Use of berberine or its salts in the preparation of a drug for preventing and / or treating diabetic encephalopathy

Abstract

This invention provides the application of berberine or its salts in the preparation of drugs for the prevention and / or treatment of diabetic encephalopathy, belonging to the field of pharmaceutical technology. The natural drug berberine provided by this invention covers multiple therapeutic targets, exhibits low drug resistance, and has few side effects, making it significant for improving diabetic encephalopathy. The berberine provided by this invention can promote the integrity of cerebral capillaries, reduce their leakage, and promote vascular repair and regeneration of cerebral capillaries.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and particularly relates to the application of berberine or its salts in the preparation of drugs for the prevention and / or treatment of diabetic encephalopathy. Background Technology

[0002] Diabetic encephalopathy is a chronic complication of diabetes, encompassing various central nervous system and cerebrovascular diseases caused by diabetes, leading to cognitive impairment. Berberine (BBR) is a natural medicine originating in China, and its efficacy in improving diabetes and other conditions has been confirmed by multiple clinical trials. It exerts its effects through various pathways, including lowering blood sugar and anti-inflammation.

[0003] Current treatments for diabetic encephalopathy are mostly symptomatic, which have drawbacks such as targeting a single point, requiring multiple drugs, and having many side effects. Currently, no drugs with multiple therapeutic targets, low drug resistance, and few side effects have been found for improving or treating diabetic encephalopathy. Summary of the Invention

[0004] In view of this, the object of the present invention is to provide the use of berberine or its salts in the preparation of drugs for the prevention and / or treatment of diabetic encephalopathy. The natural drug berberine provided by the present invention covers multiple therapeutic targets, has low drug resistance, and few side effects, and is of great significance for improving diabetic encephalopathy.

[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0006] This invention provides the use of berberine or its salts in the preparation of drugs for the prevention and / or treatment of diabetic encephalopathy.

[0007] The present invention also provides the use of berberine or its salts in the preparation of medicaments for the prevention and / or treatment of central nervous system disorders caused by diabetes.

[0008] The present invention also provides the use of berberine or its salts in the preparation of drugs for the prevention and / or treatment of cerebrovascular diseases caused by diabetes.

[0009] The present invention also provides the application of berberine or its salts in the preparation of drugs that promote the repair of cerebral capillaries.

[0010] The present invention also provides the use of berberine or its salts in the preparation of drugs that promote the regeneration of cerebral capillaries.

[0011] The present invention also provides the use of berberine or its salts in the preparation of drugs that increase the number of cerebral capillaries.

[0012] The present invention also provides the use of berberine or its salts in the preparation of drugs that increase the length of cerebral capillaries.

[0013] The present invention also provides the use of berberine or its salts in the preparation of drugs that increase the total volume of cerebral capillaries.

[0014] The present invention also provides the use of berberine or its salts in the preparation of medicaments for the prevention and / or treatment of behavioral dysfunction in diabetic encephalopathy.

[0015] Preferably, the berberine salts include berberine hydrochloride. Attached Figure Description

[0016] Figure 1 The effects of BBR treatment on body weight and fasting blood glucose in 2DEK model mice before and after treatment;

[0017] Figure 2 The effects of BBR treatment on the behavior of 2DEK model mice;

[0018] Figure 3 Representative images of capillaries in the extralateral / medial parietal cortex (LPtA / MPtA) region of mice in each group after BBR treatment;

[0019] Figure 4 A statistical diagram of blood vessel volume in the LPtA / MptA region of the brain of mice in each group after BBR treatment;

[0020] Figure 5 Statistical graph of vascular parameters in the LPtA / MptA region of the brain of mice in each group after BBR treatment;

[0021] Figure 6 Representative three-dimensional reconstructions of major brain regions in mice from groups M and BBR-H are shown.

[0022] Figure 7 Comparison of the volume of major brain regions between the M group and the BBR-H group;

[0023] Figure 8 Representative images of capillaries in the CA2 region of the hippocampus and the LPtA / MPtA region of the brain in mice of groups M and BBR-H;

[0024] Figure 9 Statistical distribution of capillary diameters in the CA2 region of the hippocampus and the LPtA / MPtA region of the brain in mice of groups M and BBR-H;

[0025] Figure 10 Statistical diagram of capillary length distribution in the CA2 region of the hippocampus and the LPtA / MPtA region of the brain in mice of groups M and BBR-H;

[0026] Figure 11 The distribution of capillary volume and number in the CA2 region of the hippocampus of mice in groups M and BBR-H is shown.

