| Objective 1.By observing the difference of the expression of Pyroptosis factors and regulatory gene NLRP3 between lupus mouse and lipopolysaccharide in lupus and lupus mice,to investigate the relationship between Pyroptosis pathway and regulatory gene NLRP3 and SLE inflammation.2.Through observe the expression of Pyroptosis factors and NLRP3 in the kidney tissue of the lupus mice under the influence of Nourishing Yin Clearing Heat Formula,to investigate the effect of Nourishing Yin Clearing Heat Formula on SLE inflammation.And to observe the effect of Nourishing Yin Clearing Heat Formula on the treatment of SLE renal tissue by electron microscopy,so as to provide ideas and methods for the study of SLE inflammation of Nourishing Yin Clearing Heat Formula.Methods 1.20 of SPF level B6.MRL-Faslpr/Jnju female rats,10 of SPF level C57BL/6JN healthy female rats,and all the rats are week age 8-10 weeks.According to the weight and growth condition,the B6.MRL-Faslpr/Jnju female rats were randomly divided into two groups:(1)LPS Group(n=10): LPS was injected intraperitoneally at a dose of 8 mg.kg-1 once every two days for 6 weeks.(2)Model Group(n=10): Intraperitoneal injected NS at an equal volume of LPS once every two days for 6 weeks.C57BL/6JN healthy rats was(3)Control Group(n=10): Intraperitoneal injected NS at an equal volume of LPS once every two days for 6 weeks.Observed and recorded each mouse body weight,activity,changes in physical appearance and other medication.After six weeks,the expression of Caspase-1,IL-1β and NLRP3 in the renal tissue of each group were detected by immunohistochemistry and fluorescence quantitative PCR.2.20 of SPF level B6.MRL-Faslpr/Jnju female rats,10 of SPF level C57BL/6JN healthy female rats,and all the rats are week age 8-10 weeks.According to the weight and growth condition,the B6.MRL-Faslpr/Jnju female rats were randomly divided into two groups:(1)TCM Group(n=10): Nourishing Yin Clearing Heat Formula by 20 g.kg-1.d-1 dose of gavage(2 times the equivalent dose)every day for 6 weeks.(2)Model Group(n=10): TCM equal volume of intragastric distilled water gavage every day for 6 weeks.C57BL/6JN healthy rats was(3)Control Group(n=10): TCM equal volume of intragastric distilled water gavage every day for 6 weeks.Observed and recorded each mouse body weight,activity,changes in physical appearance and other medication.After 6 weeks,the pathological changes of glomeruli were observed by electron microscopy,and the expression of Caspase-1,IL-1β and NLRP3 in the renal tissue of each group were detected by immunohistochemistry and fluorescence quantitative PCR.Results 1.Immunohistochemistry showed that Caspase-1 and IL-1β expression of LPS group and model group was more sites than control group,deeper dyeing;and positive lupus mice after LPS stimulation sites more than the model group;Comparison of average optical density values,the expression of Caspase-1 and l L-1β in the model group was higher than that in the control group(p<0.05),and the LPS group was higher than the model group(p<0.05),and compared with the control group Significant(p<0.01).The results of fluorescence quantitative PCR showed that the m RNA expression of Caspase-1,l L-1β and NLRP3 of renal cortical in the model group was higher than that in the control group(p<0.05),but the expression of LPS group was higher than that of the model group(p<0.05).2.The expression of IL-1β in the TCM group was significantly lower than that in the model group in the results of Immunohistochemistry(P<0.05).Comparison the results of average optical density values and fluorescence quantitative PCR,the expression of IL-1β in the TCM group was significantly lower than that in the model group(p<0.05).However,there was no significant difference between TCM group and model group of the expression of Caspase-1 in the results of average optical density values and fluorescence quantitative PCR(P<0.05),but the value in TCM group lower than the in model group,tended to decrease.The results of fluorescence quantitative PCR showed that the expression of NLRP3 m RNA in TCM group was lower than that in model group,but the difference was not statistically significant(p>0.05).The results of immunohistochemistry showed that the distribution of Nephrin expression in the TCM group was higher than that in the model group,but still lower than that in the control group,and the average optical density in the TCM group was higher than that in the model group(P<0.05).Electron microscopy showed that the TCM group could improve the glomerular and renal tubular degeneration in the renal cortex.The results of the podocyte density showed that the TCM group was higher than the model group(P<0.05).Compared with the control group,the the podocyte density of TCM group and the model group were decreased(P<0.05)also the width of the podocyte foot was significantly increased in the model group and TCM group(P<0.01),while the traditional Chinese medicine group was better than the model group(P<0.05).Conclusions 1.Pyroptosis factors and regulatory factors NLRP3 may be an important pathway for SLE inflammatory reaction.2.Nourishing Yin Clearing Heat Formula can reduce the expression of pyroptosis factors and regulatory factor NLRP3,and can reduce the renal podocyte injury and decrease podocyte pore membrane related molecules Nephrin protein expression,which may be an important pathway to control SLE inflammation of Nourishing Yin Clearing Heat Formula. |
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