| Objective:To observe the changes of autophagy-related protein levels in peripheral blood lymphocytes before and after treatment with systemic lupus erythematosus(SLE)and the recovery of clinical laboratory indicators before and after treatment with sirolimus,analysis of sirolimus The effectiveness and mechanism of action for the treatment of systemic lupus erythematosus provide more reference data for clinical application of the drug.Methods:The patients were admitted to the Pediatric Outpatient Department of the Second Hospital of Hebei Medical University and the hospitalized SLE(56 cases)from December 2016 to December 1818.No sirolimus,hormones and other immunosuppressive agents were used before the treatment.Children with SLE were randomly divided into two groups,28 in the traditional treatment group and sirolimus group.The traditional treatment group was 10-15 years old(mean age 13±1.5,male: female 3:25),and the sirolimus group was 11-16 years old(mean age 13.4±1.4,male: female 2:26).15 healthy children who were admitted to the children's growth and development clinic in the same period were collected as normal control group,aged 8-15 years(mean age 11.8±2.4,male: female 2:13).Peripheral blood of normal children,children with pre-treatment SLE,children in traditional treatment group and sirolimus treatment group were collected,lymphocytes were extracted,and LC3 and P62 protein expression levels were detected by Western blotting.Immunofluorescence was used to detect LC3,P62 expression site,through the above data to analyze changes in lymphocyte autophagy-related protein levels.The clinical efficacy of sirolimus was evaluated by observing the changes of clinical laboratory indexes,the percentage of routine lymphocytes,complement C3,complement C4,serum Anti-dsDNA and SLEDAI in the traditional treatment group and sirolimus group.Results:1.Changes in clinical laboratory indicators in children with systemic lupus erythematosus1)Compared with the traditional treatment group,the percentage of routine lymphocytes in the sirolimus group increased at the end of 3 months,and the difference was statistically significant(P<0.05).At the end of 6 months,the percentage of routine lymphocytes increased,and the difference was statistically significant(P<0.05).2)Compared with the traditional treatment group,the sirolimus group decreased the SLEDAI score at the end of 3 months,and the difference was statistically significant(P<0.05).The SLEDAI scores decreased at the end of 6 months of treatment,and the difference was statistically significant(P<0.05).At the 3rd and 6th months of treatment,the positive rate of Anti-dsDNA decreased,which was statistically significant compared with the traditional treatment group(P<0.05).3)Compared with the traditional treatment group,the sirolimus group increased the complement C3 and complement C4 at the end of 3 months,and the difference was not statistically significant(P>0.05).At the end of 6 months of treatment,both complement C3 and complement C4 were elevated,and the difference was not statistically significant(P>0.05).2.Changes in autophagy-related protein levels1)Compared with normal children,the expression of LC3 protein in peripheral blood lymphocytes was significantly higher than that in normal children([0.3686±0.832] vs [0.5175±0.0721],t=-5.428,P<0.05);peripheral blood The expression of P62 protein in lymphocytes was decreased,which was statistically significant([0.6005±0.089] vs [0.4965±0.0731],t=3.456,P<0.05).Immunofluorescence was consistent with Western blotting.2)There was no significant difference in the expression of LC3 protein in peripheral blood lymphocytes between the SLE group and the traditional treatment group after 6 months of treatment([0.5175±0.0721] vs [0.5097±0.0911],t=0.445,P>0.05);There was no significant difference in P62 protein expression in peripheral blood lymphocytes,which was not statistically significant([0.4965±0.0731] vs [0.4942±0.0688],t=0.086,P>0.05).Immunofluorescence was consistent with Western blotting.3)Compared with the traditional treatment group,the expression of LC3 protein in peripheral blood lymphocytes was increased after sirolimus group treatment for 6 months([0.6485±0.1314vs][0.5097±0.0911],t=-3.245,P<0.05);P62 protein expression in peripheral blood lymphocytes decreased,which was statistically significant([0.4028±0.0765] vss [0.4942±0.0688],t=3.068,P<0.05).Immunofluorescence was consistent with Western blotting.Conclusions:1.The pathogenesis of systemic lupus erythematosus is associated with abnormal levels of autophagy in peripheral blood lymphocytes.2.Compared with traditional treatment,sirolimus can increase the level of autophagy in peripheral blood lymphocytes,indicating that it may play a role in the treatment of systemic lupus erythematosus by regulating the level of autophagy.3.Compared with traditional treatment,sirolimus has a faster recovery of clinical laboratory indicators,a higher percentage of recovery. |
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