[0027] Figure 12 A statistical diagram showing the total volume distribution of capillaries in the CA2 region of the hippocampus in mice of groups M and BBR-H.

[0028] Figure 13 Statistical diagram of capillary volume distribution in the LPtA / MPtA region of the brain of mice in groups M and BBR-H. Detailed Implementation

[0029] This invention provides the use of berberine or its salts in the preparation of drugs for the prevention and / or treatment of diabetic encephalopathy.

[0030] In this invention, the salt of berberine is preferably berberine hydrochloride. This invention does not have a particular limitation on the source of berberine hydrochloride, but commercially available products with a purity of ≥98% are preferred.

[0031] In this invention, the diabetic encephalopathy is preferably spontaneous type 2 diabetic encephalopathy.

[0032] In this invention, the preventive or therapeutic effect of berberine hydrochloride on spontaneous type 2 diabetic encephalopathy is preferably a protective effect on cerebral blood vessels.

[0033] In this invention, the preferred dosage of berberine hydrochloride is a high dose and a low dose, wherein the high dose is preferably 200 mg / kg and the low dose is preferably 100 mg / kg.

[0034] The present invention also provides the use of berberine or its salts in the preparation of medicaments for the prevention and / or treatment of central nervous system disorders caused by diabetes.

[0035] In this invention, the symptoms of central nervous system lesions caused by diabetes preferably include behavioral dysfunction.

[0036] The present invention also provides the use of berberine or its salts in the preparation of drugs for the prevention and / or treatment of cerebrovascular diseases caused by diabetes.

[0037] In this invention, the preferred method for treating cerebrovascular lesions is to promote the integrity of capillaries in the brain, reduce their leakage, and promote vascular repair and regeneration of cerebral capillaries.

[0038] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0039] Example 1

[0040] Cerebrovascular protective effects of berberine on a mouse model of spontaneous type 2 diabetic encephalopathy

[0041] 1. Experimental Materials

[0042] 1.1 Reagents and Consumables

[0043] Berberine hydrochloride and its standard (Berberine, BBR, CAS: 633-65-8, purity: ≥98%) were purchased from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China). Deionized water for the experiment was purchased from Hangzhou Wahaha Group Co., Ltd. (Zhejiang, China). KK-Ay mouse feed No. 1042 (high-fat, high-sugar) was purchased from Beijing Huafukang Biotechnology Co., Ltd. (Beijing, China). Paraformaldehyde (CAS: 30525-89-4, purity: reagent grade, crystalline), potassium dihydrogen phosphate (CAS: 7778-77-0, purity: ≥99.0%), potassium chloride (CAS: 7447-40-7, purity: 99.0-100.5%), and sucrose (CAS: 57-50-1, purity: 99%) were purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd. (Shanghai, China). Sodium chloride (CAS: 7647-14-5, purity: AR) and sodium dihydrogen phosphate monohydrate (CAS: 10049-21-5, purity: AR) were purchased from Beijing Tongguang Fine Chemical Co., Ltd. (Beijing, China). Trichloroacetaldehyde (CAS: 302-17-0, purity: AR, >99%) was purchased from Shanghai Yi'en Chemical Technology Co., Ltd. (Shanghai, China). LR White resin embedding agent was purchased from Agar Scientific (London, UK). Lycopersicon esculentum lectin (LEL) - DyLight TM 488 was purchased from Vectorlaboratories (California, USA). All other reagents and consumables were provided by Qiancheng Mirror (Beijing) Technology Co., Ltd. (Beijing, China).

[0044] 1.2 Instruments and Equipment

[0045] Adventurer analytical electronic balance (OHAUS, China); Roche Gold Glucose Meter and test strips (Roche Diagnostics Shanghai, China); STT-2 mouse jumping platform (Institute of Materia Medica, Chinese Academy of Medical Sciences, China); BioMapping 5000 fluorescence microscopy optical section tomography system (Wuhan Woyi Biotechnology Co., Ltd., China); and other instruments and equipment were provided by Qiancheng Mirror (Beijing) Technology Co., Ltd. (Beijing, China).

