Study On The Mechanism Of Action And Clinical Efficacy Of Kidney Tonics Pill In Regulating Cellular Autophagy In The Treatment Of Steroid-induced Osteonecrosis Of The Femoral Head

Objective: In recent years,the treatment of Steroid-Induced Osteonecrosis of the Femoral Head(SONFH)is still focused on delaying the collapse of the femoral head and improving the quality of life of patients.Hip preservation is particularly important because of the difficulty of repairing osteonecrotic tissue and the high rate of disability,while the pathogenesis of SONFH is still unclear,so it is important to explore potential molecular markers for clinical diagnosis to improve the prognosis of patients.The clinical experience of the tutor,Tongue & Kidney Pill,has shown significant clinical efficacy in the treatment of SONFH,but its molecular mechanism is still unclear.In this study,we propose to screen the genes characterizing the autophagy of SONFH cells by bioinformatics methods,and investigate the clinical effects and mechanisms of SONFH treatment by Tonic Kidney Tongue Pill through cellular autophagy through clinical observation,so as to provide molecular basis and therapeutic ideas for the treatment of SONFH.Methods: 1.The SONFH dataset GSE123568 and human autophagy genes were obtained by applying searches from the GEO database and HADb database,respectively.Autophagy target genes associated with SONFH were screened by differential expression analysis,weighted gene co-expression analysis and Wayne analysis using R language software.Autophagy signature genes were further screened and identified by subject work curve analysis,and finally,real-time immunofluorescence quantitative PCR(q RT-PCR)was performed on autophagy signature genes to clarify their expression in the disease group.GO functional enrichment analysis and KEGG pathway enrichment analysis and immune cell infiltration analysis were also performed for the differential genes.Finally,q RT-PCR experiments were performed to verify the gene expression levels of autophagy signature genes in the normal and disease groups.2.Patients with SONFH who attended our hospital from September 2020 to June 2022 were selected,patient medical records were established,and those who met the inclusion criteria signed an informed consent form,and they were divided into experimental and control groups,with the experimental group given oral tonics and Tongue & Groove Pill and the control group given oral Henggu Bone The experimental group was given oral tonics and the control group was given oral healing agent for bone injury.The VAS score,Harris score and imaging evaluation were observed and analysed before and after treatment,and the differences between the two groups were compared according to statistical data.The m RNA content of the autophagy signature gene was measured by q RT-PCR before and after the treatment with Kidney Tonic Pill to illustrate the molecular mechanism of the drug.Results: The MEturquoise module of the WGCNA analysis was most closely associated with the clinical features of SONFH(number of genes 892).The results of the immuno-infiltration analysis showed a high proportion of neutrophils in the SONFH samples,suggesting that the disease was associated with inflammation,and the results of the GO and KEGG enrichment analyses showed that the differential genes were enriched in "immune response","immune response","inflammatory response" and "inflammatory response",respectively,The results of GO and KEGG enrichment analysis showed that the differential genes were enriched in "immune response","immune response","inflammatory response","chemokine signalling pathway","osteoclast differentiation","phagosome",etc.This suggests that the pathogenesis of SONFH may be related to immunity and inflammation.The ROC curves showed that STK11 and GABARAPL2 had high specificity(both AUC values > 0.9)and were selected as autophagy signature genes for subsequent clinical validation.2.q RT-PCR experiments showed that the m RNA levels of the two autophagy signature genes(STK11 and GABARAPL2)were significantly lower in the disease group than in the normal group,and there was a significant difference between them(P<0.05),which is consistent with the results expressed in the analysis of Sang Shin.3.The data on gender,age,BMI,site of disease,and ARCO stage,VAS score and Harris score of the hip joint before treatment were comparable between the two groups(P>0.05)and had no effect on the course and results of this study.4.Harris score and VAS score of the hip joint in the two groups Although the overall efficacy improved before and after treatment,the experimental group showed improvement in hip mobility and pain compared to the control group in the early and mid-treatment period for SONFH.5.During the follow-up period,the percentage of improvement was greater in the experimental group compared to the control group at the first month follow-up after treatment(71.29%).Although the difference was not significant,the experimental group had more potential therapeutic advantages and prospects than the control group in the process of femoral head repair.6.q RT-PCR results showed that in the serum samples of the experimental group treated with Kidney Tonic Pill,the expression levels of STK11 and GABARAPL2,two genes characteristic of autophagy,were gene expression levels were increased compared with those before treatment.Conclusions: 1.Two autophagy signature genes(STK11,GABARAPL2)associated with SONFH were screened by bioinformatics analysis.2.The m RNA content of the two autophagy signature genes(STK11,GABARAPL2)in the experimental group in q RT-PCR was expressed at lower levels in serum samples from patients in the disease group than in the normal group.3.In this study,the efficacy of Tonic Kidney Tongue Pill and Henggu Bone Injury Healing However,statistical analysis showed that the former was significantly more effective than the latter in treating early to mid-stage SONFH,mainly in relieving hip pain,improving hip mobility and delaying the process of femoral head collapse.4.q RT-PCR results showed that in the serum samples of the experimental group treated with Kidney Tonics Pill,STK11,GABARAPL2 and GABARAPL2 were expressed at lower levels than the normal group,The expression levels of two autophagy genes,STK11 and GABARAPL2,increased in the serum samples of the experimental group after treatment with Kidney Tonics Pill,compared with those before treatment,indicating that Kidney Tonics Pill could possibly treat SONFH by initiating cellular autophagy.It provides an effective treatment option for the early treatment of SONFH with kidney deficiency and blood stasis.