Exploring The Synergistic Mechanism Of Heat-clearing And Blood-activating Components In The Treatment Of Acute Cerebral Infarction With Fire-toxic Syndrome Based On Interleukin And Related Inflammatory Pathway

Objective:To explore the material basis of fire-toxicity syndrome of acute cerebral infarction,and to investigate the the synergistic interaction of the inflammatory response mechanisms infire-toxicity syndrome of acute cerebral infarction.Method:In clinical study:119 acute cerebral infarction patients with the fire-toxicity syndrome were admitted to hospital and were divided into 4 groups randomly by central randomization system,the basic treatment group-patients were given basic western medicine,Qing Re group-patients were given western medicine combined with Ixeri sonhifolia injection,Huo Xue group-patients were given western medicine combined withpanax notoginseng saponins,and Qing Re Huo Xue group-patients were givenwestern medicine combined with Ixeri sonhifolia injection and panax notoginseng saponins.All patients were received 14 days treatment and follow-up forl 80 days.The patients’ safety indicators were collected before and after the treatment to verify the safeness of the treatment of Qing Re Huo Xue.The NIHSS scale and fire-toxicity syndromescore were used on the 1st,3rd,5th,7th,and 14th day of the treatment.Inflammatory mediators such as IL-6 and IL-18 in the interleukins,ICAM-1 and VCAM-1 in cell adhesion molecules family,MIP-1α and MMP-1,all of these were measured on the 1st 3rd,5thand 14thday of the treatment.Analyze the regulation of Qing Re Huo Xue component on inflammatory mediators and related therapeutic indicators,the correlation between inflammatory mediators,as well as the correlation between NIHSS and inflammatory mediators.In vivo study,To investigate the effects of Qing Re Huo Xue component on neurological deficit and cerebral infarct volume in Transient Middle Cerebral Artery Occlusion(tMCAO)model rats.To investigate the regulation of the phosphorylation of p-38,p-JNK,and p-ERKl/2 in the MAPK signaling pathway and the expression of iNOS,Cox-2,PGE2,IL-6,and MMP-9 in mice brain.In vitro study,we used the primary microglia and the Oxygen Glucose Deprivation model(OGD)was establishedto investigate the expression of primary microglial M1’s marker proteins iNOS and Cox-2,and explore the regulation of Qing Re Huo Xue component on primary neurons through regulating the primary microglia.Result:In clinical study:(1)Safety indicators:the results of the blood,urine,stool routine,liver and kidney function,blood coagulation,D-dimer and electrocardiogram of the four groups after treatment showed no significant difference(P>0.05).(2)The efficacy indicators:On the 14th day of treatment,the NIHSS score in the Qing Re Huo Xuegroup was significantly lower than that in the basic treatment group(P<0.05).On the 180th day of treatment,Qing Re Huo Xuegroup had a significantly lower score than that in Huo Xue group when comparing the fire syndrome score(P<0.05).On the 7th day of treatment,Qing Re Huo Xuegroup had a significantly lower scores than that in Huo Xue group when comparing the blood stasis syndrome scores(P<0.05).(3)Effects of Qing Re Huo Xue component on inflammatory factors:On the 14th day of treatment,the Qing Re Huo Xue component could significantly down-regulated the serum level of IL-6,IL-18,ICAM-1,VCAM and MMP-9.We found that IL-6,IL-18 and adhesion factors ICAM-1,VCAM-1 are closely related through analysingthe internal correlation of inflammatory mediators,which indicating that the inflammatory mediators showed group associated changes.NIHSS score had positively correlated with Hs-CRP,ICAM-1,indicating that abnormal changes in inflammatory mediaotrs might affect the severity of acute cerebral infarction.In vivo study:(1)All treatment groups could improve the symptoms of neurological deficits caused by tMCAO and decrease the volume of cerebral infarction,but there is no difference between the Qing Re Huo Xue group and the single heat or single blood-activating group.(2)The regulation of the Qing Re Huo Xue component on the abnormally phosphorylated p-38 was more significant than that in Qing Re group(P<0.05),and there was a significant down-regulation of the abnormally phosphorylated ERK1/2.(3)In the regulation of iNOS,Cox-2 and PGE2,the the Qing Re Huo Xue component were more prominent than that in the single heat-clearing or single blood-activating regulation.In vitro study:(1)Qing Re Huo Xue componentcould decrease the expression of microglial M1-type marker protein Cox-2significantly compared with sigle blood-activating group,and the condition medium of Qing Re Huo Xue group could significantly reduce the death rate of the primary neuron.Conclusion:1 The inflammatory response after acute cerebral infarction was the pathological basis of cerebral infarction.2 Qing Re Huo Xue component could regulate acute cerebral infarction andhad a synergistic effect on the regulation of cerebral infarction.3 The possible mechanism of Qing Re Huo Xue component on fire-toxicity syndrome of acute cerebral infarctionmight be related to its regulation of the phosphorylation of p-38,ERK1/2 in the MAPK signaling pathway and the regulation of microglias.4 The cascade inflammatory response after cerebral infarction is a complex process.The regulation of Qing Re Huo Xue componenthad multiple levels,multiple targets,and correlations characters in the regulation of cerebral infarction.This also explained the complexity of the fire-toxicity syndrome of acute cerebral infarction.