| Objective: Epilepsy(EP) is a sort of chronic brain disease characterized by transient central nervous system malfunction which is result of neuron paradoxical discharge in brain. Epilepsy severely threatens human healthy and economy. At present, the therapy of epilepsy is mainly dependent on kinds of anti-epileptic drugs(AEDs), and most patients need to take AEDs for a long period of time. Our body metabolize these AEDs we eat via liver or kidney, consequently, AEDs will aggravate the burden of liver or kidney, which give the heraldings to the petiets, and at the same time duplex damage to the illness, cause relative side effect. So doctor pay close attention to hepar or renal function damage caused by AEDs. In this experiment pentetrazole(PTZ) is used to kindle Sprague Dawley(SD) rats. To interfere in epilepsia rat through valproate sodium (VPA) , phenytoin (PHT) and lamotrigine(LTG) , to observe therapeutic effect of AEDs, to detect the difference of cell apoptosis in liver or kidney between VPAgroup, PHT group, LTG group and NS group, to approach the effect and possible mechanism of cell apoptosis in liver or kidney of rats caused by VPA , PHT and LTG , to offer theory accordings to avoid the damage in liver or kidney caused by AEDs and direct doctor in choosing AEDs.Methods: 35 SD rats were randomly divided into five groups, each with seven . As a group were randomly selected as control group (NS group), the other four groups for the experimental group and then were randomly divided into epilepsy control group (PTZ group), valproate group (VPA group), phenytoin group (PHT group), Lamotrigine group (LTG group). By PTZ induced epileptic seizures in rat model, the NS group intraperitoneal injection of physiological saline, and the remaining four groups intraperitoneal injection pentylenetetrazol, sustained two weeks. NS Group and PTZ group given physiological saline gavage, VPA group, PHT group and LTG group given the medicine gavage, continues for three weeks, completes the intervention. Observation pentylenetetrazol kindled rats in the attack and carried on Racine graduation grading to it, the appraisal medicine to the epileptic paroxysm on the degree of control, to detect the structure of liver or kidney by HE staining , to detect expression by using immunohisto- chemical methods of the Bax and the Bcl-2 in liver and kidney of the rats intervened by drugs and calculate apoptosis rate.Results1 behavioral characteristics of rats kindled by PTZ: experimental rats presents 1-2 level of epileptic seizures after injection PTZ average 7 to 9 times; after 12 to 15 times, epileptic seizures intensity strengthened, the frequency increased gradually, and then enter the Stage , the attack continued for a few minutes after the short-term mitigation, afterward repeatedly attack; the whole attack lasted about 30 to 50 minutes, then eased gradually, after the cessation of attacks, the animals can move voluntarily. Rats of NS group are without any signs of attack and convulsive performance afterphysiological saline injection,their behavior is normal. 2 The control over the epilepsy after drug intervention: there is no seizure in NS group; convulsion sustained attack in PTZ group; 2 (1 / 3) sustained attack and one died in PHT group; good control of the seizure by AEDs in other 2 groups, attack frequency reduced or no seizure after treatment by gavage.3 General structure: the liver is composed of four parts, the purple red before perfusion, faint yellow after perfusion, smooth surface, no swelling, the tubercle and the goitre, animal liver ratio accounted for the body weight of each group has non-statistics significance (P>0.05).The kidney is like Vicia faba, smooth surface, without tubercle, animal kidney ratio accounted for the body weight of each group has non-statistics significance (P>0.05).4 results of HE staining: Liver tissue sections of VPA group are observed in the low power lens,there is normal structure in liver and hepatocyte, vacuolus degeneration is observed in the cytoplasm, the cell nucleus is pressed obviously to one side, the positive rate of vacuolus degeneration is obviously increasing compared with other group ,there is obviously statistics significance in the difference(P<0.05). Kidney tissue sections of VPA group are observed in the low power lens, there is normal structure in kidney and renal cells,and no abnormal compared with other groups. Liver and kidney tissue sections of other four groups are observed in the low power lens,and structure is normal.5 Immunohistochemistry results: the membrane of positive apoptotic cell is integrity and the cytoplasm was brown with a brown or yellowish brown for tiny granular or homogeneous diffuse staining, nucleus chromatin is compact which forms massive corporation nearby the nuclear membrane, the shape varies, the size different, the morphology of positive apoptotic cell still regular weakly,there are apoptotic bodies like circular or oval in shape with kytoplasm concentration and dispensable nuclear debris.6 The expression of Bax and Bcl-2 in hepatocyteThe positive rate of Bax in LTG group,PHT group,VPA group,PTZ group and NS group is 0.283±0.021,0.090±0.036,0.300±0.101,0.103±0.042 and 0.097±0.031 , there is non- statistics significance between LTG group and VPA group (P>0.05), non-statistics significance among PHT group,PTZ group and NS group(P>0.05), The positive rate of LTG group and VPA group is significantly increased compared with PHT group,PTZ group and NS group, there is remarkably statistical significance(P<0.05).The positive rate of Bcl-2 in LTG group,PHT group,VPA group,PTZ group and NS group is 0.080±0.026,0.373±0.096,0.090±0.036,0.377±0.012 and 0.327±0.060 , there is non- statistics significance between LTG group and VPA group (P>0.05), non-statistics significance among PHT group,PTZ group and NS group(P>0.05), The positive rate of LTG group and VPA group is significantly decreased compared with PHT group,PTZ group and NS group, there is remarkably statistical significance(P<0.05).7 The expression of Bax and Bcl-2 in renal cellThe positive rate of Bax in LTG group,PHT group,VPA group,PTZ group and NS group is 0.100±0.026,0.047±0.038,0.430±0.036 ,0.060±0.010 and 0.054±0.054 , there is non- statistics significance among LTG group,PHT group,PTZ group and NS group (P>0.05), but the positive rate of VPA group is significantly increased compared with other groups, there is remarkably statistical significance(P<0.05).The positive rate of Bcl-2 in LTG group,PHT group,VPA group,PTZ group and NS group is 0.360±0.030, 0.357±0.074, 0.133±0.047,0.327±0.032 and 0.370±0.010 , there is non- statistics significance among LTG group,PHT group,PTZ group and NS group (P>0.05), but the positive rate of VPA group is significantly decreased compared with other groups, there is remarkably statistical significance(P<0.05).Conclusion1 In the experiment Pentylenetetrazol is used to make rat model of epilepsy, 1-2 level of epileptic seizures presented after injection PTZ at about 1 week,5 level at 2 week, explained that the successful kindling of this model takes about two weeks time.2 Using intervention epileptic rats by drugs and discove- ring that VPA, LTG are good to control of epilepsy, PHT bad.3 HE staining is used to observe the roughly structure of liver and kidney and find that there is obviously vacuolus degeneration in hepatocyte of VPA group which explains that VPA affects lipid metabolism in the liver, it is suggested that the liver damage caused by VPA may have relationship with liver fatty degeneration of the long-term.4 By immunohistochemical method staining positive cell of Bax and Bcl-2 and calculating the rate of apoptosis, it is discovered that the rate of hepatorenal cell apoptosis is significantly increased in VPA group, the rate of hepatocyte is significantly increased in LTG group, this explain VPA promotes liver and kidney cell apoptosis , LTG promotes liver cell apoptosis, which suggests that liver damage caused by VPA and LTG may have relationship with increased apoptosis, VPA also possibly causes the kidney damage. |
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