The Mechanism Of The Drug Pair "Peach Kernel And Safflower" On Intervening The Rat’s IVDD Cells Induced By IL-1β

Objective:The degeneration of intervertebral discs is the most fundamental pathological basis for the degeneration disc disease which more common in older patients. With the adverse changes in the way people live and work, such diseases becoming more and more younger severely affecting the life quality of patients.Early studies confirmed that HSYA could delay the degeneration of intervertebral discs, Peach kernel and safflower compatibility is the most classic of TCM. It may exist the potential to delay the degeneration of intervertebral discs. The experimental study investigated the protective effect and specific signaling molecules of the drug pair "peach kernel and safflower" and each monomer on the endplate chondrocytes induced by the inflammation which has an important guiding significance for clinical medicine.Methods:The endplate chondrocytes were isolated and cultured derived from the intervertebral disc of rat,10ng/mL IL-1β was adopted to induce the normal endplate chondrocytes of rat for building the inflammatory degeneration cell model, then by cytochemical staining, CCK-8cell proliferation assay, cell immunofluorescence, Flow cytometric analysis of cell apoptosis, the gene expression was detected by RT-PCR and the protein expression by Western blot for investigating the specific mechanisms of the drug pair and each monomer.Results:The endplate chondrocytes could occur the inflammatory degeneration induced by10ng/mL IL-1β, the different concentrations of HSYA could increase the mRNA expression of Col2al, Aggrecan, Sox9while decrease the mRNA expression of Col10al, ADAMTS-5, MMP-13, IL-1β and IL-6.The protein expression of phospho-ERK1/2has a significant trend at each time point.10-2mol/L Amygdalin could inhibit the proliferation of endplate chondrocytes, Flow cytometric analysis of apoptosis was significantly increased. A certain concentration of Amygdalin could increase the mRNA expression of Aggrecan, Col2al while decrease the mRNA expression of Col10al, MMP-13, IL-1β and IL-6.10-4mol/L Amygdalin could inhibit the protein phosphorylation of ERK1/2. The pharmaco-serum of the drug pair "peach kernel and safflower" could promote the cell proliferation and decrease the cell apoptosis. It also raised the mRNA expression of Col2al, Aggrecan, Sox9, down regulation the mRNA expression of Col10al and IL-6when increasing the proteoglycan and the protein expression of type Ⅱ collagen and decreasing the protein expression of type X collagen on the endplate chondrocytes. Amygdalin combined with HSYA could raise the mRNA expression of Aggrecan, Col2al while decrease the mRNA expression of Col10al, MMP-13, IL-1β and IL-6. It also promoted the protein expression of type II collagen and decreased the protein expression of type X collagen.Conclusion:The drug pair "peach kernel and safflower" and each monomer could inhibit the degeneration of endplate chondrocytes derived from the rat which induced by IL-1β and delay the degeneration. The mechanisms maybe the inhibition on the ERK1/2protein phosphorylation of the IL-1β downstream of the cascade of signaling molecules. The pharmacological effects of Amygdalin joint HSYA was better than monotherapy.