[0046] 1.3 Laboratory Animals

[0047] Specific pathogen free (SPF)-level C57BL / 6J mice (male, 42 - 49 days old) and KK-Ay mice (male, 42 - 49 days old, SPF level) were purchased from Beijing Huafukang Bioscience Co., Ltd. (Beijing, China), license number: SCXK (Jing) 2019 - 0008, and were raised under experimental conditions (12h light / dark cycle, 8 am to 8 pm; environmental temperature 20 - 25 °C; relative humidity 40 - 70%). All animal experiments were strictly carried out in accordance with the Guidelines for the Care and Use of Laboratory Animals issued by the Institute for Animal Protection and Welfare. All animal protocols were approved by the Animal Care and Welfare Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College (Beijing, China) (No: 00005020), and were conducted according to the institutional guiding principles and ethical standards.

[0048] 2 Experimental methods

[0049] 2.1 Three-dimensional ultrastructural imaging analysis of the cerebrovasculature in diabetic encephalopathy mice

[0050] Male KK-Ay mice (6 - 7 weeks old, 80 mice) and male C57BL / 6J mice (

[0051] 6 - 7 weeks old, 15 mice) were selected. After being fed with standard feed for 1 week to adapt to the environment, the C57BL / 6J mice continued to be given standard feed, while the KK-Ay mice were given their special high-fat and high-sugar feed to further induce them into spontaneous type 2 diabetic encephalopathy (Type2diabetic encephalopathy KK-Ay, 2DEK) model mice. The body weights of each mouse were measured at a fixed time every week, and blood was taken from the tip of each mouse's tail, with a single blood volume of 5 μL, to detect blood glucose and other indicators to determine whether the model was successful. After 10 weeks, KK-Ay mice with a fasting blood glucose ≥ 11.1 mmol / L for three consecutive weeks of fasting for 6 h were selected and randomly divided into 3 groups, including a model (Model, M) group, a low-dose berberine (Low dose of berberine, BBR-L, 100 mg / kg) group, and a high-dose berberine (High dose ofberberine, BBR-H, 200 mg / kg) group, and C57BL / 6J mice were used as a control (Control, Con) group, with n = 15 in each group. Among them, the above doses were orally administered for treatment in the BBR-L group and the BBR-H group, and the Con group and the M group were given an equal volume of aqueous solution orally. During the administration period, the body weights of each mouse were measured at a fixed time every week, and blood was taken from the tip of each mouse's tail, with a single blood volume of 5 μL, to detect blood glucose and other indicators to judge the efficacy of BBR. After 10 weeks of treatment, blood glucose, body weight detection, and step-down behavioral experiments were performed on each group of mice to further evaluate the condition of diabetic encephalopathy in mice and the efficacy of BBR.

