To Investigate The Mechanism Of Miao Medicine Crossbow Acupuncture In The Treatment Of Knee Osteoarthritis In Rabbits Based On TRPV Channel Protein

Objective:Through the treatment of Knee osteoarthritis(KOA)model rabbits with miao medicine acupuncture,the effects of miao medicine acupuncture therapy on the expression of IL-1? TNF-?,and TRPV1 and TRPV4 in the synovial tissues of KOA rabbits were studied,and the mechanism of miao medicine acupuncture in the treatment of Knee osteoarthritis was further discussed.Methods:Thirty-four rabbits from New Zealand were randomly divided into 6 rabbits from normal group and 28 rabbits from model group.The knee joint cavity injection of papain and L-cysteine mixture method to establish model of KOA to randomly selected 4 only after completion of building model evaluation,and the remaining rabbit randomized,make up for with the normal group:normal group,model group,crossbows,needle,dermal needle group and daub,in 6 experimental rabbits in each group.After successful modeling,rabbits in the normal group and model group were raised normally without any intervention.The other three groups were given corresponding treatment three times a week for a total of 4 weeks.During the experiment,the general conditions of rabbits were observed and the threshold of reduced foot of mechanical stimuli was detected(von Frey test).After the treatment,the articular fluid,cartilage and synovium of the knee were taken for testing.HE staining was used to observe the morphological changes of cartilage in the knee joint of rabbits.The contents of IL-1? and tumor necrosis TNF-? were determined by enzyme-linked immunosorbent assay(ELISA).The relative expressions of TRPV1 and TRPV4 in synovial tissues were detected by Western blot.The relative expression levels of TRPV1mRNA and TRPV4mRNA in synovial tissues were detected by reverse transcription-polymerase chain reaction(RT-PCR).Results:(1)General observation:compared with the normal group,after the second injection of modeling reagent,rabbits in the model group gradually showed abnormalities,including listlessness,dull color of fur,decreased activity,lameness,decreased food intake,loose stool,irritable mood,and local redness and swelling of the right knee joint compared with the left knee joint.Compared with the model group,the mental state of the rabbits in the crossbow medicine needle group,the dermatology needle group and the smear group was between the normal group and the model group.(2)Comparison of the von Frey test:compared with the normal group,the von Frey threshold of rabbits in the model group was significantly lower(P<0.01)and continued until the end of the experiment.Compared with the model group,the von Frey threshold was significantly increased in the crossbow needle group,the dermatology needle group and the smear group(P<0.01,P<0.05).Compared with the crossbow needle group,the von Frey threshold in the dermatology needle group and the smear group was significantly reduced(P<0.01).(3)Morphological comparison of articular cartilage:the surface of articular cartilage in the normal group was not worn,smooth and smooth,with clear structure,orderly arrangement of chondrocytes,orderly tidal line and even matrix staining.The articular cartilage surface of rabbits in the model group was uneven,seriously damaged,and the structure was disordered.Chondrocytes were clustered locally,the tidal line was severely distorted,and the matrix staining was uneven or even disappeared.The crossbow needle group,the dermal needle group and the smear group all improved to a great extent,and the morphological structure was between the normal group and the model group.(4)Comparison of the contents of IL-1? and TNF-? in the joint fluid:compared with the normal group,the contents of IL-1? and TNF-? in the joint fluid of the model group were significantly increased(P<0.01).Compared with the model group,the levels of IL-1? and TNF-? in the joint fluid of the crossbow acupuncture group,the dermatology acupuncture group and the smear group were significantly decreased(P<0.01,P<0.05).Compared with the crossbow acupuncture group,the levels of IL-1? and TNF-? in the joint fluid increased(P<0.05).(5)Comparison of the relative expression levels of TRPV1 and TRPV4 proteins in synovial tissues:compared with the normal group,the relative expression levels of TRPV1 and TRPV4 proteins in synovial tissues of the model group were significantly increased(P<0.01).Compared with the model group,the relative expression levels of TRPV1 and TRPV4 in the synovial tissues of rabbits in the crossbow needle group,the dermatology needle group and the smear group were significantly decreased(P<0.01).Compared with the crossbow acupuncture group,the relative expression levels of TRPV1 and TRPV4 in the synovial tissues of the dermal acupuncture group and the smear group were increased(P<0.01,P<0.05).(6)Comparison of the relative expressions of TRPV1 and TRPV4mRNA in synovial tissues:compared with the normal group,the relative expressions of TRPV1 and TRPV4 mRNA in synovial tissues of the model group were significantly increased(P<0.01),Compared with the model group,the relative expressions of TRPV1 and TRPV4mRNA in the synovial tissues of rabbits in the crossbow medicine needle group,the dermatology needle group and the smear group were significantly decreased(P<0.01).The relative expression levels of TRPV1 and TRPV4 mRNA in the synovial tissues of the dermatology needle group were not significantly different from that of the crossbow needle group(P>0.05).The relative expression level of TRPV1 mRNA in the synovial tissues of the smear group was increased(P<0.05),and there was no significant difference between the relative expression level of TRPV4 mRNA and that of the crossbow acupuncture group(P>0.05).Conclusion:(1)Abnormal pain sensitivity exists in KOA model rabbits.In this experiment,the three intervention measures of crossbow needle,dermal needle and smear can all improve the von Frey threshold and improve pain sensitivity,which is consistent with the previous clinical results of the research group.(2)Crossbow acupuncture,cutaneous acupuncture and application therapy can improve chondropathy,inhibit the secretion of inflammatory factors,down-regulate the expression of TRPV1 and TRPV4 channel proteins,and delay the course of KOA,and crossbow acupuncture has the best effect.(3)The improvement of KOA pain sensitivity mediated by TRPV1 and TRPV4 by crossbow injection may be related to its benign regulation of inflammatory response.