Application of resolvin D1 in periodontitis systemic complications

Abstract

The invention provides application of resolvin D1 in periodontitis systemic complications, and belongs to the technical field of biological medicine. The new function of the RvD1 is found, that is, the RvD1 has the effects of relieving and / or treating periodontitis systemic complications and promoting systemic inflammation regression. As an endogenous molecule, the RvD1 has fewer side effects and can be better tolerated by a human body. The RvD1 can directly act on an inflammation regulation process, promote inflammation regression and regulate the balance of host immune response, is helpful for controlling the development of periodontitis and systemic complications and reducing repeated attack, and provides a safer, more comprehensive and more effective treatment scheme for periodontitis and systemic complications.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and particularly relates to the use of resolvin D1 in treating systemic complications of periodontitis. Background Art

[0002] Periodontitis is a chronic inflammation of the periodontal supporting tissues, initiated by plaque microorganisms and mediated by the host immune response. It is caused by multiple factors. The main clinical manifestations are gingival inflammation, periodontal pocket formation, attachment loss and alveolar bone resorption. Severe cases may lead to loose teeth and tooth loss. During the development and progression of periodontitis, plaque microorganisms and their products activate the host immune system, leading to local inflammatory reactions. As the inflammation intensifies, a large number of inflammatory cells infiltrate the periodontal tissues and release a large number of inflammatory mediators, leading to systemic complications. Although these immune responses have a killing and inhibitory effect on bacteria and their toxic products, they also cause tissue damage, leading to an imbalance between osteoclasts and osteoblasts, that is, bone resorption is greater than bone formation, and ultimately destroying the periodontal tissues. This process is closely related to the excessive inflammatory response caused by the host's pro-inflammatory / anti-inflammatory imbalance under plaque stimulation, accompanied by inflammation resolution disorders.

[0003] At present, the clinical treatment of periodontitis mainly adopts mechanical plaque removal (supragingival scaling, subgingival scaling and root planing) and drug therapy, and some patients need periodontal surgery. In terms of drug treatment, the main method is the use of antibiotics, which emphasizes further control of bacterial infection on the basis of mechanical treatment. However, the use of antibiotics has the problem of drug resistance, and antibiotics can only act on bacteria and cannot promote tissue repair. If the body's existing inflammation cannot be resolved, it will cause secondary inflammatory damage to its own tissues. At the same time, after the treatment of periodontitis, the systemic complications caused by periodontitis may not be able to be treated at the same time. Therefore, how to accurately, effectively and quickly eliminate inflammation and complications, restore the balance between pro-inflammatory and anti-inflammatory, and reduce connective tissue destruction and alveolar bone resorption are difficult and cutting-edge issues that need to be solved in clinical practice. Summary of the Invention

[0004] The present invention provides the use of resolvin D1 (RvD1) in treating systemic complications of periodontitis. RvD1 has the effect of alleviating and / or treating systemic complications of periodontitis by binding to the FPR2 receptor, providing a new target for the development of new drugs for treating periodontitis complications.

[0005] The present invention provides the use of resolvin D1 in preparing a medicine for treating and / or alleviating systemic complications of periodontitis.

[0006] In a preferred embodiment of the present invention, the systemic complications of periodontitis include complications of all organs and / or tissues in the body caused by periodontitis except periodontal tissues.

[0007] In a preferred embodiment of the present invention, the dosage form of the drug includes an oral preparation, an injection preparation, a gel preparation, a nano-encapsulated delivery preparation or a water-soluble gargle preparation.

[0008] In a preferred embodiment of the present invention, the working concentration of resolvin D1 in the drug is 1 ng / μL to 1 μg / μL based on mice.

[0009] The present invention also provides a drug for treating and / or alleviating systemic complications of periodontitis, which uses resolvin D1 as the sole active ingredient, and also includes pharmaceutically acceptable excipients, as well as chemical modifications and modifications related to resolvin D1.

