Rhodiola rosea polysaccharide, preparation method and application thereof in preparing medicine for treating chronic nephritis

Abstract

The present application discloses a Rhodiola rosea polysaccharide, a preparation method and its application in the preparation of a drug for treating chronic nephritis. The Rhodiola rosea polysaccharide is obtained by extracting, alcohol precipitation, depigmenting, deproteinizing and dialysis Rhodiola rosea. The Rhodiola rosea polysaccharide includes two molecular weight ranges of 1.05×10 3 ‑8.26×10 3 , 1.78×10 9 ‑9.45×10 9 The polysaccharide content of Rhodiola rosea polysaccharide is 76wt.%-85wt.%. Animal experimental results show that in the adriamycin-induced rat nephropathy model, the Rhodiola rosea polysaccharide can significantly improve renal tubular lesions, effectively reduce the 24h urine protein amount of rats, and reduce the degree of blood albumin reduction. It can be used to treat chronic nephritis, especially minimal change nephritis.

Description

Technical Field

[0001] The present application relates to the technical field of traditional Chinese medicine, and in particular to a Rhodiola rosea polysaccharide, a preparation method and an application thereof in the preparation of a drug for treating chronic nephritis. Background Art

[0002] Glomerular diseases are the main cause of chronic renal failure in my country, mainly affecting the glomeruli of both kidneys. Chronic primary glomerulonephritis refers to an immune inflammatory disease originating in the glomeruli caused by one or more causes. According to the results of pathological examination, it can be divided into minimal change nephropathy, membranous nephropathy, mesangial proliferative glomerulonephritis, mesangial capillary glomerulonephritis, and focal segmental sclerosis glomerulonephritis.

[0003] The prevalence of chronic kidney disease in adults in my country is 13.1%, among which the prevalence rate in women is higher than that in men. Because of its insidious onset, long course, and potentially progressive kidney damage, it eventually develops into end-stage renal disease, and must rely on dialysis or kidney transplantation to maintain life. It has become a global public health issue after cardiovascular and cerebrovascular diseases and diabetes.

[0004] At present, Western medicine mainly uses hormones and cytotoxic drugs to treat nephritis, mainly by controlling blood pressure, reducing proteinuria and other methods to delay the progression of the disease, but there are many problems such as toxic side effects, hormone dependence, excessive economic burden, and low quality of life. Traditional Chinese medicine has certain advantages in improving patients' clinical symptoms, protecting renal function, and slowing down renal fibrosis.

[0005] Rhodiola rosea L. is a perennial herbaceous plant of the Crassulaceae family, with the Tibetan name Saoromabul. Its dried rhizomes are used as medicine. It is widely distributed in Qinghai, Tibet, Xinjiang, Sichuan and other regions. Modern research shows that Rhodiola rosea contains a variety of active ingredients such as polysaccharides, glycosides, salidrosides, phenols, flavonoids, amino acids, coumarins, organic acids, steroidal compounds and alkaloids, and has a variety of potential benefits, especially in terms of human athletic ability and anti-aging, which has attracted widespread attention. Among them, Rhodiola rosea polysaccharides have the effects of anti-fatigue, anti-aging, anti-virus, hypoglycemic, immune regulation, antibacterial, anti-oxidation, hypoglycemic, regulating glucose and lipid metabolism, anti-injury, and anti-adverse stimulation. Summary of the invention

[0006] In order to solve the above-mentioned deficiencies in the art, the present application aims to provide a Rhodiola rosea polysaccharide, a preparation method and its application in the preparation of a drug for treating chronic nephritis.

[0007] According to one aspect of the present application, there is provided a use of Rhodiola rosea polysaccharide in the preparation of a drug for treating chronic nephritis, wherein the two molecular weight ranges of the Rhodiola rosea polysaccharide are: 1.05×10 3 -8.26×103 , 1.78×10 9 -9.45×10 9 ;

[0008] The polysaccharide content in Rhodiola rosea polysaccharide is 76wt.%-85wt.%.

