Application of herba artemisiae scopariae aqueous extract in enhancing liver chemotaxis of adipose tissue-derived stem cells
By using Artemisia capillaris water extract to enhance the expression of CCR1 and CXCR4 chemokines in adipose-derived mesenchymal stem cells in vitro, the problems of damage and high cost caused by cell modification in existing technologies have been solved, thereby improving liver chemotaxis and migration ability.
Patent Information
- Application Number
- CN202511829230.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-05
- Publication Date
- 2026-01-20
AI Technical Summary
Existing technologies for enhancing the liver chemotaxis of adipose-derived mesenchymal stem cells suffer from problems such as cell damage caused by gene modification, insertional carcinogenesis, and high costs, making it difficult to effectively improve their ability to treat liver injury.
Artemisia capillaris aqueous extract was used to enhance the expression level of chemokines in adipose-derived mesenchymal stem cells in vitro. By treating cells with Artemisia capillaris aqueous extract at a concentration of 0.5 mg/mL to 1 mg/mL for 24 h to 48 h, the expression of CCR1 and CXCR4 chemokines was promoted.
It enhances the liver chemotaxis and migration ability of adipose-derived mesenchymal stem cells, avoids cell damage and carcinogenesis risks caused by gene modification, and reduces costs.
Smart Images
Figure CN121362730A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The application belongs to the technical field of biotechnology, and particularly relates to application of Artemisia capillaris water extract in enhancing liver chemotaxis of adipose tissue-derived mesenchymal stem cells. BACKGROUND
[0002] At present, it is found that adipose tissue-derived mesenchymal stem cells have strong in-vitro expansion and differentiation capabilities, can be differentiated into hepatocytes under the action of different induction factors, and can treat liver injury and cirrhosis after transplantation after in-vitro expansion. However, in-vitro expansion of adipose tissue-derived mesenchymal stem cells can weaken the chemotaxis of the latter, thereby weakening the ability of the latter to treat liver injury.
[0003] The existing technology for enhancing liver chemotaxis of adipose tissue-derived mesenchymal stem cells is mainly biological engineering technology (genetic modification or surface molecule modification), that is, expressing a chemotactic factor receptor and an adhesion molecule that mediate homing on the surface of adipose tissue-derived mesenchymal stem cells, so that the adipose tissue-derived mesenchymal stem cells have a higher homing ability to target tissues. However, biological engineering technology modification often changes the gene profile of adipose tissue-derived mesenchymal stem cells, causes greater damage to the cells themselves, and is accompanied by disadvantages such as the possibility of insertional carcinogenesis, immune response side effects, and high cost. Therefore, there is an urgent need to provide a technology for enhancing liver chemotaxis of adipose tissue-derived mesenchymal stem cells and improving the ability of the adipose tissue-derived mesenchymal stem cells to treat liver injury. SUMMARY
[0004] The application provides application of Artemisia capillaris water extract in enhancing liver chemotaxis of adipose tissue-derived mesenchymal stem cells.
[0005] To achieve the above-mentioned purposes, the application adopts the following technical solutions: In a first aspect, the application provides application of Artemisia capillaris water extract in preparing a drug for enhancing liver chemotaxis of adipose tissue-derived mesenchymal stem cells.
[0006] Further, the concentration of the Artemisia capillaris water extract is 0.5 mg / mL to 1 mg / mL, and the chemotaxis time is 24 h to 48 h.
[0007] In a second aspect, the application provides application of Artemisia capillaris water extract in preparing a drug for improving expression levels of chemotactic factors of adipose tissue-derived mesenchymal stem cells.
[0008] Further, the concentration of the Artemisia capillaris water extract is 0.5 mg / mL to 1 mg / mL, and the chemotaxis time is 24 h to 48 h.
[0009] Further, the chemotactic factors are CCR1 and CXCR4.
[0010] In a third aspect, the application provides a drug for enhancing liver chemotaxis of adipose tissue-derived mesenchymal stem cells, which contains Artemisia capillaris water extract.
[0011] In a fourth aspect, the present application provides a medicine for improving the expression level of chemotactic factors of adipose-derived mesenchymal stem cells, which contains the aqueous extract of Artemisia capillaris.
[0012] Further, the chemotactic factors are CCR1 and CXCR4.
[0013] Compared with the prior art, the present application has the following advantages: The aqueous extract of Artemisia capillaris can improve the expression level of chemotactic factors of adipose-derived mesenchymal stem cells in vitro, and can intervene in the proliferation, migration and chemotactic ability of adipose-derived mesenchymal stem cells to liver cells. Compared with the existing biological engineering technology, it also has the advantages of low cost, convenient method and no adverse reactions such as insertion cancer. BRIEF DESCRIPTION OF DRAWINGS
[0014] Figure 1 It is the quality standard identification result of the aqueous extract of Artemisia capillaris.
