CN110693897B

CN110693897B - Application of oil tea flesh fruit polysaccharide in preparation of medicine or health-care product for preventing and treating type II diabetes

Info

Publication number: CN110693897B
Application number: CN201911154271.1A
Authority: CN
Inventors: 吴素珍; 李加林; 刘志平
Original assignee: Gannan Medical University
Current assignee: Gannan Medical University
Priority date: 2019-11-22
Filing date: 2019-11-22
Publication date: 2023-04-18
Anticipated expiration: 2039-11-22
Also published as: CN110693897A

Abstract

The invention discloses application of oil tea fruit polysaccharide in preparation of a medicine or a health-care product for preventing and treating type II diabetes. The oil-tea camellia succulent fruit polysaccharide is a pure natural plant polysaccharide obtained by extracting and purifying oil-tea camellia succulent fruit. Through preparing a diabetic mouse animal experiment, after the administration of the oil-tea camellia succulent fructopolysaccharides for 3 months, the blood sugar of a mouse is obviously reduced, the area under the glucose tolerance curve and the insulin tolerance curve is obviously lower, and a series of kidney pathological detection analysis of HE and PAS dyeing, immunohistochemistry, transmission electron microscope results, kidney oxidation resistance, kidney inflammation resistance, kidney hypertrophy and metabolic conditions shows that a series of pathological changes of the kidney of the diabetic mouse are all improved. Therefore, experiments prove that the oil tea fleshy fruit polysaccharide has better effects of preventing and treating type II diabetes and protecting the kidney of the diabetes.

Description

Application of oil tea flesh fruit polysaccharide in preparation of medicine or health-care product for preventing and treating type II diabetes

Technical Field

The invention belongs to the field of traditional Chinese medicines, and relates to oil-tea camellia succulent fruit polysaccharide extracted and purified by taking oil-tea camellia succulent fruits as a raw material and application of the oil-tea camellia succulent fruit polysaccharide in preparation of medicines or health-care products for preventing and treating type II diabetes.

Background

Diabetes has become the third most fatal disease following cardiovascular disease and cancer. In the population over 20 years old in China, the total prevalence rate of diabetes is 11.6%, and 1.139 hundred million adults are estimated to suffer from diabetes. The incidence rate of diabetes mellitus is gradually increased in the global scope, and the incidence rate of type II diabetes mellitus is increased most obviously. Islet beta cell dysfunction and reduced numbers are the major pathogenesis of type ii diabetes. Regeneration and restoration of islet beta cell numbers and thus islet beta cell function is fundamental to the treatment of type ii diabetes.

Diabetic Nephropathy (DN) is a common complication of diabetes, is one of the manifestations of Diabetic systemic microangiopathy, is clinically characterized by proteinuria, progressive renal impairment, hypertension and edema, and severe renal failure in the late stage, and is one of the main causes of death of Diabetic patients. The basic pathological feature of DN is that the glomerular basement membrane is uniformly thickened, and the glomerular mesangial cell matrix is increased, and the glomerular capsule and the glomerular mesangial cell are nodular thickened and have increased permeability.

The succulent fruit of the oil-tea Camellia is caused by the expansion of the oil-tea Camellia (Camellia oleifera Abel) infected by a kind of Exobasidium Vexans Massee, and can be directly eaten as a wild fruit. Researches show that the meat fruit of the oil tea is rich in nutrient components such as saccharides, lipids, proteins, amino acids, vitamins and the like, and trace elements in the meat fruit are far higher than those in common fruits, so that the meat fruit of the oil tea has a good health-care effect. Zhu Bifeng and the like find that the extract (comprising water-soluble substances and alcohol-soluble substances) of the meat fruits and the meat leaves of the oil-tea camellia can obviously reduce the blood sugar of a diabetes mouse induced by alloxan, has the effect of reducing the blood sugar equivalent to that of a traditional Chinese and western medicine combined blood sugar-reducing medicine thirst-quenching pill, can reduce the generation of liver lipid Malondialdehyde (MDA) of the diabetes mouse, and improves the activity of an antioxidant enzyme system of the diabetes mouse. The oil tea polysaccharide (CPs) extracted from other parts of oil tea comprise oil tea cake polysaccharide, oil tea leaf polysaccharide and oil tea peel polysaccharide, and oil tea shell polysaccharide, has various biological activities including bacteriostasis, immunity regulation, antioxidation, free radical removal, antitumor, alpha glucosidase inhibition and the like, and also has hypoglycemic activity.

