| Background and objective It is well known that ICH(intracerebral hemorrhage,ICH)has high morbidity and mortality.Once ICH happened,mechanical injury caused by hematoma damaged the brain firstly.Besides,inflammation and brain edema,the secondary injury,played an important role in the prognosis of ICH and were the key reason for disability as well as death.ICH characterized as suddenly happened and progressed rapidly.Although morbidity rate of ICH was lower than that of cerebral ischemia,the mortality rate was much higher than that.It is of great significance to actively looking for the treatment of cerebral hemorrhage effectively.Methods Adult male Sprague-Dawley(SD)rats were randomly divided into control group(sham group),operation group(Vehicle-ICH group)and drug treatment group(EC-ICH group).EC-ICH group rats were pretreated with(—)–Epicatechin 45mg/(kg.bt*d)by gavage for 7 days.Then collagenase Ⅶ-s was injected into the caudate nucleus of rats in the Vehicle-ICH and EC-ICH groups to build the ICH rat model.The ones with no neurological deficits,unconsciousness or coma,were not qualified,replaced with qualified ones.At day 1,day 3,day 7 and day 14 after injection,the neurological deficit score were tested on all the rats;at day 1,day 3,MRI was performed on the rats to observe and record the volume of hematoma and the degree of brain edema in the peri-hematoma areas;at 4h,day 1 and day 3,double-labeled immunofluorescence was used to detect the number of microglia,who can polarized into two different types(microglia type Ⅰ,M1;microglia type Ⅱ,M2)after ICH;At day 1,day 3,and day 7,the m RNA of IL-10,IL-6,IL-1 beta,TNF-alpha,TGF-beta were detected with q RT-PCR;at day 3,the blood-brain barrier(BBB)permeability was detected with Evans blue staining and at day 1,day 3,the albumin in the brain was also checked with immunofluorescence;at day 28 brain white matter injury were checked with MRI.Results1.The neurological deficit score of EC-ICH group rats were significantly lower than that of Vehicle-ICH group,and the neurological deficit function recovered quickly in the EC-ICH group.2.The hematoma volume of EC-ICH group(27.51 ± 2.29 mm3)at day 1 was significantly less than that of Vehicle-ICH group(44.49 ± 1.78 mm3),the hematoma volume of EC-ICH group(17.58 +1.99 mm3)at day 3 was significantly less than that of Vehicle-ICH group(29.35 + 2.29 mm3),P <0.05.3.Evans blue staining results showed that Evans blue in the EC-ICH group brain rats were obviously less than of Vehicle-ICH group;similarly,albumin was significantly less than that of the Vehicle-ICH group detected by immunofluorescence.4.In both group we found the microglia had polarized into two different types,M1 and M2.In the Vehicle-ICH group,the amount of M1 elevated from day 1,and peaked at day 3,and the M2 transiently elevated at 4 hours after ICH.While,compared with the Vehicle-ICH group at 4 hours,day 1 and day 3,the number of M1 were all decreased(P < 0.05),the number of M2 increased at 4 hours(P < 0.05).5.q RT-PCR results showed that the expression levels of IL-1,IL-6 and TNF-α were significantly decreased,and the IL-4,IL-10,TGF-βlevels were significantly increased at some time points.6.White matter injury checked by MRI showed that the white matter was significantly recovered at day 28 after ICH.Conclusion1.EC pre-treatment of in rats could inhibit the expansion of early hematoma,and improved the symptoms of neurological deficits;2.Pre-treatment of EC could reduce the damage of BBB and ameliorate the degree of brain edema;3.Pre-treatment of EC could inhibit microglia polarization to M1 and promote microglial polarization to M2 at the early stage after ICH;4.EC pre-treatment could downregulate the m RNA expression of IL-1β,TNF-α,IL-6,and upregulate IL-4,IL-10,TGF-β expression,thereby alleviating related inflammation;5.EC pre-treatment could effectively improve the white matter injury in the latter stage of hemorrhage,as well as the prognosis. |
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