Application of congee decoction in preparation of medicine for treating cognitive impairment of diabetes mellitus

Abstract

The invention provides an application of a congee decoction in preparation of a medicine for treating cognitive impairment of diabetes mellitus. On the basis of clinical application and a large number of early-stage experimental studies which are all decocted and have curative effects, bile acid and intestinal flora metabolic disorder have key effects in the pathological process of diabetes and cognitive impairment; the invention firstly proposes that the homeostasis of bile acid and intestinal microbiota can be mediated through an'intestinal flora-bile acid-brain 'way to improve the cognitive impairment of diabetes mellitus by consistent decoction.

Description

Technical Field

[0001] The invention relates to use of Yiguanjian in preparing medicine for treating diabetic cognitive impairment, and belongs to the field of medicines. Background Art

[0002] Diabetic cognitive impairment has become a major global public health issue, necessitating the urgent search for effective interventions. Traditional Chinese Medicine (TCM) considers diabetic cognitive impairment (DCI) a combination of thirst, dementia, and amnesia, a disease affecting the spleen, stomach, liver, and kidneys. Over time, the condition leads to loss of vital essence and qi, resulting in qi and yin deficiency, qi stagnation, blood stasis, and internalized toxicity, damaging brain vessels and causing encephalopathy.

[0003] Yiguanjian, originating from the book "Continuation of Famous Doctors' Cases" by the renowned Qing Dynasty physician Wei Zhiran, consists of raw Rehmannia root, Adenophora Root, Angelica Sinensis, Lycium barbarum, Ophiopogon japonicus, and Toosendan fruit. It is one of the first classic ancient prescriptions published, primarily nourishing yin and soothing the liver, and is primarily used to treat liver and kidney yin deficiency and liver qi disharmony. It is now commonly used clinically for yin deficiency and qi stagnation, including chronic gastritis, gastric and duodenal ulcers, neurosis, chronic hepatitis, and diabetes. Numerous studies have reported that Yiguanjian has significant therapeutic effects on diabetes and cognitive impairment. Wang Xiaomin et al. found that Yiguanjian effectively lowered blood glucose levels in rats with type 2 diabetes (T2DM) by reducing interleukin-6 levels. Mei Haiyun's research found that modified Yiguanjian was significantly effective in treating anxiety and depression associated with type 2 diabetes. Rao Mingyue's research suggests that modified Yiguanjian can effectively improve depression and TCM symptoms in patients with Parkinson's disease (PD) due to yin deficiency and liver stagnation. Gao Lili et al. reported significant efficacy in treating senile Parkinson's disease due to liver and kidney yin deficiency. Li Ruili uses Yi Guan Jian with added ingredients to treat various symptoms before and after menopause of liver and kidney yin deficiency, which can significantly relieve anxiety and other emotions.

[0004] In recent years, a growing number of studies have demonstrated that the development and progression of cognitive impairment are closely related to the gut microbiota, bile acids, and hepatic metabolites, such as amino acid metabolites (ammonia, indoles, and oxidols), and endotoxins. For example, when liver metabolic function is impaired, nitrogenous toxins derived from the gut cannot be broken down by the liver, resulting in large amounts of ammonia entering the brain. High concentrations of ammonia in the brain can cause brain cell inflammation, elevated brain glutamine, and astrocyte swelling, leading to brain dysfunction. The mechanism by which gut microbiota imbalance leads to cognitive impairment is primarily due to a high-fat diet that increases the intestinal LPS-to-microbiota ratio and LPS levels in T2DM animals, resulting in endotoxemia, which leads to insulin resistance, chronic low-level inflammation, and oxidative stress. Furthermore, LPS directly maintains insulin resistance and inflammation through the TLR signaling pathway. In particular, TLR4 acts as a receptor for the leukocyte differentiation antigen CD14, mediating the LPS-induced inflammatory cascade and innate immune response.

[0005] Currently, there is no literature report on the use of Yi Guan Jian in the preparation of drugs for treating diabetic cognitive impairment. Summary of the Invention

[0006] The technical solution of the present invention is to provide a new use of Yi Guan Jian, specifically, its use in preparing a medicine for treating diabetic cognitive impairment.

