Application of space lotus leaf extract

Abstract

The invention provides application of a space lotus leaf extract, and belongs to the field of traditional Chinese medicine preparations. The space lotus leaf extract is used for preparing a product with the function of improving postoperative insulin resistance, and the preparation method of the space lotus leaf extract comprises the following steps: dissolving dried space lotus leaf powder in ethanol, and carrying out alcohol extraction in an ultrasonic bath; performing centrifugation and membrane filtration treatment to obtain filter residues and supernate; repeatedly carrying out alcohol extraction, centrifugation and membrane filtration treatment on filter residues in the ultrasonic bath for at least two times; combining the supernate, performing decompression treatment, adding water, and stirring; the preparation method comprises the following steps: washing the lotus leaves with water, adsorbing by virtue of an adsorption column filled with macroporous resin, washing with water and eluting with ethanol, collecting eluent, carrying out reduced pressure treatment, and freeze-drying, so as to obtain the space lotus leaf extract. The space lotus leaf extract disclosed by the invention can improve insulin resistance and intestinal flora disorder caused by the operation, reduce the postoperative wound infection condition of a patient, reduce the occurrence probability of complications and improve the body recovery speed of the patient, and has objective clinical value and economic benefit.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine preparations, and particularly relates to the application of space lotus leaf extract. Background Art

[0002] The main treatment for gastric cancer is radical gastrectomy, which can significantly improve the survival rate of patients. However, as a powerful stressor, surgery itself may induce postoperative metabolic disorders and lead to insulin resistance (IR) in GC patients. Insulin resistance is also called insulin insensitivity or relative lack of insulin secretion. Postoperative insulin resistance is caused by the decrease in the sensitivity of the patient's body to insulin. In the early stage of insulin resistance, pancreatic β cells can still compensatory secretion of insulin to make up for its insufficient effect, and the body secretion of insulin increases, thus producing hyperinsulinemia; with the gradual decline of pancreatic β cell function, the compensatory secretion of insulin is still difficult to maintain blood sugar levels in the normal range, so blood sugar rises, and the symptoms of the body are similar to those of type 2 diabetes. The resulting hyperglycemia will also reduce body function, cause postoperative wound infection in patients, and increase the probability of postoperative complications, affecting the patient's postoperative recovery rate. In addition, surgical trauma caused by gastrectomy, digestive tract reconstruction, and prophylactic use of antibiotics can lead to severe damage to the intestinal flora.

[0003] Lotus leaf has the effects of moistening the intestine, detoxifying, reducing fat and losing weight, among which alkaloids are the main biologically active components of lotus leaf. Pharmacological research reports show that lotus leaf alkaloids have pharmacological effects such as regulating lipid metabolism and lowering blood sugar, as well as anti-cerebral ischemia, inhibiting osteoporosis, antibacterial, antiviral, anti-melanin production, and improving intestinal flora. However, no pharmacological effects of lotus leaf on postoperative insulin resistance have been found. Summary of the invention

[0004] In view of this, the purpose of the present invention is to provide an application of space lotus leaf extract, aiming to solve at least one technical problem among the background technology.

[0005] The present invention is achieved in that: The invention provides application of space lotus leaf extract, which is used for preparing a product with the effect of improving postoperative insulin resistance.

[0006] Furthermore, the post-operative procedure is gastrectomy, and the product may be a health product or a medicine.

[0007] Furthermore, the product regulates the balance of intestinal flora after surgery.

[0008] Furthermore, the product reduces postoperative fasting blood glucose levels.

[0009] Further, the product reduces postoperative fasting insulin levels.

[0010] Further, the product reduces postoperative fasting C-peptide levels.

[0011] Furthermore, the preparation method of the space lotus leaf extract comprises the following steps: The dried space lotus leaf powder is dissolved in ethanol of a set concentration, and then placed in an ultrasonic bath for alcohol extraction to obtain a preliminary extract; The preliminary extract was centrifuged and filtered to obtain the residue and the supernatant; The filter residue is subjected to the alcohol extraction in the ultrasonic bath, centrifugation and membrane filtration treatments at least twice; After the supernatants are combined, a first decompression treatment is performed to remove ethanol in the solution, and then water is added and stirred to obtain a crude extract of space lotus leaves; The crude extract of space lotus leaves flows through an adsorption column filled with macroporous resin for adsorption, then is washed with water to remove non-adsorbed matter, and then is eluted with ethanol, and the eluate is collected; The eluate is subjected to a second decompression treatment to be concentrated to obtain a clear paste, and then freeze-dried to obtain a space lotus leaf extract; the space lotus leaf extract is used to improve insulin resistance after gastrectomy.

[0012] Furthermore, in the alcohol extraction treatment, the ethanol mass concentration is 70% to 80% and the time is 45 min to 50 min.

