Research On The Mechanism Of Tongdu Tiaoshen Moxibustion Resisting Aβ Over-expression In Mci Model Rats

Objective The experiment is to establish MCI rat model, to observe the influence that the moxibustion of Tong Du Tiao Shen can make on rat behavioristic model and the expression of hippocampus Aβ, and to illustrate the mechanism of the moxibustion of Tong Du Tiao Shen anti-Aβ overexpression of its active role from aspects such as targets and anti-inflammatory neuroimmunomodulation, which can provide an important theoretical proof for clinical applications of this moxibustion therapy.Method 60 clean healthy mature male SD rat, weighing 250±50g, were selected as the sample. And then, Morris water maze screening experiment was utilized to sieve those who cannot swim or got significantly different testing result. After the test, 48 were decided to form the valid sample. Random method was employed to divide the rats into two groups. The first group was allocated with 12 rats, which was called normal group and acted as the control, and the rest 36 rats formed the other group, called model group. In model group, rats received D-galactose and NANO peritoneal injection synchronously, while the rats in normal group received 2ml normal saline peritoneal injection. After molding 40 days ago, Morris water maze was taken advantage of to undertake the neuroethology test, which can guarantee the success of the molding. In the following step, the rats in the model group were further divided into three groups, with each containing 12 rats, randomly by the computer, which were blank control group, moxibustion group and nimodipine group. In moxibustion group, the Bai Hui of the rats was pressured under Monkshood, and Moxa was hanged up and heated to cauterize Tian Men、Da Zhui. This procedure was repeated once a day, with one day off every week, and was designed to last for two weeks. In nimodipine group, rats received intragastric administration by nimodipine. The two groups were in progress at the sametime, while the rats in blank control group were only fastened on the board without any process. When the treatment was over,neuroethology test was taken and rats were in the fast for 12 hours, after which, the rats in the three groups would be undertaken collect the blood of abdominal aorta and derived the brain tissue separately.Result(1) After molding, Morris water maze was employed to do neurobehavioral test. When compared with normal group, model group got a significant rise in escape latency(P<0.01). And model group got obviously lower frequency of through the target platform than normal group did(P<0.01), which indicating the success of molding.(2) When the treatment was over, Morris Morris water maze was still utilized to do neurobehavioral test, whose results revealed that: each group with different days got significant distinction in terms of escape latency(P<0.05), and rats in all groups had spatial learning ability, with rats in normal group, moxibustion group and nimodipine group showing better learning ability than those in blank control group. And the distinction presented great importance(P<0.01). As for the frequency of through the target platform, rats in moxibustion group and nimodipine group showed obviously high level than those in blank control group(P<0.01),with those in moxibustion group displaying a special obvious rise. However, rats in nimodipine group still held a significant decrease than those in moxibustion group(P<0.05). What’s more, the comparison between rats in moxibustion group, nimodipine group and blank control group provided evidence that each treatment group had the ability to improve the learning ability of model rats remarkably. And the comparison between the treatment group s and normal group revealed a statistics importance(P<0.01), which telling that although each treatment group can improve the model rats’ learning ability observably, none can help them reach the normal level. Simultaneously, the comparison between rats in moxibustion group and nimodipine group(P<0.05)showed that therapy in moxibustion group was better that that in nimodipine group.(3) The results of the test revealed that Aβ immunoreactive staining mostly distributed in hippocampal neurons. hippocampal of rats in normal group held limited Aβ immunoreactive staining, while hippocampal of rats in blank control group held a great quantity of the Aβ immunostaining positive product(P<0.01). And the results of those of rats in moxibustion group and nimodipine group told that Aβ immunostaining positive cells and staining intensity both reduced after intervening, with those in moxibustion group more obviously(P<0.01). After moxibustion intervening, the number of Aβ immunostaining positive cells reduced and staining intensity declined. At the same time, there was the trend that Aβ accumulation was diffusing. Nevertheless, results of nimodipine group showed that the number of Aβ immunostaining positive cells declined, but still, staining intensity did not fell observably, revealing that therapy in moxibustion group was better than that in nimodipine group(P<0.05).(4) The content of PS-1、BACE1m RNA in all model group was significantly high than thatin normal group(P<0.01)with RT-PCR. The content of PS-1、BACE1m RNA in rats’ brain tissue in treatment group was lower than that in blank control group(P<0.01). And the content of PS-1、BACE-1m RNAin moxibustion group was lower than that in nimodipine group, with the two figures holding a significant difference(P<0.05). The result revealed that nimodipine and the moxibustion of Tong Du Tiao Shen can both reduce the content of PS-1 and BACE1 m RNA in brain tissue of MCI model rats,and the therapy of moxibustion group’s efficacy was much better than that of nimodipine group.(5) The level of serum IL-6 of all rats in model group was much higher than that in normal group(P<0.01)with ELISA, proving that the level of IL-6 can effect cognitive function. In all treatment group, the level of serum IL-6 in model rats can be reduced observably, and the distinction held the statistics importance(P<0.01). The level of serum IL-6 of rats in moxibustion group was markably lower than that in nimodipine group(P<0.05),revealing that the therapy in moxibustion group was much better thanthat in nimodipine group The result uncovered that the moxibustion of Tong Du Tiao Shen can terminate inflammatory response which was procued by Aβ accumulation, and then relieve and cure the damage that the inflammatory response brought about effectively, which bringing in an improvement on cognitive function.Conclusions(1) The moxibustion of Tong Du Tiao Shen can significantly improve the study ability of the rats who get mild cognitive impairment, which can be regarded as an effective treatment on mild cognitive impairment.(2) The moxibustion of Tong Du Tiao Shen can reduce the level of generation and aggregation of Aβ, which can helps terminate the pathological effect caused by Aβ accumulation.(3)The moxibustion of Tong Du Tiao Shen can make obvious reduction of the content of PS-1 and BACE1 m RNA in the brains of mild cognitive impairment model rats. BACE1 is rate-limiting enzyme produced by Aβ, and, at the same time, it is the first step for App split into Aβ. The gene of PS-1 can promote the generation of Aβ peptide, which proves that to restrain gene expression of BACE1 PS-1can terminate the generation of Aβ from the beginning and that BACE1 and PS1 are perfect target position to prevent and cure MCI.(4)The moxibustion of Tong Du Tiao Shen can also lessen the serum IL-6 levels of MCI rats and stop the inflammatory cascade effect by over-expression of Aβ,and especially weighs in MCI early intervention.