Application of N-acetylcysteine in medicine for improving diabetic hyperthyroidism complicated disease intestinal injury
By administering N-acetylcysteine to diabetic hyperthyroidism comorbidity mice, intestinal damage was improved, the problems of intestinal anti-inflammatory, antioxidant and barrier function in diabetic hyperthyroidism comorbidity were resolved, and the health status and survival rate of mice were improved.
Patent Information
- Application Number
- CN202511033586.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-25
- Publication Date
- 2025-10-31
AI Technical Summary
Current technologies lack effective measures to improve intestinal damage in patients with diabetes mellitus and hyperthyroidism complications, especially the problems of impaired intestinal anti-inflammatory, antioxidant, and barrier functions.
N-acetylcysteine (NAC) was used as a drug and administered orally to diabetic mice with hyperthyroidism to establish a specific model and expose them to a diet containing NAC, thereby improving intestinal damage.
It significantly improved intestinal anti-inflammatory, antioxidant, and intestinal barrier function in a mouse model of diabetic hyperthyroidism complications, reduced blood glucose levels, controlled excessive food and water intake, and improved the health status and survival rate of mice.
Smart Images
Figure CN120860004A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of biomedical technology, and in particular to the application of N-acetylcysteine in drugs for improving intestinal damage in diabetic hyperthyroidism. Background Technology
[0002] For a long time, clinical practice has focused primarily on glycemic control in patients with diabetes mellitus and hyperthyroidism, as well as the treatment of other emerging complications. However, a problem that is easily overlooked in this process is the intestinal issue in patients with diabetes mellitus and hyperthyroidism. Because intestinal problems are not easily detected or exposed, related reports are very few. Furthermore, because the causes and specific molecular regulatory mechanisms of intestinal damage caused by diabetes mellitus and hyperthyroidism remain unclear, there is a lack of effective measures to improve intestinal damage in these patients.
[0003] The applicant's research found that a diabetes + hyperthyroidism comorbidity model exhibited increased water intake, frequent urination, elevated blood glucose, and weight loss. Furthermore, diabetes + hyperthyroidism caused hyperglycemia, systemic inflammatory response, and decreased serum glutathione levels. Further testing revealed that diabetes + hyperthyroidism caused insufficient expression of the intestinal tight junction proteins Claudin, ZO-1, and Muc2, resulting in intestinal barrier damage. Moreover, decreased glutathione (GSH) levels in intestinal chyme were detected, which further weakens the intestinal antioxidant and anti-inflammatory levels, ultimately leading to intestinal damage. Therefore, glutathione deficiency is a key signal of intestinal damage in diabetes + hyperthyroidism comorbidity.
[0004] It is well known that when the body's glutathione levels are insufficient, the inflammatory response increases. Direct glutathione supplementation has proven to be a difficult measure to achieve the desired effect, as the bioavailability of oral glutathione supplements is poor, thus it is impossible to increase blood glutathione levels simply by swallowing pure glutathione. Over the years, manufacturers have developed specialized glutathione formulations and compounds to improve the oral bioavailability of antioxidants. Some of these glutathione formulations may be effective for most people, but high cost is a major issue. Low bioavailability of glutathione is a common problem in supplement science. Supplementing with glutathione precursors is an effective way to address this problem. Studies have consistently shown that supplementing with glutathione precursors (such as cysteine and glycine) can significantly help restore glutathione levels. If the goal is to increase glutathione levels, then taking N-acetylcysteine (NAC) is one feasible approach. Related research reports that NAC can reduce oxidative stress, thereby preventing some of the damage caused by alcohol. NAC is commonly used as a medicine to treat colds in children, so there is a safety profile for its use. However, there is no research or data available on the use of NAC to alleviate intestinal damage in patients with diabetes and hyperthyroidism complications. Summary of the Invention
[0005] The purpose of this invention is to propose the application of N-acetylcysteine in drugs for improving intestinal damage in a mouse model of diabetic hyperthyroidism, effectively alleviating abnormally high blood glucose levels in diabetic hyperthyroidism complications, improving excessive food and water intake, and alleviating intestinal damage.
