Radix morindae officinalis-inulin composition and its use in kidney fibrosis products

Abstract

The application belongs to the technical field of biological medicine, and particularly relates to a radix morindae officinalis-inulin composition and application thereof in a kidney fibrosis product. The radix morindae officinalis-inulin composition provided by the application is composed of radix morindae officinalis polysaccharide, inulin and water, can exert the synergistic effect of the two polysaccharides, improve the physical and chemical properties and biological activity of the radix morindae officinalis-inulin composition, can significantly enhance the metabolic rate of urea nitrogen creatinine of the kidney, and can significantly reduce the fibrosis positive area, and has a significant effect on the treatment of kidney fibrosis diseases. The raw materials of the composition are based on natural polysaccharides, have good safety and degradability, and meet the high safety requirements of biological materials in the medical field. Compared with the existing single component, the composite design can realize multi-target effect, significantly improve the treatment effect, and has a great market competitive advantage.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and specifically relates to a radix morindae officinalis-inulin composition and its use in a kidney fibrosis product. BACKGROUND

[0002] Renal interstitial fibrosis (RIF) is a hallmark and ultimate outcome of various progressive chronic kidney disease (CKD). Renal fibrosis is caused by the deposition of a large amount of connective tissue in the kidney tissue, which gradually leads to the destruction of the structure and function of the tubules, glomeruli and interstitium, thereby affecting the normal operation of the kidney.

[0003] In recent years, through in-depth study of the pathophysiological mechanism, it has been found that immune response plays an important role in the formation and development of RIF, and regulating immune response may be an important way to change the course of RIF. Immune imbalance is closely related to the formation of RIF. In CKD, the immune response is continuously activated, leading to a large number of inflammatory cell infiltration, kidney parenchymal cell damage or fibroblast activation, thereby promoting the persistence of renal fibrosis and eventually progressing to chronic renal failure; in the process of RIF, the immune response is often in an abnormal activated state, leading to significant changes in the immune environment of the kidney. This continuous activation of immune response is caused by pathological inflammation, and is closely related to the participation of various immune cells in the kidney. For example, the infiltration and continuous activation of T cells, B cells, macrophages and dendritic cells and other immune cells will promote the exacerbation of local inflammatory response. The infiltration of immune cells and the cytokines and chemokines secreted by them further aggravate the damage and dysfunction of kidney parenchymal cells. Therefore, regulating the immune response of the kidney becomes a potential therapeutic target for changing the course of RIF.

[0004] In recent years, domestic and foreign research has gradually focused on the effects of Morinda officinalis How and its polysaccharide components on immune regulation and kidney protection. Morinda officinalis How (MO) is a traditional Chinese medicine commonly used for tonifying kidney yang and enhancing immune function. Its active ingredients include polysaccharides, steroids, flavonoids, etc. Among them, Morinda officinalis How Polysaccharides (MOPs) as one of the main active substances, show significant immune regulation and kidney protection potential. For example, Chinese patent CN108752497A discloses a preparation of Morinda officinalis How water extract, oligosaccharides and polysaccharides and its use. Specifically, Morinda officinalis How water extract and crude polysaccharides are extracted from the roots of Morinda officinalis How by water extraction, and then oligosaccharides and polysaccharide components are separated from the crude polysaccharides and their properties are determined. Experiments show that Morinda officinalis How water extract and crude polysaccharides can promote the proliferation of mouse spleen cells and the secretion of cytokines, promote the proliferation of human liver cells and reduce the damage of toxic agents to cells, inhibit the expression of hepatitis B surface antigen and core antigen, inhibit the proliferation of liver cancer cells, and inhibit the damage of ConA to mouse liver and kidney, and have a protective effect on the kidney.