Currently, diabetes treatment methods include insulin injection and oral hypoglycemic agents, but these methods also cause some side effects during clinical application, for example, high doses of insulin and oral hypoglycemic agents may cause hypoglycemia, liver damage, lactic acidosis, diarrhea, and the like. Therefore, it is necessary to find a non-toxic antidiabetic drug existing in nature.

Disclosure of Invention

The invention aims to extract oil tea flesh fruit polysaccharide by using oil tea flesh fruits as a raw material and using a water extraction and alcohol precipitation method, and animal experiments show that the oil tea flesh fruit polysaccharide has a repairing effect on islet functions of type II diabetic mice and a protecting effect on kidneys of the diabetic mice.

In order to achieve the purpose, the invention adopts the following technical scheme:

application of oil tea succulent fruit polysaccharide in preparing medicine or health product for preventing and treating type II diabetes is provided.

Further, the medicine or health product is one of granules, pills, capsules, tablets, powder, paste and oral liquid.

Further, the preparation of the oil tea succulent fruit polysaccharide comprises the following steps:

(1) Drying and crushing the oil-tea camellia fleshy fruit, adding water for soaking overnight, extracting for 2-3 times by boiling water, and combining extracting solutions;

(2) And (3) adding ethanol into the extracting solution prepared in the step (1) for alcohol precipitation, removing protein by a Sevage method, decoloring and dialyzing to obtain the oil-tea camellia pulp fruit polysaccharide.

Furthermore, water is added in the step 1 for soaking, and the volume of the added water is 20-40 times of the weight of the oil tea fleshy fruit.

Further, the boiling water extraction in step 1 is stirring extraction or reflux extraction.

Furthermore, the boiling water extraction time in the step 1 is 1-3 h.

Furthermore, when the ethanol is precipitated in the step 2, the volume ratio of the ethanol is 2-8 times of that of the extracting solution prepared in the step 1, and the concentration of the ethanol is 75-100%.

The invention has the beneficial effects that:

compared with the prior art, the preparation method has the advantages that the oil-tea camellia flesh fruit medicinal material resources are rich, the process for preparing the oil-tea camellia flesh fruit polysaccharide is simple, the cost is low, no toxic or side effect is caused, the taste is good, and the development value is good. In addition, the invention discovers that the oil tea fruit polysaccharide is suitable for preventing and treating type II diabetes and the kidney protection effect of the oil tea fruit polysaccharide, has definite curative effect, has good medicinal value in preparing medicaments or health-care products for preventing and treating diabetes, and provides a research basis for further and deeply developing the oil tea fruit polysaccharide.

Drawings

FIG. 1 is a graph showing the change in fasting plasma glucose in mice before and 1, 2, 3 months after administration and 1 month after discontinuation.

FIG. 2 is the fasting glucose tolerance and area change under the curve in mice 1 month after drug withdrawal.

FIG. 3 is the fasting insulin tolerance and area change under the curve in mice 1 week after drug withdrawal.

FIG. 4 is a graph showing the results of HE staining of mouse renal cortex under an optical microscope (400X).

FIG. 5 is a graph showing the result of PAS staining of the renal cortex of a mouse under an optical microscope (400X).

Figure 6 is expression of mouse kidney FN.

FIG. 7 is the expression of collagen type IV in mouse kidney.

FIG. 8 is a transmission electron microscope image of mouse renal cortex.

FIG. 9 is the expression of IL-6 and alpha in mouse kidney.

Detailed Description

The present invention is further described in the following examples, which should not be construed as limiting the scope of the invention, but rather as providing the following examples which are set forth to illustrate and not limit the scope of the invention.