[0010] In a preferred embodiment of the present invention, the dosage form of the drug includes any one of the following: oral preparation, gavage preparation, injection preparation, gel preparation, nano-encapsulated delivery preparation and water-soluble gargle preparation.

[0011] In a preferred embodiment of the present invention, the working concentration of resolvin D1 in the drug is 1 ng / μL to 1 μg / μL based on mice.

[0012] In a preferred embodiment of the present invention, when the drug is in liquid form, the solvent includes physiological saline containing ethanol.

[0013] The present invention also provides a drug for treating and / or alleviating systemic complications of periodontitis, which contains resolvin D1 as a partial active ingredient and also includes pharmaceutically acceptable excipients.

[0014] In a preferred embodiment of the present invention, the dosage form of the drug includes any one of the following: oral preparation, gavage preparation, injection preparation, gel preparation, nano-encapsulated delivery preparation and water-soluble gargle preparation.

[0015] Beneficial effects: The present invention discovered a new function of RvD1, namely, RvD1 has the effect of alleviating and / or treating systemic complications of periodontitis, promoting periodontal repair and the disappearance of systemic inflammation. Compared with traditional antibiotic drugs, RvD1, as an endogenous lipid medium, its use will not lead to antibiotic resistance. As an endogenous molecule, RvD1 has fewer side effects and is better tolerated by the human body. In addition, unlike antibiotics that simply eliminate pathogens, and the immunosuppression caused by traditional anti-inflammatory drugs (hormones), RvD1 can directly act on the inflammatory regulation process, promote the disappearance of inflammation and regulate the balance of the host immune response, help control the development of periodontitis and systemic complications and reduce recurrence, and provide a safer, more comprehensive and effective treatment plan for periodontal disease and systemic complications.

[0016] In one embodiment of the present invention, a mouse model of periodontitis was constructed, exhibiting multiple systemic complications. In this model, RvD1 significantly restored weight loss in mice with periodontitis, reduced alveolar bone height loss, and promoted local alveolar bone and connective tissue repair. It also had a therapeutic effect on systemic symptoms caused by periodontitis, such as decreased femoral bone density, shortened and dysfunctional colon length, and disrupted intestinal flora. RvD1 can be used as an active ingredient in the preparation of drugs for the treatment of periodontitis and its related complications. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 is the structural diagram of RvD1;

[0018] Figure 2 The results of RvD1 treatment improving alveolar bone loss and collagen degradation in periodontitis mice are shown in the figure. In the figure, Control: healthy control group; Nacl: periodontitis model + normal saline placebo group; RvD1: periodontitis model + RvD1 treatment group; CEJ-ABC: distance from enamel-cement junction to alveolar crest; Tb.Th: trabecular bone thickness; BV / TV: bone volume fraction; *: p < 0.05; **: p < 0.01; ns: no significant difference;

[0019] Figure 3 This figure shows the results of RvD1 improving the weight loss of mice with periodontitis. In the figure, Nacl: periodontitis model + normal saline placebo group; RvD1: periodontitis model + RvD1 treatment group;

[0020] Figure 4 The results of RvD1 treatment improving colon and femoral damage in periodontitis mice are shown in the figure. In the figure, Control: control healthy group; Nacl: periodontitis model + normal saline placebo group; RvD1: periodontitis model + RvD1 treatment group; BMD: bone mineral density; Tb.Th: trabecular bone thickness; BV / TV: bone volume fraction; Colon length: colon length; globet cells / cypt: average number of crypt goblet cells; crypt depth / mucosa thickness: crypt depth / tissue thickness;

[0021] Figure 5 This is a graph showing the results of RvD1 treatment improving the disordered intestinal flora structure in mice with periodontitis. In the figure, Control: control healthy group; Nacl: periodontitis model + normal saline placebo group; RvD1: periodontitis model + RvD1 treatment group; River distribution map: schematic diagram of the changing trend of the top ten microbial groups in abundance at the phylum level; Pielou and Shannon: Alpha diversity index of the flora; PcoA analysis, inter-group differences: principal coordinate analysis of Beta diversity of the flora and its difference test. DETAILED DESCRIPTION

[0022] The present invention provides the use of resolvin D1 in preparing a medicine for treating and / or alleviating systemic complications of periodontitis.

