| Objective:To explore anti-aging effect, appropriate intervention condition, mechanism and action pathway of Artemisia burning products.Methods:(1) With time and concentration as two factors,2 x 3 factorial experiment design was applied in the experiment.70 SAMP8 mouse were randomized into 6 intervention groups and 1 model group, with 10 mice in each group. The 6 moxa smoke intervention groups included group with low concentration and 15min (Group Al), group with low concentration and 30min (Group A2), group with middle concentration and 15min (Group B1), group with middle concentration and 30min (Group B2), group with high concentration and 15min (Group Cl), and group with high concentration and 30min (Group C2).10 age-matched SAMR1 mouse were used as normal group. Fume the 6 intervention groups with Artemisia burning products (i.e. Moxa smoke) of different concentration and time for 4 weeks, and group M and Z were just done with the same grasping. The change of learning and memory behavior was tested by passive avoidance experiment and Morris water maze test. HE staining was used to observe the pathological change of lung. Contents of monoamine neurotransmitter and cytokines both in brain and serum were tested by ELISA. Antioxidant enzymes and cholinergic transmitter were tested by colorimetry. Amino acid neurotransmitter was tested by RP-HPLC. The activity of telomerase in liver was tested by PCR-ELISA. The activity of ERK, p38 MAPK and JNK in hippocampus was tested by Western-Blot.(2) 60 KM mouse were randomized into 5 groups:blank group, intervention group, sham operation group, model group and model intervention group. There were 15 mice in model group and model intervention group, and 10 in other 3 groups. Olfactory dysfunction animal model was made by intranasal administration of Zinc surfate solution. Sham operation group was administered with distilled water as control. Fume mice in model group and model intervention group with moxa smoke of middle concentration and 20min for 4 weeks. Other groups were just done with the same grasping. Olfactory dysfunction was assessed by buried food pellet test, and the contents of Monoamine neurotransmitters in brain and cytokines in serum were tested by ELISA.Results:Results of research on anti-aging effect mechanism of Artemisia burning products were as follows: (1) Animal behavior:In passive avoidance test, there was no statistically significant difference in latency and error number between group M,6 intervention groups and group Z. In place navigation test of Morris water maze, latency of group M,6 intervention groups and group Z showed a volatile downtrend with the increase of training days, but there was no statistically significant difference between 8 groups. Training day had a significant influence on latency, while the influence of time, concentration and their interaction on latency still can't be seen. For the total distance, there was a volatile downtrend with the increase of training days. Training day had a significant influence on total distance, and the interaction between time and concentration may influence total diatance.The detection indexes in spatial probe test showed no significant difference between 8 groups.(2) Cytokine:Level of Thl type cytokines in group M, like IL-4 (in brain, serum and spleen) and IL-10 (in serum and spleen), was significantly lower than that in the group Z. Level of Thl type cytokines in group M, like IFN-y (in brain, serum and spleen), IL-2 (in serum) and TNF-α(in serum and spleen), was significantly higher than that in group Z. Cytokines in 6 intervention groups were either the same with group M, or better than group M.(3) Antioxidant enzyme:Content of MDA (in brain and serum) in group M was higher than that in group Z, but activity of SOD (in serum) and GSH-Px (in brain and serum) was remarkably lower in group M than that in group Z. Indexes in 6 intervention groups were either the same with group M, or better than group M. According to factorial analysis, there were interaction between time and concentration in the influence to SOD (in serum) and GSH-Px (in brain and serum). Among 6 intervention groups, the activity of SOD (in serum) got the best result in group B2, while the activity of GSH-Px (in brain and serum) got the best result in group B1.(4) Ach:The activity of AChE in group M was higher than that in group Z, while the activity of ChAT in group M was lower than that in group Z. The activity of AChE in 6 intervention groups had no difference with that in group M. The activity of ChAT in group A2, B1 and B2 was higher than that in group M, while other groups had no difference with group M.