[0007] The present invention provides use of Yi Guan Jian in preparing a medicine for treating diabetic cognitive impairment.

[0008] Wherein, the medicine is a blood sugar lowering medicine.

[0009] Wherein, the drug is a drug for lowering glycated hemoglobin.

[0010] Wherein, the drug is a drug for lowering insulin levels.

[0011] Wherein, the medicine is a medicine that improves learning and memory.

[0012] Wherein, the drug is a drug that reduces neuronal damage.

[0013] Wherein, the drug is a drug for regulating lipid metabolism.

[0014] Wherein, the drug is a drug with specific anti-inflammatory effect.

[0015] Wherein, the drug is a drug that improves bile acid metabolism.

[0016] Based on the proven clinical efficacy of Yi Guan Jian and a large number of preliminary experimental studies, and in view of the key role of bile acid and intestinal flora metabolic disorders in the pathological process of diabetes and cognitive impairment, the present invention proposes for the first time that Yi Guan Jian may mediate the homeostasis of bile acids and intestinal microbiota through the "intestinal flora-bile acid-brain" pathway to improve diabetic cognitive impairment. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 Effects of Yi Guan Jian on blood lipids

[0018] Figure 2 HE staining results

[0019] Figure 3 Nissl staining test results

[0020] Figure 4 Glycated hemoglobin

[0021] Figure 5 Insulin levels

[0022] Figure 6 Effects of IL-1b inflammatory factor

[0023] Figure 7 Effects of IL-6 inflammatory factors

[0024] Figure 8 Effects of IL-18 inflammatory factors

[0025] Figure 9 Effects of TNF-α inflammatory factor

[0026] Figure 10 Serum bile acid content

[0027] Figure 11 Expression trends of each group

[0028] Figure 12 Cluster analysis results DETAILED DESCRIPTION

[0029] This study used a high-fat, high-sugar diet combined with STZ induction and Morris water maze testing to establish a DCI rat model. After Yi Guan Jian intervention, transmission electron microscopy was used to observe the microstructure of the hippocampal CA1 region, which can accurately evaluate the cognitive impairment of rats. The intestinal microecology and bile acid profile were studied and analyzed using intestinal flora 16S fragment sequencing and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS / MS). Gene expression microarrays were used to analyze the expression of bile acid receptors and lipid metabolism-related proteins to accurately identify subsequent detection indicators. Mechanism research, through real-time fluorescence quantitative PCR and western blot analysis, can effectively explore the potential targets and mechanisms of action of Yi Guan Jian in the treatment of diabetic cognitive dysfunction.

[0030] Experimental Example 1: Experiment on the Effect of Yiguanjian on Diabetic Cognitive Dysfunction Model Rats

[0031] 1 Construction of a diabetic rat model of cognitive impairment

[0032] After one week of adaptive feeding, SD rats were screened using a Y-maze test to identify rats with good memory function. Fasting blood glucose was measured by blood sampling from the tail vein. Rats with blood glucose levels between 3.0 and 5.0 mmol / L and normal cognitive function were included in subsequent experiments. The model group rats were fed a high-fat, high-sugar diet, while the blank group rats were fed a normal diet. Four weeks later, the model group rats were intraperitoneally injected with 30 mg kg⁻¹ of streptozotocin, while the blank group rats were intraperitoneally injected with an equal volume of citrate buffer. Seventy-two hours later, random blood glucose was measured by blood sampling from the tail vein. Rats with random blood glucose levels >16.7 mmol·L⁻¹ were considered to have successfully established a diabetic model. Random blood glucose was monitored every seven days thereafter, and rats with blood glucose levels below 16.7 mmol·L⁻¹ were excluded. Four weeks later, rats with cognitive impairment were screened using the Morris water maze.

[0033] The Morris water maze consists of a cylindrical pool, a removable cylindrical platform, an animal behavior trajectory analysis system, and a video camera system. The pool is 50 cm high and 120 cm in diameter, while the platform is 25 cm high and 8 cm in diameter. A digital camera is connected to a computer via a circular frame above the pool. Four equidistant points on the pool wall, designated N (north), E (east), S (south), and W (west), divide the pool into quadrants I, II, III, and IV. The platform is placed in quadrant III. The pool is filled with clean water to a depth of approximately 25 cm, with the water surface approximately 0.5 cm above the platform. The water temperature is maintained at (23 ± 0.5)°C using a hot water rod. A point is marked on the wall of each of the four fixed quadrants as the animal's entry point for each experiment. The platform is placed in the middle of quadrant III, opposite the entry point, and its position remains unchanged throughout the experiment.