Example 1: preparation of camellia oleifera pulp fruit polysaccharide

(1) Drying and crushing the oil tea flesh fruits for later use. Weighing 30g of dried oil-tea camellia fleshy fruit into a 2L round-bottom flask, adding 900mL of purified water, soaking at normal temperature overnight, carrying out water extraction at 99 +/-1 ℃ for 2h, filtering with four layers of gauze, repeatedly carrying out water extraction on filtered dregs at 99 +/-1 ℃ for 2h, and combining extracting solutions.

(2) Adding 500mL of absolute ethyl alcohol into the extracting solution prepared in the step 1 for precipitation, centrifugally collecting the precipitate, removing protein by a Sevage method, decoloring and dialyzing to obtain a crude product of the oil-tea Camellia Flesh Fruit Polysaccharide (CFFP), and measuring the content of the oil-tea camellia flesh fruit polysaccharide to be 40.30% by adopting an anthrone-sulfuric acid method and an ultraviolet spectrophotometer.

Example 2: mouse animal test for type II diabetes

1. Experimental protocol

1.1 pharmaceutical formulation

The prepared oil tea pulp fruit polysaccharide is taken, water is used for preparing oil tea pulp fruit polysaccharide solutions with the oil tea pulp fruit polysaccharide concentrations of 12.5mg/mL and 25mg/mL respectively, and in addition, metformin tablets are ground into powder to prepare a metformin solution with the concentration of 20 mg/mL.

1.2 establishment of type II diabetes animal model

50C 57 mice of 10 weeks old are purchased from Gannan Silike laboratory animals Co., ltd, and are bred in the center of Gannan medical institute laboratory animals, the experimental operation follows the ethical rules of animal experiments in Gannan medical institute, the mice are randomly divided into 5 groups, 10 mice are per group, 1 group is a normal control group and is fed with normal feed, the rest 4 groups are induced by high-sugar high-fat diet and small-dose Streptozotocin (STZ) to establish a type II diabetes mouse model, the fasting plasma glucose of the mouse tail tip is respectively measured 72 hours and one week after administration, and the blood glucose values of two times are all larger than 16.7mmol/L, so that the model is judged to be successful.

1.3 grouping and administration

Dividing the mice successfully molded into four groups, adding a control group to finally obtain the following 5 experimental groups of mice:

group one: normal control group (Con), gavage normal saline;

and a second group: diabetes model group (DM), gavage saline;

and (3) group III: the low dose administration group (CFFP 1, 12.5mg/ml of oil tea fruit), the stomach-filling camellia oleifera fruit polysaccharide (dose: 125mg/kg, b.w.);

group four: high-dose administration group (CFFP 2, 25mg/ml of oil-tea-containing fleshy fruit), and gavage oil tea fruit fleshy fruit polysaccharide (dose: 250mg/kg, b.w.);

group five: metformin (Met, containing metformin 20 mg/ml), metformin (dose: 200mg/kg, b.w) was gavaged.

Mice were weighed daily and dosed with different solutions for 3 months, and their random blood glucose was measured at 1 to 3 months after dosing and again at one month after 3 months of cessation of dosing.

1.4 fasting glucose tolerance test

After one month of drug withdrawal, the mice were fasted for 4h, five groups of mice were gavaged once according to the dose, and the glucose tolerance test was performed 20min later. Glucose is prepared into a glucose solution with the concentration of 25g/ml by using normal saline, and then the glucose solution is injected into the abdominal cavity according to the dosage of 2.0g/kg, namely, the volume of the glucose injected into each g of mice is 8 mul. Blood is taken at the tail parts of 0min, 30min, 60min and 120min after glucose injection, the blood sugar is measured, a curve is drawn, the area AUC under the blood sugar curve is calculated, and the calculation formula of AUC (mg. H/dL) is as follows:

AUC＝(BG 0+BG 30)×15/60+(BG 30+BG 60)×15/60+(BG 60+BG 120)×30/60,

BG 0, BG 30, BG 60, BG 120 represent blood glucose at 0min, 30min, 60min, 120min after administration of glucose, respectively.

