Chinese wolfberry fruit pigment composition for treating benign prostatic hyperplasia and application of Chinese wolfberry fruit pigment composition

Abstract

The invention discloses a fructus lycii pigment composition for treating benign prostatic hyperplasia and application. The fructus lycii pigment composition comprises lycium barbarum red, zeaxanthine, xanthophyll and beta-carotene. The fructus lycii pigment composition provided by the invention can effectively inhibit prostate stromal cell proliferation, inhibit prostate tissue hyperplasia in vivo, improve the pathological morphology of prostate tissue, regulate the androgen level, inhibit inflammatory response and relieve oxidative damage. The traditional Chinese medicine is applied to clinical treatment of prostatic hyperplasia and has the advantages of remarkable curative effect and low toxic and side effects.

Description

Technical Field

[0001] The invention belongs to the field of biomedicine, and particularly relates to a wolfberry pigment composition for treating prostate hyperplasia and an application thereof. Background Art

[0002] Benign prostatic hyperplasia (BPH) is almost universal in elderly men. Its clinical manifestations are lower urinary tract symptoms such as frequent urination, urgency, incomplete urination and increased nocturia, which seriously affect the quality of life. However, although the drugs commonly used in the clinic to treat BPH are effective, they have many side effects and adverse reactions. Therefore, it is urgent to research and develop effective innovative drugs that are efficient, cheap and low-toxic to treat BPH.

[0003] Lycium barbarum L., a plant of the Solanaceae family, is a dried mature fruit that nourishes the kidneys. Depei Materia Medica records that "wolfberry is good for urination and defecation", indicating that wolfberry can relieve symptoms such as urinary incontinence to a certain extent. The development of pure Chinese herbal natural products to treat prostate hyperplasia has broad application prospects. Summary of the invention

[0004] In order to solve the problems existing in the prior art, the present invention provides a wolfberry pigment composition having a significant effect in treating prostatic hyperplasia, and a preparation method and application thereof.

[0005] In order to achieve the above technical objectives, the technical solution adopted by the present invention is as follows:

[0006] A wolfberry pigment composition for treating prostatic hyperplasia comprises wolfberry red pigment, zeaxanthin, lutein and beta-carotene.

[0007] As a preferred embodiment, the wolfberry pigment composition for treating prostatic hyperplasia described above is composed of 25-75% wolfberry red pigment, 5-45% zeaxanthin, 4-25% lutein and 0.08-5% β-carotene by mass.

[0008] As a preferred embodiment, the wolfberry pigment composition for treating prostatic hyperplasia described above is composed of 25-60% wolfberry red pigment, 5-20% zeaxanthin, 4-15% lutein and 0.08-1.6% β-carotene by mass.

[0009] As a preferred embodiment, the wolfberry pigment composition for treating prostatic hyperplasia described above is composed of 67% wolfberry red pigment, 17% zeaxanthin, 13.6% lutein and 2.4% β-carotene by mass.

[0010] As a preferred embodiment, the wolfberry pigment composition for treating prostatic hyperplasia described above is composed of 73% wolfberry red pigment, 15% zeaxanthin, 10% lutein and 2% β-carotene by mass.

[0011] The wolfberry pigment composition for treating prostatic hyperplasia provided by the present invention comprises wolfberry red pigment, zeaxanthin, lutein and beta-carotene which can be separated and purified from wolfberry, and can also be purchased commercially.

[0012] The invention discloses an application of the wolfberry pigment composition in preparing a pharmaceutical product for treating prostatic hyperplasia.

[0013] As a preferred embodiment, the dosage form of the drug includes tablets, pills, powders, decoctions, granules, injections, capsules or decoctions, etc.