[0005] However, it is worth noting that based on the single drug treatment strategy, it is difficult to achieve the overall reconstruction of intestinal homeostasis and the continuous maintenance of long-term remission. From the perspective of pharmacology and physiology, during the transport of drugs in the gastrointestinal tract, they face extremely complex and harsh physicochemical environment challenges. The gastrointestinal tract is rich in various digestive enzymes, such as trypsin and pancreatic amylase, which can enzymatically destroy the structure of the drug; the diversity and complexity of intestinal flora cannot be underestimated, and their metabolites and bacteria themselves may interact with the drug, affecting its activity; in addition, the gastrointestinal tract presents a significant pH gradient change from the strong acidic environment in the stomach to the weak alkaline environment in the small intestine, which poses a severe test to the stability and solubility characteristics of the drug; in addition, the intestinal peristalsis is rhythmic and fast, which shortens the effective residence time of the drug in the intestine, and thus adversely affects the absorption and effect of the drug.

[0006] Therefore, providing a drug capable of regulating immune response and intestinal microbiota, in-depth analysis of the related mechanism of the key pharmacodynamic components of the drug in the process of kidney fibrosis can provide material basis and transformation value for the clinical treatment of kidney fibrosis. SUMMARY

[0007] The present application provides a Morinda officinalis How-inulin composition and its use in kidney fibrosis products. The composite gel has good biocompatibility and multifunctionality, and can simultaneously intervene in the intestinal microenvironment and kidney fibrosis, filling the gap in existing treatment methods.

[0008] To achieve the above object, the technical scheme adopted by the present application is as follows:

[0009] The present application provides a Radix Morindae Officinalis-inulin composition and its use in kidney fibrosis products.

[0010] Further, the Radix Morindae Officinalis-inulin composition consists of Radix Morindae Officinalis polysaccharide, inulin and water.

[0011] Further, the Radix Morindae Officinalis-inulin composition comprises 0.5-1.5 parts of Radix Morindae Officinalis polysaccharide, 7-12 parts of inulin and 5-15 parts of water according to weight fraction.

[0012] Further, the Radix Morindae Officinalis-inulin composition comprises 0.5-1.5 parts of Radix Morindae Officinalis polysaccharide, 7-12 parts of inulin and 10 parts of water according to weight fraction.

[0013] Further, the preparation method of the Radix Morindae Officinalis-inulin composition comprises the following steps:

[0014] S1, mixing Radix Morindae Officinalis polysaccharide and inulin according to weight fraction to obtain a mixture;

[0015] S2, heating water and adding the mixture obtained in step S1 into water according to weight fraction;

[0016] S3, stirring to homogeneous viscous state, cooling, standing and obtaining the Radix Morindae Officinalis-inulin composition.

[0017] Further, the preparation method of Radix Morindae Officinalis polysaccharide in step S1 comprises the following steps:

[0018] (1) mixing Radix Morindae Officinalis slices with water, refluxing and extracting, collecting extract A;

[0019] (2) centrifuging extract A to obtain supernatant A and Radix Morindae Officinalis residue;

[0020] (3) mixing Radix Morindae Officinalis residue with water, refluxing and extracting, collecting extract B;

[0021] (4) centrifuging extract B to obtain supernatant B, mixing supernatant A and supernatant B, adding ethanol, making the volume fraction of ethanol account for more than 80% of the total volume fraction, standing for 24-48h and collecting precipitate;

[0022] (5) treating the precipitate by washing with ethanol, dialysis, decolorization and protein removal to obtain the Radix Morindae Officinalis polysaccharide.

[0023] Preferably, the ethanol is anhydrous ethanol.

[0024] Further, the heating temperature in step S2 is 65-85℃.

[0025] Further, the temperature of the cooling in step S3 is 20-30℃.

[0026] Further, the temperature of the standing in step S3 is 0-4℃; the time of the standing is ≥12h.

[0027] Further, the renal fibrosis product includes a drug for preventing or treating renal fibrosis.

[0028] Compared with the prior art, the present application has the following beneficial effects:

[0029] (1) The radix morindae officinalis-inulin composition provided by the present application can exert the synergistic effect of the two polysaccharides, improve the physical and chemical properties and biological activity of the hydrogel, significantly enhance the metabolic rate of urea nitrogen creatinine in the kidney, and significantly reduce the fibrosis positive area, thereby having a significant effect on the treatment of renal fibrosis diseases.