(5) Monoamine neurotransmitter:Compared with group Z, contents of 3 kind of monoamine neurotransmitter (in brain and serum) in group M decreased. Compared with group M, contents of 3 kind of monoamine neurotransmitter in 6 intervention groups were either the same or much better. According to factorial analysis, time and concentration had an interaction on monoamine neurotransmitter. Among 6 intervention groups, the contents of monoamine neurotransmitter were highest in group B1.(6) Amino acid neurotransmitter:Content of GABA in group M was statistically lower than that in group Z, but contents of Glu and Asp in group M were statistically higher than that in group Z. Contents of 3 kind of amino acid neurotransmitter in 6 intervention groups were either the same with group M or much better. According to factorial analysis on GABA, there were no interaction between time and concentration, while both time and concentration had influence on GABA. Result of group B2 was the best among 6 intervention groups.(7) Pathology of lung:There were RBC and inflammatory cells infiltration in alveolar space and intracellular substance. Overall speaking, lung pathology in 6 intervention groups was better than that in group M.(8) Telomerase:Activity of liver telomerase in group M was higher than that in group Z. Among 6 intervention groups, the activity of telomerase in group B1, B2 and C1 was lower than that in group M, while other groups had no difference with group M.(9) MAPK:Grey values of hippocampus ERK, P38 and JNK in group M were higher than that in group Z and 6 intervention groups. According to factorial analysis, the appropriate intervention condition for P38 was middle concentration, while for JNK it was low concentration and 30 min.Results of research on olfactory pathway of Artemisia burning products were as follows:(1) Weight:Before the experiment, there was no difference between 8 groups; after the experiment, weight in blank group, intervention group, and sham operation group was higher than that in model group and model intervention group.(2) BFPT:Before preparing model, there were no differences in results of BFPT and control test among 8 groups. After modeling, results of BFPT and control test in model group and model intervention group were higher than that in other groups. After experiment, results of BFPT and control test in model group were higher than that in other 4 groups, and there were no difference between the 4 groups.(3) Monoamine neurotransmitter of brain:There was no difference in brain monoamine neurotransmitter between 5 groups. (4) Cytokine of serum:Contents of IFN-γand IL-2 in model group, model intervention group and sham operation group were higher than that in blank group, and there was 80% certainty on finding content of IL-10 in model intervention group lower than that in blank group. Besides, there was 80% certainty on finding content of IL-2 in intervention group higher than that in blank group, and there were no difference between other cytokines.Conclusion:Conclusions of research on anti-aging effect mechanism of Artemisia burning products are as follows:(1) Intervention by moxa smoke can't aggravate the senescence procedure of SAMP8 mouse. Instead, intervention by moxa smoke can exert anti-aging effect through the following ways:increasing contents of Ach and monoamine neurotransmitter, reducing neurotoxicity action of amino acid neurotransmitter, enhancing antioxidation, regulating the balance of Thl/Th2 cytokine and decreasing over-expression of liver telomerase, etc.(2) The appropriate intervention condition of moxa smoke varies with detection indexes. To increase content of monoamine neurotransmitter, the appropriate intervention condition is middle concentration and 15min; to enhance antioxidation, it is middle concentration and 15min or30min; to regulate the balance of Th1/Th2 cytokine, it's middle concentration.(3) The intervention of moxa smoke does no significant influence on learning and memory behavior of SAMP8 mouse.(4) The intervention of moxa smoke can work through MAPK pathway. For P38, the appropriate intervention condition is middle concentration; for JNK, it's low concentration and 30min.Conclusions of research on olfactory pathway of Artemisia burning products are as follows:(1) The model of olfactory dysfunction can be made by intranasal administration with 10μ15% ZnSO4 for 4 days. However, this model is reversible, and recovery of dysosmia can be seen in 4 weeks.(2) Intranasal administration with Zinc sulfate solution can cause stress on mice, and then results in the occurrence of inflammation.(3) This experiment can't confirm the role of olfactory pathway in the effect of Artemisia burning products.
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