[0023] The resolvin D1 (RvD1) of the present invention has Figure 1 As shown in the structure, RvD1, an endogenous lipid mediator, can regulate immune cell activity, reduce inflammatory responses, and promote tissue repair. This invention is dedicated to developing novel RvD1-based treatments to address the shortcomings of existing treatments, aid in periodontal tissue repair and reconstruction, and treat and / or alleviate systemic complications of periodontitis.

[0024] The systemic complications of periodontitis described in the present invention include complications of all organs and / or tissues of the body caused by periodontitis, except for periodontal tissues, such as: weight loss, decreased femoral bone density, shortened colon length, colon dysfunction and disordered intestinal flora structure. In one embodiment of the present invention, a periodontitis model was constructed using mice, and the model exhibited multiple complications. In the mouse periodontitis model, RvD1 was able to significantly restore the weight loss of mice with periodontitis, reduce the loss of alveolar bone height, and promote local repair of periodontitis. It has a therapeutic effect on the systemic symptoms caused by periodontitis, such as decreased femoral bone density, shortened colon length and dysfunction, and disordered intestinal flora structure. RvD1 can be used as an active ingredient in the preparation of drugs for treating systemic complications of periodontitis.

[0025] The present invention does not specifically limit the dosage form of the drug, and may include any of the following: oral formulations, oral lavage formulations, injection formulations, gel formulations, nanoencapsulated delivery formulations, and water-soluble (e.g., mouthwash) gargle formulations. In one embodiment of the present invention, the drug is administered to a mouse model by injection, and the working concentration of RvD1 during administration is 1 ng / μL to 1 μg / μL.

[0026] The present invention does not specifically limit the source of the RvD1; conventional commercially available reagents in the art may be used, such as that purchased from Cayman Chemical Company (Item No. 10012554) in one embodiment. The injection preparation method includes first preparing a 100 μg / ml stock solution using ethanol as a solvent; dissolving 10 μL of the stock solution in 1 mL of normal saline to obtain a 1 ng / μL RvD1 working solution, wherein the solvent is 1% ethanol and normal saline.

[0027] The present invention also provides a drug for treating and / or alleviating systemic complications of periodontitis, which uses resolvin D1 as the sole active ingredient and also includes pharmaceutically acceptable excipients.

[0028] In a preferred embodiment of the present invention, the drug is administered in any of the following forms: an oral formulation, a gavage, an injectable formulation, a gel formulation, a nanoencapsulated delivery formulation, or a water-soluble (e.g., mouthwash) gargle formulation. The working concentration of resolvin D1 in the drug of the present invention is 1 ng / μL to 1 μg / μL in mice.

[0029] In a preferred embodiment of the present invention, when the drug is in liquid form, the solvent includes physiological saline containing ethanol, such as the solvent in the injection solution is 1% ethanol physiological saline.

[0030] The present invention also provides a drug for treating and / or alleviating systemic complications of periodontitis, which contains resolvin D1 as a partial active ingredient and also includes pharmaceutically acceptable excipients.

[0031] In addition to RvD1 as an active ingredient, the drug of the present invention also contains other active ingredients that can be used to treat periodontitis and / or systemic complications, thereby forming a combination drug. In a preferred embodiment of the present invention, the dosage form of the drug includes any of the following: an oral preparation, a gavage, an injection, a gel preparation, a nano-encapsulated delivery preparation, or a water-soluble (e.g., mouthwash) gargle preparation. When the drug is in liquid form, the solvent includes ethanol-containing saline, such as 1% ethanol in saline in the injection solution.

[0032] To further illustrate the present invention, the use of the resolvin D1 provided by the present invention in treating systemic complications of periodontitis is described in detail below in conjunction with the examples, but they should not be construed as limiting the scope of protection of the present invention.