[0006] To achieve the above objectives, the present invention provides the application of N-acetylcysteine in drugs for improving intestinal damage in diabetic hyperthyroidism complications. N-acetylcysteine is used in the preparation of drugs for improving intestinal damage in mouse models of diabetic hyperthyroidism complications.
[0007] Preferably, intestinal damage includes intestinal anti-inflammatory, antioxidant, and intestinal barrier functions.
[0008] Preferably, N-acetylcysteine is administered orally.
[0009] Preferably, the construction of a mouse model of diabetic hyperthyroidism includes the following steps:
[0010] S1. Newborn mice were injected with streptozotocin at 3, 5 and 8 days of age, and blood glucose was monitored at 2, 3 and 4 weeks to determine whether the diabetes model was successfully established.
[0011] S2. Starting from week 2, L-thyroxine was injected daily to construct a diabetic hyperthyroidism disease model.
[0012] S3. After weaning in the third week, the pups were introduced to a diet containing N-acetylcysteine until the end of the experiment in the eighth week.
[0013] Preferably, the dosage of streptozotocin in S1 is 40-200 mg / kg body weight.
[0014] Preferably, the dosage of L-thyroxine in S2 is 0.3 mg / kg body weight.
[0015] Preferably, the S3 diet contains 0.1 g / kg of N-acetylcysteine.
[0016] The disease model mice fed NAC showed increased body weight, controlled food and water intake, and improved blood glucose levels in diabetic and hyperthyroid complications.
[0017] The beneficial effects of this invention are:
[0018] (1) This invention reveals for the first time the application of N-acetylcysteine in improving intestinal damage in a mouse model of diabetic hyperthyroidism, providing a new direction for the development of related therapeutic drugs.
[0019] (2) The present invention uses a specific method for constructing a mouse model of diabetic hyperthyroidism, which can realistically simulate the pathological state of clinical patients and is beneficial for studying the effect of drugs on improving intestinal damage.
[0020] (3) The N-acetylcysteine of the present invention has shown good effects in improving intestinal anti-inflammatory, antioxidant and intestinal barrier function damage, and is expected to bring an effective treatment to patients with diabetes and hyperthyroidism complicated with intestinal damage.
[0021] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description
[0022] Figure 1 This is a schematic diagram showing the body weight, food intake, and water consumption of the three mouse models in Example 1 of this invention. Figure 1 In the diagram, A represents weight. Figure 1 B in the diagram represents a feeding illustration. Figure 1 C in the diagram represents a drinking water intake;
[0023] Figure 2 This is a schematic diagram showing the blood glucose levels and survival rates of the three mouse models in Example 1 of this invention. Figure 2 In the diagram, A represents the blood glucose level. Figure 2 B in the diagram represents the survival rate of mice.
[0024] Figure 3 This is a schematic diagram of the intestinal morphology of the three mouse models in Example 1 of the present invention; Figure 3 In the diagram, A represents the HE staining image of the control group. Figure 3 B in the image represents the HE staining pattern of the diabetes + hyperthyroidism group. Figure 3 C in the image represents the HE staining pattern of the diabetes + hyperthyroidism + NAC group. Figure 3 In the diagram, D represents the villus height of the three mouse models. Figure 3 E in the diagram represents the crypt depth of the three mouse models. Figure 3 F in the figure is a schematic diagram of the villus / crypt ratio in the three mouse models;
[0025] Figure 4 This is a schematic diagram showing the serum interleukin levels in three groups of mouse models in Example 1 of the present invention. Figure 4 In the diagram, A represents the serum interleukin-1 level. Figure 4 B in the diagram represents the serum interleukin-6 content.