[0034] Each water maze test was conducted over six days. Each trial was limited to 60 seconds. The time required for the animal to reach the platform from the water entry point and the swimming trajectory during this time were recorded. The time required to reach the platform was used as a measure of the mouse's learning and memory function. If the platform was not found within the set time, the computer stopped tracking and the time was recorded as 60 seconds. The first five days were used for the hidden platform test: the mouse was placed into the water three times with its head facing the wall of the tub (ensuring that the mouse could not directly see the platform). The time was set to 60 seconds. If the animal found the platform within 60 seconds, its latency was recorded, and the animal was then retrieved and placed on the platform to rest for 30 seconds. If the animal still did not find the platform within 60 seconds, it was also retrieved, placed on the platform to rest for 30 seconds, and its latency was recorded as 60 seconds. The purpose of the first five days of the experiment was to familiarize the animal with the environment and the entire learning process. After each round of testing, the animal's fur was dried with a towel and returned to its home box to rest. Each day, the animal was allowed to swim in different quadrants for a total of four times. On day 6, a spatial exploration experiment was conducted, the test results of which served as a criterion for assessing the animals' learning and memory abilities. The water maze was set up as before, but the platform was removed, and the mice entered the water from the quadrant opposite the original platform. Each animal underwent only one spatial exploration experiment. The number of times the mice crossed the platform area within 60 seconds and their trajectories were recorded. The swimming distance and percentage of time spent in the target quadrant and the entry quadrant were analyzed. Data acquisition and image analysis were performed automatically by an automatic image monitoring and data processing system. Shorter latency periods and more platform quadrant crossings indicate stronger learning and memory abilities. Each Morris water maze experiment was conducted in a soundproof room. The positions of laboratory items, including the light source, water tank, computer, camera, and vivarium, remained fixed.

[0035] 2. Effect of Yiguanjian on glucose metabolism in diabetic rats with cognitive impairment

[0036] 2.1 Blood lipids

[0037] Low-dose and high-dose Yiguanjian significantly reduced triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) levels (P < 0.01), and high-dose Yiguanjian significantly increased high-density lipoprotein cholesterol (HDL-C) levels (P < 0.01). Figure 1 ).

[0038] 2.2 HE staining results

[0039] Blank: The pia mater of the brain tissue was intact, consisting of a thin layer of connective tissue with abundant blood vessels, and no obvious inflammatory exudate was observed; the cell layers in the cortical area were arranged relatively neatly, and the neuronal cell morphology was normal; the pyramidal cell layer in the hippocampus was arranged neatly, with densely packed cells. In one case (1 / 3), there was an increase in dark neurons, with smaller cell size, darker color, blurred internal cell structure, and obvious axon-like structures at the cell tail; no obvious glial cell proliferation or inflammatory cell infiltration was observed.

[0040] Model: The pia mater of the brain tissue is intact, consisting of a thin layer of connective tissue with abundant blood vessels, and no obvious inflammatory exudate is observed. The cell layers in the cortical area are arranged relatively neatly, and the neuronal cell morphology is normal. The pyramidal cell layer in the hippocampus is neatly arranged, with densely packed cells and occasional dark neurons. No obvious glial cell proliferation or inflammatory cell infiltration is observed.

[0041] Low dose: The pia mater of the brain tissue is intact, consisting of a thin layer of connective tissue with abundant blood vessels, and no obvious inflammatory exudate is observed; the cell layers in the cortical area are arranged relatively neatly, and the neuronal cell morphology is normal; the pyramidal cell layer in the hippocampus is arranged neatly, with occasional small amounts of dark neurons; no obvious glial cell proliferation or inflammatory cell infiltration is observed.