[0009] According to some embodiments of the present application, Rhodiola rosea is Rhodiola rosea.

[0010] According to another aspect of the present application, a method for preparing the above-mentioned Rhodiola rosea polysaccharide is provided, comprising:

[0011] Step 1: Extraction: extracting the Rhodiola rosea medicinal material with alcohol to obtain alcohol extraction residue and alcohol extraction filtrate;

[0012] Step 2: removing impurities: extracting the alcohol extraction residue in step 1 with water to obtain a water extract;

[0013] Step 3: Enzymatic hydrolysis: passing the aqueous extract in step 2 through a column and eluting, collecting and combining the effluent and the eluate, and concentrating to obtain a concentrate;

[0014] Step 4: alcohol precipitation: enzymatically hydrolyze the concentrated solution in step 3 and take the supernatant;

[0015] Step 5: Dialysis and drying: The supernatant in step 4 is subjected to alcohol precipitation, and the precipitate is collected, dialyzed, and freeze-dried to obtain the Chinese medicine polysaccharide.

[0016] According to some embodiments of the present application, column passing and eluting include: cooling the water extract to room temperature, passing through a macroporous resin column, and eluting with 2BV pure water;

[0017] Optionally, D101 macroporous resin is used for column passing and elution;

[0018] Optionally, the volume ratio of the loading amount of the water extract to the macroporous resin is 1:(1-3);

[0019] Further optionally, the loading flow rate is 2-3 BV / h.

[0020] According to some embodiments of the present application, dialysis comprises: washing the precipitate twice with ethanol, adding an appropriate amount of water to redissolve it, filtering it, transferring the filtrate to a dialysis bag, and dialyzing it;

[0021] Optionally, the particle size of the dialysis bag is 1000Da-85000Da, and further optionally, the particle size of the dialysis bag is 85000Da. According to some embodiments of the present application, enzymolysis comprises: adding one of papain, pepsin, trypsin, papain and endopeptidase, enzymolysis at 37-60°C for 1-1.5h, cooling and centrifuging to obtain a supernatant; optionally, the amount of enzyme used in the enzymolysis is 0.0004-0.001g / g crude drug amount.

[0022] According to some embodiments of the present application, the alcohol precipitation comprises: adding ethanol to the supernatant, adjusting the ethanol concentration to 70-95%, standing overnight, and collecting the precipitate.

[0023] According to another aspect of the present application, a traditional Chinese medicine preparation for treating chronic nephritis is provided, comprising the above-mentioned Rhodiola rosea polysaccharide, and / or the Rhodiola rosea polysaccharide prepared by the above-mentioned preparation method.

[0024] Optionally, Rhodiola rosea polysaccharide is the only active ingredient in the traditional Chinese medicine preparation; or the traditional Chinese medicine preparation also includes other active ingredients or drugs.

[0025] Optionally, the other active ingredients include nutritional supplements.

[0026] The Chinese medicine preparation may be in the form of capsules, pills, powders, syrups, tablets, granules, plasters, mixtures, pellets, suppositories, aerosols, ointments or injections.

[0027] Compared with the prior art, this application has at least the following beneficial effects:

[0028] The present application provides a traditional Chinese medicine polysaccharide - Rhodiola rosea polysaccharide, which can significantly improve renal tubular lesions, effectively reduce the amount of 24-hour urine protein in rats, and reduce the degree of reduction of blood albumin. The Rhodiola rosea polysaccharide of the present invention can be used to treat chronic nephritis, especially minimal change nephritis.

[0029] Compared with other extracted components in Rhodiola rosea medicinal materials, the Rhodiola rosea polysaccharide of the present application has a better therapeutic effect and less side effects at the same dosage; and it has diverse targets, is not easy to produce drug resistance, and has no adverse reactions or fewer adverse reactions.

[0030] The present application also provides a method for preparing Chinese medicine polysaccharides, which is simple to operate and has high implementation efficiency. The raw materials used in the operation process are safe, low in cost, and environmentally friendly, and have wide application potential and good development prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 This is the analysis spectrum of the extract of the traditional Chinese medicine polysaccharide in the exemplary embodiment of this application.