[0026] Figure 5 This is a schematic diagram showing the serum anti-inflammatory and antioxidant levels of three groups of mouse models in Example 1 of the present invention. Figure 5 In the diagram, A represents the total antioxidant capacity of serum. Figure 5In the diagram, B represents the serum malondialdehyde (MDA) content. Figure 5 The diagram shows that C represents serum glutathione levels. Figure 5 The diagram shows that D represents the serum glutathione peroxidase content.
[0027] Figure 6 This is a schematic diagram illustrating the intestinal anti-inflammatory and antioxidant levels of three mouse models in Example 1 of this invention. Figure 6 In the diagram, A represents the malondialdehyde (MDA) content in the intestine. Figure 6 B in the diagram represents the total antioxidant capacity of the gut. Figure 6 The 'C' in the diagram represents the intestinal glutathione peroxidase content. Figure 6 The diagram shows the content of glutathione in the intestine (D).
[0028] Figure 7 This is a schematic diagram of the intestinal barrier function in three mouse models in Example 1 of the present invention. Figure 7 The diagram shows the expression level of the tight junction protein Claudin. Figure 7 The diagram in B represents the expression level of the tight junction protein Zo-1. Figure 7 The diagram in C represents the expression level of intestinal mucin Muc2.
[0029] Figure 8 This is a schematic diagram illustrating the abundance analysis of bacterial communities at the phylum, genus, and species levels in three mouse models in Example 1 of the present invention. Figure 8 In the diagram, A represents a schematic representation of phylum-level bacterial abundance analysis. Figure 8 B in the diagram is a schematic representation of genus-level bacterial abundance analysis. Figure 8 C in the diagram is a schematic diagram of species-level bacterial community abundance analysis;
[0030] Figure 9 This is a schematic diagram of the abundance analysis of key bacteria at the genus level in three mouse models in Example 1 of the present invention. Figure 9 In the diagram, A represents a schematic representation of the abundance analysis of anaerobic mycoplasma. Figure 9 B in the diagram is a schematic representation of the abundance analysis of Bacteroides. Figure 9 C in the diagram represents the abundance analysis of the genus *Myxospirillum*. Figure 9 The 'D' in the diagram represents the abundance analysis of the Helicobacter genus. Figure 9 E in the diagram is a schematic diagram of the abundance analysis of Larstone. Detailed Implementation
[0031] The present invention will be further described below with reference to the accompanying drawings and embodiments. Unless otherwise defined, the technical or scientific terms used in this invention should be understood in their ordinary sense by those skilled in the art. The features mentioned above or in the specific examples mentioned in this invention can be combined arbitrarily, and these specific embodiments are only used to illustrate the invention and are not intended to limit the scope of the invention.
[0032] The mice used in this invention are ICR mice. All experimental mice were purchased from Beijing Spaford Company. The experimental subjects were newborn mice. After birth, the experimental mice were kept in a light-controlled environment (12 hours of light / 12 hours of darkness) without any special experimental requirements. All mice had free access to food and water, and the breeding environment was controlled at 20-24℃.
[0033] This invention provides the application of N-acetylcysteine in improving intestinal damage in a mouse model of diabetic hyperthyroidism. The method involves first establishing a mouse model of diabetic hyperthyroidism and administering NAC, including the following steps:
[0034] S1. Newborn mice were injected with streptozotocin (40-200 mg / kg body weight) at 3, 5 and 8 days of age, and blood glucose was monitored at 2, 3 and 4 weeks to determine whether the diabetes model was successfully established.
[0035] S2. Starting from week 2, L-thyroxine (0.3 mg / kg body weight) was injected daily to establish a diabetic hyperthyroidism disease model;
[0036] S3. After weaning in the third week, the pups were introduced to a diet containing N-acetylcysteine (0.1 g / kg) until the end of the experiment in the eighth week.
[0037] This invention resulted in increased body weight, controlled food and water intake in disease model mice fed with NAC, improved blood glucose levels in diabetic hyperthyroidism complications, and significantly improved intestinal damage in diabetic hyperthyroidism comorbidity model mice, including intestinal anti-inflammatory, antioxidant, and intestinal barrier function impairment.
