To Investigate The Effect Of Acupuncture Through "Baihui" To "Qubin"on The Mincle/syk Pathway In A Rat Model Of Intracerebral Hemorrhage

Objective:To investigate the effect of acupuncture through "Baihui"to "Qubin"on the Mincle/Syk pathway in a rat model of intracerebral hemorrhage.Method:216 experimental SD rats were randomly divided into sham group,model group and acupuncture group,and each group was divided into four sub-groups according to the time points of 6h,12h,24h and 72h after the molding.The ICH rat model was established using autohemic blood infusion.Rats in the sham group underwent all the previous operations,and were injected into the caudate nucleus without injecting blood.Rats in the sham group received both grasping and fixation but no acupuncture at the same time every day.More specifically,the acupuncture group was treated with hand-twist acupuncture of Baihui to Qubin(DU20 through GB7 on the lesion side).Modified neurological severity scores(mNSS)and brain water content was assessed after establishment of the ICH model.Expression of Mincle,Syk,and CARD9 was detected using immunohistochemistry and western blot.The levels of IL-1βmRNA were measured using a real-time PCR assay and ELISA.The histopathological changes were observed using HE staining.Result:1.mNSS score of rats in sham group was low,there was no significant difference at different time points.mNSS score of rats in model group and acupuncture group increased significantly at 6h after ICH,and the score tended to increase further with time extension,with the highest expression at 24h after ICH.Compared with sham group,mNSS score of each time point model group was significantly increased(P<0.01).Compared with the model group,mNSS score of each time point acupuncture group was significantly decreased(P<0.01).2.Brain water content of rats in the sham operation group was low,and there was no significant difference at different time points.Brain water content of model group and acupuncture group began to increase from 6h,and the content of brain water was relatively highest from 24h to 72h.Compared with sham group,the cerebral water content of rats in the model group at each time point was significantly increased(P<0.05),and the increase trend at 12,24 and 72 hours was extremely significant(P<0.01).Compared with model group,the cerebral water content in acupuncture group significantly decreased(P<0.05),while the time points of 12,24 and 72h showed a particularly significant decline(P<0.01).3.There was no obvious abnormality in brain structure and cell morphology in sham operation group.In model group,the features of lamellar hemorrhage,cell swelling,tissue edema and inflammatory infiltration were obvious.Compared with model group,the hematoma in acupuncture group was significantly reduced,with a small amount of inflammatory infiltration,and the degree of tissue destruction was significantly reduced.4.Serum IL-1β expression was low in sham group,and no significant difference was found at all time points.In model group and acupuncture group,serum IL-1β level in rats began to increase at 6h,and its expression was relatively highest at 24h,and then gradually decreased.Compared with sham operation group,IL-1β depletion in model group at each time point was significantly increased(P<0.01).Compared with model group,the content of IL-1β purge in each time point was significantly decreased(P<0.01).5.In sham group,the expression of IL-1β in the brain tissue of rats was low,and there was no significant difference between groups at different time points.In model group and acupuncture group,the expression level of IL-1β mRNA in brain tissue of rats began to increase at 6h,and was the highest at 24h,and then gradually decreased.Compared with sham group,the level of IL-1βmRNA in each time point model group was significantly increased(P<0.01).Compared with model group,the level of IL-1βmRNA in acupuncture group at each time point was significantly decreased(P<0.01).6.The expression of Mincle,Syk and CARD9 proteins in the brain tissue of rats in sham group were low,and there was no significant difference at different time points.The expression levels of Mincle,Syk and CARD9 in model group and acupuncture group began to increase 6h after the modeling,and the relative expression levels were the highest at 24h,and then gradually decreased.Mincle protein expression situation shows that:compared with sham group,model group rats at each time point group significantly increased the relative expression quantity of Mincle(P<0.01),compared with model group,acupuncture group Mincle expression significantly decreased(P<0.01).Syk protein expression situation shows that:compared with sham group,the Syk expression in model group rats had significantly higher(P<0.01),compared with model group,acupuncture group expression of Syk significantly lower(P<0.01).The protein expression of CARD9 showed that:compared with sham operation group,the content of CARD9 in the brain tissue of rats in the time point model group increased significantly(P<0.01).Compared with model group,Syk content in the acupuncture group was significantly decreased(P<0.01)7.Immunohistochemical result showed that:A small amount of Mincle,Syk and CARD9 positive cells were seen in the periphera brain tissue of rats in sham group,and the expression was no significant difference at each time point.A large number of positive cells were expressed in model group at 6h time point.The number of positive cells of Mincle,Syk and CARD9 was significantly increased at 12h and 24h time point.And the expression level was gradually decreased after that.Compared with sham group,the expression of Mincle,Syk and CARD9 were significantly increased in model group and acupuncture group(P<0.01).Compared with model group,the level of positive cells of Mincle,Syk and CARD9 was significantly reduced in acupuncture group(P<0.01).8.Correlation analysis result showed that:the expression level of IL-1β was positively correlated with that of Mincle,r2=0.6353.The expression level of Mincle was positively correlated with that of Syk,r2=0.89960.The expression level of Mincle was positively correlated with that of CARD9,r2=0.85 82.The expression level of Syk was positively correlated with that of CARD9,r2=0.7982.Conclusion:1.Acupuncture through "Baihui" to "Qubin" can significantly reduce the score of mNSS nerve function and effectively improve the neurological deficit in rats with intracerebral hemorrhage.2.Acupuncture through "Baihui" to "Qubin" can significantly reduce the cerebral edema caused by intracerebral hemorrhage.3.Acupuncture through "Baihui" to "Qubin" can significantly reduce the expression of IL-1β in the serum and brain tissues of rats with cerebral hemorrhage,and effectively inhibit the occurrence and development of inflammatory response.4.Acupuncture through "Baihui" to "Qubin" may inhibit inflammatory responses through Mincle/Syk signaling pathways.