The Study On The Distribution Of IL-1β,C5a And Claudin-5 In The Brain Tissue Of Lupus Mice And The Intervention Of Nourishing Yin And Clearing Heat Formula

Objective 1.To observe the different expressions of cytokine IL-1β,complement factor C5 a,and blood brain barrier connection protein Claudin-5 in brain parenchyma and meninges of different weeks lupus mice,and investigate the dynamic laws of brain damage of lupus mice.2.By observing the expression changes of the cytokine IL-1β and complement factor C5 a under the influence of nourishing Yin Clearing heat Formula,to explore the therapeutic effect and mechanism of nourishing Yin Clearing heat Formula on NPSLE.Methods 1.10 of 6-week-old SPF level B6.MRL-Faslpr/Jnju female rats as small weekly model group,weighing 15.4 to 19.4(average 18.4)g;10 of 18-week-old SPF level B6.MRL-Faslpr/Jnju female rats as old weekly model group,weighing 19.6 to 23.6(average21.6)g;10 of6-week-old SPF level C57BL/6JN healthy female rats as small weekly normal group,weighing15.4 to 19.4((average18.4)g;10 of 30-week-old SPF level C57BL/6JN healthy female rats as old weekly normal group,weighing 29.5 to 35.1(average32.3)g.Comparing the express characteristics of IL-1β,C5 a,and Claudin-5 in brain parenchyma and meninges of different weeks of MRL mice by RT-PCR and immunohistochemistry.2.20 of 6-week-old SPF level B6.MRL-Faslpr/Jnju female rats,weighing 15.3 to 19.8(average 18.4)g,10 of 6-week-old SPF level C57BL/6JN healthy female rats,weighing15.5 to 19.4((average18.4)g;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/d 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.After 6 weeks,Comparing the express characteristics of IL-1β and C5 a in meninges of all mice by RT-PCR and immunohistochemistry.Results 1.The immunohistochemistry result showed that: In 6-week-old MRL mice,IL-1β was mainly expressed in the perivascular space of the meninges and cerebral microvascular endothelial cells;C5a was mainly expressed in brain parenchyma and meningeal vascular tissue;Claudin-5 was mainly expressed in the perivascular space of meninges and cerebral microvascular endothelial cells.RT-PCR showed,1.As the expression of IL-1β: It has no significant difference in brain parenchyma in four groups mice(P>0.05);in meninges,18-week-old MRL mice was significant higher than 6-week-old C57 mice(P=0.00),30-week-old C57 mice(P=0.00)and 6-week-old MRL mice(P=0.00),the 6-week-old MRL mice was significant higher than 6-week-old C57 mice(P=0.04).As the comparison in the same group between brain parenchyma and meninges:The expression of IL-1β in meninges were all significant higher than that of brain parenchyma in 30-week-old C57 mice(P=0.00),6-week-old MRL mice(P=0.01)and 18-week-old MRL mice(P=0.00);In 6-week-old C57 mice,the expression of C5 a in meninges has no significant differencewith that of brain parenchyma(P>0.05).As the expression of C5a: In brain parenchyma,6-week-old MRL mice was significant higher than 6-week-old C57 mice(P=0.00),30-week-old C57 mice(P=0.00)and 18-week-old MRL mice (P=0.00);In meninges,6-week-old MRL mice was significant higher than 6-week-old C57 mice(P=0.00),30-week-old C57 mice(P=0.00)and18-week-old MRL mice(P=0.00);As the comparison in the same group between brain parenchyma and meninges:The expression of C5 a in meninges were all significant higher than that of brain parenchyma in 30-week-old C57 mice(P=0.00),6-week-old MRL mice(P=0.03)and 18-week-old MRL mice(P=0.01);In 6-week-old C57 mice,the expression of C5 a in meninges has no significant difference with that of brain parenchyma(P>0.05).As the expression of Claudin-5: In brain parenchyma,18-week-old MRL mice was significant higher than 6-week-old C57 mice(P=0.01),30-week-old C57 mice(P=0.00)and 6-week-old MRL mice(P=0.00);In meninges,6-week-old MRL mice was significant higher than 6-week-old C57 mice(P=0.00),30-week-old C57 mice(P=0.01)and 18-week-old MRL mice(P=0.01);As the comparison in the same group between brain parenchyma and meninges:The expression of Claudin-5 in meninges were all significant higher than that of brain parenchyma in 30-week-old C57 mice(P=0.00),6-week-old MRL mice(P=0.00)and 18-week-old MRL mice(P=0.04);In 6-week-old C57 mice,the expression of Claudin-5 in meninges has no significant difference with that of brain parenchyma(P>0.05).2..After the intervention of Nourishing Yin Clearing Heat Formula,the immunohistochemical staining showed that the expression of IL-1β and C5 a in TCM group were lower than that in the model group,and the positive site decreased.RT-PCR showed,(1)As the expression of IL-1β in meninges: Model group was higher than TCM group but has no significance(P=0.535);Model group(P=0.00)and TCM group(P=0.01)were all significant higher than control groups.(2)As the expression of C5 a in meninges: Model group was significant higher than TCM group(P=0.00)and control group(P=0.00);TCM group was significant higher than control group(P=0.001).Conclusions 1.According to the express characteristics of IL-1β and C5 a in the brain parenchyma and meningeal tissue of lupus mice,we hypothesized that immune vasculitis may be the main pathogenic mechanism of NPSLE.According to theexpress characteristics of C5 a and Claudin-5 in the brain parenchyma and meningeal tissue of lupus mice,we believe that thedynamic change ruleof C5 a and Claudin-5 is of exploring significancefor the study of BBB mechanism in the incidence of NPSLE.2.Nourishing Yin Clearing Heat Formula can reduce the expression of IL-1βand C5 a in meninges of lupus mice,and we speculate that Nourishing Yin Clearing Heat Formula can ease the vasculitis in the brain of lupus,thus alleviating the process of NPSLE.