[0030] (2) The radix morindae officinalis-inulin composition provided by the present application has good biocompatibility and multifunctionality, and has potential efficacy in intestinal barrier repair, inflammation regulation, microbial regulation and inhibition of renal fibrosis, thereby providing a new treatment option for the clinic; the composition can simultaneously intervene in the intestinal microenvironment and renal fibrosis, thereby filling the gap in the existing treatment methods.

[0031] (3) The radix morindae officinalis-inulin composition provided by the present application is based on natural polysaccharides, has good safety and degradability, and meets the high safety requirements of the medical field for biological materials. Compared with the existing single-component hydrogel, the composite design can realize multi-target action, significantly improve the treatment effect, and has a great market competitive advantage. BRIEF DESCRIPTION OF DRAWINGS

[0032] Figure 1 is a glucose standard curve graph.

[0033] Figure 2 is a characterization graph of the radix morindae officinalis-inulin composition of Example 1; A in the graph is the phenomenon of the radix morindae officinalis-inulin composition injected into water by a syringe; B is the phenomenon of water at an angle of 45°; C is the phenomenon of the radix morindae officinalis-inulin composition at an angle of 45°.

[0034] Figure 3 is an FT-IR spectrum graph of the radix morindae officinalis-inulin composition of Example 1.

[0035] Figure 4 is a SEM scanning result graph of the radix morindae officinalis-inulin composition of Example 1.

[0036] Figure 5 is a rheological property test result graph of the radix morindae officinalis-inulin composition of Example 1.

[0037] Figure 6 Figure 9 is a graph of body weight growth trend and kidney index chart; A in the figure is the body weight growth trend from the beginning of modeling to sampling; B is the body weight change of mice at the time of sampling compared with the beginning of modeling; C is the kidney index at the time of sampling; ns in the figure represents no significant difference between groups; * represents a significant difference p<0.05 compared with the RIF model group; ** represents a significant difference p<0.01 compared with the RIF model group; *** represents a significant difference p<0.001 compared with the RIF model group; **** represents a significant difference p<0.0001 compared with the RIF model group; # represents a significant difference p<0.05 compared with the Radix Morindae Officinalis polysaccharide group; ### represents a significant difference p<0.001 compared with the Radix Morindae Officinalis polysaccharide group.

[0038] Figure 7 Figure 10 is a graph of serum biochemical indicators; A in the figure is a serum creatinine indicator chart; B is a serum urea nitrogen indicator chart; ns in the figure represents no significant difference between groups; * represents a significant difference p<0.05 compared with the RIF model group; # represents a significant difference p<0.05 compared with the Radix Morindae Officinalis polysaccharide group; ### represents a significant difference p<0.001 compared with the Radix Morindae Officinalis polysaccharide group.

[0039] Figure 8 Figure 11 is a graph of urine biochemical indicators; A in the figure is a urine protein indicator chart; B is a urine creatinine indicator chart; C is a urea nitrogen indicator chart; ns in the figure represents no significant difference between groups; * represents a significant difference p<0.05 compared with the RIF model group; ** represents a significant difference p<0.01 compared with the RIF model group; *** represents a significant difference p<0.001 compared with the RIF model group; # represents a significant difference p<0.05 compared with the Radix Morindae Officinalis polysaccharide group.

[0040] Figure 9 Figure 12 is a graph of histomorphology and pathology results, in which the blue color in Masson staining is the fibrosis positive area, and the arrow points to the glomerular morphology. DETAILED DESCRIPTION

[0041] The present application will be described in detail below through specific embodiments, so that the technical scheme of the present application is easier to understand and master, but the present application is not limited thereto, and the described embodiments are only a part of the embodiments of the present application, not all the embodiments.

[0042] The endpoints of the ranges and any values disclosed herein are not limited to the precise values stated. The endpoints of the ranges are included in the range unless it states otherwise. For ranges comprising a single number, the endpoints are included in the range, unless it states otherwise. The disclosure of a single value of a parameter or characteristic is not intended to be a disclosure of only that single value but rather is meant to be a disclosure of a range ending with that single value. Unless otherwise indicated, the singular forms "a", "an", and "the" include plural referents. Numerical ranges include all values from and including the lower and the upper values, in increments of one unit. In this application, the use of "adapted" means the entity is merelv modified as appropriate to the context of use.