[0013] Furthermore, the temperature of the first decompression treatment and the second decompression treatment is 60° C. and the pressure is 0.085 MPa.

[0014] Compared with the prior art, the present invention has the following beneficial effects: 1. The present invention uses space lotus as plant raw material, extracts its effective ingredients, and develops a product that regulates the balance of intestinal flora after gastrectomy and improves postoperative insulin resistance, reduces postoperative wound infection in patients, reduces the probability of complications, and improves the patient's physical recovery speed, which has objective clinical value and economic benefits.

[0015] 2. The present invention has good effects on alleviating and repairing the severe destruction of intestinal microbiota and damage to the intestinal mucosal barrier caused by surgical trauma of gastrectomy, digestive tract reconstruction and prophylactic use of antibiotics. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 This is a PCoA analysis diagram of the intestinal flora of each group of rats at the end point of the experiment in Example 5 of the present invention. DETAILED DESCRIPTION

[0017] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific implementation cases described herein are only used to explain the present invention and are not used to limit the present invention.

[0018] Example 1 This embodiment is to prepare a space lotus leaf extract for improving postoperative intestinal flora and insulin resistance, comprising the following steps: S1. Dissolving the dried space lotus leaf powder in ethanol of a set concentration, and subjecting the mixture to alcohol extraction in an ultrasonic bath to obtain a preliminary extract; Specifically: use the leaves of the space lotus variety produced in Guangchang County, Jiangxi Province as raw materials, crush the dried space lotus leaves, and pass them through an 80-mesh sieve for later use; take 100g of space lotus leaf powder, suspend it in 2L 70% ethanol, dissolve it in a dry conical flask, run it at a frequency of 40kHz in a 600W ultrasonic bath at a temperature of 45°C for 50 minutes, and obtain a preliminary extract.

[0019] S2, the preliminary extract is subjected to centrifugation and membrane filtration to obtain a filter residue and a supernatant; Specifically, the preliminary extract was centrifuged at 5000×g for 15 minutes and filtered through a 0.45 μm nylon syringe membrane to collect the supernatant.

[0020] S3, repeat steps S1 and S2 twice for the filter residue.

[0021] S4, after combining the supernatants, perform a first decompression treatment to remove ethanol in the solution, then add water and stir to obtain a crude extract of space lotus leaves; Specifically, the three supernatants obtained in steps S2 and S3 are combined, the filtrate is decompressed and ethanol is recovered under the conditions of a temperature of 60° C. and a pressure of 0.085 MPa, water is added and stirred to obtain a crude extract of space lotus leaves.

[0022] S5, the crude extract of space lotus leaves flows through an adsorption column equipped with macroporous resin for adsorption, then is washed with water to remove non-adsorbed matter, and then is eluted with ethanol, and the eluate is collected; Specifically: the crude extract of space lotus leaves is adsorbed using an adsorption column equipped with AB-8 macroporous resin, first rinsed with water and the water washing liquid discarded, then eluted with 60% ethanol, and the eluate was collected.

[0023] S6. The eluate is subjected to a second decompression treatment to be concentrated to obtain a clear paste, which is then freeze-dried to obtain the space lotus leaf extract.

[0024] Specifically: the eluent is concentrated under reduced pressure at a temperature of 60°C and a pressure of 0.085 MPa to a clear paste, and then freeze-dried to obtain the space lotus leaf extract; calculated by weight, the yield of the space lotus leaf extract is measured to be 6.94%.

[0025] Example 2 This Example 2 is for preparing a space lotus leaf extract for improving postoperative intestinal flora and insulin resistance. The only difference between it and Example 1 is that the ethanol concentration of the alcohol extraction in step S1 is adjusted to 80%, and the other preparation steps and reaction conditions are the same as those in Example 1. The final yield of the space lotus leaf extract is 6.83%.

[0026] Example 3 This Example 3 is for preparing a space lotus leaf extract for improving postoperative intestinal flora and insulin resistance. The only difference between this example and Example 1 is that the operation time of the ultrasonic bath in step S1 is adjusted to 45 minutes, and the other preparation steps and reaction conditions are the same as those in Example 1. The final yield of the space lotus leaf extract is 6.72%.

[0027] Example 4 This Example 4 is for preparing a space lotus leaf extract for improving postoperative intestinal flora and insulin resistance. The only difference between it and Example 1 is that the ethanol concentration of the alcohol extraction in step S1 is adjusted to 80% and the ultrasonic bath operation time is adjusted to 45 minutes. The other preparation steps and reaction conditions are the same as those in Example 1. The final yield of the space lotus leaf extract is 6.67%.