[0038] Example 1
[0039] This invention provides the application of N-acetylcysteine in the preparation of drugs to improve intestinal damage in a mouse model of diabetic hyperthyroidism:
[0040] Constructing the NAC group (diabetes + hyperthyroidism + NAC) includes the following steps:
[0041] S1. Newborn mice were injected with streptozotocin (40-200 mg / kg body weight) at 3, 5 and 8 days of age, and blood glucose was monitored at 2, 3 and 4 weeks to determine whether the diabetes model was successfully established.
[0042] S2. Starting from week 2, L-thyroxine (0.3 mg / kg body weight) was injected daily to establish a diabetic hyperthyroidism disease model;
[0043] S3. After weaning in the third week, the pups were introduced to a diet containing N-acetylcysteine (0.1 g / kg) until the end of the experiment in the eighth week.
[0044] Construct a control group
[0045] The newborn mice received no treatment and were fed a regular diet. No drugs were administered during the experiment.
[0046] Constructing a diabetes mellitus-hyperthyroidism complication group (diabetes + hyperthyroidism) includes the following steps:
[0047] S1. Construct a mouse model of diabetic hyperthyroidism. Newborn mice were injected with streptozotocin (40-200 mg / kg body weight) on days 3, 5, and 8 of age, and blood glucose was monitored at weeks 2, 3, and 4 to determine whether the diabetic model was successfully constructed.
[0048] S2. Starting from week 2, mice were injected daily with L-thyroxine (0.3 mg / kg body weight) until the end of the experiment to establish a diabetic hyperthyroidism model; during this period, the mice were fed a regular diet.
[0049] Indicator detection and result analysis:
[0050] Weight, feed intake, and water intake statistics
[0051] Thirty-two newborn mice were used in each group, with four mice per cage. Eight parallel cage replicates were performed. The weight of each mouse, the weekly food intake per cage (four mice), and the daily water intake per cage (four mice) were recorded weekly. The feeding situation of the mice during the experiment was recorded and analyzed.
[0052] Figure 1 This is a schematic diagram showing the body weight, food intake, and water consumption of the three groups of mouse models in Example 1 of this invention, as shown below. Figure 1 As shown, compared with the control group, the mice in the diabetes + hyperthyroidism group experienced a decrease in body weight, while the mice in the diabetes + hyperthyroidism + NAC group showed a recovery in body weight after consuming an NAC diet. Figure 1 As shown in B and C, the food intake and water consumption of mice in the diabetes + hyperthyroidism group were significantly higher than those in the control group, while the symptoms of excessive food intake and water consumption in the NAC group were alleviated, and the water intake and food consumption were closer to those of normal mice. The blood glucose level was closer to that of the control group than that of the diabetes + hyperthyroidism group.
[0053] Blood glucose testing
[0054] Eight mice were randomly selected each week from the control group, the diabetes + hyperthyroidism group, and the diabetes + hyperthyroidism + NAC group to test blood glucose levels. At the end of the 8th week, the mice were euthanized by cervical dislocation in accordance with animal ethics requirements, and blood samples were collected for serum biological analysis.
[0055] Figure 2 This is a schematic diagram showing the blood glucose levels and survival rates of the three mouse models in Example 1 of this invention, as shown below. Figure 2 As shown, the blood glucose level of mice in the diabetes + hyperthyroidism group increased. The blood glucose level of mice in the diabetes + hyperthyroidism + NAC group was controlled after feeding NAC diet, and was lower than that of mice in the diabetes + hyperthyroidism group at the same time point. Furthermore, the health status of mice fed NAC diet was significantly improved and the mortality rate decreased.
