Application of forsythin in preparation of medicine for preventing oedema caused by rosiglitazone

Abstract

The invention discloses application of forsythin in preparation of a medicine for preventing oedema caused by rosiglitazone, and belongs to the technical field of biological medicine. Aiming at the problem that the rosiglitazone can generate the side effect of edema when treating the type 2 diabetes mellitus, the forsythin and the rosiglitazone are matched for use, and the rosiglitazone can effectively reduce the edema caused by the rosiglitazone while relieving the type 2 diabetes mellitus, so that the side effect of the rosiglitazone medicine can be remarkably reduced, and the dosage of the rosiglitazone medicine can be reduced. The invention provides a novel safe and effective pharmaceutical composition for treating and preventing insulin resistance diseases, and also provides theoretical guidance for preventing and treating oedema side effects of rosiglitazone.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to the application of forsythin in the preparation of a medicine for preventing edema caused by rosiglitazone. Background Art

[0002] The incidence of type 2 diabetes is increasing year by year, posing a serious threat to human health and quality of life. Insulin resistance is the primary pathological characteristic of type 2 diabetes and a key factor in the metabolic disorders caused by type 2 diabetes. Rosiglitazone, a thiazolidinone insulin sensitizer and an agonist of the nuclear receptor PPARγ, primarily produces insulin sensitization by stimulating PPARγ in adipose tissue, which can delay the progression of type 2 diabetes. However, long-term use of rosiglitazone can produce the adverse reaction of edema. Therefore, there is an urgent need to develop a drug that can effectively reduce the edema side effect of rosiglitazone, reduce its dosage, and maximize its role in preventing and treating islet resistance diseases.

[0003] The Traditional Chinese Medicine (TCM) forsythia suspensa has the effects of clearing heat, detoxifying, reducing swelling and dispersing nodules. Forsythiaside is the main pharmacologically active ingredient in TCM forsythia suspensa and is also a quality control indicator component of TCM forsythia suspensa as specified in the Chinese Pharmacopoeia. Studies have shown that forsythiaside can improve insulin resistance. Given the different mechanisms of action of rosiglitazone and forsythiaside, and the complexity of the mechanisms of insulin resistance, it is unclear whether the combination of rosiglitazone and forsythiaside can produce a synergistic effect in the treatment of insulin resistance while simultaneously reducing the edema side effect caused by rosiglitazone. Summary of the Invention

[0004] In response to the above-mentioned problems, the present invention aims to provide the use of forsythiaside in the preparation of a medicament for preventing edema caused by rosiglitazone. Forsythiaside and rosiglitazone are used in combination to treat insulin resistance. By utilizing the synergistic effect between the two, the course of type 2 diabetes can be delayed through rosiglitazone, and edema caused by rosiglitazone can be effectively prevented.

[0005] In order to achieve the above object, the technical solution adopted by the present invention is as follows: In one aspect, the present invention provides the use of forsythiaside in the preparation of a medicament for preventing edema caused by rosiglitazone.

[0006] In another aspect, the present invention further provides a pharmaceutical combination for preventing and / or treating insulin resistance, comprising forsythiaside and rosiglitazone.

[0007] Preferably, the mass ratio of forsythin to rosiglitazone is 25:0.6-40.

[0008] Preferably, the mass ratio of forsythin to rosiglitazone is 25:0.6-10.

[0009] Furthermore, the drug combination also includes a pharmaceutically acceptable carrier.

[0010] Furthermore, the dosage form of the drug combination includes tablets, capsules, pills, granules, solutions or suspensions.

[0011] On the other hand, the present invention also provides use of the aforementioned drug combination in the preparation of a drug for treating type 2 diabetes.

[0012] The beneficial effects of the present invention are: The present invention, through animal experiments, has found that combining forsythiaside and rosiglitazone has a more effective protective effect against insulin resistance than either forsythiaside or rosiglitazone alone. By leveraging the synergistic effect between forsythiaside and rosiglitazone, the course of type 2 diabetes can be delayed with rosiglitazone. Furthermore, the addition of forsythiaside can effectively reduce rosiglitazone-induced edema, significantly reducing the side effects of rosiglitazone and reducing its dosage. This provides a new, safe and effective drug combination for the treatment and prevention of insulin resistance and offers theoretical guidance for preventing and treating the edema side effect of rosiglitazone. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 These are the results of the effects of different treatments on glucose tolerance in experimental mice.

[0014] Figure 2 These are the results of the effects of different treatments on the blood glucose levels of experimental mice.

[0015] Figure 3 These are the results of the effects of different treatments on serum insulin levels in experimental mice.

