The Effect Of Gubi Mixture On Expression Of NF-?B And Bad Protein Downstream PI3K/AKT In Chondrocytes Of Rat Knee Osteoarthritis

Objective: The rat knee osteoarthritis(KOA)model was made by intraparenchymal injection of papain in the knee joints of rats.HE staining was then performed on paraffin sections and laboratory models were successfully established in open field experiments.Finally,the joints of the knee joints were removed.Cartilage tissue was used to detect the expression of genes related to Bad(Bcl-2-associated death promoter)and NF-?B(nuclear factor kappa-light-chain-enhancer of activated B cells)proteins in cartilage tissue by western blot and q-PCR.Then we observed the effect of Gu Bi Mixture on chondrocyte apoptosis in rat knee osteoarthritis.At the same time,explore the possible mechanism of the treatment of osteoarthritis of the knee with Gu Bi mixture.Methods: Ten-week-old male SD rats(300-325 g;10-12 weeks old;n=32)were placed in a ventilated animal room at a controlled temperature(16°C)and humidity(29 RH%),and the pressure difference in the cage(25 Pa).Ventilation frequency(050 N)Wind power 99,air intake power 96.12 hours light/dark cycle,free to provide food and water.Pentobarbital sodium was intraperitoneally anesthetized,and 50 ?l of 8% papain was injected into the joint cavity of the right knee joint of each SD rat using a 100 ?l syringe to prepare rat knee osteoarthritis.The movement of the rats was continuously observed during the modeling.The rats were randomly divided into three groups(10 in each group):normal saline group,aminomethine group,and Gu Bi mixture group.According to the Meeh-Rubner formula,rats in the normal saline group were given intragastric administration of gavage,and rats in the Gu Bi mixture were fed with Gu Bi Mixture(provided by Yunnan Traditional Chinese Medicine Hospital).Rats in the positive control group were given intragastric administration for each administration of lycosaminoglycin(approved symbol:Guo Yaozhun H20051760 provided by the Yunnan Provincial Hospital of Traditional Chinese Medicine),and each group had intragastric administration for four weeks.Then each rat was subjected to open-field test,HE staining,western blot and q-PCR experiments to detect the transcription and protein expression of NF-?B and bad proteins and related genes downstream of the PI3K/AKT signaling pathway.Result: In the open field test and rat knee cartilage tissue section and HE staining,it can be seen that the number of upright occurrences and the exaggeration activity of Gu Bi Mixture group was significantly better than that of saline group(P values of both weeks and weeks were less than 0.05,existed(Statistical differences):At the same time,the number and arrangement of cartilage cells in the Gu Bi Mixture group were significantly improved compared with the normal saline group,and the cartilage extracellular matrix was also significantly increased compared with the saline group.In the Western Blot trial,the NF-?B protein in the cartilage of the knee joint in the Gu Bi mixture group was significantly higher than that in the normal saline group and the aminosulfurin group,and the Bad protein was significantly higher in the cartilage than in the ampicillin group and the saline group.The reduction.Finally,in q-PCR experiments,the transcription of NF-?B protein-related genes was increased compared to the saline group,but slightly lower than that of the aminomycin group.The transcription of related genes of the Bad protein was significantly lower than that of the saline group and the amimycin group.Conclusion: The Gu Bi mixture increased the expression of NF-?B protein in cartilage cells in the rat knee osteoarthritis model,while the bad protein also decreased.Due to the increase of NF-?B protein in rat KOA knee cartilage tissue,it promotes the cell Addition and differentiation,while the reduction of Bad protein can reduce the apoptosis of chondrocytes.It can be concluded that Gu Bi Mixture can regulate the expression of NF-?B and Bad protein in the downstream of PI3K/AKT signaling pathway and the transcription of protein-related genes in rat knee cartilage cells,promote the proliferation of cartilage cells,and inhibit cartilage.Apoptosis of the cells improves the symptoms of rat knee osteoarthritis.