| AIM: To observe the effects of human umbilical cord stem cells(hUCMSCs)on bone tissue structure,bone metabolism markers and bone tissue-related gene expression in type 1 diabetic mice,and to explore the protective effect of human umbilical cord mesenchymal stem cells on bone tissue in type 1 diabetic mice.Methods: Twenty-eight healthy male non-obese diabetic(NOD)6-8-week-old mice,after 9 weeks of adaptive feeding,were diagnosed with type 1 diabetes in 20 mice and unaffected in 8 mice.Eighteen randomly selected NOD mice were divided into blank group(n = 6),insulin group(n = 6),umbilical cord stem cell group(stem cell group,n =6),and 6 randomly selected NOD mice without disease were used as the control group(normal group,n = 6).Blank group: no treatment except normal feeding.Insulin group:daily subcutaneous injection of appropriate amount of insulin glargine ingection.Stem cell group: daily subcutaneous injection of appropriate amount of insulin glargine ingection,hUCMSCs were given on the third day after the onset of illness.The latter 1 ×10 ^ 6 first needed to be added with 0.3 ml of PBS solution,and then injected into the tail vein of mice.Normal group: the same normal feeding,no need to inject other drugs,no special treatment.The grouped experimental animals were kept for 8 weeks.After 8weeks,all mice were subjected to internal canthal vein puncture,venous blood was taken,serum was obtained,and then tested by ELISA to determine the content of acid phosphatase.After removing the neck,the experimental animals were sacrificed,and the femur was taken for subsequent experiments.The right femur was examined with Micro-CT to determine the microstructure of the femoral tissue.Prepare 10%ethylenediaminetetraacetic acid,use it to treat the left femur,decalcify,and make paraffin sections.The above sections were stained with alkaline phosphatase.The bone tissue was tested,and the expression levels of related genes were determined by RT-PCR.Results: Compared with mice in the blank group,TRACP levels were relatively lower in the insulin group and stem cell group,with a significant difference(P < 0.05).Compared with the normal group,TRACP levels were significantly increased in the blank group and insulin group(P < 0.05),and TRACP was also increased in the stem cell group(P = 0.45).The TRACP level in the stem cell group was 13.26 ± 0.91 ?g/mL,which was lower than that in the insulin group(15.17 ± 0.89 ?g/mL),and the BMC,BMD,Calib.Tb.Th.3D,BV/TV,Tb.N,SMI,Calib.Tb.Sp.3D,BS/BV increased.Compared with the normal group,the BMC,BMD,Calib.Tb.Th.3D,BV/TV,Tb.N decreased,however SMI,Calib.Tb.Sp.3D,BS/BV increased.Compared with the blank group,the BMC,BMD,Calib decreased in the stem cell group and the insulin group.Tb.Th.3D,BV/TV,Tb.N,SMI,Calib.Tb.Sp.3D and BS/BV increased in blank group.Diabetic osteoporosis isa systemic metabolic bone disease that is caused by diabetes and is characterized by a decrease in bone mass,destruction of the fine structure of the bone,and an increase in the brittleness of the bone.It is a combination of genetic and environmental factors.ALP staining of bone tissue sections showed that the number of osteoblasts in the blank group was significantly lower than that in the normal group,and the number of bone cells composed of stem cells was lower than that in the normal group,but higher than that in the insulin treatment group and the blank group.Compared with the normal group,the expression levels of RUNX2 and OCN in the stem cell group,the insulin group and the blank group were decreased,and the expression levels of RUNX2 and OCN in the stem cell group and the insulin group were increased compared with the blank group(all P<0.05).Compared with the normal group,the blank group and other groups showed increased relative expressions of bone-related genes TRAP and PPAR-g.Compared with the blank group,expressions of TRAP and PPAR-G in the stem cell group,insulin group and insulin group were decreased,and the differences were statistically significant(P<0.05).The relative expression of RUNX2 in the stem cell group(0.70±0.15)was significantly higher than that in the insulin group(0.47±0.14)(P<0.05).The relative expression level of PPAR-G in the stem cell group(1.60±0.42)was lower than that in the insulin group(2.30±0.42),and the difference was statistically significant.CONCLUSION: In this study,the application of umbilical cord stem cell therapy can significantly improve the serum tartrate-resistant acid phosphatase(TRACP)level and femoral structure in type 1 diabetic mice and delay the occurrence of osteoporosis,which may be mediated by up-regulating the expression of RUNX2 and OCN genes and inhibiting the expression of TRAP and PPAR-G genes,thereby promoting bone formation and inhibiting bone destruction. |
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