[0032] Figure 2 This application illustrates the effects of the four extracts of Example 1 on weekly urine protein in nephritic rats.

[0033] Figure 3 This is an illustration of the effects of the four extracts of Example 1 of the present application on urine indicators of nephritis rats.

[0034] Figure 4This is an illustration of the effects of the four extracts of Example 1 of the present application on serum indicators of nephritis rats.

[0035] Figure 5 This is an illustration of the effects of the four extracts of Example 1 of the present application on renal pathological indicators. DETAILED DESCRIPTION

[0036] The technical solution of the present application will be clearly and completely described below in conjunction with the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present application.

[0037] It is particularly important to point out that similar substitutions and modifications made to the present application are obvious to those skilled in the art, and they are all deemed to be included in the present application. Relevant personnel can obviously modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit and scope of the present application to implement and apply the technology of the present application. Obviously, the described embodiments are only some embodiments of the present application, not all embodiments.

[0038] If no specific conditions are specified in this application, the preparation shall be carried out under conventional conditions or the conditions recommended by the manufacturer. The raw materials or excipients used, as well as the reagents or instruments used, for which the manufacturers are not specified, are all conventional products that can be obtained commercially.

[0039] In the present application, the terms “further”, “furthermore”, “particularly”, etc. are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of protection of the present application.

[0040] In the present application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0041] In the present application, the percentage content, unless otherwise specified, refers to mass percentage for solid-liquid mixing and solid-solid mixing, and refers to volume percentage for liquid-liquid mixing.

[0042] In this application, when it comes to percentage concentration, unless otherwise specified, it refers to the final concentration. The final concentration refers to the proportion of the added component in the system after the addition of the component.

[0043] In the present application, wt.% means weight percentage, and vol% means volume percentage.

[0044] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application belongs. The terms used herein in the specification of this application are only for the purpose of describing specific embodiments and are not intended to limit this application.

[0045] The application is described in detail below.

[0046] Rhodiola rosea is a rare medicinal plant that contains more than 40 chemical components and is widely distributed in Qinghai, Tibet, Xinjiang, Sichuan and other regions.

[0047] Modern research shows that Rhodiola rosea contains a variety of active ingredients, including polysaccharides, glycosides, tyrosol, salidroside, phenols, flavonoids, amino acids, coumarins, organic acids, mineral elements, steroidal compounds and alkaloids. It has many potential benefits, especially in terms of human athletic ability and anti-aging, which has attracted widespread attention. Among them, Rhodiola rosea polysaccharides have the effects of anti-fatigue, anti-aging, anti-virus, hypoglycemic, immune regulation, antibacterial, anti-oxidation, hypoglycemic, regulating sugar and lipid metabolism, anti-injury, and anti-adverse stimulation.

[0048] This application is a study on Rhodiola rosea medicinal materials, in order to find active ingredients that are effective in treating chronic nephritis. During the experiment, it was found that the polysaccharide part of Rhodiola rosea has a good therapeutic effect; and further research is conducted on the extraction process and ingredients.

[0049] The Rhodiola rosea polysaccharide of the present application is prepared from Rhodiola rosea by alcohol extraction followed by water extraction, macroporous resin impurity removal, alcohol precipitation, enzymatic hydrolysis and dialysis. The polysaccharide content is 76wt.%-85wt.%, and the molecular weight ranges are: 1.05×10 3 -8.26×10 3 , 1.78×10 9 -9.45×10 9 .

[0050] The present application also provides a method for preparing Rhodiola rosea polysaccharide, comprising:

[0051] Step 1: Extraction: extracting the Rhodiola rosea medicinal material with alcohol to obtain alcohol extraction residue and alcohol extraction filtrate; extracting the alcohol extraction residue with water again,

[0052] Step 2: Impurity removal: The water extract is cooled to room temperature, passed through a macroporous resin column, eluted with 2BV of pure water, the effluent and the water wash are collected and combined, and concentrated to 1 g / mL of crude drug.