[0052] Based on the results of platform jumping behavior, mice in the median data groups were selected. Three-dimensional ultrastructural imaging and analysis of the cerebral blood vessels were performed by Qiancheng Mirror (Beijing) Technology Co., Ltd. (Beijing, China). The specific method is as follows: After weighing, the mice were anesthetized by intraperitoneal injection of 3.5% trichloroacetaldehyde at a dose of 100 g / ml. LEL-Dylight 488 (140 μl mL / mouse) was injected via the tail vein. Three minutes later, the mice were fixed on the dissection table, and the thoracic cavity was opened to expose the heart. The constant flow pump was turned on, and the tubing and needle were flushed with 0.01M PBS. After confirming patency, the pump was turned off. The heart was held with hemostatic forceps in the left hand, and the injection needle was inserted into the left ventricle from the apex of the heart in the right hand. The needle position was fixed, the right atrial appendage was cut open, the constant flow pump was turned on, and 50 mL of 0.01M PBS was used for perfusion. After the liver quickly turned white, it was fixed with 100 mL of 4% paraformaldehyde solution. After the mouse tail twitched and gradually stiffened, the brain tissue was harvested and transferred to a centrifuge tube containing 4% paraformaldehyde solution (40 mL / brain tissue). The tissue was fixed at 4°C for 24 hours. The brain tissue was then transferred to a centrifuge tube containing pre-chilled PBS solution (40 mL / brain tissue) for rinsing at 4°C for 24 hours. The PBS was discarded, and any remaining solution was removed with a pipette. A gradient dehydration process was then performed by placing 50 mL centrifuge tubes in 40 mL of 50%, 75%, and 100% ethanol at 4°C for 2 hours, followed by overnight incubation in 100% ethanol. After dehydration, the samples were infiltrated with 50%, 75%, and 100% LR White resin embedding agents at 4°C for 2 hours, respectively. After infiltration with 100% LR White resin embedding agent for 48 hours, the brain tissue samples were placed into gelatin capsules (size 000), filled with 100% filtered LR White resin embedding agent, and polymerized using a gradient process at 45°C for 2 hours, 48°C for 4 hours, and 50°C for 3 hours. Microscopic optical tomography was performed on each group of samples using the fMOST system, achieving an imaging resolution of 0.35μm × 0.35μm × 2μm, enabling high-resolution imaging of the entire brain tissue's blood vessels. The excitation light and channel were Ex = 488nm, with LEL-Dylight 488 specifically labeling the blood vessels. The raw image data were preprocessed using Imaris (X649.0.0) and Woyi Data preprocessing software (v0.82), including seamless image stitching, brightness uniformity correction, and image defect elimination. Image optimization is achieved through steps such as background correction, noise reduction and contrast enhancement, and image rotation correction. Every 50 images are subjected to projection processing and three-dimensional reconstruction.

[0053] 3 Experimental Results

[0054] 3.1 Effects of berberine on body weight and blood glucose in diabetic encephalopathy mice

[0055] The results are as follows Figure 1As shown, at week 10 of modeling (week 0 of treatment), the body weight and fasting blood glucose of mice in group M were significantly higher than those in group Con (***P<0.001, Figure 1 (A, D) and during the subsequent 10-week treatment period, the weight and fasting blood glucose of group M showed an increasing trend, while the weight and fasting blood glucose of groups BBR-L and BBR-H continuously decreased to a steady state. Figure 1 B, E), at 10 weeks of treatment, the body weight and fasting blood glucose levels in both the BBR-L and BBR-H groups were significantly lower than those in the M group (***P<0.001), and the BBR-H group required a shorter time to reach steady-state body weight, with its fasting blood glucose levels approaching those of the Con group at the end of treatment (B, E). Figure 1 C, F).

[0056] Figure 1 (A) Comparison of body weight of mice in each group after modeling; (B) Changes in body weight of mice in each group before and after 10 weeks of BBR treatment; (C) Comparison of body weight of mice in each group at week 10 of BBR treatment; (D) Comparison of fasting blood glucose of mice in each group after modeling; (E) Changes in fasting blood glucose of mice in each group before and after 10 weeks of BBR treatment; (F) Comparison of fasting blood glucose of mice in each group at week 10 of BBR treatment; n=10, ***P<0.001; Con, control group; M, model group; BBR-L, low-dose BBR (100mg / kg) group; BBR-H, high-dose BBR (200mg / kg) group, with the same labeling for subsequent images.

[0057] 3.2 Effects of berberine on the behavior of diabetic encephalopathy mice

[0058] To explore the effects of BBR on behavioral aspects such as learning, recognition, and memory in 2DEK model mice, we used a step-down test to evaluate behavioral function after BBR treatment. The results are as follows: Figure 2 As shown, in the step-down test, compared with group M mice, the step-down latency of mice in all groups was significantly increased after BBR treatment (*P<0.05, **P<0.01). Figure 2 A) The number of errors in stepping down was significantly reduced (*P<0.05, **P<0.01), and the BBR-H group showed better behavioral performance. Figure 2 B).

[0059] Figure 2 (A) Comparison of the latency period for mice to fall off the platform in the platform jumping test; (B) Comparison of the number of incorrect steps to fall off the platform in the platform jumping test, n=10; *P<0.05, **P<0.01.