1.5 fasting insulin tolerance test

After one week of drug withdrawal, animals were fasted for 4h and subjected to insulin tolerance test. Subcutaneously injecting 0.4U/Kg of insulin, taking blood at the tail of 0min, 40min and 90min after injection, measuring blood sugar, drawing a curve, and calculating the AUC (mg. H/dL) of the area under the blood sugar curve as follows:

AUC = (BG 0+ BG 40) x 20/60+ (BG 90+ BG 40) x 25/60, BG 0, BG 40, BG 90 represent blood sugar 0min, 40min,90min after insulin injection, respectively. During the experiment, whether the mouse has the hypoglycemia reaction or not is observed, and if the hypoglycemia reaction of the mouse is found, the mouse is immediately injected with a glucose solution.

1.6 Kidney histological examination

After termination of the mouse biopsy experiment, mice were sacrificed and dissected, kidney tissue was taken and fixed in 10% formalin for histological examination. The kidney tissue was embedded in paraffin, cut into slices having a thickness of 4 μm, stained with Hematoxylin and Eosin (HE) and periodic acid-Schiff (PAS), respectively, and observed under an optical microscope.

1.7 immunohistochemical detection of renal tissue

The 4 μm thick sections of protocol 1.6 were deparaffinized for immunostaining and observed for accumulation of extracellular matrix Fibronectin (FN) and collagen type IV (Col IV) in the renal cortex of mice.

1.8 Transmission Electron microscopy of renal cortex

End smallAfter the test experiment of the living body of the mouse, the mouse is sacrificed and dissected, the kidney is taken out quickly, the medulla is removed, the cortex is taken out, and the renal cortex is trimmed to 0.5-1mm ³ Volume, PBS rinsed clean, blotted dry with absorbent paper, and soaked in 2.5% glutaraldehyde. Embedding the tissue and ultrathin section by glutaraldehyde-osmic acid double fixation method. And finally, observing the change conditions of the glomerular basement membrane and the podocyte of the mouse under a transmission electron microscope.

1.9 Oxidation resistance test of the Kidney

After the test experiment of the mouse living body is finished, the mouse is sacrificed and dissected, the kidney is quickly taken out, the content of superoxide dismutase (SOD), glutathione (GSH) and Malondialdehyde (MDA) in the kidney of the mouse is detected by a commercial kit, and the antioxidation of the oil tea flesh fruit polysaccharide in the kidney is researched.

1.10 anti-inflammatory assay of the kidney

After the end of the mouse in vivo test experiment, the mice were sacrificed and dissected, the renal cortex was isolated, and the anti-inflammatory effect of the oil tea fruit polysaccharide in the kidney was studied by detecting the secretion of inflammatory mediators of IL-6 and TNF α by immunohistochemistry and real-time qPCR.

1.11 renal hypertrophy and Metabolic experiment

Before the animal is sacrificed, the weight (BW) is weighed, the mouse is sacrificed and then the material is rapidly obtained, and the method comprises the following steps of firstly collecting blood from the heart, centrifuging (5000rpm, 4 ℃ and 10 min), separating serum, preserving at-80 ℃ and measuring blood indexes such as Blood Urea Nitrogen (BUN), blood creatinine (Scr) and the like. Secondly, the kidneys were removed, fresh kidney tissue was removed, washed clean in PBS, dried with absorbent paper and accurately weighed (KW), and the kidney weight ratio (KW/BW) was calculated, i.e. KW/BW = kidney weight (mg)/weight (g).