[0053] The macroporous resin model is selected from one of D312, LSA-30, LX-68G, AB-8 and D101.

[0054] Step 3: Enzymatic hydrolysis: add papain, pepsin, trypsin or endopeptidase to the concentrate, perform enzymatic hydrolysis at 37-60°C, boil, inactivate the enzyme for 10 minutes, cool to room temperature, centrifuge and take the supernatant.

[0055] Step 4: Alcohol precipitation: Add ethanol to the supernatant obtained in the previous step, adjust the ethanol concentration to 70-95%, let the alcohol precipitation stand overnight, and collect the precipitate.

[0056] Step 5 Dialysis and drying: Wash the precipitate twice with ethanol. Add appropriate amount of water to re-dissolve, filter, transfer the filtrate to a dialysis bag, dialyze, concentrate the retentate to 1g / mL crude drug, pre-freeze at -30℃, and freeze-dry to obtain.

[0057] Among them, the ratio of the sample loading amount and the macroporous resin in the impurity removal step is 1:(1-3);

[0058] The sample loading flow rate in the impurity removal step is 2-3BV / h.

[0059] The particle size of the dialysis bag is 1000Da-85000Da; optionally, the particle size of the dialysis bag is 85000Da.

[0060] The above preparation method also includes:

[0061] The alcohol extraction uses 60vol%-95vol% ethanol aqueous solution;

[0062] In the alcohol extraction step, the ethanol solution is refluxed for extraction at least twice, each time for 0.5-3 hours; the solid-liquid ratio of each alcohol extraction is 1: (8-20), and the ratio relationship is g / mL;

[0063] In the water extraction step, water is added and boiled at least twice, each time for 0.5-2 hours; the material-liquid ratio of each water extraction is 1:(10-20), and the ratio relationship is g / mL;

[0064] Enzymatic hydrolysis uses papain;

[0065] In the enzymatic hydrolysis step, the amount of papain used is 0.0004-0.001 g / g crude drug amount;

[0066] In the enzymatic hydrolysis step, the enzymatic hydrolysis temperature is 55-60° C. and the enzymatic hydrolysis time is 1-1.5 h.

[0067] After the centrifugation step, the solid is washed at least once with an ethanol solution to obtain a precipitate; the concentration of the ethanol solution is 80 vol%-95 vol%.

[0068] The pharmaceutical preparation of the present application comprises a preventive and / or therapeutically effective amount of Rhodiola rosea polysaccharide;

[0069] Optionally, the Rhodiola rosea polysaccharide is the only active ingredient in the traditional Chinese medicine preparation; or the medicine preparation also includes other active ingredients or drugs; other active ingredients include nutritional supplements.

[0070] The Chinese medicine preparation may be in the form of capsules, pills, powders, syrups, tablets, granules, plasters, mixtures, pellets, suppositories, aerosols, ointments or injections.

[0071] The technical solution of the present application is further described below in conjunction with specific embodiments.

[0072] Example Preparation of Rhodiola rosea polysaccharide of the present application.

[0073] Step 1 Extraction: Take 500g of Rhodiola rosea, crush through No. 1 sieve, add 10 times and 8 times of 95% ethanol to extract twice, each time for 1 hour, filter, leave the residue for 24 hours, add 12 times and 10 times of water, decoct twice, extract for 1 hour each time, filter, collect the filtrate, and discard the residue. Take 1 / 3 of the water extract, concentrate it, and dry it under reduced pressure.

[0074] Step 2: Impurity removal: The filtrate in step 1 was cooled to room temperature, passed through a D101 macroporous resin column (500 g wet resin) at a flow rate of 2 BV / h, and the effluent was concentrated to 1 g / mL crude drug amount; the D101 macroporous resin was further eluted with 4 BV of 95% ethanol, 95% of the eluate was collected, concentrated and dried under reduced pressure.

[0075] Step 3: Enzymatic hydrolysis: add 0.2 g papain to the concentrated solution of 1 g / mL crude drug, perform enzymatic hydrolysis at 60°C for 1 h, boil, inactivate the enzyme for 10 min, cool to room temperature, centrifuge at 4000 r / min for 10 min, and take the supernatant.

