| Purpose:1 The clinical efficacy of the in-hospital preparation "Buzheng Xugu Pill" combined with artificial bone grafting in the treatment of early and mid-stage femoral head necrosis(ONFH)was analyzed retrospectively.A treatment method may prolong the survival time of the femoral head and provide objective clinical evidence.2 To explore the possible targets and signaling pathways of Buzheng Xugu Pill in the treatment of femoral head necrosis based on network pharmacology,and apply Buzheng Xugu Pill combined with artificial bone grafting to intervene in early and mid-stage femoral head necrosis in dogs to verify its effect and advantages.3 To explore the possible mechanism of Buzheng Xugu Pill combined with artificial bone grafting in the treatment of avascular necrosis of the femoral head at the level of molecular biology.Material and method:1 Analysis of the effect and curative effect of Buzheng Xugu Pill combined with artificial bone graft in the treatment of early and mid-stage femoral head necrosis through a clinical retrospective study.The cases in this study were collected from patients with early and mid-stage ONFH admitted to the Second Orthopedics Department of the Affiliated Hospital of Liaoning University of Traditional Chinese Medicine.A total of 95 cases(113 hips)were selected from September 2017 to May 2021 for retrospective analysis.Divided into exposure group(Buxing Xugu pill + artificial bone grafting)and control group(shock wave + TCM physiotherapy treatment),collect the basic information of patients: gender,age,BMI index,ARCO stage,TCM syndrome classification;collect treatment information : Hip Harris score,VAS score,hip replacement time,survival time before and after treatment and statistical analysis.Objectively evaluate its clinical efficacy.2 Based on network pharmacology and experimental verification to explore the effect of Buzheng Xugu pills combined with artificial bone grafting in the repair of avascular necrosis of the femoral head in dogs Through the traditional Chinese medicine system pharmacology database and analysis platform(TCMSP)and data platform(Batman-tcm),the chemical components of various traditional Chinese medicines in Buzheng Xugu Pills were retrieved,and the active components of the drugs were screened.The corresponding targets were screened out through the Uniprot database and a data set of drug active ingredient targets was established,and GO enrichment analysis and KEGG enrichment analysis were performed.It is speculated that the relevant pathways and targets of Buzheng Xugu pills and femoral head necrosis.For animal experiments,12 healthy adult beagle dogs with closed epiphysis were selected,6 males and 6 males,18 to 24 months old and 12 to 18 kg in weight.The patients were divided into normal control group,model group,Buzheng Xugu pill + artificial bone support group and Xianling Gubao capsule group + artificial bone support group by random number table method.The model of avascular necrosis of the femoral head was made by the improved liquid nitrogen freezing method.After the model was successfully established,the corresponding intervention methods(surgical intervention and oral administration of traditional Chinese medicine)were performed respectively.During the period,the general condition of the dog was observed to verify whether the model was successful.After 12 weeks of intervention The animals were sacrificed in batches,the general morphology of the femoral head was observed,the area of femoral head necrosis was observed at the imaging level(DR and MRI),and the femoral head of each group was scanned by micro-CT to observe the bone changes.In terms of pathology,HE staining was used to observe the histopathological changes of the dog’s femoral head,and ELISA was used to detect the expression levels of VEGF,b FGF and IGF-1 in the dog’s serum,and to explore the blood supply status of the vascular repair in the femoral head.3 Based on the endoplasmic reticulum stress PERK/eIF2α/ATF4 pathway to explore the mechanism of Buzheng Xugu pills combined with artificial bone grafting to repair avascular necrosis of the femoral head in dogs.Based on clinical experience and experimental research,it is concluded that Buzheng Xugu Pill combined with artificial bone grafting has a definite curative effect and effect on repairing femoral head necrosis.Network pharmacology was used to screen out the effective targets and pathways of Buzheng Xugu Pill,and it was found that the endoplasmic reticulum should Stimulation of PERK/e IF2α/ATF4 pathway may play an important role in the process