2. Results of the experiment

2.1 fasting glucose tolerance test results

Fasting plasma glucose changes in mice before and 1, 2, 3 months and 1 month after drug withdrawal, P <0.05 compared to control; # denotes P <0.05 compared to the diabetes model group. The blood sugar of the control group mouse is always maintained at a stable baseline level, and the blood sugar of the diabetes model group mouse is always more than 20mmol/L; before administration, the blood sugar of the mice in the diabetes model group, the low dose group, the high dose group and the metformin group has no obvious difference and is obviously higher than that of the control group, and the blood sugar level of the mice shows that the diabetes mice are successfully modeled; after 1 month of administration, the blood sugar of each group of mice has no obvious change before administration; after 2 months of administration, the blood glucose of mice in the low dose group, the high dose group and the metformin group was significantly reduced compared to the diabetes model group, but still significantly higher than that of the control group; after 3 months of administration, the blood sugar of the mice in the low-dose group is still higher than that of the control group but is reduced compared with that of the mice in the 2 months of administration, and the blood sugar of the mice in the high-dose group and the metformin group is reduced to a normal level; after 1 month of drug withdrawal, the blood sugar of mice in each group of the drug administration group is not obviously increased. The result shows that the camellia oleifera flesh fruit polysaccharide has an obvious blood sugar reducing effect like metformin, particularly has a more obvious blood sugar reducing effect in a high-dose group, and blood sugar hardly rebounds after administration is stopped for one month, which shows that the camellia oleifera flesh fruit polysaccharide has a repairing effect on the damage of the islet function of type II diabetic mice.

2.2 fasting insulin tolerance test results

The results of the fasting glucose tolerance test in mice 1 month after drug withdrawal are shown in fig. 2. Panel (a) fasting glucose tolerance in mice; graph (B) calculates the area under the curve result; in the figure, P <0.05 compared to the diabetes model group is indicated. As can be seen from the graph (A), the blood glucose levels of the mice in each group increased and then decreased. Blood sugar of the control group is always maintained at a normal level and reaches a peak value at 30 min; the blood sugar of the diabetes model group is always obviously higher than the normal level and reaches the peak value at 60 min; the blood sugar of the low-dose group, the high-dose group and the metformin reaches a peak value at 30min and then gradually decreases, the blood sugar of the low-dose group, the high-dose group and the metformin is obviously lower than that of the diabetes model group at 60min, and the decrease degree of the high-dose group and the metformin is equivalent. As can be seen from the graph (B), the AUC of the area under the curve is calculated, and the AUC of the control group, the low dose group, the high dose group and the metformin group is significantly lower than that of the diabetes model group, and the reduction degree of the high dose group is equivalent to that of the metformin group. The result shows that the oil tea fruit polysaccharide has the effect of improving the sugar tolerance of the type II diabetes mice.

2.3 mice fasting insulin tolerance results

Fasting insulin tolerance in mice 1 week after drug withdrawal, the results are shown in figure 3. Panel (a) fasting insulin tolerance in mice; graph (B) calculates the area under the curve result; in the figure, P <0.05 compared to the diabetes model group is indicated. As can be seen from the graph (a), after the injection of insulin, the blood glucose levels of the mice in each group were first decreased and then increased, and at each time point, the control group, the low dose group, the high dose group, and the metformin group were all significantly lower than those in the diabetes model group, and the decrease degree of the high dose group was comparable to that of the metformin group. The area under the curve is calculated, and as can be seen from the graph (B), the AUC of the control group, the low dose group, the high dose group and the metformin group is significantly lower than that of the diabetes model group, and the reduction degree of the high dose group is equivalent to that of the metformin group. The result shows that the oil tea fruit polysaccharide has the effect of improving the insulin tolerance of the type II diabetic mice.

2.4 renal histology examination results

(1) Staining of mouse renal cortex HE

The morphological change of the glomeruli of the mice is observed by HE staining, the magnification of an optical microscope is multiplied by 400, and the result is shown in figure 4, the glomeruli of a diabetes model group is shrunk, the glomerular basement membrane is thickened, while the control group, the low dose group, the high dose group and the metformin group are obviously better than the diabetes model group, and the reduction degree of the high dose group and the metformin group is equivalent. The result shows that the oil tea fruit polysaccharide has a protective effect on the glomerular morphology of type II diabetic mice.