[0076] Step 4: alcohol precipitation: add ethanol to the supernatant obtained in step 3 to adjust the ethanol concentration to 80%, let the alcohol precipitation stand overnight, centrifuge, collect the precipitate, and freeze-dry.

[0077] Step 5: Dialysis and drying: Wash the precipitate obtained in step 3 twice with ethanol, add appropriate amount of water to re-dissolve, filter, and place the filtrate in dialysis bags with particle sizes of 1000 Da, 14000 Da, 50000 Da, and 85000 Da, respectively. Dialyze for 24 hours and concentrate to 1 g / mL crude drug; pre-freeze at -30°C and freeze-dry to obtain extract 1, extract 2, extract 3, and extract 4, respectively.

[0078] Experimental example

[0079] 1. Analysis of content and molecular weight of extract 4 in Example 1

[0080] The polysaccharide content of Rhodiola rosea polysaccharide was determined by the phenol-sulfuric acid method, and D-glucose was used as the standard monosaccharide composition analysis of extract 4. The standard curve was y = 9.985x - 0.0023, R² = 0.9934, and the proportion of polysaccharides in extract 4 was 86.33 ± 0.023%. The standard curve of molecular weight retention time was y = -0.39755x + 9.67152, R² = 0.9942, as measured by HPGPC according to the Shodex STANDARD P-82 standard. Figure 1 As shown, the calculated molecular weight distribution of extract 4 is 1.05×10 3 , 1.78×10 9 .

[0081] II. Therapeutic effects of different extracts in Example 1 on rats with chronic nephritis induced by tail vein injection of doxorubicin

[0082] 1. Experimental Materials

[0083] Animals: SPF male SD rats weighing 180-220 g were kept in a clean environment with a temperature of 23±2°C, a humidity of 50%-60%, and a 12-hour day and night light cycle. The rats were fed ad libitum and the drinking water was filtered and sterilized before use.

[0084] Drugs and reagents: Doxorubicin hydrochloride (Shanghai McLean Biochemical Technology Co., Ltd.); Prednisone acetate tablets (Tianjin Xinyi Jinjin Pharmaceutical Co., Ltd.)

[0085] Instruments: Refrigerated centrifuge (ThermoFisher Scientific, USA); vortex mixer (Qilin Bell Instrument Manufacturing Co., Ltd.); electronic balance (Startorius, USA); pathology slicer (Shanghai Leica Instrument Co., Ltd.); upright optical microscope (Nikon, Japan)

[0086] 2. Dosage design: The dose of extract 1 for rats is 100 mg / kg / day, the dose of extract 2 for rats is 100 mg / kg / day, the dose of extract 3 for rats is 100 mg / kg / day, and the dose of extract 4 for rats is 100 mg / kg / day. The positive drug is prednisolone acetate, and the dosage is 0.9 mg / kg / day.

[0087] 3. Experimental methods

[0088] (1) Initial screening: After the adaptive feeding of rats was completed, all rats were fasted but not watered and placed in metabolic cages to collect urine. Urine protein was measured using urine protein test strips. Rats with negative urine protein were selected as rats for subsequent experiments, and 6 rats were randomly selected as the blank group.

[0089] (2) Modeling: The remaining rats were modeled in the following manner:

[0090] First modeling: Doxorubicin was prepared into a 2 mg / mL solution with normal saline. Except for the blank group, all other rats were injected with 4 mg / kg of doxorubicin through the tail vein;

[0091] Second modeling: 7 days after the first injection, the second tail vein injection was performed. Doxorubicin was first prepared into a 1 mg / mL solution with normal saline. Except for the blank group and the rats with tail ulcers in the first modeling, all rats were injected with 2 mg / kg of doxorubicin through the tail vein.

[0092] Ten days after the second doxorubicin modeling was completed, all rats were fasted but not watered and placed in metabolic cages for 24 hours to collect urine. Urine protein was measured using a urine protein quantitative test kit to verify whether the model was successfully established.

