A Study On The Influence Of Gu Bi Zhi Tong Solution To The Exprssion Of TGF-?1 And Cbfa1 In The Cartilage Cells Of Rats Knee Osteoarthritis

Objective: In this study,Westar rats were selected as the research objects to observe the impact of GuBiZhiTong(GBZT)solution on TNF-??IL-6 in serum and TGF-?1 ? Cbfa1 in articular cartilage of rats with knee osteoarthritis through the animal experiment,so as to further explore the mechanism of articular cartilage repair and to provide theoretical basis for clinical application of GBZT liquid.Methods: 90 Wistar rats of SPF grade,45 males and 45 females,weight 280�20g,were randomly divided into six groups: sham operation group,model group,glucosamine sulfate group,high dosage group of GBZT,middle dose group of GBZT and low dose group of GBZT.There are 15 rats in each group,both male and female,and they are kept in cages between groups.The rats in group A were treated with skin incision and suture,the remaining 5 groups were treated with modified Hulth method to make models of knee osteoarthritis.After successful modeling,the serum and specimen was extracted after 8 weeks of continuous intervention of the drug:The morphological changes of cartilage were observed in the eyes;TNF-? and IL-6 in serum by ELISA;TGF-?1 mRNA and Cbfa1 mRNA in cartilage by RT-PCR,;and TGF-?1,Cbfa1 protein was detected by immunohistochemistry.Results:(1)Macroscopic observation:The articular cartilage in group A was smooth,transparent,lustrous,intact,no ulcer and no obvious change of the articular surface;The surface of the joints in group B was rough,the color was dark,and the transparency of cartilage was obviously decreased.,Cartilage erosion,ulcers formation,even exfoliation,defects and subchondral bone exposure;The cartilage transparency,colour and lustre of group F were better than that of group B,the surface of cartilage was rough,and the overall curative effect was a little worse than that of group E.(2)ELISA detection:Compared with group A,the level of serum TNF-a?IL-6 in group B was significantly higher than that in group A(P < 0.05);Compared with group B,the levels of TNF-a?IL-6 in serum of the C,D,E,F four groups were all lowerthan those of group,but the decreased was more obvious in group C and D(P <0.05);Compared with the D,E,F three groups,the group D was decreased more obviously,and the difference was statistically significant(P<0.05);There was no significant difference between group C and group D(P > 0.05).(3)PCR detection:The results of TGF-?1 m RNA detected by PCR showed that:The expression of TGF-?1 mRNA in articular cartilage of group B was significantly lower than that of group A(P<0.05);The expression of TGF-?1 mRNA in articular cartilage of the C,D,EF four groups was higher than that of group B,but the increase was more obvious in group C and D(P<0.05);Comparison of the expression of TGF-?1 mRNA in articular cartilage of D,E,F three groups,the group D was increased more obviously and the difference was statistically significant(P<0.05);There was no significant difference between group C and group D(P > 0.05).The results of Cbfa1 mRNA detected by PCR showed that:The expression of TGF-?1 mRNA in articular cartilage of group B was significantly higher than that of group A(P<0.05);The expression of Cbfa1 mRNA in articular cartilage of the C,D,E,F four groups was lower than that of group B,but the decrease was more obvious in group C and D(P<0.05);Comparison of the expression of Cbfa1 mRNA in articular cartilage of D,E,F three groups,the group D was decreased more obviously and the difference was statistically significant(P<0.05);There was no significant difference between group C and group D(P > 0.05).(4)Immunohistochemical detection of TGF-? 1,Cbfa1 protein expressionImmunohistochemical detection of TGF-? 1 protein expression:In group A,the expression of TGF-?1 was high in brown granules of articular cartilage.Compared with group A,the brown granules in articular cartilage tissue cytoplasm or nucleus of rats in group B were significantly lower than those in group A(P<0.05).Compared with the group B,the expression of cytoplasm and nuclear brown granules in articular cartilage tissue of C,D,E,F four rats increased,but the expression of C and D was increased more obviously(P<0.05).Comparison of the expression of TGF-?1 amongD,E,F three groups,the light density of D group increased significantly(P<0.05).There was no significant difference between group C and group D(P > 0.05).Immunohistochemical detection of Cbfa1 protein expression:The expression of Cbfa1 in articular cartilage of group A was less or no.Compared with group A,the brown granules in articular cartilage tissue cytoplasm or nucleus of rats in group B were significantly higher than those in group A(P<0.05).Compared with the group B,the expression of cytoplasm and nuclear brown granules in articular cartilage tissue of C,D,E,F four rats reduced,but the expression of C and D was decreased more obviously.(P<0.05).Comparison of the expression of Cbfa1 among D,E,F three groups,the light density of D group decreased significantly(P<0.05).There was no significant difference between group C and group D(P > 0.05).Conclusions:(1)GBZT could decrease the level of TNF-a?IL-6 in serum,increase the expression of TGF-?1 mRNA and TGF-?1 protein in articular cartilage,and decrease the expression of Cbfa1 mRNA and Cbfa1 protein in articular cartilage of KOA rats.(2)The expression of various factors was more obvious in the high-dose group and glucosamine sulfate group,which confirmed the therapeutic effect of GBZT on KOA,and confirmed that the curative effect of the high-dose GBZT group was more significant.