| ObjectiveTo discuss the variation rule of the inflammatory reaction of the brain tissue around the acute stage of cerebral hemorrhage rats and to study the affection on the expression of inflammatory reaction treated by the different frequency of waves of electro-acupuncture.MethodsWe totally used 136 Sprague-Dawley, which was to abbreviate SD, male rats. We used random digits table to divided them into 5 big groups, that was the control group, the sham operated group, the model group, the intracerebral hemorrhage plus low frequency waves of electro- acupuncture group and the intracerebral hemorrhage plus high frequency waves of electro- acupuncture group. As to the control group, it took 8 rats. As to the other four groups, each group took 32 rats and divided into the 6 hours group, the 24 hour group, the 72 hours group and 7 days group. As to these small groups, each group took 8 rats.We used stereo taxis instrument to inject 0.6U collagenase VII, its concentration was 0.2 U per microlitre, into the right caudate nucleus of rats to establish the model of cerebral hemorrhage. As to the sham operated group, we injected 3 microlitre of physiological saline. No operation for the control group. No treatment for the sham operated group and model group. The acupuncture-point was Dazhui, Fengfu and Shuigou. The Dazhui and Fengfu were connected with electric acupuncture apparatus, which brand was Huatuo.As to the intracerebral hemorrhage plus low frequency waves of electro- acupuncture group, we used 0.175 millisecond wave width, 2 hertz frequency and 1.0 mill ampere electric current, 1 volt electric voltage. As to the intracerebral hemorrhage plus high frequency waves of electro- acupuncture group, we used 0.175 millisecond wave width, 100 hertz frequency and 1.0 mill ampere electric current and 1 volt electric voltage. Each time for electro-acupuncture was 30 minutes and rats were treated once a day.The method of ethnology, immunohistochemistry and pathology was used. We observed the variation rule of the neurological deficit score, interleukin-1β, interleukin-6 and Interleukin-10 in different point of time. We studied the effects on those indexes above by the treatment of different frequency of waves of electro-acupuncture.ResultsAs to the neurological deficit score, results were as follows:In the model groups, the neurological deficit could be seen in the 6 hours group. Then the neurological deficit score attended its peak in the 48 hours group and lasted for 72 hours. The neurological function began to recover after one week. The variation rule of the neurological deficit in both the intracerebral hemorrhage plus low frequency waves of electro-acupuncture groups and the intracerebral hemorrhage plus high frequency waves of electro-acupuncture groups was similar to that in the model groups. But in those two groups, the neurological function began to recover after 48 hours and was recovered to normal after 7 days. To compare with the model groups, the neurological impairment score of the 48 hours and 72 hours both low and high frequency waves of electro-acupuncture groups was significantly lower than that at the corresponding time point (P<0.05 or P<0.01) . At the same time, the comparison between the low frequency waves of electro-acupuncture groups and high frequency waves of electro-acupuncture groups show no significantly difference in the neurological impairment score (P>0.05) .It showed that there was less neurological impairment in the two frequency waves of electro-acupuncture groups than that in the model groups. And rat with intracerebral hemorrhage recovered earlier in the two frequency waves of electro-acupuncture groups than that in the model groups. As to the case fatality, the model group took 10 dead rats. But the low frequency waves of electro-acupuncture group took 4 and the the high frequency waves of electro-acupuncture group took 6. Most death cases were occurred within 48 hours. The case fatality in 3 big groups above was 31.25 %, 12.50 % and 18.75 % respectively. There was no significantly difference in the case fatality within these 3 big groups (P>0.05) . Pathomorphologyical changes of the brain tissue in rats were of the model groups as follows:Under the light microscope, widespread bleeding, severely damaged nerve cell and histolysis could be seen in the 6 hours model group. Red cell lysis in the hematoma, considerable swollen cells around the hematoma and obvious dropsy outside of the cells and around the vessels could be seen in the 24 hours model group. In this group, abscess-formation, a great quantity of heterophil granulocyte infiltration at the edge of hematoma also could be found. In the 72 hours model group, the red blood cell began to absorb in the centre of hematoma and the peripheral edema was still exist. Cellular proliferation of gitter cell and widespread infiltration of heterophil granulocyte also could be found in this group. As to the 7 days model group, the dropsy was obviously resolved, and the cellular swelling was relieved. The leukomonocyte related chronic inflammation and desmoplasia was the main change in this group.As to the affection on the cell factor of intracerebral hemorrhage model rats by the different frequency of waves of electro-acupuncture, the results were as follows:Firstly, both the low and high frequency of waves of electro-acupuncture could suppress the expression of Interleukin-1βand interleukin-6, which were two inflammatory factors, in the intracerebral hemorrhage model rats.The method of immunohistochemistry showed that there was slight amber positive expression of interleukin-1βand interleukin -6 in nerve cells of the control group. As to the model groups, the hypochromatic yellow positive expression of interleukin-1βin nerve cells could be seen around the hematoma in the 6 hours model group. It reached to its peak in the 24 hours model group and the trachychromatic yellow brown positive expression of interleukin-1βin nerve cells could be seen. The positive expression of interleukin-1βin nerve cells in the 7 days model group began to drawdown. There was slight amber positive expression of interleukin-6 in nerve cells around the hematoma in the 6 hours model group. The trachychromatic yellow positive expression of interleukin-6 in