[0056] HE staining
[0057] After the mice were sacrificed, intestinal tissue was collected and fixed with paraformaldehyde for 48 hours. The intestines were then dehydrated, embedded, paraffin-embedded, and stained with hematoxylin and eosin (HE) according to the established procedure. Figure 3 This is a schematic diagram of the intestinal morphology of three mouse models in Example 1 of the present invention, as shown below. Figure 3 As shown, the intestinal villus height and villus-to-crypt depth were significantly decreased in the diabetes + hyperthyroidism group, while the intestinal villus height and villus-to-crypt depth increased after the diabetes + hyperthyroidism + NAC group was fed an NAC diet. Therefore, NAC can improve the intestinal morphological and pathological damage induced by the diabetes + hyperthyroidism model.
[0058] Biochemical indicator testing
[0059] Serum, intestinal tissue, and chyme samples were prepared and tested according to the instructions of biochemical indicator kits such as GSH. Figure 4 This is a schematic diagram showing the serum interleukin levels in three mouse models in Example 1 of the present invention, as shown below. Figure 4 As shown, compared with the control group, the interleukin-1 and interleukin-6 levels in the diabetes + hyperthyroidism group were significantly increased, while the serum interleukin-1 and interleukin-6 levels in the diabetes + hyperthyroidism + NAC group were significantly decreased after consuming NAC diet.
[0060] Total serum antioxidant capacity measures the total antioxidant capacity in mouse serum. Malondialdehyde (MDA) is a product of lipid peroxidation, and its content reflects the degree of oxidative stress. Glutathione is an important antioxidant, and glutathione peroxidase is an important component of the body's antioxidant enzyme system. Figure 5 This is a schematic diagram showing the serum anti-inflammatory and antioxidant levels of three mouse models in Example 1 of the present invention, as follows: Figure 5As shown, the control group had relatively high serum total antioxidant capacity, low malondialdehyde (MDA) content, and high levels of glutathione and glutathione peroxidase. In contrast, the diabetes + hyperthyroidism group had significantly lower total antioxidant capacity than the control group, significantly higher MDA content, and lower levels of glutathione and glutathione peroxidase. The diabetes + hyperthyroidism + NAC group showed some recovery in total antioxidant capacity after consuming NAC diet, with a decrease in MDA and a recovery in glutathione and glutathione peroxidase levels.
[0061] Figure 6 This is a schematic diagram illustrating the intestinal anti-inflammatory and antioxidant levels of three mouse models in Example 1 of the present invention, as shown below. Figure 6 As shown, the glutathione content in the intestine of the diabetes + hyperthyroidism + NAC group was significantly higher than that of the diabetes + hyperthyroidism group. Compared with the diabetes + hyperthyroidism group, the glutathione peroxidase and antioxidant levels in the intestine of the diabetes + hyperthyroidism + NAC group were significantly higher, while the malondialdehyde content in the intestine was significantly lower.
[0062] This indicates that NAC treatment significantly improves the total antioxidant capacity of serum and intestine, increases the levels of glutathione and glutathione peroxidase in serum and intestine, and reduces the level of malondialdehyde in serum. It also has a positive effect on antioxidant-related indicators in mice with diabetes and hyperthyroidism.
[0063] Gene and protein expression detection
[0064] RNA and protein extraction reagents were used to extract RNA and protein from intestinal tissues, and relevant tests were performed. To further confirm the beneficial role of NAC in intestinal barrier function, intestinal barrier-related tight junction proteins were detected at the gene and protein levels.
[0065] Figure 7 This is a schematic diagram of the intestinal barrier function of three mouse models in Example 1 of the present invention, as shown below. Figure 7 As shown, the relative expression of tight junction proteins Claudin, Zo-1, and intestinal mucin Muc2 in the diabetes and hyperthyroidism groups was lower than that in the control group. The expression levels in the diabetes + hyperthyroidism + NAC group were higher than those in the diabetes + hyperthyroidism group and also higher than those in the control group. This indicates that NAC can promote the expression of intestinal tight junction-related genes and proteins, such as Claudin, Zo-1, and Muc2, and improve intestinal function and morphology. NAC improves the intestinal barrier function by promoting the expression of intestinal tight junction proteins.