(2) PAS staining results of mouse renal cortex

PAS staining was performed to observe mesangial matrix accumulation in mouse glomeruli, and the results are shown in FIG. 5. FIG. (A) shows the accumulation of mesangial matrix in glomeruli at magnification of 400 with an optical microscope; panel (B) quantitative analysis of mesangial matrix in glomeruli; graph indicates P <0.05 compared to control; # denotes P <0.05 compared to the diabetes model group. As can be seen, significant mesangial matrix accumulation was seen in glomeruli of the diabetes model group, while the control, low, high and metformin dose groups were significantly better than the diabetes model group, and the high dose group was slightly better than the metformin group. The result shows that the camellia oleifera fleshy fruit polysaccharide has an improvement effect on mesangial matrix accumulation in the glomerulus of type II diabetic mice.

2.5 results of immunohistochemical examination of renal cortex

Immunohistochemistry was performed to observe the accumulation of extracellular matrix Fibronectin (FN) and collagen type iv (Col iv) in the renal cortex of mice, as shown in fig. 6. FIG. (A) immunohistochemical detection of renal tissue FN protein levels at light microscope magnification x 400; FIG. (B) analysis of FN protein expression in immunohistochemistry; FIG (C) Westernblotting to observe the effect of CFFP on FN protein expression; FIG. (D) FN protein quantitation; FIG. (E) real-time quantitative PCR for detecting the effect of CFFP on FN gene expression; graph indicates P <0.05 compared to control; # denotes P <0.05 compared to the diabetes model group. It can be seen from the figure that there was more accumulation in the diabetes model groups FN and Col iv, while the control group, low dose group, high dose group and metformin group were significantly better than the diabetes model group, and the high dose group was better than the metformin group and closer to the control group. The result shows that the oil tea fruit polysaccharide has a protective effect on kidney cortex immunohistochemistry of type II diabetic mice.

Immunohistochemistry mice were observed for expression of kidney type IV collagen, as shown in figure 7. FIG. (A) immunohistochemistry for detection of renal type IV collagen levels; FIG. (B) type IV collagen quantification in immunohistochemistry; FIG. C is a Westernblotting method for detecting the effect of CFFP on type IV collagen expression; graph (D) change in type IV collagen expression levels compared to control; FIG. (E) collagen fibers were detected by Masson staining; panel (F) statistical analysis of collagen fibers accumulated in renal cortex; FIG. G is a graph showing the effect of CFFP on type IV collagen gene expression using real-time PCR; graph indicates P <0.05 compared to control; # denotes P <0.05 compared to the diabetes model group. It can be seen from the figure that there was a greater accumulation of type IV collagen in the diabetic model group, while the control, low, high and metformin doses were significantly better than in the diabetic model group, and the high dose was better than in the metformin dose and closer to the control group. The result shows that the camellia oleifera flesh fruit polysaccharide has an effect of improving the accumulation of type IV collagen of type II diabetic mice.

2.6 Transmission Electron microscopy of renal cortex

Changes in mouse Glomerular Basement Membrane (GBM) and podocytes were observed by transmission electron microscopy, as shown in fig. 8. FIG. (A) shows a representative micrograph at magnification × 10,000; panel (B) measures GBM thickness (nm) and is presented as a bar graph. Graph indicates P <0.05 compared to control; # denotes P <0.05 compared to the diabetes model group. As can be seen from fig. 8 and fig. 4, the glomerular basement membrane of the mice in the diabetes model group was locally thickened, part of the podocytes were exfoliated, and the podocytes were fused, whereas the control group, the low dose group, the high dose group and the metformin group were significantly better than the diabetes model group, and the high dose group was better than the metformin group and was closer to the control group. The results show that the oil tea fruit polysaccharide improves Glomerular Basement Membrane (GBM) and foot-process exudation of type II diabetic mice.

2.7 Oxidation resistance test results of Kidney

The oil tea fleshy fruit polysaccharide reduces the accumulation of Malondialdehyde (MDA) in the kidney of a mouse by increasing products of superoxide dismutase (SOD) and Glutathione (GSH), and relieves the oxidative stress of the kidney of a diabetic mouse. As shown in Table 1, the GSH and SOD activities of the oil tea fleshy fruit polysaccharide group and the metformin group are obviously higher than those of the diabetes model group, the MDA level is lower than that of the diabetes model group, and the effects of the high-dose group and the metformin group are equivalent to those of the control group. The result shows that the camellia oleifera fleshy fruit polysaccharide enhances the antioxidation effect of the kidney on the type II diabetic mice.

