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Application of PEDF in preparing medicine for diabetes bone tissue complication

A technology for bone tissue and complications, which is applied in the application field of preparing medicines for treating diabetic bone tissue complications, and can solve the problems of accelerating the osteogenesis process and the like

Inactive Publication Date: 2017-04-26
XINXIANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, it may also accelerate the osteogenesis process by stimulating blood vessel signals

Method used

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  • Application of PEDF in preparing medicine for diabetes bone tissue complication
  • Application of PEDF in preparing medicine for diabetes bone tissue complication
  • Application of PEDF in preparing medicine for diabetes bone tissue complication

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1 Isolation and culture of osteoblasts

[0016] Bone waste samples were collected from foot amputee patients and divided into diabetic and non-diabetic groups. Tissue samples were provided by Penn State Hershey College of Medicine. After the collected tissue samples were rinsed with PBS for 3 times, the connective tissue and adipose tissue on the surface of the bone tissue were removed, and the cells were treated with alcohol for 15 seconds (to remove the fibroblasts on the surface of the bone tissue). Digest the bone tissue with collagenase at a concentration of 4 mg / ml, collect the supernatant of the digestion solution every 45 minutes, centrifuge at 1000 rpm for 5 minutes, and keep the precipitate for three consecutive times. After the fourth cell digestion for 45 minutes, collect the bone tissue and single cell pellet by centrifugation. Spread them evenly into 100mm cell culture dishes for culture. The composition of the culture medium is a-MEM, 10% FBS, 1...

Embodiment 2

[0017] Example 2 PEDF up-regulates the level of VEGF in diabetic osteoblasts

[0018] According to the culture conditions of Example 1, 250ng / ml of PEDF was added to the osteogenic culture medium, and the level of VEGF was detected by conventional qPCR technology. The melting curve was 95°C 15s (100%); 65°C 1min, 95°C 15s (100%), +0.3°C / circle, b-actin was used as internal reference. The results showed that the expression of VEGF was up-regulated after the cells were treated with 250ng / ml PEDF for 7 days. like figure 2 shown.

Embodiment 3

[0019] Example 3 PEDF up-regulates the expression levels of OCN and Runx2 genes

[0020] OCN and Runx2 are marker genes of osteoblasts. According to the culture conditions of Example 1, 250 ng / ml of PEDF was added to the osteogenic culture medium, and the expression levels of OCN and Runx2 genes were detected by conventional qPCR technology. ℃ 30s), the melting curve was 95℃ 15s (100%); 65℃ 1min, 95℃ 15s (100%), +0.3℃ / circle, b-actin was used as internal reference. The results showed that the expression of OCN and Runx2 genes was up-regulated after 250ng / ml PEDF treated the cells for 7 days. like image 3 shown.

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Abstract

The invention discloses application of PEDF in preparing medicine for diabetes bone tissue complication, and most diabetes patients have bone tissue complication, including osteoporosis, bone healing difficulty, gangrene and the like,. When PEDF expression levels of an osteoblast and osteogenic capability of a diabetes patient and a non-diabetes patient are compared, it is found that PEDF expression level of the non-diabetes patient is higher, and the osteogenic differentiation ability, the osteogenic efficiency and the calcium deposition level of the non-diabetes patient are higher than those of the diabetes patient (p<0.01). Exogenetic PEDF can up-regulate expression of osteogenic differentiation related genes OCN and Runx2. Compared with VEGF expression levels of the osteoblast of the two groups of the patients, it is found that the overall VEGF level of the non-diabetes patient is higher than that of a diabetes patient, and the exogenetic PEDF can up regulate expression of VEGF. PEDF can improve osteogenic differentiation ability of the osteoblast and calcium deposition level. PEDF can be applied to treating bone tissue complication caused by diabetes.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the application of PEDF in the preparation of medicines for treating diabetic bone tissue complications. Background technique [0002] Diabetes not only causes abnormalities in the three major nutrients including sugar, fat, and protein, but also involves metabolic disorders such as electrolytes and bone minerals. Therefore, complications caused by diabetes are the biggest problem at present, such as long-term non-healing of wounds in patients, and other symptoms. Bone tissue is also a tissue that is greatly affected by diabetic complications. Studies have shown that diabetics are prone to osteoporosis due to the decrease in calcium deposition concentration, and patients with fractures are not easy to heal. It has been reported that the bone formation and accumulation in diabetics during the development period is lower than that of normal people, and there will be obstacle...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/57A61P19/08A61P3/10
Inventor 宋娜王海军钟加滕赵铁锁邵明龙郭胜齐金博
Owner XINXIANG MEDICAL UNIV
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