[0014] Beneficial effects: The present invention provides a medlar pigment composition that is both medicinal and edible. Experiments have shown that the composition can inhibit the proliferation of prostate stromal cells, improve the hyperplastic prostate tissue morphology, organ index, and prostate tissue pathological morphology, reduce the levels of androgens and inflammatory factors in the body, and relieve oxidative stress, and has a significant effect in treating prostate hyperplasia. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 The results of the in vitro inhibition of prostate stromal cells WPMY-1 by the wolfberry pigment composition. A-CCK-8 was used to detect the cytotoxicity of the wolfberry pigment composition, B-CCK-8 was used to detect the inhibitory effect of the wolfberry pigment composition on the proliferation of WPMY-1 cells induced by testosterone propionate (TP), C-EDU was used to detect the anti-proliferation ability, D-trend graph of the proportion of EDU-positive cells, E-type II 5α-reductase (5-AR2) level in the cell supernatant, F-dihydrotestosterone (DHT) level in the cell supernatant

[0016] Figure 2 The results of wolfberry pigment composition improving prostate tissue phenotype in rats with prostate hyperplasia are shown in Figure 1. A-tissue phenotype diagram, B-prostate wet weight, C-prostate index.

[0017] Figure 3 The results of the wolfberry pigment composition improving the histopathological morphology and epithelial thickness of rats with prostatic hyperplasia are shown.

[0018] Figure 4 The results of the effect of the wolfberry pigment composition on the androgen levels in rat serum and prostate tissue are shown in Figure 2. AD represents the levels of testosterone (T), DHT, 5-AR2 and prostate-specific antigen (PSA) in serum, and EH represents the levels of T, DHT, 5-AR2 and PSA in prostate tissue.

[0019] Figure 5 The figure shows the effect of the wolfberry pigment composition on the levels of inflammatory factors in rat serum and prostate tissue. AC represents the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) in serum, and DF represents the levels of TNF-α, IL-6 and IL-1β in prostate tissue, respectively.

[0020] Figure 6 The results of the effect of the wolfberry pigment composition on oxidative stress factors in rat serum and prostate tissue are shown in Figure 5. AC represents the levels of malondialdehyde (MDA), glutathione (GSH) and superoxide dismutase (SOD) in serum, and DF represents the levels of MDA, GSH and SOD in prostate tissue. DETAILED DESCRIPTION

[0021] It should be noted that the following detailed descriptions are illustrative and are intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used in the present application have the same meanings as those commonly understood by those skilled in the art to which the present application belongs.

[0022] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments according to the present application. As used herein, unless the context clearly indicates otherwise, the singular form is also intended to include the plural form. In addition, it should be understood that when the terms "comprise" and / or "include" are used in this specification, it indicates the presence of features, steps, operations, devices, components and / or combinations thereof.

[0023] Example 1

[0024] A wolfberry pigment composition for treating prostatic hyperplasia, the composition consists of 67% of wolfberry red pigment, 17% of zeaxanthin, 13.6% of lutein and 2.4% of beta-carotene in mass percentage.

[0025] Example 2

[0026] A wolfberry pigment composition for treating prostatic hyperplasia, the composition consists of 73% by mass of wolfberry red pigment, 15% by mass of zeaxanthin, 10% by mass of lutein and 2% by mass of beta-carotene.

[0027] Example 3

[0028] A wolfberry pigment composition for treating prostatic hyperplasia, the composition consists of 60% by mass of wolfberry red pigment, 20% by mass of zeaxanthin, 15% by mass of lutein and 5% by mass of beta-carotene.

[0029] Example 4 Lycium barbarum pigment composition inhibits prostate WPMY-1 cell proliferation in vitro

[0030] 1. Cell culture and cytotoxicity assay

[0031] Human normal prostate stromal immortalized cells (WPMY-1) were purchased from Wuhan Pronocell Life Science Co., Ltd. and cultured in complete medium (5% FBS + 1% P / S DMEM medium) in an incubator at 37°C, 5% CO2, and 90% relative humidity. 4 The cells were inoculated at a density of 100 μL / mL in a 96-well cell culture plate, and 100 μL was added to each well. After culturing for 24 h, different concentrations of the wolfberry pigment composition of Example 1 (31.3, 62.5, 125, 250, 500, 1000 μg / mL) were added respectively. After incubation for 24 h, complete culture medium containing 10% CCK-8 was added to each well to measure the absorbance. The concentration at which there was no significant difference in cell survival rate between the drug administration group and the control group was selected as the drug safety concentration.

[0032] 2. Antiproliferation assay

[0033] The cells were inoculated according to the above method. After 24 hours, complete culture medium was added to the control group, testosterone propionate (TP, 3 μM) was added to the model group and the drug-treated group, and different concentrations of the wolfberry pigment composition (GQ) of Example 1 were added to the drug-treated group. Each group was cultured in 6 replicate wells in parallel for 24 hours. The culture medium was discarded, and complete culture medium containing 10% CCK-8 was added to each well. After incubation for 2 hours, the absorbance of each well was measured at 450 nm to detect the cell survival rate.