[0043] All other embodiments obtained by a person of ordinary skill in the art based on the embodiments of the present application without creative labor shall fall within the scope of protection of the present application. The experimental methods described in the following examples are all conventional methods unless otherwise specified; the reagents and materials described are all commercially available unless otherwise specified.

[0044] Example 1

[0045] 1. Preparation of Radix Morindae Officinalis polysaccharide:

[0046] According to the ratio of ultrapure water (100 g) to Radix Morindae Officinalis (sliced) (100 g), mix and reflux extract at 90 °C for 2 h. Centrifuge the extraction solution at 4000 rpm for 20 min, and harvest the supernatant. Repeat the process for secondary extraction and centrifugation, and combine the supernatants after multiple filtrations. Treat the combined mixture with anhydrous ethanol to a content of 80% and precipitate at 4 °C for 24 h to obtain crude polysaccharide.

[0047] Wash the crude polysaccharide twice with 95% ethanol, then dialyze with distilled water (using a molecular weight cut-off of 3.5 kDa), decolorize with activated carbon, and remove protein by the Savage method (Sevage reagent (n-butanol: chloroform in a volume ratio of 1:4) and 500 mg / ml of polysaccharide solution in a volume ratio of 1:2, oscillate for 20 min, centrifuge at 3000 rpm for 3 min, and retain the supernatant). Obtain purified polysaccharide. Freeze-dry the purified polysaccharide, and obtain Radix Morindae Officinalis polysaccharide in the form of off-white freeze-dried powder.

[0048] 2. Preparation of Radix Morindae Officinalis-inulin composition:

[0049] Mix 10 g of inulin (purchased from Macron Fine Chemicals, item number I811905-10 g) and 0.8 g of Radix Morindae Officinalis polysaccharide to obtain a mixture of 10.8 g. Heat 10 g of water to 72 °C, and slowly add the mixture. Mix by magnetic stirring until homogeneous and viscous. After the sample cools to 26 °C, stand at 4 °C overnight (12 h). Obtain the Radix Morindae Officinalis-inulin composition.

[0050] Figure 2 Gel form, injectability and angle stability of the Radix Morindae Officinalis-inulin composition of Example 1 were implemented. It can be seen that the Radix Morindae Officinalis-inulin composition has good injectability under visual observation, remains no deformation at 45° angle, and has high viscosity.

[0051] Example 2

[0052] 1. Preparation of Radix Morindae Officinalis polysaccharide: the same as Example 1.

[0053] 2. Preparation of Radix Morindae Officinalis-inulin composition:

[0054] 8 g of inulin and 1.5 g of Radix Morindae Officinalis polysaccharide were mixed to obtain a mixture of 9.5 g. 8 g of water was heated to 85°C, and the mixture was slowly added. The mixture was mixed by magnetic stirring until it became homogeneous and viscous. After the sample was cooled to 20°C, it was left to stand at 4°C overnight. The Radix Morindae Officinalis-inulin composition was obtained.

[0055] Example 3

[0056] 1. Preparation of Radix Morindae Officinalis polysaccharide: the same as Example 1.

[0057] 2. Preparation of Radix Morindae Officinalis-inulin composition:

[0058] 12 g of inulin and 0.5 g of Radix Morindae Officinalis polysaccharide were mixed to obtain a mixture of 15.5 g. 15 g of water was heated to 65°C, and the mixture was slowly added. The mixture was mixed by magnetic stirring until it became homogeneous and viscous. After the sample was cooled to 30°C, it was left to stand at 4°C overnight. The Radix Morindae Officinalis-inulin composition was obtained.

[0059] Effect example

[0060] 1. Preparation of glucose standard curve

[0061] 10 mg of glucose reference substance was accurately weighed, dissolved in distilled water, and then transferred to a 100 mL volumetric flask for constant volume to prepare a glucose reference substance stock solution with a concentration of 0.100 mg / mL.