[0015] Figure 2 It is the influence of different concentrations of the aqueous extract of Artemisia capillaris on the proliferation ability of adipose-derived mesenchymal stem cells.
[0016] Figure 3 It is the influence of different concentrations of the aqueous extract of Artemisia capillaris on the migration ability of adipose-derived mesenchymal stem cells.
[0017] Figure 4 It is the influence of different concentrations of the aqueous extract of Artemisia capillaris on the surface chemotactic factors of adipose-derived mesenchymal stem cells. DETAILED DESCRIPTION
[0018] In order to further illustrate the technical scheme of the present application, the present application will be further illustrated by examples. EXAMPLE
[0019] (1) Overall research idea: Extract the aqueous extract of fresh Artemisia capillaris freeze-dried powder, prepare 10 mL of mother liquor with a concentration of 100 mg / mL for 1 g of freeze-dried powder, and prepare different aqueous extract concentration gradients to intervene in adipose-derived mesenchymal stem cells, and detect the influence of different concentrations of Artemisia capillaris aqueous extract on the proliferation, migration ability and chemotactic factor expression of adipose-derived mesenchymal stem cells.
[0020] (2) Condition optimization: First, compare the toxic effects of different concentrations of Artemisia capillaris aqueous extract on adipose-derived mesenchymal stem cells to determine the optimal concentration and intervention time of Artemisia capillaris aqueous extract; then, use the optimal aqueous extract to intervene in adipose-derived mesenchymal stem cells, and use cell scratch, cell migration, PCR and other experimental methods to detect the proliferation, migration and chemotactic factor expression of adipose-derived mesenchymal stem cells.
[0021] Specific content: 1. Extraction of Artemisia capillaris aqueous extract 80 g Artemisia capillaris was decocted 3 times, the decoction was filtered through a vacuum filter, concentrated to 100 mL by an electromagnetic oven, and then concentrated to about 40 mL by a rotary evaporator. The extract was dried in a vacuum drying box to obtain an extract. The yield was calculated based on a concentration of 100%. The water extract was determined by high performance liquid chromatography (General Rule 0512) to determine the content of chlorogenic acid. The determined content of chlorogenic acid was used as a standard to control the Artemisia capillaris extract used in subsequent experiments. 1 g of the water extract was dissolved in 10 mL of ultrapure water, centrifuged, and the supernatant was filtered through a 0.22 μm microporous filter to obtain a mother liquor with a concentration of 100 mg / mL, which was stored in a -20°C refrigerator.
[0022] 2. Culturing of fat mesenchymal stem cells intervened by Artemisia capillaris water extract Rat fat mesenchymal stem cells (Meilunbio, PWC-RA010) were cultured, and the culture medium was changed every 3-4 days. The concentration of the Artemisia capillaris water extract for intervention was as follows: (1) Normal group: high-sugar DMEN containing 10% fetal bovine serum, cultured for 24 h; (2) 0.25 mg / mL Artemisia capillaris water extract intervention ADSCs, 10% fetal bovine serum high-sugar DMEM, cultured for 24 h; (3) 0.5 mg / mL Artemisia capillaris water extract intervention ADSCs, 10% fetal bovine serum high-sugar DMEM, cultured for 24 h; (4) 1 mg / mL Artemisia capillaris water extract intervention ADSCs, 10% fetal bovine serum high-sugar DMEM, cultured for 24 h; When the cell confluence was 70-80%, the supernatant was collected, the cells were digested with 0.25% trypsin digestion solution containing EDTA, then centrifuged at 2000 r / min for 3-5 min, and the cells were collected and subcultured.
[0023] 3. Detection of the proliferation, migration and chemotaxis of fat mesenchymal stem cells intervened by Artemisia capillaris water extract EdU incorporation experiment: each group of ADMSCs was inoculated in a 24-well plate containing 10% FBS culture medium, and after 24 h of stabilization, 5-ethynyl-2'-deoxyuridine (EdU) labeling reagent was added. After 6 h of action, the culture medium was discarded, the cells were fixed with paraformaldehyde for 15 min, washed 3-5 times, 0.3% Triton X-100 was added to PBS and reacted for 15 min, then Click reaction solution was added according to the instructions, and the fluorescence signal was observed under 488 nm excitation light. The proliferation of fat mesenchymal stem cells was detected.
[0024] Cell scratch test: select logarithmic growth phase cells, with 10% fetal bovine serum in the medium, inoculated into 6 well plates, when the cell confluence reached 90%-100%. With 10 μL gun head vertically in the cell layer, draw a straight way, PBS wash 3 times to remove cell debris. With fetal bovine serum-free medium, at 0h, 24h, 48h, the same field under the microscope to take pictures. Use Image software to analyze the blank area of cells at different time points. Observe the migration ability of adipose mesenchymal stem cells.