[0042] High dose: The pia mater of the brain tissue was intact, consisting of a thin layer of connective tissue with abundant blood vessels, and no obvious inflammatory exudate was observed; the cell layers in the cortical area were arranged relatively neatly, and the neuronal cell morphology was normal; the pyramidal cell layer in the hippocampus was arranged neatly, with densely packed cells, and a very small number of dark neurons increased in one case (1 / 4); no obvious glial cell proliferation or inflammatory cell infiltration was observed.

[0043] Positive: The pia mater of the brain tissue is intact, consisting of a thin layer of connective tissue with abundant blood vessels, and no obvious inflammatory exudate is observed; the cell layers in the cortical area are arranged in a relatively neat manner, and the neuronal cell morphology is normal; the pyramidal cell layer in the hippocampus is arranged in a neat manner, and the cells are densely arranged; no obvious glial cell proliferation and inflammatory cell infiltration are observed (see Figure 2 ).

[0044] 2.3 Nissl staining test results

[0045] The Nissl bodies of neurons in the blank group were normally distributed in the dendrites and cell bodies of neurons; compared with the control group, the Nissl bodies of neurons in the model group were significantly reduced or even disappeared, and the cells showed obvious damage; compared with the model group, the Nissl bodies of neurons in the Yiguanjian group and the positive drug group were restored, and the neuronal damage was alleviated (see Figure 3 ).

[0046] 2.4 Glycated hemoglobin

[0047] After the diabetic cognitive dysfunction model was successfully established, the drug was administered orally for 8 weeks. The results showed that the low-dose and high-dose groups could significantly reduce the glycated hemoglobin (HbA1c) of the model, with statistical significance (P < 0.01) (see Figure 4 ).

[0048] 2.5 Insulin Levels

[0049] After the diabetic cognitive dysfunction model was successfully established, the drug was administered orally for 8 weeks. The results showed that the low-dose and high-dose groups could significantly reduce the insulin level of the model, with statistical significance (P < 0.01) (see Figure 5 ).

[0050] 2.6 Inflammatory Factors

[0051] 2.6.1IL-1b

[0052] Interleukin 1b (IL-1b) is an inflammatory cytokine located upstream in the cytokine cascade reaction. It is widely believed to be necessary for the effective initiation of innate immunity and the formation of adaptive immune responses to resolve inflammation. It is an amplifier of the immune response. At the same time, it is also the central mediator of cell-to-cell interactions in the inflammatory microenvironment. After the successful modeling, the IL-1b levels in the hippocampus and serum of the model group rats were significantly higher than those in the blank group (P < 0.01). After the intervention of Yi Guan Jian, the levels in the low-dose group and the high-dose group were significantly lower than those in the model group. This shows that Yi Guan Jian can significantly regulate the IL-1b levels in the hippocampus and serum of rats, and play a certain anti-inflammatory role (see Figure 6 ).

[0053] 2.6.2IL-6

[0054] Interleukin 6 (IL-6) is an important member of the cytokine network. It plays a central role in the inflammatory response and can stimulate the production of C-reactive protein and fibrinogen. When infection and inflammation occur, IL-6 is produced first and its level rises rapidly, reaching a peak in 2 hours. The level of increase is consistent with the severity of inflammation. After successful modeling, the IL-6 levels in the hippocampus and serum of the model group rats were significantly higher than those in the blank group (P < 0.01). After the intervention of Yi Guan Jian, the levels in the low-dose group and the high-dose group were significantly lower than those in the model group. This shows that Yi Guan Jian can significantly regulate the IL-6 levels in the hippocampus and serum of rats and play a certain anti-inflammatory role (see Figure 7 ).

[0055] 2.6.3IL-18

[0056] Interleukin 18 (IL-18) is a potent pro-inflammatory cytokine that can induce IFNγ, GM-CSF, TNFα and IL-1 in immune-active cells, activate lymphocyte killing, and upregulate the expression of certain chemokine receptors. IL-18 is essential for the host's defense against severe infections. After successful modeling, the IL-18 levels in the hippocampus and serum of the model group rats were significantly higher than those in the blank group (P < 0.05). After the intervention of Yi Guan Jian, the IL-18 levels in the hippocampus of the high-dose group were significantly lower than those in the model group (P < 0.01). This indicates that high-dose Yi Guan Jian can significantly regulate the IL-18 in the rat hippocampus and play a certain anti-inflammatory role (see Figure 8 ).