[0062] The prepared glucose reference substance stock solution was diluted to a series of concentrations of 0.01 mg / mL, 0.02 mg / mL, 0.03 mg / mL, 0.04 mg / mL, 0.05 mg / mL, 0.06 mg / mL, 0.08 mg / mL, and 0.10 mg / mL, and then 1.0 mL was taken into a test tube, 1.0 mL of 5% phenol reagent and 5.0 mL of 98% concentrated sulfuric acid were added, and it was shaken and boiled for 30 min. The absorbance (A) was measured at 490 nm by ultraviolet spectrophotometry. The standard curve was drawn with the glucose concentration (C) as the abscissa and the absorbance (A) as the ordinate, and the standard curve was as followsFigure 1 as shown.

[0063] 2. Determination of the content of polysaccharides in Radix Morindae Officinalis

[0064] Take the polysaccharides in Radix Morindae Officinalis, and quantitatively re-dissolve in ultrapure water. Determine the content of polysaccharides in the polysaccharides in Radix Morindae Officinalis, and the content of polysaccharides is > 85% to be qualified. The specific steps are as follows:

[0065] Accurately weigh 0.2 g (B) of polysaccharides in Radix Morindae Officinalis prepared in Example 1 into a 100 mL volumetric flask, dissolve with distilled water, and then re-dissolve to the calibration line. Take 10 mL into a 100 mL volumetric flask, and then re-dissolve to the calibration line with distilled water. Then take 1.0 mL and inject it into a test tube, add 1.0 mL of 5% phenol reagent and 5.0 mL of concentrated sulfuric acid, shake well and boil for 30 min. Measure the absorbance (A1) at 490 nm by ultraviolet spectrophotometry. Put the A1 value into the linear regression equation of the standard curve to obtain the polysaccharide concentration (C1).

[0066] Repeat the determination of the absorbance A1 three times to take the average value 1.406, and put it into the linear regression equation of the standard curve to obtain the polysaccharide concentration C1 of 0.1882 mg / mL. It is calculated that the polysaccharide content of the polysaccharides in Radix Morindae Officinalis is 94.1%. It can be seen that the polysaccharide content of the polysaccharides in Radix Morindae Officinalis prepared by the present application is > 85%, which is qualified.

[0067] 3. FT-IR spectrum measurement of Radix Morindae Officinalis-inulin composition

[0068] The experiment was performed using KBr-disk method with a Nicolet iZ-10 spectrometer (Thermo Nicolet, USA). First, potassium bromide (KBr) powder was pre-dried at 104°C for 12 hours. Then, the Radix Morindae Officinalis-inulin composition (MOPs-Inulin Gel, MI Gel) sample prepared in Example 1 was mixed with the pre-dried KBr powder (100:1) in a corundum mortar, and was pressed into 1 mm pellets. The spectrum was collected in the range of 400 to 4000 cm -1 .

[0069] The FT-IR spectrum of the MI Gel sample is shown as Figure 3 , and the test shows that:

[0070] 3368 cm -1 corresponds to the stretching vibration absorption peak of O-H, and a wide absorption band is formed;

[0071] 2931 cm -1 corresponds to the stretching vibration peak of CH3, CH2 or CH;

[0072] 1419 cm -1corresponding to the variable angle vibration peak of CH3, CH2 or CH;

[0073] 1031 cm -1 corresponding to the stretching vibration peak of C-OH;

[0074] 936 cm -1 corresponding to the symmetric stretching vibration peak of C-O-C;

[0075] 1636 cm -1 corresponding to the variable angle vibration peak of OH, indicating that the sample contains a small amount of water;

[0076] From the above characteristic peaks, it can be judged that the MI Gel sample contains the presence of saccharide compounds.

[0077] 4. SEM scanning of Radix Morindae Officinalis-inulin composition

[0078] The Radix Morindae Officinalis-inulin composition sample prepared in Example 1 was coated with gold under vacuum and imaged using a scanning electron microscope at a magnification of 100 to 4000 times using an acceleration voltage of 5.0 kV.

[0079] The scanning electron microscope (SEM) results are shown in Figure 4 It can be seen that the Radix Morindae Officinalis-inulin composition has a loose and porous network of tight connection structure, which meets the general characteristics of polysaccharide gel.

