| Objective: to study the effect of the Qilong capsule on rats with acute cerebral hemorrhage at different time points and to determine the best time of medication.On this basis,to study whether Qilong capsule can play an anti-inflammatory effect by inhibiting P2X7R/NLRP3-IL-1 β signal pathway so as to improve the prognosis of rats with intracerebral hemorrhage and provide new ideas for clinical treatment.Methods: 1.SPF male SD rats(300g ±20)were randomly divided into a sham operation group,model group,pre-administration group,6-hour administration group,and24-hour administration group.The rats in the sham operation group were injected with normal saline into the striatum,and the other groups were injected with collagenase Ⅳ to establish the model of a cerebral hemorrhage.Before installing the model,the pre-administration group was given a Qilong capsule of 250mg/kg for seven days,and the other groups were given saline for seven days.After the establishment of the model,the rats in the sham operation group and the model group were given normal saline,the pre-administration group and the 6-hour administration group were given Qilong capsule250mg/kg after 6 hours,and the 24-hour treatment group was given Qilong pill 250mg/kg after 24 hours.The rats in each group were given medicine every 24 hours,once a day.The modified neurological deficit score(m NSS)was used to detect the neurological deficit of rats in each group on the 1st and 3rd days after modeling.The behavior of rats was recorded and evaluated,the changes in body mass of rats were detected,the changes in brain water content were detected by dry and wet weight method,and Image J calculated the volume of a cerebral hemorrhage.2.SPF male SD rats(300g ±20)were randomly divided into sham operation group,model group,positive drug group,high-dose Qilong capsule group,middle-dose Qilong capsule group,and low-dose Qilong capsule group.The rats in the sham operation group were injected with normal saline into the striatum,and the rats in the other groups were injected with collagenase Ⅳ to establish the model of a cerebral hemorrhage.Twenty-four hours after the establishment of the model,the rats in the positive drug group were given12.5mg/kg,while the rats in the high,middle,and low dose groups of Qilong capsule were given Qilong capsule 375mg/kg,250mg/kg and 125mg/kg for three days.The neurological deficit was detected by modified neurological deficit score(m NSS)at 1 and 3 days after modeling.Dry and wet weight methods noticed changes in brain water content.The expression of Iba1 and NLRP3 in brain tissue was detected by immunofluorescence staining.The area of cerebral hemorrhage was calculated by HE staining,and the changes of P2X7 R,NLRP3,ASC,caspase-1,and IL-1 β protein content were detected by Western blot technique.Results: 1.Qilong capsule intervention 24 hours after acute cerebral hemorrhage can effectively improve the prognosis of a cerebral hemorrhage.The results of the neurological function score showed that the scores of the model group(P < 0.01),pre-administration group(P < 0.01),6-hour administration group(P < 0.01),and 24-hour administration group(P < 0.01)were higher than those of sham operation group at one day after modeling.Still,there was no significant difference between the treatment and model groups.Three days after modeling,compared with the sham operation group,the neurological function scores in the model group(P < 0.01),pre-administration group(P < 0.05),and 6-hour administration group(P < 0.01)were higher than those in the model group.In contrast,those in the 24-hour administration group were lower than those in the model group(P <0.05).Compared with one day after modeling,the neurological function scores decreased in a 3-day pre-administration group(P < 0.01),6-hour administration group(P < 0.05),and24-hour administration group(P < 0.01).The calculation results of body mass changes showed that compared with the sham operation group,the body mass changes of rats in the model group(P < 0.01),pre-administration group(P < 0.01),and 6-hour administration group(P < 0.05)were increased.In contrast,those in the 24-hour administration group were lower than those in the model group(P < 0.05).The cerebral hemorrhage volume calculation results showed that compared with the model group,the cerebral hemorrhage volume in the 24-hour administration group decreased(P < 0.05).The brain water content results showed no significant difference between each treatment group and the model group(P > 0.05).2.Qilong capsule can improve the neuroinflammatory response after intracerebral hemorrhage by inhibiting P2X7R/NLRP3-IL-1 β signal pathway.The results of the neurological deficit score showed that on the first day after modeling,compared with the sham operation group,the scores of the model group(P < 0.01),positive drug group(P <0.01),high dose group(P < 0.01),middle dose group(P < 0.01)and low dose group(P <0.01)increased.Three days after modeling,compared with the sham operation group,the score of each administration group increased(P < 0.01),and compared with the model group,the middle dose group of the Qilong capsule could significantly reduce the neurological function score(P < 0.05).Compared with one day after modeling,the scores of the positive drug group(P < 0.01),high dose group(P < 0.05),middle dose group(P <0.01),and low dose group(P < 0.01)decreased after three days.The results of brain water content showed no significant difference in brain water content between each treatment group and the model group(P > 0.05).The results of HE staining showed that the area of cerebral hemorrhage could be effectively reduced in the middle-dose group(P < 0.01)and low-dose group(P < 0.01)of Astragalus capsules compared with the model group rats.The results of immunofluorescence labeling of NLRP3 in the tissues surrounding cerebral hemorrhage showed that the expression was increased in the model group compared with the sham-operated group(P < 0.05)and decreased in the positive drug group(P < 0.05)and the mid-dose group of Astragalus capsules(P < 0.01)compared with the model group.Western blot assays showed that the expression of P2X7R(P < 0.01),NLRP3(P < 0.01),ASC(P < 0.01),caspase-1(P < 0.01),and IL-1β(P < 0.01)protein contents were significantly increased in the model group compared with the sham-operated group,and P2X7R(P < 0.01),NLRP3(P < 0.05),ASC(P < 0.01),caspase-1(P < 0.01),and IL-1β(P< 0.01)protein contents were significantly increased in the high-dose group of Astragalus capsules-1(P < 0.01),P2X7R(P < 0.01),caspase-1(P < 0.01)in the low-dose group of Astragalus capsules;compared with the model group,the content of P2X7R(P < 0.01)and caspase-1 protein in the brain tissue of rats in the positive drug group was reduced(P <0.01),and P2X7R(P < 0.01)and NLRP3(P < 0.01)in the brain tissue of rats in the middle-dose group of Astragalus capsules.0.01),NLRP3(P < 0.05),ASC(P < 0.05),caspase-1(P < 0.05),and IL-1β(P < 0.05)protein contents were reduced in the brain tissue of rats in the middle dose group of Astragalus capsules,and P2X7R(P < 0.05)in the low dose group of Astragalus capsules.Conclusion: the intervention of Qilong capsule 24 hours after acute cerebral hemorrhage can effectively improve the prognosis of cerebral hemorrhage,which may play an anti-inflammatory effect by inhibiting the P2X7R/NLRP3-IL-1 β signal pathway. |
PDF Full Text Request