TABLE 1

* P <0.05 indicates P <0.05 compared to the control group and # indicates P <0.05 compared to the diabetes model group.

2.8 anti-inflammatory assay results for Kidney

Hyperglycemia with diabetes also causes inflammatory reactions that then damage the kidneys. After 3 months of administration with the oil tea succulent fruit, renal cortex was isolated, and secretion of inflammatory mediators of IL-6 and TNF α was detected by immunohistochemistry and real-time qPCR, as shown in fig. 9. FIG. A shows the detection of the IL-6 protein level in renal tissue by immunohistochemistry; panel (B) quantitative analysis of renal cortical IL-6; FIG. C shows the detection of the expression of mRNA for IL-6 in renal cortex by RT-qPCR; FIG. (D) immunohistochemistry for detection of renal tissue TNF α protein levels; panel (E) quantification of TNF α content in renal cortical immunohistochemistry; panel (F) detection of renal cortical tumor necrosis factor (α) mRNA expression using RT-qPCR; graph indicates P <0.05 compared to control; # denotes P <0.05 compared to the diabetes model group. As can be seen from the graphs, the expression of IL-6 and TNF α was much higher in the diabetes model group than in the control group, whereas the camellia oleifera fleshy fruit group and metformin group were lower than in the diabetes model group, in which the high dose group and metformin group were close to the control group. The result shows that the oil-tea camellia pulp fruit polysaccharide enhances the anti-inflammatory effect of the kidney on type II diabetic mice.

2.9 renal hypertrophy and Metabolic parameter test results

The kidney/body weight ratio is considered to be one of the important indicators of kidney hypertrophy, and serum creatinine (Scr) and urea nitrogen (BUN) reflect kidney functions, and the kidney/body weight ratio, scr and BUN indicators of each group of mice were measured, and the results are shown in table 2. As can be seen from table 2, the kidney/body weight ratio, scr, and BUN indices of the camellia oleifera fleshy fruit group and the metformin group were all lower than those of the diabetes model group, in which the camellia oleifera fleshy fruit high dose group and the metformin group were close to those of the control group. The results show that the oil-tea camellia succulent fruit polysaccharide can relieve renal hypertrophy and can reduce the content of Scr and BUN for type II diabetic mice.

TABLE 2

* Indicating P <0.05 compared to the control group. In the figure, # indicates that P <0.05 compared to the diabetes model group.

Claims

1. The application of the oil tea fruit polysaccharide in preparing the medicine for preventing and treating type II diabetic nephropathy comprises the following steps:

(1) Drying and crushing the oil-tea camellia fleshy fruit, adding water for soaking overnight, extracting for 2-3 times by boiling water, and combining the extracting solutions;

(2) And (2) adding ethanol into the extracting solution prepared in the step (1) for alcohol precipitation, removing protein by a Sevage method, decoloring and dialyzing to obtain the oil-tea camellia pulp fruit polysaccharide.

2. Use according to claim 1, characterized in that: the medicine is one of granules, pills, capsules, tablets, powder, paste and oral liquid.

3. Use according to claim 1, characterized in that: and (2) soaking in water in the step (1), wherein the water adding amount is 20-40 times of the weight-volume ratio of the oil tea fleshy fruits.

4. Use according to claim 1, characterized in that: the boiling water extraction mode in the step (1) is stirring extraction or reflux extraction.

5. Use according to claim 1, characterized in that: the extraction time of the boiling water in the step (1) is 1-3 h.

6. Use according to claim 1, characterized in that: when the ethanol is precipitated in the step (2), the amount of the ethanol is 2-8 times of the volume ratio of the extracting solution prepared in the step (1), and the concentration of the ethanol is 75-100%.