[0016] Figure 4 The results show the effects of different treatment methods on the hematocrit of experimental mice. DETAILED DESCRIPTION

[0017] In order to enable those skilled in the art to better understand the technical solution of the present invention, the technical solution of the present invention is further described below in conjunction with the accompanying drawings and embodiments.

[0018] It should be understood that the terminology used herein is intended only to describe particular embodiments and is not intended to limit the present invention. Furthermore, for numerical ranges used herein, it should be understood that each intermediate value between the upper and lower limits of that range is specifically disclosed. Intermediate values within any stated value or range, as well as each smaller range between any other stated value or intermediate value within that stated range, are also encompassed within the present invention. The upper and lower limits of such smaller ranges may independently be included or excluded within the range. Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention pertains. Although preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of this invention. Numerous modifications and variations can be made to the specific embodiments described herein without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments will be apparent to those skilled in the art from this description. The present description and examples are illustrative only. As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to mean "including, but not limited to."

[0019] 1. Materials and Methods 1.1 Research subjects and experimental reagents Male C57BL / 6J mice, SPF grade, were purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd., Laboratory Animal Production License No. SCXK (Su) 2018-0008. The experimental animals were housed in an SPF-grade environment in accordance with the requirements for laboratory animal husbandry in my country. The environment was well ventilated, with a room temperature of 21–23°C and a relative humidity of 45%–55%. The light and dark cycles were alternating 12 h each. Mice were housed 5 per cage with free access to food and water.

[0020] Forsythin was purchased from Chengdu Lemeitian Pharmaceutical Technology Co., Ltd., rosiglitazone was purchased from Shanghai MacLean Biochemical Technology Co., Ltd., D-glucose was purchased from Shanghai Sangon Biotechnology Co., Ltd., insulin enzyme-linked immunosorbent assay (ELISA) kit was purchased from Shanghai Sangon Biotechnology Co., Ltd., and blood glucose test strips were purchased from Roche. Hirschmann heparinized capillary tubes (60 μL) were purchased from Hirschmann Laboratory Instruments, Germany. 60% fat content high-fat feed was purchased from Nanjing Harmony Biotechnology Co., Ltd., and insulin ELISA kit was purchased from Shanghai Sangon Biotechnology Co., Ltd.

[0021] 1.2 Experimental groups and treatments Mice were acclimated for one week and then randomly divided into a normal group, a high-fat diet (HFD) group, a Phil group (25 mg / kg), a Rosi1 group (40 mg / kg), a Rosi2 group (10 mg / kg), a Rosi3 group (2.5 mg / kg), a Rosi4 group (0.6 mg / kg), a Phil + Rosi1 group (25 + 40 mg / kg), a Phil + Rosi2 group (25 + 10 mg / kg), a Phil + Rosi3 group (25 + 2.5 mg / kg), and a Phil + Rosi4 group (25 + 0.6 mg / kg), with 10 mice in each group. The normal group received a standard diet, while the other groups received a high-fat diet with 60% fat content and free access to water. Forsythin and rosiglitazone were suspended in a 1% CMC-Na aqueous solution, each containing a 3-day dose. Mice in each group were orally administered the corresponding drug solution daily for 17 weeks. Mice in the normal and high-fat diet groups received an equal volume of vehicle as a control. Note: Phil: forsythiaside; Rosi: rosiglitazone.

[0022] 1.3 Oral glucose tolerance test Mice were fasted for 6 hours starting at 7:00 AM. Fasting blood glucose was measured by tail tip blood sampling, which was used as the 0-minute blood glucose value. Subsequently, mice in each group were gavaged with 2 g / kg glucose solution. Blood glucose levels were measured using a glucometer 10, 20, 30, 60, 90, and 120 minutes after gavage. GraphPad Prism software was used for data analysis, and the area under the curve (AUC) was calculated.

[0023] 1.4 Non-fasting and fasting blood glucose testing Blood was collected from the orbital venous plexus of mice in each group, or the mice in each group were fasted overnight and blood was collected from the tip of the tail of the mice, and blood glucose was measured using a blood glucose meter.

[0024] 1.5 Non-fasting and fasting serum insulin level testing Blood was collected from the orbital venous plexus of mice in each group, or the mice in each group were fasted overnight and blood was collected from the orbital venous plexus. The blood samples were centrifuged at 4°C and 10,000 rpm for 10 min to separate serum. The non-fasting and fasting serum insulin levels were detected by ELISA.

[0025] 1.6 Hematocrit test Packed Cell Volume (PCV) determination: Blood was collected from the orbital vein using a Hirschmann heparinized capillary tube and centrifuged. The PCV was expressed as the height of the blood cells divided by the length of the whole blood column. Hemolyzed samples were excluded.

[0026] 1.7 Statistical methods All data are expressed as mean ± SEM. GraphPad Prism 9.0 software was used for data analysis or t-test. Statistical significance was considered to be P < 0.05.