[0066] 16S sequencing
[0067] The detection was performed by Beijing Skybio Biotechnology Co., Ltd., which involved extracting total DNA from the sample and then selectively amplifying a portion of the bacterial 16S rRNA gene using PCR technology.
[0068] Figure 8This is a schematic diagram of the abundance analysis of bacterial communities at the phylum, genus, and species levels in three mouse models in Example 1 of the present invention, as shown below. Figure 8 As shown, the gut microbiota structure in the diabetes + hyperthyroidism group was disordered. After the diabetes + hyperthyroidism + NAC group was fed an NAC diet, the gut microbiota structure at the phylum, genus and species levels was similar to that of the control group, indicating that NAC can improve gut microbiota stability.
[0069] Figure 9 This is a schematic diagram of the abundance analysis of key bacteria at the genus level in three mouse models in Example 1 of the present invention, as shown below. Figure 9 As shown, the abundance of some microorganisms detrimental to gut health, such as anaerobic mycoplasma, *Helicobacter*, and *Helicobacter*, increased in the diabetes + hyperthyroidism group. However, in the diabetes + hyperthyroidism + NAC group, the abundance of these genera decreased after consuming an NAC diet, approaching that of the control group. Conversely, the abundance of beneficial bacteria, *Larstonia*, decreased in the diabetes + hyperthyroidism group, but increased in the diabetes + hyperthyroidism + NAC group after consuming an NAC diet. Analysis of gut microbiota stability, at the phylum, genus, and species levels, indicates that NAC can improve gut microbiota stability and the abundance of beneficial bacteria.
[0070] Therefore, the N-acetylcysteine of this invention alleviated the symptoms of excessive food and water intake in a diabetic hyperthyroidism comorbidity mouse model, improved blood glucose levels, increased anti-inflammatory and antioxidant levels in serum and intestines, enhanced intestinal barrier function, improved the health status of the diabetic hyperthyroidism comorbidity mouse model, and increased the survival rate of the mice. These findings have significant clinical value in the treatment of diabetic hyperthyroidism comorbidity.
[0071] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.
Claims
1. Application of N-acetylcysteine in the preparation of drugs to improve intestinal damage in diabetic hyperthyroidism complications.
2. The application of N-acetylcysteine according to claim 1 in drugs for improving intestinal damage in diabetic hyperthyroidism complications, characterized in that: Intestinal damage includes intestinal anti-inflammatory, antioxidant, and intestinal barrier functions.
3. The application of N-acetylcysteine according to claim 1 in drugs for improving intestinal damage in diabetic hyperthyroidism complications, characterized in that: N-acetylcysteine is administered orally.
4. The application of N-acetylcysteine according to claim 1 in drugs for improving intestinal damage in diabetic hyperthyroidism complications, characterized in that: The construction of a mouse model for diabetic hyperthyroidism complications includes the following steps: S1. Newborn mice were injected with streptozotocin at 3, 5 and 8 days of age, and blood glucose was monitored at 2, 3 and 4 weeks to determine whether the diabetes model was successfully established. S2. Starting from week 2, L-thyroxine was injected daily to construct a diabetic hyperthyroidism disease model. S3. After weaning in the third week, the pups were introduced to a diet containing N-acetylcysteine until the end of the experiment in the eighth week.
5. The application of N-acetylcysteine according to claim 4 in drugs for improving intestinal damage in diabetic hyperthyroidism complications, characterized in that: The dosage of streptozotocin in S1 treatment is 40-200 mg / kg body weight.
6. The application of N-acetylcysteine according to claim 4 in drugs for improving intestinal damage in diabetic hyperthyroidism complications, characterized in that: The dosage of L-thyroxine in S2 is 0.3 mg / kg body weight.
7. The application of N-acetylcysteine according to claim 4 in drugs for improving intestinal damage in diabetic hyperthyroidism complications, characterized in that: The S3 diet contains 0.1 g / kg of N-acetylcysteine.