[0033] 1. Preparation of RvD1 injection: RvD1 was purchased from Cayman Chemical Company; the stock solution was 100 μg / ml, and the solvent was ethanol. 10 μL of the stock solution was dissolved in 1 mL of normal saline to obtain a 1 ng / mL RvD1 working solution, and the solvent was 1% ethanol and normal saline.

[0034] 2. Experimental Animals and Housing: Male C57BL / 6 mice, 6–7 weeks old, were purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd. They were housed at the Experimental Animal Center of Nanjing Agricultural University (SPF grade) under the following housing conditions: temperature of 22–24°C, humidity of 40–70%, alternating light and dark cycles with a lighting period of 12 h, and free access to water and food.

[0035] 3. Construction of mouse periodontitis model:

[0036] (1) Mice were acclimatized in an SPF-grade standardized animal center for 1 week.

[0037] (2) The second molars of mice were ligated with 4-0 silk sutures and coated with 1 μg / μL bacterial lipopolysaccharide (LPS), 15-20 μL per side. The sutures were removed after 1 week to establish a mouse periodontitis model.

[0038] Example 1 Therapeutic effect of RvD1 on mouse periodontitis model

[0039] 1. RvD1 administration method

[0040] After suture removal, the treatment group (RvD1) received 100 ng of RvD1 via intraperitoneal injection every other day for 4 weeks. The solvent was 1% ethanol in saline. The disease group (NaCl) received a placebo (1% ethanol in saline), and the control group (Control) received no treatment.

[0041] 2. Mouse alveolar bone measurement

[0042] MicroCT morphological observation of alveolar bone loss: Maxillary bones were isolated from each group and fixed in 4% histological fixative at 4°C for 48 hours. MicroCT scanning was performed. The cementoenamel junction to alveolar crest (CEJ-ABC) distance was measured at three locations on the buccal and palatal mesiodistal surface of the second molar alveolar bone (a total of six locations bilaterally) to assess the amount of alveolar bone height loss in each group. The bone volume fraction (BV / TV) was also measured, and the amount of alveolar bone loss was calculated for each group.

[0043] The results are as follows Figure 2 As shown, RvD1 mitigates periodontitis-induced alveolar bone height loss, resulting in a decrease in the CEJ-ABC distance. Bone density analysis revealed a significantly lower BV / TV ratio in the periodontitis group compared with the control group, while no significant difference was observed between the RvD1-treated group and the control group. This suggests that RvD1 can promote tissue repair in periodontitis and mitigate periodontitis-induced alveolar bone loss and collagen degradation.

[0044] 3. Mouse Weight Measurement

[0045] The body weights of the mice were measured on days 4, 7, 11, 15, 19, 23, 27, 31, and 35 after periodontitis modeling, and a line graph of the percentage changes in body weight was drawn.

[0046] The results are as follows Figure 3 As shown, periodontitis causes weight loss in mice, and RvD1 can improve the amount of weight loss in periodontitis mice, enabling them to restore weight earlier and enhance their physical fitness.

[0047] 4. Measurement of Mouse Colon Length and Functional Assessment

[0048] The entire colon of the mice was removed and measured, and the colon length values of the mice in each group were compared. Colon sections were stained with AB-PAS to assess the number of colonic goblet cells and crypt length.

[0049] The results are as follows Figure 4 As shown, RvD1 treatment ameliorates colonic damage in mice with periodontitis. The colon of mice with periodontitis is significantly shortened, whereas RvD1 treatment increases colon length. AB-PAS staining revealed a significant decrease in goblet cell number and crypt length in the colon of the periodontitis group, whereas RvD1 treatment significantly increases goblet cell number and crypt length.

[0050] 5. Mouse Femur Measurement

[0051] MicroCT morphological observation of femoral bone loss: Femora were isolated from each experimental group and fixed in 4% histiocytic fixative at 4°C for 48 hours. MicroCT scanning was performed. Bone mineral density (BMD), bone volume fraction (BV / TV), and trabecular thickness (Tb / Th) were calculated by measuring trabeculae 1-2 mm below the growth plate.