[0060] 3.3 Preliminary evaluation of the effects of berberine on the brain capillary structure of diabetic encephalopathy mice

[0061] Three-dimensional ultrastructural imaging of cerebral blood vessels was performed on mice in each group, and the results are as follows: Figure 3 As shown in the two-dimensional projection map, compared to the Con group, the M group showed significant dye leakage, indicating disruption of cerebral vascular integrity and increased vascular permeability; while the dye leakage around the cerebral blood vessels in all groups of mice treated with BBR was significantly reduced, and the overall images were clearer. Figure 3 A, B); From the three-dimensional reconstruction images, it can be seen that group M has the fewest cerebral blood vessels and the sparsest distribution of blood vessels. However, BBR treatment can significantly promote cerebral vasodilation, increase the number of blood vessels, and make the distribution denser. Among them, group BBR-H is almost comparable to the control group. Figure 3 (C, D, E). Based on the above three-dimensional reconstruction map, the lateral parietal association cortex (LPtA) and medial parietal association cortex (MPtA) regions, located 1.70-1.85 mm from the Bregma point in the brain, were used for three-dimensional quantitative statistical analysis of vascular-related parameters. The results are as follows: Figure 4 As shown. In terms of vascular volume, compared with the Con group, the total vascular volume of the M group decreased, the number of large-volume vessels decreased, and the number of small-volume vessels increased significantly; after BBR treatment, the total vascular volume of all groups increased significantly, with the total vascular volume of the BBR-L group being almost more than twice that of the M group, and the BBR treatment group had fewer small-volume vessels and more large-volume vessels.

[0062] Figure 3 (A) Two-dimensional overall projection of the whole brain blood vessels of each group of mice, with the yellow box indicating the subsequent magnified local view; (B) Two-dimensional projection magnified local view of the brain capillaries of each group of mice; (C) Three-dimensional reconstruction of the brain capillaries of each group of mice; (D) Surface algorithm construction diagram of the brain capillaries of each group of mice; (E) Fiment algorithm construction diagram of the brain capillaries of each group of mice; Scale bar is 100μm.

[0063] Figure 4 (A) Comparison of total volume of brain capillaries in each group of mice; (B) Distribution of brain capillary volume and number in each group of mice.

[0064] Further evaluation of vessel diameter led to the classification of cerebral capillaries into two categories: microcapillaries (0-3.0 μm) and small capillaries (3.0-13.0 μm). The results are as follows: Figure 5As shown in Figure A, regarding microcapillaries, the vessel diameter in group Con was concentrated between 0.8 and 2.8 μm, with a peak value of 1.8 μm. In group M, compared to group Con, the peak diameter of the vessel diameter distribution curve shifted to the left of the peak value, concentrated between 0.5 and 3.0 μm, with the peak value shifting to 1.0 μm. The number of vessels increased significantly, approximately four times that of group Con, indicating a decrease in the diameter and a substantial increase in the number of microcapillaries in the brain of the 2DEK model mice. In group BBR-L, the vessel diameter was concentrated between 1.0 and 3.0 μm, with a peak value of 2.0 μm. Compared to group M, the peak value of the vessel diameter distribution curve shifted to the right, and the curve shifted downwards, indicating a decrease in the number of microcapillaries. In group BBR-H, the vessel diameter was concentrated between 0.5 and 3.0 μm, with a peak value of 1.2 μm. Compared to group M, the peak value of the vessel diameter distribution curve shifted slightly to the right. The curve shifted significantly downwards, approaching the distribution of the Con group curve, indicating a decrease in the number of microcapillaries. Regarding small capillaries, the vessel diameter values ​​in the Con group were concentrated between 4.0-11.0 μm, with a peak of 7.4 μm. In contrast, the peak diameter of the vessel diameter distribution curve in the M group was concentrated between 3.0-6.0 μm, shifting to 4.6 μm, indicating a significant downward shift in the number of vessels. The number of small capillaries in the Con group was approximately twice that of the M group, suggesting a decrease in both the diameter and number of small capillaries in the brain of the 2DEK model mice. In the BBR-L group, the vessel diameter values ​​were concentrated between 3.0-13.0 μm, with a peak of 9.0 μm. Compared to the M group, the peak of the vessel diameter distribution curve shifted significantly to the right, while the curve also shifted significantly upwards, indicating a substantial increase in both the diameter and number of small capillaries. The vessel diameter values ​​in the BBR-H group were concentrated between 3.0 and 9.0 μm, with a peak value of 6.2 μm. Compared to the M group, the peak vessel diameter value shifted to the right, and the curve shifted significantly upward, second only to the BBR-L group. The diameter and number of small capillaries increased. Similarly, vessel length was evaluated, and the results were as follows... Figure 5As shown in B. Regarding microcapillaries, the length of microcapillaries in each group was concentrated between 4.0 and 36.0 μm. The peak value of the Con group was 21.0 μm. Compared to the Con group, the peak value of the curve in the M group shifted to the left, and the vessel length curve shifted significantly upward, approximately twice that of the Con group. The curve of the BBR-L group was almost identical to that of the M group. Regarding small capillaries, the length of microcapillaries in each group was concentrated between 36.0 and 250.0 μm. The peak value of the Control group was 72.0 μm. In the Model group, after the first peak of 18.0 μm, its... The capillary length gradually decreased, and the vessel length-number distribution curve shifted to the left overall, indicating that the number of microcapillaries in the brain of 2DEK model mice increased significantly while their length decreased, resulting in a decrease in the number and shortening of small capillaries. In the BBR-L group, although the number of microcapillaries did not decrease significantly, the number of small capillaries increased significantly, and the overall vessel length increased. In addition, the number of microcapillaries in the BBR-H group decreased significantly, while the number of small capillaries increased moderately, and the vessel length-number distribution curve shifted to the right overall.