[0028] Example 5 1. Experimental subjects Forty-eight male SD rats aged 6 to 8 weeks, weighing 180 g to 200 g, were selected and provided by Sibeifu (Beijing) Biotechnology Co., Ltd. The animals were housed in the Animal Center of Nanchang University, and an ethical approval letter for animal experiments had been obtained.

[0029] The breeding environment is: under standard environmental conditions (SPF animal room, that is, a specific pathogen-free animal room), all rats are adaptively fed; the culture conditions are: temperature 22℃±2℃, humidity 55%~60%, L / D=12h / 12h, and food and water are provided.

[0030] Before surgery, rats were divided into 4 groups according to the different food they were given, with n=12 in each group. After 3 weeks of feeding, except for the control group, gastrectomy models were constructed in the model group, experimental group, and comparison group: (1) Control group, given free access to drinking water and food; (2) Model group, intragastric administration of normal saline daily; (3) The experimental group was gavaged with 1 ml of space lotus leaf extract every day. The space lotus leaf extract was prepared in Example 1 and dissolved in physiological saline. The dosage was adjusted according to the weight of the rats. The daily dosage for 1 kg rat was 60 mg. (4) The control group was gavaged with 1 ml of common lotus leaf extract per day. The common lotus leaf extract was dissolved in physiological saline and the dosage was adjusted according to the body weight of the rats. The daily dosage standard for 1 kg rats was 60 mg. The preparation method of the common lotus leaf extract was the same as that in Example 1, except that conventional white lotus leaves (produced in Guangchang County, Jiangxi Province) were used as raw materials.

[0031] 2. Establishment of Gastrectomy Model (1) Gastrectomy Gastrectomy was performed in a sterile environment. Rats in the model group, experimental group, and control group were strictly fasted for 12 hours before surgery and had free access to water. 2.5% Avertin was injected intraperitoneally 10 minutes before surgery to induce anesthesia, and the abdomen was shaved and disinfected with povidone-iodine. When anesthesia was completed, a midline incision of the xiphoid process was made 3 cm below the rat, and the duodenum was cut about 0.5 cm proximal to the pylorus. The incision was disinfected with an iodine cotton swab, and the stomach was removed about 0.5 cm from the cardia. Through skilled surgical techniques, most of the gastric tissue of all rats was successfully removed, and the abdominal incision in the layer was sutured with 3.0 nylon thread. After gastrectomy, 20 mL of saline was injected subcutaneously into the rat, and it was resuscitated in a 40°C rewarming device and fed without bedding. After 2 weeks of the experiment, the animals were killed.

[0032] (2) Sample collection: Venous blood was collected from the inferior vena cava of fasting rats one day before and one day after surgery, centrifuged at 1000 g for 20 min at 4°C, and then the supernatant was carefully removed and stored at -80°C.

[0033] (3) Testing and data analysis: The fasting blood glucose (FBG) level (mmol / L), fasting insulin (FIns) level (μIU / mL), and fasting C-peptide (ng / mL) levels in the blood were tested, and the insulin resistance index HOMA-IR was calculated, HOMA-IR = FIns level × FBG level / 22.5. In addition, feces were subjected to 16S rRNA high-throughput sequencing.

[0034] 3. Effect of space lotus leaf extract on insulin resistance IR in rats after gastrectomy The preoperative test results of the control group, model group, experimental group and comparison group are shown in Table 1, the postoperative test results of the model group, experimental group and comparison group are shown in Table 2, the comparative data analysis of the preoperative test results of each group is shown in Table 3, the comparative data analysis of the postoperative test results of each group is shown in Table 4, and the preoperative and postoperative comparative analysis of the data of the model group, experimental group and comparison group is shown in Table 5. In Tables 1 to 5, the data of quantitative variables are expressed as mean ± standard deviation; independent sample t test or nonparametric test is used to evaluate the measurement comparison between two groups; analysis of variance is used for comparison among multiple groups; P value <0.05 is statistically significant.

[0035] Table 1

[0036] Table 2

[0037] Table 3

[0038] Table 4

[0039] Table 5

[0040] From the data in Tables 1 and 3, it can be seen that the FBG, FIns, HOMA-IR, and Fasting C-peptide indicators of each group before surgery were all compared with P>0.05, that is, there was no difference in the indicators before surgery.

[0041] From the data in Table 2 and Table 4, it can be seen that: compared with the experimental group, the FBG, FIns, HOMA-IR, and Fasting C-peptide indices of the model group were P < 0.05; compared with the control group, the FBG and Fasting C-peptide indices of the model group were P < 0.05; the FIns and HOMA-IR indices were P > 0.05, but the model group was still greater than the control group, with the differences of -2.42±11.03 and -1.61±4.36 respectively; compared with the experimental group, the FBG and Fasting C-peptide indices were P > 0.05, indicating that there was no significant difference between the two groups; the HOMA-IR and FIns indices were P < 0.05, which were statistically significant, and the effect of the experimental group was better than that of the control group.