[0093] The rats with successful modeling were randomly divided into a blank control group (Control), a model group (Model), an extract group 1 (100 mg / kg / day), an extract group 2 (100 mg / kg / day), an extract group 3 (100 mg / kg / day), an extract group 4 (200 mg / kg / day) and a positive drug group (prednisone acetate, 0.9 mg / kg / day), with 6 rats in each group.

[0094] (3) Observation indicators and efficacy evaluation methods

[0095] Blood sample collection: Rats were anesthetized by intraperitoneal injection of 20% urethane solution (0.7 mL / 100 g), and blood was collected from the abdominal aorta. The blood samples were allowed to stand at room temperature for 2 h, centrifuged in a low-temperature high-speed centrifuge for 10 minutes (4°C, 3000 rmp), and the supernatant was divided into sterile EP tubes and stored in a -80°C refrigerator for later use. After blood was collected from the abdominal aorta, the right kidney was quickly removed, rinsed with saline, and the peripheral tissues and renal capsule were cleaned. The kidney tissue was placed on ice, a small incision was made in the middle of the kidney with a scalpel, and fixed in 4% paraformaldehyde solution for light microscopy examination.

[0096] The various indicators tested are as follows:

[0097] Serum: total cholesterol (TC), triglyceride (TG), serum creatinine (Scr), blood urea nitrogen (BUN), albumin (ALB);

[0098] Urine: urine protein (measured every week), urine creatinine (Ucr), urea nitrogen (BUN), albumin (ALB);

[0099] The operation process was completely in accordance with the instructions of each kit. GraphPad Prism 9 software was used to draw graphs for inter-group difference analysis.

[0100] 4. Experimental results

[0101] (1) Effect of weekly rat urine protein

[0102] Week 0 is the day before drug administration, as shown in Table 2 and Figure 2 It can be seen that during the entire medication period, the 24-hour urine protein of rats in the model group was significantly higher than that in the blank group, indicating that the model was successfully established.

[0103] During the 0-6 week dosing period, the 24-hour urine protein content of extract group 1 and extract group 2 was generally higher than that of the model group; the 24-hour urine protein of extract group 3 and extract group 4 was lower than that of the model group, and the reduction in extract group 4 was significantly higher than that in extract group 3, indicating that Rhodiola rosea polysaccharide has a certain effect in reducing the proteinuria of rats with nephrotic rats, and the effect is increased after purification.

[0104] Table 1 Effects of different experimental groups on urine protein content in rats

[0105]

[0106] (2) Effects on urine parameters of rats with nephritis:

[0107] Depend on Figure 3 It can be seen (Note: # represents the comparison between the model group and the normal group, P < 0.05, * represents the comparison between the drug-treated group and the model group, P < 0.05, ** represents the comparison between the drug-treated group and the model group, P < 0.01), the urine albumin (ALB) level of the rats in the model group was significantly higher than that in the normal group, and the extract groups 3 and 4 had a correction effect; there was no significant difference in the urine Cr level of the rats in each group; the ACR value (urine ALB / Cr ratio), another core indicator in the diagnosis and treatment guidelines, was significantly higher in the model group than in the normal group, and the extract group 4 had a correction effect, indicating that the model was successfully established, and Rhodiola rosea polysaccharide can reduce the amount of albumin and ACR value in the urine of rats with nephrotic syndrome, and the effect is increased after purification.

[0108] (3) Effects on serum parameters of rats with nephritis:

[0109] Depend on Figure 4 It can be seen (Note: # represents the comparison between the model group and the normal group, P < 0.05, * represents the comparison between the drug administration group and the model group, P < 0.05, ** represents the comparison between the drug administration group and the model group, P < 0.01), the serum albumin (ALB) level of the rats in the model group was significantly lower than that in the normal group, the extract group 3 had a certain correction trend, but there was no significant difference compared with the model group; the extract group 4 had an obvious correction trend, which was statistically different from the model group, the serum creatinine (Cr) level of the rats in the model group was higher than that in the normal group, and the extract groups 3 and 4 had a downward trend; after purification, the effect increased. There was no significant difference in the urine nitrogen (BUN) of the rats in each group.