of femoral head necrosis.At the molecular biology level,Real-time PCR,immunohistochemistry and Western blot were used to detect PERK/e IF2α/ATF4 pathway and its downstream The expression of factors CHOP,Bc L-2,BAX and Caspase-3 were detected,and the expression of apoptosis-regulating genes CHOP,Bcl-2,BAX and Caspase during the repair of avascular necrosis of the canine femoral head were detected,so as to explore the combination of Buzheng Xugu Pills Mechanism of artificial bone graft in repair of avascular necrosis of the femoral head in dogs.Result:1 A retrospective clinical study of corrective bone pills combined with artificial bone grafting for the treatment of necrosis of the femoral head in the early and medium termThrough years of clinical application experience,after summarizing and summarizing,using retrospective analysis to explore the effect of Buzheng Xugu pills combined with artificial bone support bone grafting in the treatment of early and mid-stage femoral head necrosis,a total of 95 cases(113 hips)were included,of which the exposure group 45 cases(55 hips)and 50 cases(58 hips)in the control group.There was no significant difference in baseline data such as age,gender,VAS score and Harris score before treatment between the two groups,and they were comparable.In terms of the total time of artificial hip replacement in the two cohorts: the average replacement time of the exposure group(Buizheng Xugu Pill+ artificial bone graft)was 375.86±119.52 days,and the average replacement time of the control group(shock wave + TCM physiotherapy treatment)was 167.33±126.66 days,the difference was statistically significant(t=3.877,p=0.001 < 0.05).In terms of the overall survival time of the femoral head of the two groups of patients: the average survival time of the femoral head in the exposed group was 624.73(days),and the survival time of the femoral head in the non-exposed group was 384.29(days).There was a statistical difference between the two groups(χ2=10.60,P = 0.001 < 0.05).Comparison of the time of artificial hip replacement in different ARCO(I-III)stages in the two cohorts: the average replacement time of the ARCOII exposure group: 380.75±153.90 days;the average replacement time of the control group: 178.88±109.78 days.have a statistical difference.The average replacement time of ARCOIII exposure group: 363.33±84.94 days;the average replacement time of control group: 111.10±39.94 days,with statistical difference.Finally,through COX regression coefficient analysis: according to different covariates(different ARCO stages,gender,Harris score,VAS score,etc.),the COX risk regression model was introduced to obtain the exposure group(artificial bone graft + correction Xugu pill)and different stages of the femoral head.There was a definite effect on the outcome event(hip replacement)That is to say,the application of artificial bone grafting and supplementing Xugu pills to intervene in ARCO(I-III)stage of femoral head necrosis can delay femoral head necrosis and delay the time of hip replacement.(χ2=1.782,P<0.05;χ2=2.435,P<0.05.Risk factor: RR(different groups)=0.168;RR(different stages)=0.188).2 Based on network pharmacology and experimental verification to explore the effect of Buzheng Xugu pills combined with artificial bone grafting in the repair of avascular necrosis of the femoral head in dogsBased on previous clinical experience,it was concluded that Buzheng Xugu Pills combined with artificial bone grafting can significantly improve the symptoms of femoral head necrosis in the early and middle stages,and delay the time of hip replacement.In order to further observe its curative effect,based on network pharmacology research,it was found that there are 137 active ingredients in Buzheng Xugu Pills,1089 Chinese medicine targets,and 140 disease targets.In the enrichment pathway speculation,apoptosis proteins BCL-2,Bax,and Caspase3 are closely related to the endoplasmic reticulum stress PERK/e IF2α/ATF4 signaling pathway.We used liquid nitrogen freezing to prepare canine femoral head necrosis model,and observed the changes of femoral head in each group.The femoral head in the core decompression group was irregular,with collapse,cartilage destruction,and severe hyperplasia of surrounding tissues.In the bone grafting + Buzheng Xugu Pills group,the femoral head was roughly spherical,with uneven distribution of cartilage,ducts in the greater trochanter,no collapse of the femoral head,and hyperplasia of connective tissue at the base.In the bone graft + Xianlinggubao group,the cartilage on the surface of the femoral head was