and there was a very significant difference with the model group. The number of fecal pellets of the model group and the blank group was also very significantly different. The fecal weight of the constipated mice in the blank group was the largest, and there was a significant difference with the fecal weight of the constipated mice in the model group. The fecal weight of the medium and high dose groups of the alcohol-soluble fraction of water-extracted Solanum nigrum was significantly different from that of the model group (P<0.05). The fecal water content of the constipated mice in the positive group and the constipated mice in the low dose group of Solanum nigrum was very significantly different from that of the model group. The fecal water content of the constipated mice in the medium and high dose groups of Solanum nigrum was significantly different from that of the model group. The above shows that the functional constipation mouse model was successfully established in this experiment, and the alcohol-soluble components of the water-extract of Solanum nigrum have an improving effect on functional constipation in mice.

[0063] Effects of alcohol-soluble fractions extracted from Solanum nigrum on small intestinal peristalsis in constipated mice

[0064] Depend on Figure 7 As shown in Figure 3, the small intestine length of the constipated mice in the blank group was the longest, and the small intestine length of the constipated mice in the model group was the shortest. There was a significant difference in the small intestine length of the constipated mice in the blank group compared with that in the model group (P<0.05). The small intestine length of the constipated mice in the low, medium and high dose groups of the alcohol-soluble components of the water extract of Solanum nigrum increased with the increase of the dose, but there was no significant difference compared with the model group. It can be seen that phenolphthalein and the low, medium and high dose groups of the alcohol-soluble components of the water extract of Solanum nigrum have no significant effect on the small intestine length of constipated mice.

[0065] The higher the ink propulsion rate, the better the small intestinal peristalsis function of the mice. The movement rate of the substance in the small intestine, i.e., the gastrointestinal transit rate, is an important reference for whether the test substance has the effect of moistening the intestine and relieving constipation. The ink propulsion rate and small intestine length can clarify the effect of the alcohol-soluble fraction of the water extract of Solanum nigrum on the gastrointestinal transit rate of constipated mice. The ink propulsion rate of constipated mice in the low-dose group of the alcohol-soluble fraction of the water extract of Solanum nigrum was the lowest. The ink propulsion rate of constipated mice in the blank group was higher than that in the model group, with a very significant difference (P<0.001). The ink propulsion rate of constipated mice in the positive group was higher than that in the model group, and there was a very significant difference between the two groups (P<0.01). The ink propulsion rate of constipated mice in the medium-dose group of the alcohol-soluble fraction of the water extract of Solanum nigrum was higher than that in the model group, and there was a significant difference (P<0.05). The ink propulsion rate of constipated mice in the high-dose group was higher than that in the model group, and there was a very significant difference (P<0.01). In summary, the medium dose of alcohol-soluble components of water extract of Solanum nigrum has a significant effect on the ink propulsion rate of constipated mice, the high dose groups of phenolphthalein and Solanum nigrum have a very significant effect on the ink propulsion rate of constipated mice, and the medium and high dose groups of alcohol-soluble components of water extract of Solanum nigrum have a promoting effect on the intestinal peristalsis of constipated mice.

[0066] Example 3: Laxative effect test of water-extracted alcohol sediment

[0067] This example uses animal experiments to clarify the specific laxative site of the water extract of Solanum nigrum on constipated mice. The water-extracted alcohol precipitate obtained in Example 1 was used to verify the laxative effect. The operation method is the same as that of Example 2. The animal grouping and administration method are shown in Table 1.

[0068] Table 1 Animal grouping and drug administration

[0069]

[0070] Effects of water-extracted and ethanol-precipitated components of Solanum nigrum on body weight, drinking water and diet in constipated mice

[0071] like Figure 8As shown in the figure, the mice were weighed for 7 consecutive days and the weight changes were recorded. From the data, it can be seen that the weight of the mice fluctuated, and the fluctuation range generally showed an upward trend, but did not exceed the normal weight range, indicating that after being affected by different doses of the control group and the test substance, the weight of the mice increased normally.

[0072] like Fig. 9 As shown in the figure, in the experiment, there was no significant difference in the amount of water drunk by each group of mice (p>0.05), which indicates that in the experiment, the test substances did not have a significant effect on the amount of water drunk by mice; compared with the blank control group, the food intake of the model control group was reduced by 1.43g / d / mouse, and the difference between the two groups was extremely significant (p<0.01), indicating that the test substances in the model control group would affect the food intake of mice and cause a decrease in their appetite.

[0073] Effects of water-extracted and ethanol-precipitated components of Solanum nigrum on defecation parameters in constipated mice

[0074] The time when mice excrete the first black stool is one of the most important indicators of bowel movement and laxative effect. It indicates the time it takes for ink to be discharged from the entire mouse intestine. The faster the ink is discharged, the better the digestive function of the mouse. Fig.10 As shown in Figure A, the time of the first black stool in the model control group was extended by 56 minutes compared with the blank group, and the difference between the two groups was extremely significant (p<0.01), indicating that the construction of the constipation model in this experiment was successful. The time of the first black stool in the water-extracted alcohol-soluble group was 62 minutes earlier than that in the model control group, and there was a very significant difference between the two groups (p<0.001), while there was no significant difference between the low, medium and high dose groups and the model control group (p>0.05).

[0075] like Fig.10 As shown in B, the number of fecal pellets in the model group was 4 less than that in the blank group at 6 hours, and the difference between the two groups was significant (p<0.05); compared with the model group, the number of fecal pellets in the water-extract alcohol-soluble group increased by 11 at 6 hours, and the difference between the two groups was extremely significant (p<0.001). The low-dose group increased by 7 pellets, the medium-dose group increased by 9 pellets, and the high-dose group increased by 14 pellets. As the dose increased, the number of fecal pellets in constipated mice also increased, indicating that the dose of the water-extract alcohol-precipitated components of Solanum pauciflorum will affect the number of defecation in mice.