[0065] Figure 5 (A) Distribution of capillary diameter and number in the brain of mice in each group; (B) Distribution of capillary length and number in the brain of mice in each group.

[0066] 3.4 Comprehensive evaluation of the effects of berberine on the brain capillary structure of diabetic encephalopathy mice

[0067] Based on the above results, the images collected from mice in the M group and BBR-H group were further preprocessed and reconstructed in three dimensions. The volumes of five brain regions—the whole brain, right cortex, right hippocampus, right striatum, and right midbrain—were specifically calculated. The reconstructed images are shown below. Figure 6 As shown. The volumes of five brain regions in the two groups of mice were calculated and compared. It was found that the volumes of the hippocampus, midbrain, striatum, and whole brain were slightly larger in the M group before and after high-dose BBR treatment. Figure 7 However, there was no significant difference.

[0068] Figure 6 (A) Three-dimensional reconstruction results of the whole brain volume of a mouse; (B) Three-dimensional reconstruction results of the right cortex volume of a mouse; (C) Three-dimensional reconstruction results of the right hippocampus of a mouse; (D) Three-dimensional reconstruction results of the right striatum of a mouse; (E) Three-dimensional reconstruction results of the right midbrain of a mouse; Scale bar is 800 μm.

[0069] Figure 7 In the middle brain, n=3; RC, right cortex; RH, right hippocampus; RM, right midbrain; RS, right striatum; WB, whole brain.

[0070] In addition, the LPTA / MPtA region of the brain and the CA2 region of the hippocampus were excised, and the volume, diameter, and number of capillaries were statistically analyzed. The excised volume was 400×400×400μm. 3 The location of the intercepted area is as follows Figure 8 As shown in Figure A, in the two-dimensional projection images, partial dye leakage was observed in the LPTA / MPtA region and the CA2 region of the hippocampus in group M, resulting in slightly blurred overall images of the cerebral vessels; while the overall images of the cerebral vessels in group BBR-H were clearer, and no dye leakage was observed. Figure 8 B); From the three-dimensional reconstruction images, it can be seen that the M group has fewer cerebral blood vessels in the LPTA / MPtA region and the CA2 region of the hippocampus, and the blood vessel distribution is also more sparse; the BBR-H group has an increased number of cerebral blood vessels and a denser distribution, which is more obvious in the CA2 region of the hippocampus. Figure 8 C, D, E).

[0071] Figure 8 (A) Two-dimensional overall projection of the whole brain blood vessels of each group of mice, with the yellow box indicating the subsequent magnified local view; (B) Two-dimensional projection magnified local view of the brain capillaries of each group of mice; (C) Three-dimensional reconstruction of the brain capillaries of each group of mice; (D) Surface algorithm construction diagram of the brain capillaries of each group of mice; (E) Fiment algorithm construction diagram of the brain capillaries of each group of mice; Scale bar is 50μm.

