| ObjectivesTo Study Apigenin with Hydrostatic Pressure Mediates the Change of Proteoglycan Synthesis and core protein and link protein related gene expression in the Human Lumbar Intervertebral. Disc Nucleus Pulposus, and analyze the data obtained, if Apigenin can improve the synthesis of proteoglycan and increase core protein with link protein gene expression,suggesting that apigenin can slow down the process of disc degeneration, which open up a new way to prevent and delay the clinical lumbar intervertebral disc degeneration.In-depth understanding the relationship between proteoglycans synthesis metabolism in lumbar intervertebral disc degeneration and the various Hydrostatic Pressure,clearing the proteoglycan core protein,link protein gene expression,and revealing lumbar intervertebral disc degeneration's molecular mechanisms, which have an important significance to lumbar intervertebral disc degenerative disease's prevention,diagnosis and treatment.Methods1.Apigenin's dose-dependent study on the intervertebral disc nucleus pulposus proteoglycan synthesis.12 samples of intervertebral disc nucleus pulposus,which are obtained from the volunteer patients due to lumbar disc herniation discectomy operation in the Department of Beijing University of Chinese Medicine Dongzhimen Hospital orthopedics,aged 50-60 years,mean 54.3 years of age. Add a medium low or high doses of apigenin (3.125-125μmol/L),to study the best dose apigenin with latm mediate the proteoglycan synthesis.2.Apigenin with Hydrostatic Pressure Mediates the Change of Proteoglycan Synthesisand core protein and link protein related gene expression in the Human Lumbar Interverte-bral Disc 32 samples of intervertebral disc nucleus pulposus,aged 40-60 years old with a mean age of 51.6 years.In which 40-50 years old male and female of all 8 cases,51-60-years old male and female of all 8 cases.Each is divided into nine cases of nucleus pulposus group(blank latm group,blank 3atm group,blank 30atm group,L-NMMA3atm group,L-NMMA 30atm group, apigenin 3atm group,apigenin 30atm group,SNAP3atm group,SNAP30atm group).cultured for 2 hours in the hydrostatic pressure devices,then measured the proteoglycan content in the supernatant and statistically analyzed the resul-ts.Eachgroup in the nucleus pulposus embedded in paraffin,sliced,forHE,Masson staining,immunohistochemistry and in situ hybridization,through the organizational and morphological analysis to study the apigenin on the intervertebral disc nucleus pulposus proteoglycan core protein and link protein gene transcription in vitro.Results1.Apigenin can inhibit nitric oxide release and promote synthesis of proteoglycan of the intervertebral disc nucleus pulposus,and there are dose-dependent manner,proteoglycan synthesis higher than other groups when adding 62.5μmol/L apigenin in the medium.2.To 9 group,apigenin 3atm group and L-NMMA 3atm group have the highest rate of proteoglycan synthesis,there was no significant difference between the two groups (P>0.05), there was significant difference compared with other groups(P<0.05),SNAP 30atm group have the lowest rate of proteoglycan synthesis,there were significantly different compared with the various groups (P＜0.01). In Control group, L-NMA group, SNAP group and apigenin group under the 3atm proteoglycan synthesis rates are higher than under the 30atm,compared with a significant difference (P<0.05); apigenin 3atm group and the L-NMMA 3 atm group proteoglycan core protein by immunohistochemistry and in situ hybridization results showed the largest number of positive nucleuspulposus cells,with a largest average optical density (MOD).and proteoglycan link protein's imm-unohistochemistry results showed the largest link protein's area percentage.Conclusions1.3atm's hydrostatic pressure can reduce exogenous nitric oxide(NO) the ratio of output and increase proteoglycan (PG) synthesis ratio of the intervertebral disc nucleus pulpo-sus,30atm's hydrostatic pressure can can increase exogenous nitric oxide(NO) the ratio of output and reduce proteoglycan(PG)synthesis ratio of the intervertebral disc nucleus pulposus.2.Apigenin can increase proteoglycan synthesis ratio of the intervertebral disc nucleus pulposus under 3atm's hydrostatic pressure,and reduce the 30atm's hydrostatic pressure's inhibition on proteoglycan synthesis ratio.3.This study reveals the part of molecular mechanisms of disc degeneration from the genetic level.Data and imagation analysis showed that for the degeneration intervertebral disc nucleus pulposus,Apigenin plays an important role in the promotion of proteogl-ycan core proteins and link protein mRNA gene expression,so we guess Apigenin may increase the extracellular matrix protein synthesis by increasing gene transcription levels of the nucleus, and can restruct proteoglycan core protein and link protein in the biochemical composition.Apigenin may take an action by stabilizing chondrocyte phenotype,it is necessary to study gene transcription fragment of the degeneration intervertebral disc nucleus pulposus cells,and thoroughly clear the mechanism of apigenin. |
