The Effect Of Sparc On The Proliferation And Apoptosis,and The Expression Of Collagen Type Ⅰ And Collagen Type Ⅲ Of Human Keloid Fibroblasts And Normal Fibroblasts

Objective This study is intended to investigate the effect of Sparc on the proliferation and apoptosis of human keloid fibroblasts and normal skin fibroblasts at the cellular level,as well as its effect on the expression of collagen type Ⅰand collagen type Ⅲ,so as to confirm the important role of Sparc in the formation of keloid.Methods Ten cases of keloid and five cases of normal skin were collected and cultured respectively,so as to obtain primary human keloid fibroblasts and normal human fibroblasts.When the cells were in the logarithmic growth phase,the cell status was observed under a microscope,and the subsequent dosing treatment can be performed.The two types of cells were divided into experimental group and control group,experimental group to join DMEM with 2 μg/ml Sparc protein of incubation in 72 h,the control group to join the equivalent DMEM.CCK-8 cell proliferation assay was used to detect the effect of SPARC protein on cell proliferation in each group,and flow cytometry was used to detect the effect of SPARC protein on cell apoptosis in each group.WB and RT-PCR detect the expression of collagen type Ⅰand collagen type Ⅲ in each group.Results 1.CCK-8 cell proliferation assay showed that the proliferation of cells was more obvious in the treatment groups with SPARC protein than in the control group.2.Flow cytometry showed that growth of cells was inhibited in the SPARC group compared with the control group.3.Western blot experiments suggest that the treatment groups with SPARC protein showed more obvious expression of collagen type Ⅰ than the control group,keloid fibroblasts experimental group showed the higher expression of collagen type Ⅲthan the control group,but the expression of collagen type Ⅲ in normal skin fibroblasts experimental group showed no significance difference compared with the control group.4. Real-time reverse transcription polymerase chain reaction prompt the higher expression of collagen type Ⅰ and collagen type Ⅲ in each experimental group.Conclusion Sparc proteins can significantly promote the proliferation in each experimental group,inhibit its apoptosis.Sparc proteins can significantly promote the expression of collagen type Ⅰ and collagen type Ⅲ in each experimental grouop.Therefore,SPARC protein may be an important protein to promote keloid formation,or may be targeted to regulate and inhibit keloid formation.