摘要
Objective:To observe the effects of kidney-tonifying and blood circulation-promoting recipes and platelet-rich plasma (PRP) on the activity and expression of type I collagen in co-culture system.Methods:The subcutaneous adipose tissue and degenerative nucleus pulposus tissue of patients with nucleus pulposus removal under lumbar intervertebral disc degeneration were isolated and cultured by multiple enzyme digestion methods to establish a co-culture system of ADSCs/NPCs. Patients with lumbar intervertebral disc degeneration were enrolled for 2 weeks.The venous blood was taken 50mL before the first dose and one hour after the last dose. Platelet-rich plasma was prepared by secondary centrifugation and cultured with different plasma. The experiment was divided into five groups: platelet-rich plasma group (PRP), platelet-poor plasma group (PPP), drug-rich platelet-rich plasma group (M-PRP) and drug-poor platelet plasma group (M-PPP), fetal bovine serum group (FBS). The morphological changes of the cells in each group were observed under an inverted microscope. The changes of type I collagen activity were detected by immunofluorescence assay. The expression of CCOL1A1 gene was detected by RT-PCR two weeks after the intervention.Results:In this study, the results of kidney-tonifying and blood circulation-promoting recipes and PRP group intervention were shown. The results of the group intervention showed that the M-PRP group and the PRP group had clearer cell contours and better refractive index than the other groups, and there was no significant difference between the two groups. The results of immunofluorescence staining showed that the number of cells in PRP group and M-PRP group was higher, but the staining intensity of single cells was significantly lower than that in FBS group. RT-PCR results showed that the expression of COL1A1 mRNA in PPP group, PRP group, M-PRP group and M-PPP group was significantly down-regulated (P<0.05), respectively: 0.343±0.040, 0.294±0.018, 0.310±0.022, 0.430±0.020.Conclusion:Kidney-tonifying and blood circulation-promoting recipes combined with platelet-rich plasma can decrease the activity and exhibit a tendency to inhibit the expression of type I collagen.
基金
Key project at central government level:The ability establishment of sustainable use for valuable Chinese medicine resources(2060302)
the National Natural Science Foundation of China(No.81674005,81904230,81804120,81302992)
the Fundamental Research Funds for the Central public welfare research institutes(ZZ10-015)
Beijing Natural Science Foundation Project(No.7164313)
China Postdoctoral Science Foundation(No.2017M611125,No.2016M591364).