destroyed,and the femoral head did not collapse.In terms of micro-CT scan,the results showed that the subchondral bone structure of the normal group was complete,and the bone density was higher.The support body + traditional Chinese medicine oral group observed that the bone structure was basically complete,the subchondral bone was not damaged,and the bone microstructure was slightly damaged.In the core decompression group and the model group,the subchondral bone was severely damaged,even collapsed,the bone density was low,the distribution was uneven,and the microstructure was severely damaged.In terms of pathology,HE staining showed that the rate of empty bone lacuna in the normal control group was 9.87±3.21%,and the rate of empty bone lacuna in the model group was 31.43±6.54%.The lacuna rate was 15.78±4.23%,and the empty bone lacuna rate in the Xianlinggubao combined with artificial bone support bone graft group was 11.22±2.31%;among them,4groups had statistical significance(P<0.05);Compared with the treatment group(Buzheng Xugu Pills,Xianling Gubao Capsules combined with artificial bone support bone grafting),the difference was statistically significant(P<0.05);but there was no significant difference between the two groups in the treatment group,and there was no statistical significance(P>0.05).In terms of serum detection,Buzheng Xugu pills combined with artificial bone grafting can increase the expressions of VEGF,b FGF,and IGF-1,which indicates that it is beneficial to angiogenesis and further realizes the repair of blood supply in the femoral head.3 Based on the endoplasmic reticulum stress PERK/e IF2α/ATF4 pathway to explore the mechanism of Buzheng Xugu pills combined with artificial bone grafting to repair avascular necrosis of the femoral head in dogsBased on the previous results,it is concluded that Buzheng Xugu Pill combined with artificial bone graft can repair femoral head necrosis.Therefore,we speculate that Buzheng Xugu Pill combined with artificial bone graft may regulate endoplasmic reticulum stress PERK/e IF2α/ATF4 pathway and relieve endoplasmic reticulum stress.Network stress,and then anti-apoptosis,repair femoral head necrosis.The results of Western blot showed that in the treatment group,after the treatment of femoral head necrosis with Buzheng Xugu pills +artificial bone support,Xianling Gubao capsule + artificial bone support,the protein expressions of PERK,e IF2α,ATF4,and BAX were down-regulated;The protein expression of apoptotic protein Bc L-2 was indeed up-regulated.In terms of immunohistochemistry,BAX and caspase 3 were observed to be expressed in the nuclei of osteocytes in each group,and the positive expression was light yellow or light brown.Strong positive responses were observed in the model group.BCL-2 was positively expressed in the nucleus,and the positive expression in the model group was lower than that in the normal group.By up-regulating the expression of Bcl-2 in canine necrotic femoral head,down-regulating the expression of Bax and Caspase-3,reducing cell apoptosis,prolonging cell survival time,further inhibiting the activity of osteoclasts,and realizing the repair of femoral head.Buzheng Xugu pills combined with artificial bone support bone grafting can affect the PERK/e IF2α/ATF4 pathway,thereby regulating angiogenesis and apoptosis,and alleviating the process of femoral head necrosis.Therefore,the PERK/e IF2α/ATF4 signaling pathway has potential application value in the development of drugs for the clinical treatment of necrosis of the femoral head.Conclusion:1 Buzheng Xugu Pills combined with artificial bone grafting is effective in clinical treatment,and may delay the time of hip replacement in patients with ARCOI-III stage necrosis of the femoral head.2 The artificial bone support has good osteoinductivity and mechanical properties matching with human bone,which can provide good support and avoid the possibility of femoral head collapse after lesion removal.3 The endoplasmic reticulum stress(PERK-e IF2α-ATF4)pathway is closely related to the pathological changes of the femoral head and may be a potential mechanism for the treatment of femoral head necrosis.4 Buzheng Xugu Pills combined with artificial bone support bone grafting can up-regulate the m RNA and protein expression levels of BCL-2 in the femoral head,and down-regulate the m RNA and protein levels of PERK、e IF2α、ATF4、CHOP、BAX and Caspase-3.The mechanism may be as follows.By relieving endoplasmic reticulum stress(PERK-e IF2α-ATF4),alleviating apoptosis is achieved. |
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