[0027] 2. Experimental Results 2.1 Effects of rosiglitazone combined with forsythiaside on glucose tolerance The results of the effects of rosiglitazone combined with forsythiaside on glucose tolerance are shown in the attached figure. Figure 1 As shown in the attached Figure 1 In the figure, a is the oral glucose tolerance curve of mice in each group, and b is the area under the glucose tolerance curve.

[0028] As attached Figure 1 As shown in Figures a and b, compared with normal mice, mice in the HFD group developed significant glucose intolerance. The Phil, Rosi1, Rosi2, and Rosi3 groups all significantly improved glucose intolerance in HFD mice, indicating that forsythin and 2.5, 10, and 40 mg / kg of rosiglitazone, when used alone, significantly improved glucose intolerance in high-fat mice. Furthermore, the Phil + Rosi2 group (forsythin combined with 10 mg / kg rosiglitazone) further improved high-fat-induced glucose intolerance compared to either forsythin or 10 mg / kg rosiglitazone alone.

[0029] The Phil + Rosi 4 group (forsythin combined with 0.6 mg / kg rosiglitazone) showed a trend of further reducing glucose intolerance in HFD mice compared with 0.6 mg / kg rosiglitazone alone ( p =0.06). This indicates that rosiglitazone combined with forsythin achieves a synergistic effect in the treatment of insulin resistance in obesity, and has better effects than either drug alone.

[0030] 2.2 Effects of rosiglitazone combined with forsythiaside on blood glucose The results of the effects of rosiglitazone combined with forsythin on blood glucose in mice are shown in the attached figure. Figure 2 As shown, a is the fasting blood glucose test result, and b is the non-fasting blood glucose test result.

[0031] The fasting blood glucose test results of experimental mice are shown in the attached Figure 2As shown in middle a, compared with normal mice, the fasting blood glucose level of HFD mice was significantly increased; rosiglitazone at doses of 2.5, 10, and 40 mg / kg (Rosi3, Rosi2, and Rosi1 groups) alone significantly reduced the fasting blood glucose level of HFD mice; and the combination of forsythin and 10 mg / kg rosiglitazone (Phil + Rosi 2 group) further reduced the fasting blood glucose level of HFD mice compared with forsythin (Phil group) or 10 mg / kg rosiglitazone (Rosi2 group) alone; the combination of forsythin and 0.6 mg / kg rosiglitazone (Phil + Rosi 4 group) showed a trend of further reducing the fasting blood glucose level of HFD mice compared with 0.6 mg / kg rosiglitazone (Rosi4 group) ( p =0.054).

[0032] The results of non-fasting blood glucose test of experimental mice are shown in the attached Figure 2 As shown in middle b, compared with normal mice, the non-fasting blood glucose levels of HFD mice were significantly increased; the non-fasting blood glucose levels of mice in the forsythin (Phil group) and rosiglitazone dose groups (Rosi1 group, Rosi2 group, Rosi3 group, Rosi4 group) were not significantly different from those of HFD mice; however, the non-fasting blood glucose levels of mice in the forsythin combined with 10 mg / kg rosiglitazone (Phil + Rosi 2 group) showed a trend of further decrease compared with the mice in the 10 mg / kg rosiglitazone (Rosi2 group) group ( p =0.058); moreover, compared with HFD mice, the non-fasting blood glucose levels of mice in the forsythiaside combined with 10 mg / kg rosiglitazone group (Phil + Rosi group 2) and the forsythiaside combined with 2.5 mg / kg rosiglitazone group (Phil + Rosi group 3) were significantly reduced.

[0033] The above results fully demonstrate that rosiglitazone combined with forsythiaside in the treatment of obesity-induced insulin resistance achieves a synergistic effect and has better effects than the two single drugs.

[0034] 2.3 Effects of rosiglitazone combined with forsythiaside on serum insulin levels The results of the effects of rosiglitazone combined with forsythiaside on serum insulin levels in mice are shown in the attached figure. Figure 3 As shown in the attached Figure 3 In the figure, a is the fasting serum insulin test result, and b is the non-fasting serum insulin test result.

[0035] The results of fasting serum insulin test of experimental mice are shown in the attached Figure 3As shown in center a, compared with normal mice, serum fasting insulin levels in HFD mice were significantly increased, confirming that HFD mice developed insulin resistance. Both forsythiaside (Phil group) and the 40 mg / kg rosiglitazone group (Rosi1 group) significantly reduced fasting serum insulin levels in HFD mice. There were no significant differences in serum fasting insulin levels between mice in the 10, 2.5, and 0.6 mg / kg rosiglitazone groups (Rosi2, Rosi3, and Rosi4 groups) and those in the HFD group. However, compared with the HFD group, fasting serum insulin levels were significantly reduced in mice in the forsythiaside combined with 40 mg / kg rosiglitazone group (Phil + Rosi1 group), the forsythiaside combined with 10 mg / kg rosiglitazone group (Phil + Rosi 2 group), and the forsythiaside combined with 2.5 mg / kg rosiglitazone group (Phil + Rosi 3 group).