[0042] From the data in Table 5, we can see that: the comparison of each data before and after surgery in the model group was P < 0.05, indicating that gastrectomy caused insulin resistance in rats; the comparison of each data before and after surgery in the control group was P < 0.05, indicating that rats still had insulin resistance after taking ordinary lotus leaf extract, but the postoperative insulin resistance was alleviated compared with the model group; and the comparison of each data before and after surgery in the experimental group showed that FBG, FIns, and HOMA-IR indicators were P > 0.05, and Fasting C-peptide index was P < 0.05, indicating that taking space lotus leaf extract can alleviate the insulin resistance of rats after gastrectomy.

[0043] 4. Effect of space lotus leaf extract on intestinal flora diversity in rats after gastrectomy At the end of the experiment, rat feces were collected and 16S rRNA high-throughput sequencing was performed to observe the effects of space lotus leaf extract on the α-diversity and β-diversity of rat intestinal flora.

[0044] Among them, α diversity refers to the diversity of the flora within a community or biological environment. The number and distribution of the flora are calculated based on OTU data. The number of flora species in a community reflects the richness, and the number and distribution of each flora reflect the uniformity. Specifically, it includes: the Shannon index for evaluating the richness and uniformity of the flora composition in the sample; the Goods_coverage index that reflects whether the sequencing results represent the true situation of the sample, and the Chao1 index that estimates the number of OTUs contained in the sample. The results are shown in Table 6. The Shannon index and Chao1 index increase with the increase of flora richness and uniformity; the Goods_coverage index is the flora coverage rate. The higher its value, the lower the probability that the new flora in the sample has not been detected.

[0045] Table 6

[0046] It can be seen from the data in Table 6 that the α diversity index of the model group rats was significantly lower than that of the control group rats. After intervention with the control group of ordinary lotus leaf extract and the experimental group of space lotus leaf extract, the richness and uniformity of the intestinal flora can be restored to a certain extent, and the effect of the experimental group is more obvious.

[0047] The β diversity index of the intestinal flora of each group of rats was further analyzed. Specifically, in this example, PCoA analysis of the intestinal flora was performed at the OTU level, that is, principal coordinates analysis. The results are as follows: Figure 1 As shown. Figure 1It was found that the community structure of intestinal flora in the model group rats showed significant changes compared with the control group rats. After intervention, the control group of ordinary lotus leaf extract and the experimental group of space lotus leaf extract could help restore the normal community structure of intestinal flora, and the effect of the experimental group was more obvious.

[0048] The above-mentioned embodiments only express several implementation methods of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the patent of the present invention. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. The application of space lotus leaf extract is characterized by: The space lotus leaf extract is used for preparing a product with the effect of improving postoperative insulin resistance.

2. The use of the space lotus leaf extract according to claim 1, characterized in that: The postoperative period is after gastrectomy; the product is a health product or a medicine.

3. The use of the space lotus leaf extract according to claim 1, characterized in that: The product regulates the balance of intestinal flora after surgery.

4. The use of the space lotus leaf extract according to claim 1, characterized in that: The product reduces postoperative fasting blood sugar levels.

5. The use of the space lotus leaf extract according to claim 1, characterized in that: The product reduces postoperative fasting insulin levels.

6. The use of the space lotus leaf extract according to claim 1, characterized in that: The product reduces postoperative fasting C-peptide levels.

7. The use of the space lotus leaf extract according to claim 1, characterized in that: The preparation method of the space lotus leaf extract comprises the following steps: The dried space lotus leaf powder is dissolved in ethanol of a set concentration, and then placed in an ultrasonic bath for alcohol extraction to obtain a preliminary extract; The preliminary extract was centrifuged and filtered to obtain the residue and the supernatant; The filter residue is subjected to repeated alcohol extraction in an ultrasonic bath, centrifugation and membrane filtration treatments at least twice; After the supernatants are combined, a first decompression treatment is performed to remove ethanol in the solution, and then water is added and stirred to obtain a crude extract of space lotus leaves; The crude extract of space lotus leaves flows through an adsorption column filled with macroporous resin for adsorption, then is washed with water to remove non-adsorbed matter, and then is eluted with ethanol, and the eluate is collected; The eluate is subjected to a second decompression treatment and used for concentration to obtain a clear paste, which is then freeze-dried to obtain the space lotus leaf extract.

8. The use of the space lotus leaf extract according to claim 7, characterized in that: In the alcohol extraction process, the ethanol mass concentration is 70% to 80% and the time is 45 min to 50 min.

9. The use of the space lotus leaf extract according to claim 7, characterized in that: The temperature of the first decompression treatment and the second decompression treatment is 60° C. and the pressure is 0.085 MPa.

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