[0052] The results are as follows Figure 4 As shown, Tb, Th, and BMD in the femoral cancellous bone of the periodontitis group were significantly decreased compared to the control group, but RvD1 treatment reversed this trend. Although the difference in BV / TV was not significant, it showed a similar trend, indicating that RvD1 can treat the periodontitis-induced decrease in femoral cancellous bone density and trabecular thickness, thereby improving femoral damage in periodontitis mice.

[0053] 6. Determination of mouse intestinal flora structure

[0054] 16S rRNA sequencing analysis of mouse cecal contents: The cecal contents of mice were collected into sterile EP tubes, snap-frozen in liquid nitrogen for 20 seconds, and stored at -80°C. 16S rRNA gene sequencing and bacterial flora analysis were performed on the cecal contents.

[0055] The results are as follows Figure 5As shown in the figure, the relative abundance of the bacterial communities at the phylum level shows a sharp decrease in the relative abundance of Bacteroidota, Firmicutes_A, and Firmicutes_D, while the relative abundance of Desulfobacterota increases. However, after RvD1 treatment, the proportions of Bacteroidota, Firmicutes_A, and Firmicutes_D rapidly recovered, while the abundance of Desulfobacterota decreased. This suggests that RvD1 effectively improves the community structure imbalance caused by periodontitis. The Tukey-HSD test revealed significant differences in alpha diversity overall: the overall Pielou index was p = 0.0371, and the overall Shannon index was p = 0.0192. Further pairwise comparisons revealed that the Pielou index in the periodontitis group was significantly lower than that in the control and RvD1-treated groups (p < 0.05), while no significant difference was found between the control and RvD1-treated groups. The Shannon index also showed a similar trend. Bray-Curtis principal coordinate analysis (PCoA) revealed some overlap in the microbial composition between the RvD1 group and the control group, while no overlap was observed between the periodontitis group and either group, indicating a trend of separation. Similar trends were observed in between-group differences in Beta diversity. These results suggest that RvD1 treatment improves the disturbed intestinal microbial composition in mice with periodontitis.

[0056] Although the above embodiment provides a detailed description of the present invention, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the scope of protection of the present invention.

Claims

1. Use of resolvin D1 in the preparation of a drug for treating and / or alleviating systemic complications of periodontitis.

2. The application according to claim 1, characterized in that The systemic complications of periodontitis include complications in all organs and / or tissues of the body caused by periodontitis except for periodontal tissues.

3. The application according to claim 1, characterized in that The dosage form of the drug includes oral preparations, injection preparations, gel preparations, nano-encapsulated delivery preparations or water-soluble gargle preparations.

4. The application according to claim 3, characterized in that Based on mice, the working concentration of resolvin D1 in the drug is 1 ng / μL to 1 μg / μL.

5. A drug for treating and / or alleviating systemic complications of periodontitis, comprising resolvin D1 as the sole active ingredient, characterized in that: It also includes pharmaceutically acceptable excipients, as well as chemical modifications and modifications related to resolvin D1.

6. The drug according to claim 5, characterized in that The dosage form of the drug includes any one of the following: oral preparation, gavage preparation, injection preparation, gel preparation, nano-encapsulation delivery preparation and water-soluble gargle preparation.

7. The medicine according to claim 5, characterized in that Based on mice, the working concentration of resolvin D1 in the drug is 1 ng / μL to 1 μg / μL.

8. The medicine according to claim 5, characterized in that When the drug is in liquid form, the solvent includes physiological saline containing ethanol.

9. A drug for treating and / or alleviating systemic complications of periodontitis, characterized in that: The invention contains resolvin D1 as a partial active ingredient and also includes pharmaceutically acceptable excipients.

10. The drug according to claim 9, characterized in that The dosage form of the drug includes any one of the following: oral preparation, gavage preparation, injection preparation, gel preparation, nano-encapsulation delivery preparation and water-soluble gargle preparation.

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