Based On The Zinc-ZIP8-MTF1 Axis To Study The Mechanism Of Du Huo Ji Sheng Decoction In The Treatment Of Knee Osteoarthritis

Objective:By establishing a C57BL/6 mouse knee osteoarthritis（Knee osteoarth ritis,KOA）animal model,and giving the intervention of Duhuojijie Decoction,it is ev aluated that Duhuojijie Decoction regulates the Zinc-ZIP8-MTF1 axis,produces articu lar cartilage protection and prevents KOA.To explore the mechanism of action of Duh uojii Decoction in the treatment of KOA,and provide laboratory evidence for Duhuoji i Decoction in the treatment of KOA.Method:54 C57BL/6 mice were selected,and 18 mice were randomly selected a s the blank group（Control group）.The remaining mice were used Destabilization of th e medial meniscus（DMM）to construct a mouse knee osteoarthritis model After confir ming that the model was successfully established,the experimental mice were random ly divided into the model control group（Model group）,the Duhuojijitang group（DHJ ST group）,with 18 mice in each group;the DHJST group was given 18.6g/kg Duhuoji jitang by gavage,Model The group was given an equal volume of normal saline by ga vage,the control group was not given;once a day,after 4 weeks of continuous admini stration,the mice were sacrificed,and the intact knee joints were taken for Safranin-O staining to observe the pathological changes of the knee joint cartilage of mice in each group,Use Fluo Zin-3 AM to detect the Zn²⁺concentration in chondrocytes;Take the knee joint cartilage,use RT-q PCR to detect cartilage ZIP8,ZNT9,MTF1,Mt1,Mt2,MMP3,MMP13,COL2A1,LC3 m RNA expression,use Western blotting to detect ca rtilage ZIP8,ZNT9,MTF1,MT,MMP3,MMP13,COL2A1,LC3 expression.Results:Compared with the Control group,the mouse cartilage histopathology s core in the Model group was significantly increased（P<0.05）,and the difference bet ween the two groups was statistically significant.Compared with the Model group,the mouse cartilage histopathology score in the DHJST group Significantly decreased（P<0.05）,the difference between the two groups was statistically significant;Compared with the Control group,the Zn²⁺content in the cartilage cells of the Model group was significantly increased.Compared with the Model group,the Zn²⁺content in the cartil age cells of the DHJST group was greatly reduced.Compared with the Control group,the expression of ZNT9,MTF1,MMP3,MM P13,and LC3 m RNA in the knee cartilage tissue of the Model group increased signifi cantly（P<0.05）,and the difference was statistically significant.Compared with the Co ntrol group,the Model group ZIP8,The expression of COL2A1 m RNA was significan tly reduced（P<0.05）,and the difference was statistically significant.Compared with t he Control group,the expression of Mt1 and Mt2 m RNA in the Model group had no si gnificant difference（P>0.05）,and the difference was not statistically significant;com pared with the Model group Compared with the DHJST group,the expression of ZIP8,ZNT9,MTF1,MMP3,and MMP13 m RNA in the cartilage tissue of the knee joint of mice in the DHJST group was significantly reduced（P<0.05）.Compared with the M odel group,the expression of Mt1,Mt2,COL2A1,LC3 m RNA expression increased significantly（P<0.05）,the difference was statistically significant.Compared with the Control group,the expressions of ZIP8,MTF1,MMP3,MMP13,and LC3 protein in the knee cartilage tissue of the Model group increased significa ntly（P<0.05）,and the difference was statistically significant.Compared with the Cont rol group,the Model group mice The expression of COL2A1 protein in the cartilage ti ssue of the knee joint was significantly reduced（P<0.05）,and the difference was statis tically significant.Compared with the Control group,the expression of ZNT9 and MT protein in the cartilage tissue of the knee joint of mice in the Model group had no sign ificant difference（P>0.05）,The difference is not statistically significant;compared w ith the Model group,the expression of ZIP8,MTF1,MMP3,and MMP13 proteins in t he cartilage tissue of the knee joint of the DHJST group was significantly reduced（P<0.05）.Compared with the Model group,the DHJST group mice The expression of C OL2A1,MT,LC3 protein in the cartilage tissue of the knee joint increased significantl y（P<0.05）,and the difference was statistically significant.Compared with the Model group,the expression of ZNT9 protein in the cartilage tissue of the knee joint of mice in the DHJST group had no significant difference（P>0.05）,not statistically significan t.Conclusion:1.Duhuojijietang can reduce the expression of Zn²⁺,thereby inhibiti ng the expression of MTF1,MMP3,MMP13,COL2A1,and ultimately protect the kne e joint cartilage,revealing the mechanism of Duhuojijietang for treating knee osteoart hritis and Zinc-ZIP8-MTF1 There is a correlation between the axes.2.Duhuojijietang has no obvious effect on the key links of Zn²⁺entering and leaving cartilage cells.Duh uojijietang can protect knee articular cartilage by reducing Zn²⁺,which may be related to increasing the expression of articular cartilage LC3.