[0080] 5. Rheological test of Radix Morindae Officinalis-inulin composition

[0081] The Radix Morindae Officinalis-inulin composition of Example 1 was subjected to amplitude test at fixed frequency using a rheometer; linear change of shear rate; temperature scan with fixed strain and frequency. The results are shown in Figure 5 Temperature scan was performed between 0°C and 90°C. Strain-dependent oscillatory shear rheology, fixed frequency of 1 rad / s. Viscosity measurement at shear rate of 0.01 to 100 s −1 The above is the rheological characteristics of the Radix Morindae Officinalis-inulin composition (reference is deionized water).

[0082] 6. Therapeutic effect of Radix Morindae Officinalis-inulin composition on renal fibrosis (RIF)

[0083] (1) Experimental animals

[0084] The experimental animals were 8-week-old male C57BL / 6J mice, 64 in number, purchased from Guangdong Rige Biological Technology Co., Ltd. The mice were raised in the SPF environment of the animal experiment center of Guangdong Pharmaceutical University, with a 12-hour light / dark cycle, a temperature of 20-25°C, and a humidity of 60 ± 5%. The animals could freely eat and drink water. All experimental mice were adaptively fed for 1 week before the experiment. The experimental scheme and procedure were approved by the Animal Experiment Ethics Committee of Guangdong Pharmaceutical University on October 16, 2023, in accordance with the Guidelines for the Management and Use of Laboratory Animals.

[0085] (2) Induction of RIF model and grouping and administration

[0086] The experiment was divided into 8 groups, 8 mice in each group: healthy control group (Control, Ctrl), RIF model group (RIF), positive control group (losartan, LOS), Example 1 group (MI Gel), and 2 single-drug control groups (treated with polysaccharides from Radix Morindae Officinalis and inulin, respectively).

[0087] From weeks 1 to 5, all groups except the blank control group were given a diet supplemented with 0.25% adenine. From weeks 6 to 11, all groups except the blank control group were given a diet supplemented with 0.10% adenine, and the treatment of mice began.

[0088] The RIF model group and the healthy control group were given pure water by gavage (0.1 mL / d) (the administration volume of all treatment groups was controlled to be 0.1 mL).

[0089] The positive control group was given 100 μL / D of a losartan aqueous solution with a concentration of 30 mg / kg.

[0090] Example 1 group: 100 μL / D of the Radix Morindae Officinalis-inulin composition prepared in Example 1 was given by gavage.

[0091] Polysaccharides from Radix Morindae Officinalis group (MOPs): 100 μL / D of an MOPs aqueous solution (MOPs aqueous solution concentration: 200 mg / kg) was given by gavage.

[0092] Inulin group (Lnulin): 100 μL / D of an Lnulin aqueous solution, i.e., 3125 mg / kg / d of Lnulin, was given by gavage.

[0093] The mice were euthanized at week 11.

[0094] (3) Experimental results - body weight change rate and kidney index

[0095] The body weight growth trend of the experimental animals from the beginning of modeling to the time of sampling and the kidney index at the time of sampling were recorded. The determination results are shown in Table 1. Figure 6 Figure 6 ​In the figure, A represents the weight growth trend from the beginning of modeling to the time of sample collection; B represents the weight change of mice at the time of sample collection compared to the beginning of modeling; and C represents the kidney index.

[0096] Except for the healthy control group mice, the body weight of mice in other groups showed a decreasing trend in the first 5 weeks. After drug treatment, the body weight of mice in the Example 1 group, Example 2 group, Example 3 group, Morinda officinalis polysaccharide group and inulin group continued to recover, indicating that the Morinda officinalis-inulin combination, Morinda officinalis polysaccharide and inulin can all improve the weight loss caused by RIF, among which the Morinda officinalis-inulin combination has the most obvious improvement effect.

[0097] The kidney index of mice in the RIF model group was significantly lower than that of healthy control mice (p < 0.0001), indicating that the renal fibrosis model was successfully established. The kidney index of mice in the Morinda officinalis-inulin combination group, Morinda officinalis polysaccharide group, and inulin group all increased to varying degrees, with the Morinda officinalis-inulin combination group showing the best improvement in kidney index.

[0098] (4) Experimental results - biochemical indicators

[0099] Twelve hours after the last administration of the drug to mice, blood was collected by enucleation. The blood samples were centrifuged at 3000 rpm for 15 min to obtain serum. Serum urea nitrogen (BUN) and creatinine (Scr) were measured using a biochemical assay kit.