[0076] like Fig.10As shown in C and D, compared with the model group, the total weight of feces in the water-extracted alcohol-soluble group increased significantly, and the feces water content also increased significantly, and the difference between the two was extremely significant (p<0.001); the difference between the high-dose group and the model group was not very significant in the total weight of feces (p=0.0505), indicating that the high-dose group had an effect on the total weight of feces of constipated mice but it was not obvious. The feces water content of the model group was significantly reduced compared with the blank group, and the difference between the two was extremely significant (p<0.001), indicating that the feces of the constipated mice induced by loperamide hydrochloride would become dry and hard, causing the feces water content to decrease.

[0077] Effects of water-extracted and ethanol-precipitated components of Solanum nigrum on small intestinal peristalsis in constipated mice

[0078] The small intestinal length and ink propulsion rate can clarify the role of the water-extracted and alcohol-precipitated components of Solanum nigrum in the gastrointestinal transit rate of constipated mice. Fig.11 As shown in the figure, the small intestine length of mice in the high-dose group decreased by 0.16 cm compared with the model control group during the experiment, and the difference between the two groups was extremely significant (p<0.01). Compared with the blank control group, the ink propulsion rate of mice in the model control group decreased by 20.54%, and the difference between the two was extremely significant (p<0.001), which once again proved that the constipation model of this experiment was successfully constructed. The ink propulsion rate of mice in the water-extracted alcohol-soluble group increased by 13.74% compared with the model control group, and the difference between the two groups was significant (p<0.05). Compared with the model control group, the ink propulsion rate of the high-dose group also increased by 12.57%, and there was a significant difference between the two groups (p<0.05), indicating that high doses of water-extracted alcohol-precipitated components can promote small intestinal peristalsis in constipated mice.

[0079] In summary, the present invention uses Solanum nigrum sparse flower crude powder to prepare Solanum nigrum sparse flower water extract dry powder by hot water extraction method, and ultrapure water and 70% ethanol are added to the water extract dry powder in a ratio of 1:24:56. After centrifugation, the supernatant is rotary evaporated, and freeze-dried to obtain the Solanum nigrum sparse flower water extract alcohol-soluble component, and the precipitate after centrifugation is freeze-dried to obtain the alcohol precipitation component. The present invention uses loperamide hydrochloride to induce constipation mouse model, and uses the Solanum nigrum sparse flower water extract alcohol-soluble component as a low, medium and high dose group to intervene in constipated mice. The experimental results show that the first black stool time of constipated mice intervened with the Solanum nigrum sparse flower water extract alcohol-soluble component is earlier than that of the model group, the number of fecal particles increases, the fecal water content increases, and the ink propulsion rate of the small intestine increases, indicating that the Solanum nigrum sparse flower alcohol-soluble component has a good improvement effect on the constipation of mice. Compared with the alcohol-precipitated component of Solanum nigrum, the alcohol-soluble component of Solanum nigrum water extract is more effective. After alcohol treatment and centrifugation, the large molecules are precipitated and the small molecules are retained in the supernatant. The small molecules in the alcohol-soluble component play a key role in the study of intestinal lubrication and laxative effects on mice.

[0080] The above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the present invention.

Claims

1. Application of Solanum nigrum extract in the preparation of medicines or health foods for treating constipation.

2. The use of the Solanum nigrum extract according to claim 1 in the preparation of a medicine or health food for treating constipation, characterized in that: The Solanum nigrum extract is a water-extracted alcohol-soluble product.

3. The use of the Solanum nigrum extract according to claim 2 in the preparation of a medicine or health food for treating constipation, characterized in that: The preparation method of the water-extracted alcohol-soluble product is as follows: Add the crude powder of Solanum nigrum leaf to boiling water, stir while boiling, filter after boiling for several minutes, repeat the above operation 2-3 times for the residue; combine the filtrate, centrifuge after cooling, freeze-dry the supernatant to obtain the dry powder of Solanum nigrum water extract; The obtained dry powder of Solanum nigrum water extract is dissolved in water, ethanol is slowly added, and the mixture is stirred until flocs appear, then sealed with plastic wrap, placed in a cold storage for one day, and the liquid is placed in a centrifuge for centrifugation. After the centrifugation operation, the supernatant is collected; the mixture is concentrated by rotary evaporation, and the collected liquid is placed in a freeze dryer for freeze drying to obtain the alcohol-soluble dry powder of Solanum nigrum water extract.

4. The use of the Solanum nigrum extract according to claim 3 in the preparation of a medicine or health food for treating constipation, characterized in that: The preparation process of the water-extracted alcohol solution is as follows: the mass volume ratio of the Solanum nigrum leaf powder to water is 1:9-15; the mass volume ratio of the Solanum nigrum water extract dry powder, water and ethanol is 1:15-25:45-60; the concentration of the ethanol is 50%-80%.

5. The use of the Solanum nigrum extract according to claim 3 in the preparation of a medicine or health food for treating constipation, characterized in that: In the preparation method, the centrifugal speed is 3500-4500 r / min and the centrifugal time is 8-15 min.

6. The use of the Solanum nigrum extract according to claim 1 in the preparation of a medicine or health food for treating constipation, characterized in that: The dosage of the Solanum nigrum extract is 150-600 mg / kg.

7. The use of the Solanum nigrum extract according to claim 1 in the preparation of a medicine or health food for treating constipation, characterized in that: The constipation is functional constipation.

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