[0057] 2.6.4TNF-α

[0058] Tumor necrosis factor α (TNF-α), as one of the mediators of neuroinflammation, can stimulate brain pericytes to secrete matrix metalloproteinase-9 (MMP-9), leading to basement membrane rupture, pericyte migration, and blood-brain barrier damage. After successful modeling, the TNF-α levels in the hippocampus and serum of the model group rats were significantly higher than those in the blank group (P < 0.01). After the intervention of Yi Guan Jian, the TNF-α levels in the hippocampus and serum of the low-dose and high-dose groups were significantly lower than those in the model group (P < 0.05). This indicates that high-dose Yi Guan Jian can significantly regulate IL-18 in the rat hippocampus and play a certain anti-inflammatory role (see Figure 9 ).

[0059] 2.7 Bile acid metabolism level

[0060] 2.7.1 Serum bile acid content

[0061] Bile acids are mainly converted from cholesterol in the liver, and are modified by intestinal flora to produce secondary bile acids, which return to the blood through active transport or passive diffusion, and flow throughout the body through blood circulation and body fluid circulation. Bile acids play a major regulatory and signal transduction function in the gut-liver-brain axis, affecting and regulating signal transduction in the brain. This may suggest the association between changes in bile acid and changes in cognitive ability. After successful modeling, the serum bile acid levels of rats in the model group were significantly higher than those in the blank group (P < 0.05). After intervention with Yi Guan Jian, the serum bile acid levels in the low-dose and high-dose groups were significantly lower than those in the model group (P < 0.05). This indicates that both low-dose and high-dose Yi Guan Jian can significantly reduce the serum bile acid levels of the model and improve cognitive levels (see Figure 10 ).

[0062] 2.7.2 Fecal bile acid metabolism

[0063] 2.7.2.1 Expression trend of each group

[0064] UPLC-MS / MS multicomponent content determination was used to quantitatively analyze 33 bile acid components. As shown in the stacked bar chart above, compared with the blank group, the model group had a higher proportion of four metabolites: HDCA, DCA, r-MCA, and α-MCA. Compared with the blank group, the high-dose group had two major metabolites: DCA and HDCA. Compared with the blank group, the low-dose group had DCA and HDCA. Compared with the model group, the low-dose group had HDCA, DCA, and LCA. Compared with the model group, the high-dose group had LCA and DCA. Compared with the high-dose group, the low-dose group had DCA and HDCA. This indicates that HDCA and DCA are the main metabolites in fecal bile acid metabolites in each group.

[0065] Further analysis of expression differences was performed. As shown in the box plot, the r-MCA and a-MCA components in the model group were significantly different from those in the control group (P < 0.05). The r-MCA content in the low- and high-dose groups of Yi Guan Jian was significantly different from that in the model group. This suggests that Yi Guan Jian may improve bile acid metabolism by regulating r-MCA (see Figure 11 ).

[0066] 2.7.2.2 Cluster analysis results

[0067] The expression levels of all 33 bile acid metabolites were calculated using a distance matrix, and cluster analysis was performed using hierarchical clustering to more comprehensively and intuitively display the differences in the expression patterns of metabolites in different samples. The cluster analysis of the blank group, model group, high-dose group, and low-dose group is shown in the figure. The redder the color, the higher the expression level of the metabolite, and the bluer the color, the lower the expression level of the metabolite (see Figure 12 ).

Claims

1. Use of Yi Guan Jian in preparing medicine for treating diabetic cognitive impairment.

2. The use according to claim 1, characterized in that: The medicine is a blood sugar lowering medicine.

3. The use according to claim 1, characterized in that: The drug is a drug for lowering glycated hemoglobin.

4. The use according to claim 1, characterized in that: The drug is a drug that lowers insulin levels.

5. The use according to claim 1, characterized in that: The medicine is a medicine that improves learning and memory.

6. The use according to claim 1, characterized in that: The medicine is a medicine for alleviating neuronal damage.

7. The use according to claim 1, characterized in that: The medicine is a medicine for regulating lipid metabolism.

8. The use according to claim 1, characterized in that: The drug is a drug with specific anti-inflammatory effect.

9. The use according to claim 1, characterized in that: The medicine is a medicine for improving bile acid metabolism.

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