[0100] Urine was collected from mice 24 hours prior to the last administration using metabolic cages. Urine protein, blood urea nitrogen, and creatinine were measured using biochemical assay kits. The kits used were: blood urea nitrogen kit (purchased from Nanjing Jiancheng, catalog number C013-2-1); creatinine kit (purchased from Nanjing Jiancheng, catalog number C011-2-1); and urine protein kit (purchased from Nanjing Jiancheng, catalog number C035-2-1).

[0101] Serum biochemical indicators such as Figure 7 As shown, urine biochemical indicators are as follows: Figure 8 As shown in the figure. The results showed that the Morinda officinalis-inulin combination significantly reduced serum creatinine and urea nitrogen levels; inhibited the increase of 24-hour proteinuria; and significantly enhanced the renal metabolic rate of urea nitrogen and creatinine.

[0102] (5) Experimental results – histomorphology and pathology

[0103] Representative photographs of kidney and colon tissues collected from mice euthanized on the last day of week 11 were taken, fixed overnight in 4% paraformaldehyde solution at 4°C, then embedded in paraffin and cut into 4 μm sections. Hematoxylin-eosin (HE) staining was used to observe glomerular structure. Masson staining was used to show the degree of renal fibrosis.

[0104] The results are as followsFigure 9 As shown, the kidney of RIF model mice was significantly atrophic and fibrotic compared to the healthy control group, and the acorus- inulin composition group had a trend of improvement. H&E staining showed that the glomerulus of RIF mice was atrophic, which was alleviated by acorus- inulin composition treatment. In Masson staining, the acorus- inulin composition group significantly reduced the fibrosis positive area.

[0105] Finally, it should be noted that the above is only used to illustrate the technical solutions of the present application, and is not a limitation on the scope of protection of the present application. Simple modifications or equivalent replacements of the technical solutions of the present application made by those skilled in the art do not deviate from the essence and scope of the technical solutions of the present application.

Claims

1. Use of a Radix Morindae Officinalis-inulin composition in the preparation of a medicament for preventing or treating renal fibrosis, the Radix Morindae Officinalis-inulin composition consisting of 0.5-1.5 parts of Radix Morindae Officinalis polysaccharide, 7-12 parts of inulin and 5-15 parts of water by weight; The preparation method of the Radix Morindae Officinalis polysaccharide comprises the following steps: (1) After slicing, the Radix Morindae Officinalis is mixed with water, refluxed and extracted, and the extract A is collected; (2) The extract A is centrifuged to obtain supernatant A and Radix Morindae Officinalis residue; (3) The Radix Morindae Officinalis residue is mixed with water, refluxed and extracted, and the extract B is collected; (4) The extract B is centrifuged to obtain supernatant B, and the supernatant A and the supernatant B are mixed, and then ethanol is added to a volume fraction of more than 80% of the total volume fraction, and the precipitate is collected after standing for 24-48 h; (5) The precipitate is washed with ethanol, dialyzed, decolorized and deproteinized to obtain the Radix Morindae Officinalis polysaccharide.

2. Use according to claim 1, characterized in that: The Radix Morindae Officinalis-inulin composition consists of 0.5-1.5 parts of Radix Morindae Officinalis polysaccharide, 8-12 parts of inulin and 10 parts of water by weight.

3. Use according to claim 1, characterized in that: The preparation method of the Radix Morindae Officinalis-inulin composition comprises the following steps: S1, mixing Radix Morindae Officinalis polysaccharide and inulin according to weight parts to obtain a mixture; S2, heating water and adding the mixture obtained in step S1 to the water according to weight parts; S3, stirring until homogeneous and viscous, cooling, standing and obtaining the Radix Morindae Officinalis-inulin composition.

4. Use according to claim 3, characterized in that: The heating temperature in step S2 is 65-85°C.

5. Use according to claim 3, characterized in that: The cooling temperature in step S3 is 20-30°C.

6. Use according to claim 3, characterized in that: The standing temperature in step S3 is 0-4°C, and the standing time is ≥12 h.

Images