| Objective: To explore the effect of Huayu Qushi Decotion on the expression level of the key genes of necroptosis in KOA(Knee Osteoarthritis)by using bioinformatics technology and clinical intervention test,in order to provide a new way to explain the pathogenesis of KOA;At the same time,it also provides theoretical basis for revealing the treatment of KOA with the prescription for Huayu Qushi Decotion at the molecular level.Methods:(1)Bioinformatics: KOA-related genes and necroptosis-related genes were screened from the GEO database and the Gene Card database respectively,and then the related genes of necroptosis in KOA were obtained by the intersection of the two,and the PPI network was further constructed.The related Hub genes of necroptosis in KOA was obtained by the five calculation methods of the Cytoscape software,and then SD rats were selected to model,and the relative expression of the Hub gene was detected by RT-PCR after extraction,Finally,the first three genes with the most significant differential expression in rat KOA model were selected as the Key genes(target genes)of KOA necroptosis.(2)Clinical intervention test part:after screening the target gene,42 outpatients with KOA who met the criteria were randomly divided into two groups: the test group(Huayu Qushi Decotion group)and the control group(Irecoxib group),with 21 patients in each group;The VAS score and WOMAC score were evaluated before and 2wee Ks after treatment,and the expression level of target gene was detected by ELISA with peripheral venous blood,and then the inter-group and intra-group comparisons were made and the overall treatment rate was counted.Results:(1)Bioinformatics:990 KOA-related genes and 690 necroptosisrelated genes were screened,and 38 related genes of necroptosis in KOA were obtained after intersection;Cytascape’s five different algorithms further screened five Hub genes: CYBB,FADD,IL-1β,MYC and EGFR;And RT-PCR detection indicated that in rat KOA model IL-1β,MYC and EGFR are the first three genes with the most significant differential expression(P<0.05),namely IL-1β,MYC and EGFR are the key genes of necroptosis in KOA.(2)Clinical intervention test part: 20 subjects in the test group(1case dropped off),and 21 subjects in the control group participated in the test process completely;General data(gender,age,course of disease,K-L grade)and observation indicators(VAS score,WOMAC score and the expression levels of IL-1β,MYC and EGFR)had no significant difference(P>0.05),which was comparable;Compared with before treatment,the observation indexes(VAS score,WOMAC score and and the expression levels of IL-1β,MYC and EGFR in control and test group after treatments)were statistically significant(P>0.05);After treatment,The each observation index(VAS score,WOMAC score and and the expression levels of IL-1β,MYC and EGFR)was statistically significant of the comparison between.And through the comparison before and after treatment,in terms of VAS score,the difference in the control group is greater than that in the test group,but in terms of WOMAC score and and the expression levels of IL-1β,MYC and EGFR,the difference degree of the test group was greater than that of the control group;In addition,the overall treatment rate was 80% in the test group and 74.1% in the control group,but there was no significant difference between the two groups(P>0.05).Conclusion(s):(1)IL-1β,MYC and EGFR are the key genes of necroptosis in KOA.(2)Huayu Qushi Decotion can play a similar role in the treatment of KOA as iricoxib,and may affect The expression level of IL-1β,MYC and EGFR in KOA through necroptosiss pathway;Huayu Qushi Decotions regulatory effect of IL-1β,MYC and EGFR may be stronger than that of iricoxib.(3)The analgesic effect(VAS score)of Huayu Qushi Decotion is not as good as that of iricoxib,but it is better than that of iricoxib in terms of pain relief,joint stiffness and improvement of joint activity(WOMAC score),and the treatment rate of Huayu Qushi Decotion is higher than that of iricoxib. |
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