Graded and time effects of huoxue huayu tang on anti-injury of cerebral ischemic reperfusion in rats / 中国组织工程研究

ABSTRACT

BACKGROUND: It has been known that re-recovery of blood flow after cerebral ischema may aggravate brain injury. It has been verified in some researches that the components for activating blood circulation and removing stasis in Chinese herbs can resist cerebral ischemic reperfusion injury.OBJECTIVE: To observe 4 most basic Chinese herbs for activating blood circulation and resolving stasis [honghua (Flos Carthami), taoren (Semen Persicase), chuanxiong (Rhizoma ligustici Chuanxiong) and chishao (Radix Paeoniae Rubra)] as the exertive factors on their influences on immune index changes in IgG, C-reactive protein (CRP) and complements of serum and homogenate in brain tissue so as to verify their graded and time effects.DESIGN: Randomized control experiment and single blind method.SETTING: Guangzhou Red Cross Hospital and Department of Neurology of the Fourth Hospital Affiliated to Jina University.MATERIALS The experiment was performed from January 2001 to Decemb er 2002 in Experimental Room of Traumatic Institute of Guangzhou Red Cross Hospital. Totally 138 SD female adult rats were employed, mass weighted varied from 280 to 300 g, provided from Experimental Animal Center of Guangzhou University of Chinese Medicine. Honghua (Flos Carthami), taoren (Semen Persicase), chuanxiong (Rhizoma ligustici Chuanxiong)and chishao (Radix Paeoniae Rubra)] were the concentrated granules individually. At the ratio of 1112, they were prepared into 2.5 g/mL raw herbal decoction (huoxue huayu tang).INTERVENTIONS: Thread embolic method was used to prepare cerebral medium sized artery obstruction model in rats (reperfusion for 24 hours after cerebral medium sized artery obstruction for 2 hours). Totally 138 rats were randomized into 6 groups, 23 rats in each. Sham operation group ligature was done in each vessel without blocking medium sized artery of cerebrum.First perfusion group 30 minutes before operation, gastric peffusion was done with 2 g/kg huoxue huayu tang. Second peffusion group 30 minutes before operation, gastric perfusion was done with 2.5 g/kg huoxue huayu tang. Third perfusion group before operation, gastric perfusion was done with 2 g/kg huoxue huayu tang continuously for 7 days. Fourth perfusion group before operation, gastric perfusion was done with 2.5 g/kg huoxue huayu tang continuously for 7 days. In the control, physiological saline of all of the groups, when rats woke up, after 24 hours reperfusion and 2 hours ischemia, neural functional deficits were evaluated (5-score system, 0-1After 24 hours reperfusion, 10 rats were randomized from each group for determination of the contents of CRP in homogenate and serum, complehours reperfusion, 10 rats were randomized from each group. After anesthetized, they were sacrificed rapidly to collect brains. Dried over heat in oven at 110℃ till constant weight, the water content of brain tissue was domized from each group to observe ultrastructure of brain tissue under transmission electron microscope (TEM) of rats in each group.and homogenate, complement C3 and C4 as well as serum IgG in rats of of severe neural functional deficits after 24 hours ischemic reperfusion in rats of each peffusion group was lower remarkably than the control (P ＜ 0.01),in which, that in fourth peffusion group was lower than second group (13%,group and each perfusion group was lower than the control (P ＜ 0.05), in which, that in third peffusion group was lower than first group (P ＜ 0.01).ment C3 That in sham operation group and each perfusion group was lower sham operation group and each perfusion group was lower than the control ( P ＜ 0.01), in which, that in third perfusion group was lower than first group (P ＜ 0.01); That in fourth perfusion group was lower than second and group and each perfusion group was lower than the control (P ＜ 0.01), in which, that in fourth perfusion group was lower than second group(P ＜ 0.05).each perfusion group was lower than the control (t=5.626-17.929, P ＜ 0.01),in which, that in third perfusion group was lower than first group (P ＜ 0.01),complement C3 in brain homogenate That in sham operation group and each perfusion group was lower than the control (P ＜ 0.05-0.01), in which, third perfusion group was lower than first group (P ＜ 0.01), that in fourth group brain homogenate That in sham operation group and each perfusion group was lower than the control ( P＜ 0.05-0.01), in which, that in third perfusion group was lower than first group (P ＜ 0.01), that in fourth group was lower tissue The inflammatory reaction in the control was obvious. Brain edema and pathological damage in perfusion groups were milder than the control.Ulztrastructure was normal in sham operation group. The edema was obvious in cells, capillaries and myelin sheath in cortical necrosis margin area and neuronal cell organs were decreased in the control. In third and fourth peffusion groups, cellular membrane was clear in boundary line,integrative in structure, rich in mitochondria and even in size and the morphology of medullated fibers was normal. The situation of first and second perfusion was between two of them.deficits in the evaluation of rats and the score with long medication was lower, suggesting that long medication can even better improve neural funcplement C3 and C4 in brain bomogenate and serum and decreases serum IgG content, explaining that the medicine alleviates brain tissue injury by startup complement system. The remarkable decrease of C-reactive protein further suggests the inhibition of such medicine on inflammatory reaction.jury is, and the better result is achieved at dosage of 2.5 g/L.