| Objective:Along with the progress of the society and the improvment of health care level, how to anti-aging has become the focus in the21st century. Aging has become an unprecedented important research subj ect human facing. Seek drugs and methds which can slow the aging and prolong life, having important economic and social benefits. Acu-puncture has characteristics and advantages in anti-aging aspect. This study adopt Shuangguyitong (tonifying congenital and acquired essence, eliminating pathogen) acupuncture therapy, to investigate the influence of spleen lymphocytes apoptosis and its related gene on the aging rats. Furthermore, it provides a certain experiment basis on electroacupuncture regulation of aging.Methods:Forty Sprague-Dawley rats (female),5-month old, were divided into normal control (Nc),aging model group (Model), shuangyuyitong acupuncture group (EA) and a control acupuncture group (EAc),10rats in each group. The model of aged rats was established by D-galactose in the latter three groups except the control group. The acupoints of "Guanyuan (GV4)"、"Housanli (ST36)"、"Baihui (GV20)’ were used for electroacupuncture treatment. The acupoints of "Zhongji(GV3)"、"Yinl inquan (SP9)" and "Yintang (GV29)" were used for electroacupuncture control treatment, whice was near the con trol poin selection,electrically (2Hz,1mA),15minis everyday,six times per week for4weeks. The escape latency was verified by Morris water maze experiment. Chemluminescence deteckd SOD activity. Ly mphocyte apoptosis was tested by Annex in V double staining flow cytometry. Real-time quantitative RT-PCR to test the expression of pro-apoptotic genes Fas mRNA、Bax mRNA and anti-apoptosis gene Bcl-2mRNA in aging-rats.Result:1. Escape latency to find the platform in Nc group was45s and that was63s in model group, which was obvious different from each other (P<0.01). In EA and EAc group, escape latency to find the platform was51s and53s respectively, with no significant difference between them. However, there was different from those of the model group (both P<0.05).2. Serum superoxide dismutase SOD level of aging model rats in Nc group was84.3±8.82NU/ml and that was19.83±3.95NU/ml, which was significantly decreased compared with Nc group (P<0.01) SOD level in EA and EAc group was33.93±2.81NU/ml and35.31±4.42NU/ml, with no obvious difference between them. However, there was significantly different from that of the model group (both P<0.05).3. In comparison of the control rats (8.5±2.22)%, the model rats expressed with the lower level of Lymphocyte apoptosis (28.94±3.68)%, which was different from each other (P<0.01).In (13.9±2.92)%and EAc (21.24±3.7)%, the level of spleen cell apoptosis was cut down, which was different from those of the model group (both P<0.05). Furthermore, the therapeutic effect of the EA group is different from that of EAc group (P<0.05).4. The expresstion of anti-apoptotic gene Bcl-2mRNA was (0.481±0.12)%and apoptosis-related genes Bcl-2/Bax ratio (0.966±0.018) in the NC group, those were (0.247±0.031)%,(0.323±0.041)in the model group,which were both significantly lowered compared with Nc group (P<0.01). The EA group were (0.539±0.018)%and (0.538±0.039), differences were statistically significant between the model group (P<0.05).5. The expresstion of apoptotic gene Fas mRNA was (0.487±0.03)%in the Nc group, which was different from the model group (0.961±0.21)%(P<0.01). It was (0.942±0.16)%in electroacupuncture on "Shuangguyitong" acupoints group(EA), but there was no obvious significance between them.Conclusion:1. Compared with Nc group, spatial learning and memory capacity and serum superoxide dismutase SOD levels of aging-rats was significantly decreased (P<0.01).Electroacupuncture can not only improve the decline of spatial learning and memory, but also the content of SOD in serum and thus enhance the body response to oxidative stress, Aging model rats induced by D-galactose showed that the model was successful in the behavioral and biochemical indication.2. Spleen lymphocyteapoptosis rate of the model group decreased obviously, intervening excessive apoptosis of lymphocytes by electroacupuncture by means of improving the lymphocytes function, which can regulate immune function,for the purpose of delaying the immunosenescence.3. Electroacupuncture can enhance the level of Bcl-2gene and Bcl-2/Bax ratio expresstion except Fas mRNA in aging-rats, with coordination and integration related pro-apoptotic genes and anti-apoptoticgene expression. It could be regulation patterns of gene-related apoptosis in aging rats, which was effected by electroacupuncture " Shuangguyitong " acupotints. Therapeutic effect of the regulation of electroacupuncture at "Shuanggu yitong" acupoints on apoptosis of lymphocytes in aging rats, which was better than EAc group.4. The effectiveness of regulating Lymphocyte apoptosis in shuangyuyitong treatment group with "Guanyuan’、"Housanli’、"Baihui" which was much better than that was in the conventional distribution point group with "Zhongji"." Yingl ingquan"、"Yintang", revealing the superiority of increaseing zhengqi in combating the aging process. |
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