Application of TGF-beta inhibitor in preparation of medicine for treating diabetic foot

A technology of β inhibitors and diabetic foot, applied in the field of tissue repair drugs, can solve the problems that TGF-β inhibitors affect the growth activity of endothelial cells, and the proliferation activity and angiogenesis function of endothelial cells have not been seen with TGF-β inhibitors. Reduce the level of intracellular reactive oxygen species, improve the function of angiogenesis, and improve the effect of treatment

Pending Publication Date: 2022-04-29
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no report about the effect of TGF-β inhibitors on the growth activity of endothelial cells, and there is no report that TGF-β inhibitors positively regulate the proliferation activity and angiogenesis of endothelial cells under high-glucose and low-nutrient conditions. Reports on the application of small molecules in the treatment of diabetic foot

Method used

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  • Application of TGF-beta inhibitor in preparation of medicine for treating diabetic foot
  • Application of TGF-beta inhibitor in preparation of medicine for treating diabetic foot
  • Application of TGF-beta inhibitor in preparation of medicine for treating diabetic foot

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 Screening for Small Molecular Substances That Can Induce Endothelial Cells to Grow and Passage Under High Glucose and Low Nutrient Conditions

[0053] The vascular endothelial cells used in this example are umbilical vein endothelial cells (hUVECs), isolated from umbilical cord veins preserved in West China Hospital of Sichuan University, and have obtained ethical approval.

[0054] The specific steps of recovery and subculture of hUVECs are as follows:

[0055] a. Cell resuscitation: Take 1 tube of frozen hUVECs, thaw quickly at 37°C, transfer the cells to a 15ml centrifuge tube, add 8ml of complete medium for dropwise recovery; the complete medium used in this example contains 5% FBS and growth factors EGM2 complete medium. Centrifuge at 1200rpm for 3min, remove the supernatant, add 5ml of complete medium to resuspend; inoculate each tube of cells into a T75 culture flask, add complete medium to 15ml, and place in CO 2 Incubator (37°C, 5% CO 2 , saturated...

Embodiment 2

[0066] Concentration screening when adopting SB431542 domestication culture in embodiment 2

[0067] Experiments were performed by adding different concentrations of SB431542, the concentrations were: 2 μM, 5 μM, 10 μM, 20 μM, and 40 μM. The result is as Figure 5 As shown, the addition of 2-10 μM SB431542 can maintain the survival and proliferation of hUVECs under the condition of high glucose and low nutrition, but if the concentration is too high, it will affect the state of the cells and reduce the ability of the cells to survive and proliferate. In the present invention, our preferred additive dose is 5 μM.

Embodiment 3

[0068] Example 3 Using SB431542 of the present invention to induce culture to obtain FE-ECs

[0069] The formula of the induction medium used in the embodiment is: 500ml endothelial basal medium (Sciencell company, product number 1001), 25ml FBS, do not add endothelial cell growth supplement, add 75mM concentration glucose to prepare high glucose culture fluid, add 5 μ M SB431542 Small Molecules.

[0070] The specific preparation method is as follows: Take the preparation of 100ml volume induction medium as an example, pipette 95ml endothelial cell culture medium, add 5ml FBS, and do not add endothelial cell growth supplement. Then weigh 1.2 g of glucose (Sigma, Cat. No. V900392) and dissolve it in the above 100 ml basal medium to form a high-sugar and low-nutrition endothelial cell culture medium with a sugar content of 75 mM, and add 5 μM SB431542 small molecule at the same time.

[0071] The specific operation steps are as follows:

[0072] a. Select hUVECs of passage P2 ...

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Abstract

The invention belongs to the technical field of tissue repair medicines, and particularly relates to application of a TGF-beta inhibitor in preparation of a medicine for treating diabetic foot. Aiming at the problems that cells are easy to apoptosis and the treatment effect of cell transplantation is not lasting when the cells are used for treating the diabetic foot, the invention provides the application of the TGF-beta inhibitor in preparing the medicine for treating the diabetic foot, and the TGF-beta inhibitor is proved to be capable of promoting the cells to maintain the mitochondrial membrane potential to be stable under the conditions of high glucose and low nutrition; and the intracellular active oxygen level is reduced, so that the cells maintain good proliferative activity and vascularization function. The function-enhanced endothelial cells obtained through induction and domestication culture of the TGF-beta inhibitor can survive for a long time in limbs of the ischemic diabetic foot of a mouse and form new vessels, and the treatment effect of endothelial cell transplantation on the ischemic diabetic foot is improved. According to the invention, the application range of the TGF-beta inhibitor is widened, a more effective method is provided for the cell therapy of diabetic foot, and the TGF-beta inhibitor has a good prospect.

Description

technical field [0001] The invention belongs to the technical field of tissue repair medicines, and in particular relates to the use of a TGF-β inhibitor in the preparation of medicines for treating diabetic foot. Background technique [0002] Diabetic foot disease (Diabetic foot, DF) refers to the clinical syndrome of gangrene, ulceration or infection in the distal limbs of diabetic patients due to limb atherosclerosis, microcirculation, and neurotrophic disorders. It is the most common clinical syndrome in diabetes. One of the complications of the disease has a very high rate of disability and mortality. With the gradual increase in the incidence of diabetes worldwide, DF will also become the main social and economic burden of chronic disease treatment in my country. [0003] In recent years, cell therapy has developed rapidly. By supplementing proliferative endothelial cells (ECs), promoting bypass angiogenesis and improving blood circulation in affected limbs, it has br...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K31/4439A61K31/517A61K31/4709A61P17/02A61P3/10
CPCA61K45/00A61K31/4439A61K31/517A61K31/4709A61P17/02A61P3/10
Inventor 邓洪新成福义
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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