Correlative Study On The Expression Of ULK1? Beclin1 And LC3 In Articular Cartilage Of Knee Osteoarthritis With Yang-deficiency Knee

Objective: To investigate the distribution of TCM constitution types in patients with Knee Osteoarthritis(KOA)using epidemiological investigation methods,based on a clear hypothesis that KOA has a high constitution of Yang deficiency,Molecular biology method for the study of fatty acid synthase in lipid metabolism related proteins,and correlation between sterol regulatory element binding protein-1c(SREBP-1c)and ULK1,Beclin1,and LC3 in yangdeficient KOA articular cartilage.The in-depth analysis of the pathogenesis of KOA patients with yang deficiency in Gansu area provides an objective basis for the diagnosis and treatment of yang-deficient KOA patients,and provides new ideas and therapeutic targets for the prevention and treatment of KOA in traditional Chinese medicine.Methods: Refer to the "Chinese Constitution Classification and Criteria for Criteria" to develop a case survey form and print and bound together.From September 2016 to July 2017,a questionnaire survey was conducted on KOA patients admitted to the Department of Orthopaedic Inpatients of Hospital of Traditional Chinese Medicine in Gansu Province,and medical records,serum and cartilage specimens were collected in a total of 152 cases,including cases of Yang deficiency KOA patients.A total of 57 specimens were collected;95 cases of non-positive knee osteoarthritis patients were included in the data and specimens.During the same period,non-blood femoral neck fractures were collected for hip replacement surgery as a control group.76 non-osteoarthritis cartilage specimens of serum and femoral head were collected.,and then a database of KOA patient cases was established.Evidence-based method was used to select the indexes closely related to lipid metabolism and articular cartilage metabolism during the pathogenesis of KOA.The contents of FAS and SREBP-1c in serum of each group were detected by ELISA.The expression of autophagy-related genes ULK1,Beclin1,and LC3 A was detected by q-PCR in each group of cartilage.The relative expression of Beclin1 and LC3 B proteins in cartilage was measured by Western Blot,and analyzed by SPSS20.0 software.Results: a)A total of 152 cases of KOA patients,including 23 males and 129 females;There were 76 cases in the control group,including 29 males and 47 females.b)In a survey of 152 patients with KOA,it was found that yang deficiency,qi deficiency,and yin deficiency were the common biased constitutions of KOA,of which 57 were yang deficiency accounting for 37.50%,and 12 were yin deficiency,accounting for 7.89%.Qi deficiency was 16 cases,accounting for 10.52%.c)The results of ELISA showed that there were significant differences in serum FAS and SREBP-1c between groups(P<0.05).d)The results of 4q-PCR showed that the Beclin1,LC3 A and ULK1 genes were significantly different between KOA patients and controls(P<0.05).e)The results of 5WB showed that the average expression of Beclin1 protein in Yang deficiency group was lower than that in non-yang deficiency group and control group(P<0.05).The average expression of LC3 B protein in Yang deficiency group was higher than that in non-yang deficiency group.Compared with the control group,the difference was statistically significant(P<0.05).Conlusion: a)It was confirmed again that the yang-deficient substance indicated by the research group in the early stage was the type of high constitution of KOA.b)The work environment,nature of work,and education level are related to the occurrence of KOA.c)The gender(female),body mass index,and age were risk factors for the onset of KOA.d)FAS and SREBP-1c were highly expressed in KOA patients,and Yang deficiency group was higher than non-yang deficiency group,but there was no correlation between FAS and SREBP-1c in each group.e)Beclin1,LC3 A,and ULK1 were highly expressed in articular cartilage of KOA patients,suggesting that autophagy activity was higher in the articular cartilage of KOA patients,but there was no difference in the expression of the three genes between the Yang deficiency group and the non-yang deficiency group.f)FAS and SREBP-1c were negatively correlated with autophagy-related genes Beclin1,LC3 A,and ULK1 in yang-deficient KOA patients,suggesting that abnormal lipid deposition may affect the occurrence of KOA due to autophagy activity.