Discussion Tripterygium Medication At Different Times Of The Collagen–Induced Arthritis In Rats With Liver Injury

Objectives To investigate the effect of tripterygium glycoside on liver function in collagen–induced arthritis rats(CIA)at different times and the related mechanism,and to observe the therapeutic effect of CIA rat joint imaging,the tripterygium glycosides provide time toxicological evidence for the time treatment of rheumatoid arthritis.Methods 96 Wistar rats,females,weighing 180 ± 20 g.Wistar rats were placed in the time biology laboratory,fixed light(L): dark(D)time(L: D = 12:12,period 07:00 ~ 19: 00,dark 19: 00 ~ 07: 00),room temperature 23±2?,relative humidity of 60± 10%,free drinking water,to adapt to 10 days after the experiment.96 rats were randomly divided into normal control group(n = 16),model group(n = 80),the CIA model was established after grouping(except for the normal control group),the modeling results were observed and evaluated on day 21 of the first immunization,the successful model of the rats were randomly divided into CIA model group,tripterygium group(TG group),tripterygium polyglycoside + total glucoside group(TG+TGP group).TG group and TG +TGP group were randomly divided into two subgroups of ZT6 group and ZT18 group according to the time of drug intervention.From the first day of intervention,TG group(40mg / kg / d),TG+TGP group(TG40mg / kg / d,TGP200 mg / kg / d)were given to the CIA group,the normal control group and the model group were given the same amount of normal saline,were 1 time per day,continuous administration of 42 days.One day before sacrifice,the rat ankle,foot joint were taken with X–ray machine,and the imaging was observed and analyzed.After the end of the administration,take blood,take liver and lesion joints.The expression level of alanine aminotransferase(ALT),aspartate transaminase(AST),glutathione peroxidase(GSH–Px),superoxide dismutase(SOD)and tumor necrosis factor–?(TNF–?)in serum were detected.The expression of interleukin–6(IL–6),interleukin–18(IL–18),CLOCK and BMAL1 in synovial cells were detected by immunohistochemistry;Pathological examination of liver and joint tissues in rats by HE staining.The normal distribution of measurement data are the mean standard deviation((?)±s)said.The variance analysis of multi sample mean was used to judge the difference between the two groups,and the t test was used to judge the difference between the two groups.Test level ? =0.05.Results 1 Changes of liver function and serum SOD,GSH–Px and TNF–? in CIA rats:Compared with the normal control group,the levels of serum ALT,AST and SOD activity and GSH–Px in the model group did not show significant difference(P>0.05),the TNF–? content in the model group was significantly higher than that in the normal control group(P < 0.05).The levels of serum ALT and AST in TG group and TG+TGP group were significantly higher than those in normal control group and model group(P < 0.05)after 42 days of continuous administration.The levels of SOD activity and GSH–Px in TG group and TG + TGP group were significantly lower than those in normal control group and model group(P<0.05).Compared with the model group,the content of serum TNF–? in TG group and TG + TGP group decreased(P<0.05),and the content of TNF–? in TG + TGP group was significantly lower than that in TG group(P < 0.05).Compared with TG group,serum ALT and AST levels in TG+TGP group were significantly decreased(P < 0.05),serum SOD activity and GSH–Px content were significantly increased(P < 0.05).Compared with TG–ZT6 group,serum ALT and AST levels were lower,SOD activity and GSH–Px content were higher and TNF–? content was lower(P<0.05)in TG–ZT18 group.Compared with TG+TGP–ZT6 group,serum ALT and AST levels were lower,SOD activity and GSH–Px content were higher and TNF–? content was lower(P<0.05)in TG + TGP–ZT18group.2 The circadian rhythm of liver function and serum SOD,GSH–Px and TNF–?levels in CIA rats: The serological indexes of each group showed a certain rhythm change,ALT and AST generally showed a trend of high daytime low night,SOD and GSH–Px showed a trend of low daytime high night.Where the peak of ALT appeared at ZT18,the valley appeared at ZT6;AST peak reached the peak at ZT14,and the valley appeared at ZT22.When the peak of SOD appeared at ZT6,the valley appeared at ZT18 And the trend of ALT fluctuation is roughly the opposite;GSH–Px peak at ZT2,the valley appeared at the ZT14.The changes of TNF–? in the rats in the model group,TG group and TG+TGP group were larger than those in the control group,and the peak appears at ZT22 and reaches the valley at ZT14.3 The effects of day and night at different times application the TG and TGP on joint imaging in CIA rats: Through the X–ray can be observed in the normal group of rat ankle articular surface clear and etatarsophalangeal joint cortical were completed,there was no abnormality in the intervertebral