Experimental Study Of The Relationship Between The Cyclins During The G₁Phase In Osteonlast And The Remodeling Of Subchondral Bone In Knee Osteoarthritis And The Effect Of Touguxiaotong Granula On The Cyclins

1ObjectiveTo reveal the relationship between Cyclin D1、CDK6、p21and the remodeling of Subchondral Bone in Knee osteoarthritis, and research the effects of Touguxiaotong granul a (TGG) on the cycli ns during the G1phase i n osteobl ast of Subchondral Bone, SO that to explore the mechanism of TCM on the role of osteoarthritis(OA).2Methods2.1Animal experiment2.1.1Grouping and treatment with medicine:96rabbits were randomly divided into3groups:sham group (32rabbits), model group (32rabbits), experimental group(32rabbits).All groups do routine preoperative preparation and according the improvement Hulth method made mode to them except the sham group. One week after operti on, we driven all animals2times everyday, each time were30minutes, the model group and sham group were respectively perfused with normal saline10ml.d-1,the experimental group perfused with TGG10ml.d-1(contained medcine1.12g).After operation,8rabbits each of the rabbits remaded belong to the3group were sacrificed respectively in3,6,9,12weeks.2.1.2Detection:Through the X-ray radiography observation of macroscopical structure changes,and observed subchondral bone changes with light microscope i n the rabbi t knee. The subchondral bone mineral density changes were measured by dual-energy X-ray absorptiometry(DxA)for the rabbit knee,and Used RT-PCR method to detect the expression of CyclinD1CDK6and p21mRNA in osteoblast of subchondral bone.2.2Cinical researchAccording to our selection standards,60patients, they treated in the traditional Chinese medicine hospital of zhangzhou Fuj i an from October2010to July2011. Selected for this test and were randomly divided i nto two groups. Group A:taking TGG6g,2times a day for six weeks. Group B:taking glucosamine750mg,2times a day forsix weeks.The WOM AC Scori ng was adopted to evaluate the clinical outcomes of the two groups after0.4.8weeks they were treat.To Watch the knee pain symptoms and X-ray changes happened i n the two groups.3Result3.1The results of animal research3.1.1The results measured by X-ray:3weeks After operation, there were no obvious difference in three groups Comparison with each other,and the sham group have no obvious difference in all time points. By the end of the sixth week after operation, the medial knee joint space was narrow joint edge of a slight osteophyte formation in the model group and experimental group.and became more worse with the time, all has a gnifi cant differences Comparison with sham group (P<0.05); the experimental group higher than sham group, but low than model group, And also have a gnifi cant differences between them。3.1.2The results measured by Light microscopy:Three groups Comparison with themself on different time after opertion,thesham group had no change, there were cartilage degeneration and subchondral bone sclerosis gradually deteriorated in the model group with thetime, compared with sham group hasagnificantly difference,the experi mental group were less serious than those i n the model group,but higher than sham group;and by the end of thethreeth week after operation.Three groups have no difference.3.1.3The results measured by Bone mineral density:Three groups Comparison with themself on different time after operation, subchondral bone density in the sham group had no change, but in the model group increased agnificantly; and subchondral bone density i n the experi mental group were hi gher than the sham group, but lower than the model group。By the end of the sixth week after operation, three groups Compari son with each other have differences.3.1.4The results measured by RT-PCR:CyclinD1、CDK6、p21mRNA of subchondral bone in sham group have no change, but Cyclin D1、CDK6mRNA of subchondral bone in model group was significantly higher than that in sham group on different ti me after operation.On the contrary, p21mRNA of model group was significantly lower than that in sham group,and the experi mental group between them.3.2The results of clinical research3.2.1The results measured by WOMAC score:Two groups of WOMAC total score Were reduced significantly with the time, compared with themself has significantly difference, but did not seethe marked differences between the two groups.3.2.2The results measured by X-ray:therewere no obvious difference in two groups Comparison with each other or themselves.4Conclusion4.1The Cyclin D1、CDK6i n osteoblast of Subchondral Bone i n the process of Knee osteoarthri ti s i n model rabbi ts are hyper-expressed, but p21mRNA was On the contrary, and speedi ng up the cell mul ti pi i cati on and bone formation; So that the relative increase of Subchondral Bone cause the high transform osteoarthritis4.2TGG can increase the expression of p21expres-sion in osteoblast of Subchondral Bone i n Knee osteoarthriti s model rabbits, so that to sti mul ate cell prolif erati on of Subchondral Bone.It is perhaps one of mechani sms that TGG can cure osteoarthritis.4.3TGG can effective treatment for Knee osteoarthritis...