[0034] 3. EDU detection

[0035] The cells were divided into 5 × 10 4 The cells were inoculated at a density of 100 μg / mL in a 12-well plate and cultured for 24 hours. The culture medium was discarded. Except for the control group, each group was added with a culture medium containing 3 μM TP. The positive drug group was given 50 μM finasteride (Fi), and the drug-treated group was added with 100 μg / mL wolfberry pigment composition (GQ). The cells were cultured in an incubator for 24 hours, the culture medium was discarded, and 10 μM EdU was added to each well for 3 hours. The remaining steps were performed according to the instructions for the kit. The cells were photographed under an inverted fluorescence microscope, and at least 3 random fields of view were photographed in each well.

[0036] 4. Determination of Type II 5α-reductase (5-AR2) and dihydrotestosterone (DHT) levels in cells

[0037] According to the method in "EDU detection", the cultured cells were inoculated and treated with drugs, and the cell supernatant was collected after 24 hours of incubation and centrifuged (3000 rpm / min, 4°C, 20 min). According to the operating procedures of the kit, the levels of 5-AR2 and DHT were measured respectively.

[0038] 5. Experimental Results

[0039] like Figure 1 As shown in A, the wolfberry pigment composition with a concentration of 31.3 to 1000 μg / mL had no obvious cytotoxicity to WPMY-1 cells, and its inhibitory effect on TP-induced WPMY-1 cell proliferation was further evaluated. Compared with the model group, the wolfberry pigment concentration ≥ 100 μg / mL significantly inhibited TP-induced WPMY-1 cell proliferation (P < 0.05) ( Figure 1 B), and the area of ​​the EDU-positive proliferation region is comparable to that of the positive drug finasteride ( Figure 1 C and 1D).

[0040] ELISA to detect 5-AR and DHT levels Figure 1 As shown in E and F, the levels of 5-AR and DHT in the cell supernatant increased significantly after TP induction (P<0.05), and decreased significantly after administration of wolfberry pigment and finasteride (P<0.05), indicating that wolfberry pigment can improve prostate cell proliferation by inhibiting androgen levels.

[0041] Example 5 In vivo treatment of TP-induced prostatic hyperplasia in rats with wolfberry pigment composition

[0042] 1. Experimental animals and drug administration

[0043] Thirty SPF male SD rats, weighing 180-220g and aged 6-8 weeks, were purchased from Shanghai Slake Experimental Animal Co., Ltd. (SCXK (Hu) 2022-0004) and were raised in the barrier environment of the Experimental Animal Center of Nanjing University of Chinese Medicine, with a temperature of (22±2)°C, a relative humidity of (55±10)%, and a light exposure of 12h alternating between light and dark, and free access to water and food. After adaptive feeding, the animals were divided into a control group (Con), a model group (TP), a low-dose group of wolfberry pigment composition in Example 1 (GQ-L, 70mg / kg / d), a high-dose group of wolfberry pigment composition (GQ-H, 105mg / kg / d), and a finasteride group (Fi, 1mg / kg / d). Except for the control group, the remaining rats were subcutaneously injected with 5mg / kg / d of testosterone propionate dissolved in corn oil to induce a rat prostatic hyperplasia model for 4 consecutive weeks, and oral administration and modeling were performed simultaneously in each group.

[0044] 2. Sample collection and testing

[0045] 24 hours after the last administration, rats were anesthetized by intraperitoneal injection of 1.2 g / kg urethane. After the pain reflex of the rats disappeared, blood was collected from the abdominal aorta, and 5 mL of serum was collected by centrifugation. The prostate tissue of the rats was removed by dissection and placed in 4% paraformaldehyde fixative for pathological section analysis. ELISA kits were used to detect the levels of testosterone (T), prostate-specific antigen (PSA), 5-AR2, DHT, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) in rat serum and prostate tissue, and biochemical kits were used to detect the levels of glutathione (GSH), superoxide dismutase (SOD), and malondialdehyde (MDA).