[0036] The results of non-fasting serum insulin test in experimental mice are shown in the attached Figure 3 As shown in middle b, the serum non-fasting insulin level of HFD mice increased significantly, indicating that HFD mice developed insulin resistance; Forsythin and rosiglitazone alone in each dose group could significantly reduce the non-fasting serum insulin level of HFD mice; Forsythin combined with 10 mg / kg rosiglitazone dose group (Phil + Rosi 2 group) further reduced the serum non-fasting insulin level of mice compared with the 10 mg / kg rosiglitazone single-dose group (Rosi2 group) ( p =0.06), the serum non-fasting insulin level of mice in the group of forsythin combined with 0.6 mg / kg rosiglitazone (Phil + Rosi 4 group) was further reduced compared with the group of 0.6 mg / kg rosiglitazone alone (Rosi 4 group) ( p =0.06).

[0037] The above results show that rosiglitazone combined with forsythiaside in the treatment of obesity-induced insulin resistance achieves a synergistic effect and has better effects than the two single drugs.

[0038] 2.4 Rosiglitazone combined with forsythin significantly reduced the side effect of edema The results of the effects of rosiglitazone combined with forsythin on edema are shown in the attached Figure 4 As shown. Figure 4It can be seen that compared with the normal group, the HFD group and the forsythin group (Phil group) had no significant effect on the hematocrit of mice; the 40 mg / kg, 10 mg / kg and 2.5 mg / kg rosiglitazone dose groups (Rosi1 group, Rosi2 group and Rosi3 group) could significantly reduce the hematocrit of HFD group mice, and the 0.6 mg / kg dose of rosiglitazone (Rosi 4 group) had no effect on the hematocrit of HFD mice. The forsythiaside combined with 40 mg / kg rosiglitazone dose group (Phil + Rosi1 group) still significantly reduced the hematocrit of HFD mice, while the forsythiaside combined with 0.6 mg / kg rosiglitazone dose group (Phil + Rosi 4 group) had no effect on the hematocrit of HFD mice; the forsythiaside combined with 10 mg / kg rosiglitazone dose group (Phil + Rosi 2 group) and the forsythiaside combined with 2.5 mg / kg rosiglitazone dose group (Phil + Rosi 3 group) had no effect on the hematocrit of HFD mice.

[0039] The above results show that the use of rosiglitazone alone at a dose higher than (including) 2.5 mg / kg can significantly reduce the hematocrit of mice and produce the side effect of edema; while forsythiaside combined with 0.6-10 mg / kg rosiglitazone does not produce edema when treating obesity and insulin resistance; forsythiaside combined with 40 mg / kg rosiglitazone reduces the side effect of edema when treating obesity and insulin resistance.

[0040] The basic principles, main features, and advantages of the present invention are shown and described above. Those skilled in the art should understand that the present invention is not limited to the foregoing embodiments. The foregoing embodiments and descriptions are merely illustrative of the principles of the present invention. Various changes and modifications may be made to the present invention without departing from the spirit and scope of the present invention. Such changes and modifications are intended to fall within the scope of the present invention. The scope of protection claimed in the present invention is defined by the appended claims and their equivalents.

Claims

1. Application of forsythiaside in the preparation of a drug for preventing edema caused by rosiglitazone.

2. A pharmaceutical combination for preventing and / or treating insulin resistance, characterized in that: The drug combination comprises forsythiaside and rosiglitazone.

3. The pharmaceutical combination for preventing and / or treating insulin resistance according to claim 2, characterized in that: In the drug combination, the mass ratio of forsythiaside to rosiglitazone is 25:0.6-40.

4. The pharmaceutical combination for preventing and / or treating insulin resistance according to claim 3, characterized in that: In the drug combination, the mass ratio of forsythiaside to rosiglitazone is 25:0.6-10.

5. The pharmaceutical combination for preventing and / or treating insulin resistance according to claim 2, characterized in that: The pharmaceutical combination further includes a pharmaceutically acceptable carrier.

6. The pharmaceutical combination for preventing and / or treating insulin resistance according to claim 2, characterized in that: The dosage forms of the drug combination include tablets, capsules, pills, granules, solutions or suspensions.

7. Use of the drug combination according to any one of claims 2 to 6 in the preparation of a medicament for treating type 2 diabetes.

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