[0025] Chemokine receptor detection: extract total RNA from each group of ADSCs, PCR detection of each chemokine on the surface of ADSCs, such as CCR1, CCR7, CX3CR1, CXCR4, CXCR6. According to the PCR results, determine the next step of chemokine protein level detection.
[0026] Transwell chamber with 8 μm pore polycarbonate membrane 12 well plate, add ADSCs to the upper chamber, add the same complete medium to the upper and lower chambers, place in 37℃ cell incubator for about 6h to observe cell adhesion, discard the culture medium, wash gently with PBS for 2 times, replace the upper chamber with complete medium containing different doses of drugs, add complete medium containing chemokines (CCR1, CXCR4) to the lower chamber, continue to culture until the cells cover the chamber, use a cotton swab to wipe off the cells in the upper chamber of Transwell, add 1 mL 40 g / L paraformaldehyde to the lower chamber for 20 min at room temperature, after PBS washing, add 1 ml 0.1% crystal violet staining solution to each well for 30 min, and take pictures of the results.
[0027] 4. Experimental results: (1) The product contains not less than 0.50% of chlorogenic acid (C16H18O9) calculated as dry product, that is, the product is considered to be qualified. The results show that (Table 1) Figure 1 ), the content of chlorogenic acid in Yinchen powder is 0.90%, and the extraction of Yinchen water extract reaches the standard.
[0028] (2) Compared with the normal control group, Yinchen water extract 1mg group can significantly promote the proliferation of adipose mesenchymal stem cells at 24h and 48h, and Yinchen water extract 0.5mg group can significantly promote the proliferation of adipose mesenchymal stem cells at 48h (P<0.05) Figure 2 ), suggesting that Yinchen water extract can promote the migration ability of adipose mesenchymal stem cells. Transwell chamber experiment results show that compared with the normal group, Yinchen water extract 1mg group can significantly promote the migration ability of adipose mesenchymal stem cells (P<0.05) Figure 3 ).
[0029] (3) PCR detection of the influence of different concentrations of Artemisia capillaris water extract on the expression of chemotactic factor receptors CCR1, CCR7, CX3CR1, CXCR4 and CXCR6 of adipose-derived mesenchymal stem cells. Compared with the normal control group, Artemisia capillaris water extract 1mg group and Artemisia capillaris water extract 0.5mg group can significantly improve the expression of chemotactic factor receptors CCR1 and CXCR4 of adipose-derived mesenchymal stem cells. CCR1 and CXCR4 can promote the chemotaxis of adipose-derived mesenchymal stem cells to damaged hepatocytes. Figure 4 ).
[0030] In summary, Artemisia capillaris water extract can improve the expression level of chemotactic factor of adipose-derived mesenchymal stem cells in vitro, and Artemisia capillaris water extract can intervene the proliferation, migration and chemotaxis of adipose-derived mesenchymal stem cells to hepatocytes.
[0031] The above shows and describes the main features and advantages of the present application, and it is obvious to those skilled in the art that the present application is not limited to the details of the above exemplary embodiments, and the present application can be implemented in other specific forms without departing from the spirit or essential characteristics of the present application. Therefore, from any point of view, the embodiments should be regarded as exemplary and non-limiting, and the scope of the present application is defined by the appended claims rather than the above description, and it is intended to encompass all changes falling within the meaning and scope of the equivalent elements of the claims.
[0032] In addition, it should be understood that although the present specification is described in terms of embodiments, not every embodiment contains only one independent technical solution, and the description manner of the specification is only for clarity, and those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can be properly combined to form other embodiments that those skilled in the art can understand.
Claims
1. Application of Artemisia capillaris water extract in the preparation of a drug for enhancing chemotaxis of adipose tissue-derived mesenchymal stem cells to liver.
2. Use according to claim 1, characterized in that, The concentration of the Artemisia capillaris water extract is 0.5 mg / mL-1 mg / mL, and the chemotaxis time is 24 h-48 h.
3. Application of Artemisia capillaris water extract in the preparation of a drug for increasing expression level of chemotactic factors of adipose tissue-derived mesenchymal stem cells.
4. Use according to claim 3, characterized in that, The concentration of the Artemisia capillaris water extract is 0.5 mg / mL-1 mg / mL, and the chemotaxis time is 24 h-48 h.
5. Use according to claim 3, characterized in that, The chemotactic factors are CCR1 and CXCR4.
6. A medicament for enhancing the liver chemotaxis of adipose tissue-derived mesenchymal stem cells, characterized by, The Artemisia capillaris water extract is contained.
7. A medicine for enhancing the expression level of chemotactic factor of fat mesenchymal stem cells, characterized by, The Artemisia capillaris water extract is contained.
8. The medicament according to claim 7, characterized in that, The chemotactic factors are CCR1 and CXCR4.