[0110] (4) Impact on renal pathological indicators:

[0111] Depend on Figure 5 It can be seen that the model was established by injecting doxorubicin twice through the tail vein. The main lesions in the model group were degeneration of renal tubular epithelial cells, transparent casts in the tubular lumen, increased mesangial matrix in some glomeruli, and suspected inflammatory cell infiltration. There was a statistical difference between the model group and the normal group, indicating that the doxorubicin nephritis model was successfully established. After the use of the extract, the renal tubular lesions were alleviated, with the best result in extract 4 group, followed by extract 3 group.

[0112] In summary, this modeling was successful, and the extract groups 4 were able to significantly reduce the amount of urine protein, albumin and ACR value in rats with renal disease, and significantly increase the albumin content in blood samples of rats with renal disease. Combined with the pathological results, it was shown that the Rhodiola rosea polysaccharide of the present application could improve the permeability of renal tubules in rats with renal disease, and alleviate glomerular lesions in rats with renal disease.

[0113] The description of the above embodiments is only used to help understand the method and core idea of ​​the present application. It should be noted that, for ordinary technicians in this technical field, several improvements and modifications can be made to the present application without departing from the principles of the present application, and these improvements and modifications also fall within the scope of protection of the claims of the present application.

Claims

1. A use of Rhodiola rosea polysaccharide in the preparation of a drug for treating chronic nephritis, characterized in that: The molecular weight distribution of the Rhodiola rosea polysaccharide is 1.05×10 3 , 1.78×10 9 ; The polysaccharide content in the Rhodiola rosea polysaccharide is 76wt.%-85wt.%; Wherein, the Rhodiola rosea polysaccharide is extracted from Rhodiola rosea, and the preparation method of the Rhodiola rosea polysaccharide comprises: Step 1: Extraction: extracting the Rhodiola rosea medicinal material with alcohol to obtain alcohol extraction residue and alcohol extraction filtrate; Step 2: Extraction: extracting the alcohol extraction residue in step 1 with water to obtain a water extract; Step 3: removing impurities: passing the aqueous extract in step 2 through a column and eluting, collecting and combining the effluent and the eluate, and concentrating to obtain a concentrate; Step 4: enzymatic hydrolysis: enzymatically hydrolyze the concentrated solution in step 3 and take the supernatant; Step 5: alcohol precipitation, dialysis and drying: subjecting the supernatant in step 4 to alcohol precipitation, collecting the precipitate, dialysis and freeze-drying to obtain the Rhodiola rosea polysaccharide; The dialysis comprises: washing the precipitate twice with ethanol, adding an appropriate amount of water to redissolve it, filtering it, transferring the filtrate to a dialysis bag, and dialyzing it; the particle size of the dialysis bag is 85000Da.

2. The use according to claim 1, characterized in that: The column and elution include: cooling the water extract to room temperature, passing through a macroporous resin column, and eluting with 2BV pure water; The column and elution adopt D101 macroporous resin; The volume ratio of the water extract to the macroporous resin is 1:(1-3); The loading flow rate was 2-3 BV / h.

3. The use according to claim 1, characterized in that: The enzymatic hydrolysis comprises: adding one of papain, pepsin, trypsin and endopeptidase, performing enzymatic hydrolysis at 37-60° C. for 1-1.5 hours, cooling and centrifuging to obtain a supernatant; Wherein, the dosage of the enzyme in the enzymolysis is 0.0004-0.001 g / g of crude drug.

4. The use according to claim 1, characterized in that: The alcohol precipitation comprises: adding ethanol to the supernatant, adjusting the ethanol concentration to 70-95%, standing overnight, and collecting the precipitate.

5. A Chinese medicine preparation for treating chronic nephritis, characterized in that: The invention comprises the Rhodiola rosea polysaccharide used in any one of claims 1 to 4.

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