[0072] The number of blood vessels in these two regions was further statistically analyzed from the perspectives of capillary diameter and length. For example... Figure 9 As shown in Figure A, the capillary diameters in the CA2 region of the hippocampus in group M were mostly distributed between 3-9 μm, with a peak value of 7 μm. After BBR treatment, the capillary diameters in this region were mostly distributed between 3-8 μm, with a peak value of 5 μm, representing an increase in number of capillaries by approximately 42%. In the LPtA / MPtA region, the capillary diameters in group M were mostly distributed between 3-8 μm, with a peak value of 7 μm. After BBR treatment, the capillary diameters in this region were mostly distributed between 4-9 μm, with a peak value of 8 μm, representing an increase in number of capillaries by approximately 60%. Figure 9 B).

[0073] Figure 9 (A) Distribution of capillary diameter and number in the CA2 region of the hippocampus of mice in each group; (B) Distribution of capillary diameter and number in the LPtA / MPtA region of the brain of mice in each group; n = 3; *P < 0.05,

[0074] ***P<0.001.

[0075] Regarding capillary length, such as Figure 10As shown, the capillary lengths in the CA2 region of the hippocampus were mostly distributed between 0 and 5 μm in both the M and BBR-H groups. However, the number of capillaries in this range was significantly higher in the BBR-H group than in the M group (***P<0.001), and the number of capillaries longer than 5 μm was also greater in the BBR-H group than in the M group. Figure 10 A); while in the LPtA / MPtA region, although the number of capillaries in the 0-10μm range of the BBR-H group was similar to that of the M group, the number of capillaries distributed in the 10-22μm range was significantly higher in the BBR-H group than in the M group (*P<0.05). Figure 10 B).

[0076] Figure 10 (A) Distribution of capillary length and number in the CA2 region of the hippocampus of mice in each group; (B) Distribution of capillary length and number in the LPtA / MPtA region of the brain of mice in each group; n = 3; *P < 0.05,

[0077] ***P<0.001.

[0078] Regarding capillary volume, although the number of capillaries in the CA2 region of the hippocampus was less in the M group after BBR treatment ( Figure 11 However, the total capillary volume in group M was mostly concentrated between 0 and 1 × 10⁶ μm³, while the total capillary volume in group BBR-H was mostly concentrated between 4 × 10⁵ and 2.2 × 10⁶ μm³, and the total volume was approximately 67.4% higher than that in group M. Figure 12 (A, B). Furthermore, similar results were observed in the capillaries of the LPtA / MPtA region after BBR treatment, such as... Figure 13 As shown in Figure A, the number of capillaries in the BBR-H group was less than that in the M group, but the total capillary volume was between 0 and 4 × 10⁻⁴. 5 μm 3 and 1.5×10 6 -2.2×10 6 μm 3 The ranges were all significantly higher than those in group M, and the capillary volume in group M was relatively flat throughout the entire volume distribution range. Figure 13 B, C).

[0079] Figure 12 (A) Distribution of capillary volume-total volume in CA2 region of hippocampus in mice of each group; (B) Comparison of total capillary volume in CA2 region of hippocampus in mice of each group, n=3.

[0080] Figure 13(A) Distribution of capillary volume and number in the LPtA / MPtA region of the brain of mice in each group; (B) Distribution of capillary volume and total volume in the LPtA / MPtA region of the brain of mice in each group; (C) Comparison of total capillary volume in the LPtA / MPtA region of the brain of mice in each group, n=3; *P<0.05, ***P<0.001.

[0081] The berberine of this invention can promote the integrity of capillaries in the brain, reduce their leakage, and have a vascular repair and regeneration effect on brain capillaries.

[0082] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Application of berberine salts in the preparation of drugs for the prevention and / or treatment of cerebrovascular diseases caused by diabetes.

2. The application according to claim 1, characterized in that, Berberine salts promote the repair of cerebral capillaries.

3. The application according to claim 1, characterized in that, Berberine salts promote the regeneration of cerebral capillaries.

4. The application according to claim 1, characterized in that, Berberine salts increase the number of brain capillaries.

5. The application according to claim 1, characterized in that, Berberine salts increase the length of cerebral capillaries.

6. The application according to claim 1, characterized in that, Berberine salts increase the total volume of cerebral capillaries.

7. The application according to any one of claims 1 to 6, wherein the berberine salts include berberine hydrochloride.

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