Diurnal Variation Of The Effects Of Tripterygium Glycosides And Tumor Necrosis Factor Inhibitors On Collagen-induced Arthritis

Objectives To observe the effect of Tripterygium glycosides(TG)and Tumor necrosis factor(TNF)inhibitors on collagen-induced arthritis(CIA)-related inflammatory cytokines Impacts provide laboratory evidence for the treatment of clinical rheumatoid arthritis.Methods 1 CIA modeling and grouping: Wistar rats,80 males,were reared in a time biology laboratory,with a bright period of 08: 00 to 20: 00 and a dark period of 20: 00 to 08:00.After 10 days of acclimatization,CIA models were made and models were successfully established.Rats were randomly divided into two groups: early 8:00(ZT0)and late 8:00(ZT12)groups;TNF inhibitors were early 8:00(ZT0)and late 8:00(ZT12).TG group was given intragastrically(10mg/kg)for 1 day;TNF inhibitor group was subcutaneously injected(6mg/kg),1/week,continued for 28 days;equal amount of carboxymethyl cellulose in the blank group and model group.Sodium gavage.After the intervention,blood was collected and joints were taken.2 Experimental method: After the beginning of treatment,the volume of joints in rats was measured weekly using a toe volume meter.After 28 days of treatment,the rats were photographed with X-rays of both feet in an orthotopic position.The rats were sacrificed and the serum was collected to detect TNF-? and Interleukin-6(IL-6)expression levels;Ankle joints of rats in each group were taken and pathological morphology of synovial tissue of the sacroiliac joints of rats was observed under a microscope.The synovial membrane TNF-? was detected by immunohistochemical method.The expression levels of TNF-?,IL-6,IL-17 A,BMAL1,and CLOCK were measured.After time treatment was completed,the experimental data were analyzed statistically.3 Statistical analysis: Analyze using IBM SPSS21.0 software,which is in accordance with the normal distribution measurement data,expressed as mean±standard deviation(x ±s).One-way ANOVA was used for comparison between groups;for comparison,the LSD method was used to test for variance;when the variance was not uniform,Dunnett's T3 method was used for testing,and the test standard was ?=0.05.Results 1 Changes of Toe Volume in Rats: There was no significant change in the toe volume in the blank group,and the toe volume of the model group increased.Compared with the TG group,the toe volume of the TG-ZT12 group was lower than that of the TGZT0 group.After 4 weeks of treatment,the toe volume of the TG-ZT12 group was smaller than that of the model group(P<0.05);the TNF inhibitor-ZT0 group had a toe volume Less than TNF inhibitor-ZT12 group,after 4 weeks of treatment,toe volume of TNF inhibitorZT0 group was smaller than model group(P<0.05).2 X-ray Observation of Ankle Joints in Rats in Each Group: There was no abnormal X-ray in the ankle of the blank group.In the model group,the bone surface of the bilateral sacroiliac joint surface was uneven,and obvious defects were seen.The metacarpophalangeal joint showed a massive bone defect,and the joint space became narrower.Compared with TG group,ZT12 group had less joint damage than T0 group.Compared with TNF inhibitors in the two groups,ZT0 group had less joint damage than ZT12 group.3 Serum TNF-? and IL-6 levels in rats in each group: After the intervention,compared with the model group,serum TNF-? and IL-6 levels in the TG group and the TNF inhibitor group were significantly decreased(P<0.05).Serum TNF-? and IL-6 levels in TG-ZT12 group were significantly lower than those in TG-ZT0 group(P<0.05).Serum TNF-? and IL-6 levels in the TNF inhibitor-ZT0 group were significantly lower than those in the TNF inhibitor-ZT12 group(P<0.05).4 The pathological observation of the sacroiliac joints of CIA rats in each group: There was no abnormality in pathological staining of the sacroiliac joint in the blank group.In the model group,pathological staining of the ankle joint showed severe synovial cell hyperplasia and extensive infiltration of inflammatory cells and papillary processes.Compared with TGZT0 group,TG-ZT12 group had less infiltration of inflammatory cells and less proliferation of synovial cells.Compared with TNF inhibitor ZT12 group,TNF inhibitor ZT0 group had less infiltration of inflammatory cells and less proliferation of synovial cells.5 Immunohistochemical detection of TNF-?,IL-6,IL-17 A,BMAL1 and CLOCK in rat ankle joints: Immunohistochemistry showed that the expression of inflammatory cytokines TNF-?,IL-6 and IL-17 A in the synovial cells of the model group was significantly higher than that of the blank group;meanwhile,the TG-ZT12 group was also found.Compared with TG-ZT8 group and TNF inhibitor ZT0 group,the expression of TNF-?,IL-6 and IL-17 A in synoviocytes was decreased in different degrees compared with TNF inhibitor ZT12 group.The expression of circadian rhythm protein BMAL1 and CLOCK was similar to that of inflammatory cytokines,and the expression of TG-ZT12 group was lower than that of TG-ZT0 group and TNF inhibitor ZT0 group compared with TNF inhibitor ZT12 group.Conclusions 1 Both TG and TNF inhibitors can effectively reduce joint swelling,inhibit inflammatory cytokines,reduce imaging progression of the sacroiliac joint in CIA rats,and reduce expression of circadian rhythm proteins BMAL1 and CLOCK.2 There is a difference between the optimal time of treatment of multi-target anti-rheumatic drug-TG and single-target anti-rheumatic drug-TNF inhibitor in the sooner or later.