| ObjectiveFrom the theory of "toxin infringe upon the brain, the brain is damaged, kidney-essence is inadequate, the brain's support is inadequate",to explore the etiological factor of Alzheimer's disease (AD), follows the principle of " treating the asthenia-syndrome by therapy of invigoration" and "treating the sthenia-syndrome by therapy of purgation", and" treating a disease by looking into both its root cause and symptoms", we advance the therapy of detoxifying and replenishing kidney-essence is operative on treating AD. In the experiments, the model rats were administered D-gal injection and Aβ25-35 into hippocampus, establish a complex rat model of AD. To observe the effects of detoxifying and and replenishing kidney-essence therapy on learning memory, pathological, the content of IL-1β, IL-6, TNF-α, CRP, the positive expression on NF-κB and iNOS for analyzing the therapeutic mechanism of action of AD. To provide traditional Chinese medicine with experimental accordance and clinical guidance.MethodsThe normal 90 male 8-month-old SD rats were randomly divided into six groups:normal group, blank group, AD model group,piracetam treated group, high-dose of Qiancengta Decoction (HQD for short), low-dose of Qiancengta Decoction(LQD for short), there are 15 rats in every group. After feeding in ordinary ways for one week, except the normal group, AD model group, piracetam treated group, HQD group and LQD group received neck back subcutaneous injection of D-galaetose (150mg/kg. d) for six months, and the blank group rats with lavage. At the beginning of the seventh week, the four group rats were injected Aβ25-35 in the bilateral Hippocampus. The blank group rats were injected the same dose of lavage. After 3 days, every group rats were treated with different madicine for four weeks.Then used Morris water maze(MWM) to observe the ability of space learning memory. Used the light microscope and electron microscope to observe the changes of morphology and structure in the hippocampus. Radioimmunoassay method was used to detecting the IL-1β, IL-6, TNF-α.ELISA was used to detecting CRP. Immunohistochemistry method was used to detecting the positive expression of NF-κB and iNOS.Results1. In the behavior experiment, the average incubation period,20%,40% aera of every group rats in the learning ability gain, the average incubation period, percentage of quadrant, the frequency in span of platform localization in space research. The results show that have significant differences between the model group and normal group, black group (p< 0.01), it indicate that the AD model method can decrease the AD rat's ability of learning gain; there are significant differences between the HQD and model group (p< 0.01), it indicate that the HQD can improve the AD rat' s ability of learning; there are insignificant differences between theLQD group and piracetam treated group(p> 0.05); there are significant differences between the HQD and piracetam treated group (p< 0.01). These results indicate that the AD model in this experiment is successful, the therapy of detoxifying and replenishing kidney-essence can remarkable improve the AD rat' s ability of learning, its effect better than the piracetam treated group.2. By the light microscope and electron microscope observation, the sum neuron of model group was less than that of normal group, death cell could be seen clearly, the neuron decreased and deformed in model group rats. Pellet vacuole denaturation, cell nucleus lessen, the mitochondria swelling and vacuole, the volume of mitochondria was bigger.The endoplasm reticulum dilatation, amount of synapses decrease. HQD can ameliorate these pathological changes obviously, there are significant differences between the HQD and model group, there are significant differences between the HQD and piracetam treated group. These results indicate that the therapy of detoxifying and replenishing kidney-essence can ameliorate these pathological changes obviously.3. The detecting results of the IL-1β, IL-6, TNF-α, the content of IL-1β, IL-6, TNF-αin the model rats higher remarkably than normal group,black group (p< 0.01). There are significant differences between the HQD and model group (p< 0.01), there are significant differences between the HQD and piracetam treated group (p< 0.01), the effect of HQD better than the piracetam treated group, the LQD group (p< 0.01). These results indicate that detoxifying and replenishing kidney-essence can remarkable decrease the content of AD model rats' s IL-1β, IL-6, TNF-αin the hippocampus.4. The detecting results of the content of CRP, the content of CRP has significant differences between the normal group, black group (p<0.01), there are significant differences between the HQD and model group (p<0.01), there are significant differences between the HQD and piracetam treated group (p<0.01), there are significant differences between the LQD and piracetam treated group (p<0.01). These results indicate that detoxifying and replenishing kidney-essence can remarkable decrease the content of AD model rats' s blood CRP.5. The detecting results of the NF-κB, iNOS. Viewing under the light microscope, the most brown and yellow pellets can be seen in the model group. It indicate that the most positive expression of NF-κB and iNOS in the model group, the brown and yellow pellets decrease between the each treated group and the model group, it indicate that the each treated group can inhibition the positive expression of NF-κB and iNOS. In the four treated groups, a very few brown and yellow pellets can be seen, there are significant differences between the HQD and model group (p<0.01), there are significant differences between the HQD and piracetam treated group (p<0.01), there are insignificant differences between the LQD group and piracetam treated group (p>0.05). These results indicate that detoxifying and replenishing kidney-essence can remarkble inhibition the positive expression of NF-κB and iNOS, obtain yield notable results.Conclusion1. The theory of "toxin infringe upon the brain, the brain is damaged, kidney-essence is inadequate, the brain's support is inadequate" is the basic etiological factor for AD, the therapy of detoxifying and replenishing kidney-essence is a effective and new cure to treat AD.2. The model group rats were administered D-gal injection and Aβ25-35 into hippocampus, establish a complex rat model of AD, it not only mock the action changes of AD, but also mock the changes of pathological, immunity and so on, it is a reliable animal model.3. The therapy of detoxifying and replenishing kidney-essence can remarkble improve the learning ability of AD rats, ameliorate the pathological changes, remarkable decrease the content of AD model rats's IL-1β, IL-6, TNF-αin the hippocampus, remarkable decrease the content of AD model rats' s blood CRP, remarkble inhibition the positive expression of NF-κB and iNOS.4. From the above-mentioned experiment results,we can say the therapy of detoxifying and replenishing kidney-essence has the curative effect of clearing heat and detoxicating removing phlegm, promoting blood circulation and removing blood stasis, replenishing kidney-essence, replenishing brains marrow, it has the treat effect of anti inflammation, it can decrease the inflammation reaction of the AD model rats, ameliorate the pathological changes, improve the learning ability of AD rats. It is a effective and new cure to treat AD. Qiancengta Decoction is a effective traditional Chinese medicinal recipe to treat AD.
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