space.In the CIA model group,the ankle joint was blurred and the bone defect was obvious.The metatarsophalangeal joint showed large bone defect and severe interphalangeal joint fusion.Compared with the model group,TG treatment group significantly reduced the degree of joint disease.In the TG–ZT6 group,the ankle joint surface was blurred,and the ankle joint defect was seen.The muscular defect was found in the metatarsophalangeal joint,and the interphalangeal joint was mildly merged.The articular surface of the ankle joint of TG–ZT18 group was clear,Less damage,mild changes in joint space.Compared with TG group,TG and TGP combined with the treatment group of mild degree of joint disease.TG+TGP–ZT6 group,the articular surface of the ankle joint was clear,but still a small number of bone defects,joint space slightly narrow;TG+TGP–ZT18 group of rats ankle joint surface is clear,metatarsophalangeal joint and joint space no obvious abnormal changes.4 The effects of day and night at different times application the TG and TGP on Liver Pathology in CIA rats: In the normal control group,the size of the hepatocytes was consistent,the structure was complete,the uniform and regular arrangement,no degeneration and necrosis of the liver cells.and the structure of the hepatic lobules was complete.Compared with the normal control group,the pathological changes of the liver in the model group were not obvious.TG–ZT6 group of rats with severe liver cell degeneration,part of the nucleus dissolved and dispersed,liver lobule structure damage,there are multiple points of foci necrosis,hepatic sinus widening and inflammatory cells;TG–ZT18 group of rats liver cell edema,steatosis,mild inflammatory cell infiltration,mild hepatic lobular destruction,part of the visible patchy necrotic area,can be observed interstitial fibrous tissue hyperplasia.TG+TGP–ZT6 group had mild edema of liver cells,occasional steatosis,liver lobular structure was normal,occasional inflammatory cell infiltration,mild interstitial fibrosis;TG + TGP–ZT18 group of liver cells no obvious edema degeneration,compared with the normal group of rats,slightly sinus sinus,even visible inflammatory cells.5 The effects of day and night at different times application the TG and TGP on joint pathology in CIA rats:The results showed that the synovial surface of the ankle joint in the normal control group was smooth,no papillary processes and hyperplasia,no multinucleated cell infiltration.CIA model group rats ankle synovial cell hyperplasia,showing several layers of synovial cell accumulation,synovial membrane can be seen at the edge of papillae bulge and a large number of inflammatory cell infiltration.Compared with the model group,TG treatment group significantly reduced the degree of joint disease.TG–ZT6 group also showed mastoid bulge and synovial cell hyperplasia,inflammatory cell infiltration;TG–ZT18group of rats ankle synovial hyperplasia is mild,still visible papillae bulge and a small amount of inflammatory cell infiltration.Compared with TG group,TG and TGP combined with the treatment group of mild degree of joint disease.TG + TGP–ZT6 group,the ankle synovial cell hyperplasia was mild,a small amount of papillae bulge,inflammatory cell infiltration;TG+TGP–ZT18 group of rats ankle synovial smooth,no significant proliferation and papillae bulge,a small amount of inflammatory cell infiltration.6 The effects of day and night at different times application the TG and TGP on expression of IL–6,IL–18 and Clock CLOCK and BMAL1 in CIA rats: The expression of IL–6,IL–18 and CLOCK and BMAL1 in the model group,TG time group and TG and TGP group were higher than those in the blank group.Compared with the model group,the expression of IL–6,IL–18 and CLOCK and BMAL1 were significantly decreased in TG treatment group and TG and TGP combined therapy group.The expression of IL–6,IL–18and clock CLOCK and BMAL1 in ZT18 group was significantly lower than that in ZT6 group.Conclusions 1 CIA rats liver function,liver pathology and normal rats compared to no significant abnormal changes;TG can lead to CIA rat liver function and liver pathology to varying degrees of injury.2 The levels of serum ALT,AST,SOD,GSH–Px in normal control group and CIA model group showed some circadian rhythm.The effect of TG on liver function of CIA rats was also showed diurnal changes,ZT18 administration group(nocturnal administration)was lighter than that of ZT6 group(daytime administration).TGP showed protective effect on hepatic injury induced by TG in CIA rats.3 TG group and TG and TGP combined group had significant inhibitory effect on joint injury and TNF–? level of serum in CIA rats,that showed some diurnal changes,and the effect of nocturnal administration was better.4 The diurnal differences of TG caused liver injury in CIA rats and its protective effect on CIA rat joints,may be related to its influence on the expression of CLOCK and BMAL1.