[0046] 4. Experimental Results

[0047] like Figure 2 As shown in the results, the wet weight of prostate in TP group [(1157.63±71.51) mg] was significantly higher than that in control group [(540.68±46.90) mg] (P<0.01). After drug intervention, each group had different degrees of improvement on prostate tissue, down-regulating the wet weight of prostate tissue and prostate index (P<0.05), indicating that the wolfberry pigment composition can improve prostate hyperplasia in vivo.

[0048] The results of H&E staining of rat prostate tissue are as follows Figure 3 As shown in the results, compared with the control group, the glandular cavity of the prostate in the TP group was significantly expanded, the glandular shape was irregular, and glandular epithelial cell hyperplasia was observed. The acinar epithelium thickened, forming many wrinkles and protrusions, and the number of papillary protrusions increased significantly. In addition, the thickness of the prostate epithelium in the TP group was (81.07±8.15) μm, which was significantly higher than that in the control group (31.97±10.17) μm. After administration, the above phenomena were significantly alleviated, and the thickness of the prostate tissue epithelium was significantly reduced (P<0.05), indicating that the Lycium barbarum pigment composition can improve the histopathology of prostate hyperplasia.

[0049] The expression levels of T, DHT, 5-AR and PSA were detected by ELISA. Figure 4 As shown, the levels of various indicators in serum and prostate tissue of the TP group were significantly higher than those of the control group (P<0.01), and the androgen levels were reduced to varying degrees after drug intervention, indicating that the wolfberry pigment composition improves prostate hyperplasia by inhibiting the androgen pathway.

[0050] The results of inflammatory factor determination are shown in Figure 5 As shown, the expression levels of IL-6, IL-1β, and TNF-α in the serum and prostate tissue of the model group were significantly increased (P<0.01), indicating that the model rats were in an inflammatory state, and the wolfberry pigment composition could inhibit the expression of inflammatory factors, showing a good anti-inflammatory effect.

[0051] By detecting the expression of three oxidative stress factors in rat serum and prostate tissue, we found Figure 6 As shown, the expression of GSH and SOD activity in the serum and prostate tissue of the model group rats was significantly lower than that of the control group (P<0.01), and the expression of MDA was significantly increased (P<0.01). Each administration group can improve the SOD activity and GSH expression in the serum and prostate tissue of the model rats to varying degrees, and reduce the expression of MDA, indicating that the wolfberry pigment composition can alleviate the oxidative stress state of rats with prostatic hyperplasia.

[0052] The above examples are only used to illustrate the technical solution of the present invention and are not intended to limit it. Although the present invention is described in detail with reference to the given examples, a person of ordinary skill in the art may modify or replace the technical solution of the present invention as required without departing from the spirit and scope of the technical solution of the present invention.

Claims

1. A wolfberry pigment composition for treating prostatic hyperplasia, characterized in that: The composition comprises lycium barbarum, zeaxanthin, lutein and beta-carotene.

2. The wolfberry pigment composition for treating prostatic hyperplasia according to claim 1, characterized in that: The composition consists of 25-75% of lycium barbarum red pigment, 5-45% of zeaxanthin, 4-25% of lutein and 0.08-5% of beta-carotene in mass percentage.

3. The wolfberry pigment composition for treating prostatic hyperplasia according to claim 2, characterized in that: The composition consists of 25-60% of lycium barbarum red pigment, 5-20% of zeaxanthin, 4-15% of lutein and 0.08-1.6% of beta-carotene in mass percentage.

4. The wolfberry pigment composition for treating prostatic hyperplasia according to claim 2, characterized in that: The composition consists of 67% of lycium barbarum red pigment, 17% of zeaxanthin, 13.6% of lutein and 2.4% of beta-carotene in percentage by weight.

5. The wolfberry pigment composition for treating prostatic hyperplasia according to claim 2, characterized in that: The composition consists of 73% of lycium barbarum red pigment, 15% of zeaxanthin, 10% of lutein and 2% of beta-carotene in mass percentage.

6. Use of the wolfberry pigment composition according to any one of claims 1 to 5 in the preparation of a pharmaceutical product for treating prostatic hyperplasia.

7. The use according to claim 6, characterized in that: The wolfberry pigment composition and a pharmaceutically acceptable carrier are prepared into tablets, pills, powders, decoctions, granules, injections, capsules or decoctions.

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