nerve cells around the hematoma could be seen in the 24 hours model group. This peak lasted for 72 hours and the positive expression of interleukin-6 began to decrease in the 7 days model group.Statistics showed that there was significant expression of Interleukin-1βin the 6 hours model group. This expression was at its peak in the 24 hours model group and began to go down in the 72 hours model group. It went down to normal level in the 7 days.As to the comparison in group, the expression of interleukin-1βin the 7 days model group was significantly lower than that in the 6 hours, 24 hours and 72 hours model group(P<0.01).There was increased expression of interleukin-6 in the 6 hours model group. This expression was at its peak in the 24 hours model group and lasted for 72 hours. Then it began to go down in the 7 days model group. It was still higher than normal level even in the 7 days model group. As to the comparison in group, the expression of interleukin-6 in the 24 hours, 72 hours model group was significantly higher than that in the 6 hours model group (P<0.01). The expression of interleukin-6 in the7 days model group was significantly lower than that in the 24 hours model group (P<0.01). The expression tendency of interleukin- 1βin both the low and high frequency of waves of electro-acupuncture group was similar to that in the model group. The expression of interleukin-1βin all time points of model groups, 6 hours, 24 hours and 72 hours low and high frequency of waves of electro-acupuncture groups was higher than that in the control group (P<0.05 or P<0.01). The expression of interleukin-1βin 6 hour, 72 hours low frequency of waves of electro-acupuncture groups and 72 hours high frequency of waves of electro-acupuncture groups was significantly lower than that in the corresponding time point of the model group (P<0.05) .At the same time, there was no significantly difference between all time points of low and high frequency of waves of electro-acupuncture groups (P>0.05) .The expression tendency of interleukin-6 in both the low and high frequency waves of electro-acupuncture groups was similar to that in the model group. But it went down to normal level in the 72 hours of both low and high frequency waves of electro-acupuncture groups.There was no significantly difference between the 72 hours of both low and high frequency of waves of electro-acupuncture groups and the control group for the expression of interleukin-6 (P>0.05). As to the comparison in groups, the expression of interleukin-6 in all time points of model groups, the 6 hours, 24 hours of both low and high frequency waves of electro-acupuncture groups was significantly higher than that in the control group (P<0.01 or P<0.05). The expression of interleukin-6 in 72 hours and 7 days both low and high frequency waves of electro-acupuncture groups was significantly lower than that in the corresponding time point of the model groups (P<0.01) . At the same time, there was no significantly difference between all time points of low and high frequency waves of electro-acupuncture groups (P >0.05) .Secondly, both the low and high frequency waves of electro-acupuncture could promote the expression of interleukin-10, which was an anti-inflammatory factor, in the intracerebral hemorrhage model rats.The method of immunohistochemistry showed that there was much yellow positive expression of interleukin-10 in nerve cells of the control group. As to all time points of the model groups, a small quantity of the amber positive expression of interleukin-10 in nerve cells could be seen around the hematoma. A large quantity of trachychromatic yellow positive expression of interleukin-10 in nerve cells could be seen around the hematoma in the 6 hours, 24 hours and 72 hours both low and high frequency waves of electro-acupuncture groups.As to statistics results, there was no significantly difference of the expression of interleukin-10 in nerve cells among all time points of the model groups (P>0.05) .The expression of interleukin-10 in nerve cells began to increase in the 6 hours both low and high frequency waves of electro-acupuncture groups. And it began to draw down in the 72 hours groups and decrease to the normal level in the 7 days both low and high frequency waves of electro-acupuncture groups. There was no significant difference between the 6 hours, 24 hours and 72 hours model groups and the control group in the expression of interleukin-10 in nerve cells(P>0.05). The expression of interleukin-10 in nerve cells in the 7 days model group was significantly lower than that in the control group (P<0.05). The expression of interleukin-10 in nerve cells in the 6 hours, 24 hours and 72 hours both low and high frequency waves of electro-acupuncture groups was significantly higher than that in the control group (P<0.01 or P<0.05) . The expression of interleukin-10 in nerve cells all time points of both the low and high frequency waves of electro-acupuncture groups was significantly higher than that in the corresponding time points model groups (P<0.01 or P<0.05) . But there was no significant difference between all time points of the low frequency waves of electro-acupuncture groups and corresponding time points high frequency waves of electro-acupuncture groups (P >0.05) .ConclusionResults of pathobiology and immunohistochemistry showed that there did have inflammatory reaction, which was character for the increasing of leukocyte and gitter cell, around the hematoma of the intracerebral hemorrhage in the acute stage. At the same time, the expression of interleukin-1βand interleukin-6 was high. Both the low and high frequency waves of electro-acupuncture could restrain the expression of interleukin-1βand interleukin-6 and increase the expression of interleukin-10 to some extent. This might relief the secondary lesion of the brain tissue that caused by inflammatory factor and increase the protection of the anti-inflammatory factor for brain. There was no significant difference between the low and high frequency waves of electro-acupuncture in the aspects of improving the neurological impairment, restraining the expression of interleukin-1βand interleukin-6 and increasing the expression of interleukin-10.
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