[Influence of electroacupuncture and moxibustion and their treated mouse serum on the proliferation of the cultured splenetic CD4+ CD25+ regulatory T cells of tumor-bearing mice].
OBJECTIVE
To observe the effect of electroacupuncture (EA) and moxibustion of "Dazhui" (GV 14) on the proliferation levels of the splenetic CD4+ CD25+ regulatory T cells (Tregs) of H22 tumor-bearing mice in vitro.


METHODS
Forty eight Balb/c mice were randomized into control, model, moxibustion and EA groups, with 12 cases in each. H22 tumor-bearing model was set up by hypodermic injection of H22 tumor cells (0.2 ml, 1 x 10(7) cells/ml). EA (2 Hz, 2 mA) was applied to "Dazhui" (GV 14) and left "Huantiao" (GB 30) for 20 min, and moxibustion was applied to "Dazhui" (GV 14) 2 moxa-cones every time. The treatment was given from the 2nd day on after innoculation of tumor cells, once every other day, 6 times altogether. After the treatment, the mice were killed by peeling off the eyeball and blood samples were collected to be separated into serum. Then, Tregs of the spleen tissues of Balb/c mice in different groups were isolated by using megnetic activated cell sorting (MACS) system to be cultured independently, and co-cultured with EA-treated serum and moxibustion-treated serum separately in culture fluid for 96 h, added with 3H-tritiate thymidine (TdR) in the culture-fluid 12 h before the end of culture, followed by collecting the cells and detecting their proliferation levels (count per minute, cpm) by using a lipid scintillation device.


RESULTS
The proliferation level of Tregs in model group was elevated significantly compared to normal control group (P < 0.05), while in comparison with model group, those of Tregs of EA and moxibustion groups decreased considerably (P < 0.01). After separate application of the diluted acupuncture-treated serum and moxibustion-treated serum at 1 : 1 and 1 : 8 (not 1 : 16 and 1 : 32) to the cultured Tregs, their proliferation levels (cpm) in EA and moxibustion groups were obviously upregulated in comparison to those of normal control group (P < 0.05), and the cpm in EA group was significantly higher than that in model group (P < 0.05), suggesting a different action mechanism between acupuncture-moxibustion treatment and serum stimulation.


CONCLUSIONS
EA of "Dazhui" (GV 14) and "Huantiao" (GB 30) and moxibustion of "Dazhui" (GV 14) can effectively downregulate the proliferation level of the cultured splenetic Tregs of the tumor bearing mice. EA-treated serum and moxibustion-treated serum diluted at 1 : 1 and 1 : 8 can evidently upregulate the proliferation level of Tregs in vitro.
Liu
Zhi-Dan
ZD
Department of Acu-moxibustion, Longhua Hospital Affiliated to Shanghai University of Chinese Medicine, Shanghai 200032, China. liu54643@163.com
Pei
Jian
J
Fu
Qin-Hui
QH
Li
Hai-Yan
HY
Yu
Qi-Wen
QW
Zhang
Ji-Ying
JY
Zhang
Dong-Qing
DQ
D015669:Acupuncture Points; D000818:Animals; D015704:CD4 Antigens / Q000378:metabolism*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015671:Electroacupuncture; D005434:Flow Cytometry; D053645:Interleukin-2 Receptor alpha Subunit / Q000378:metabolism*; D008280:Magnetics; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009071:Moxibustion; D009369:Neoplasms / Q000276:immunology*; D044967:Serum; D013154:Spleen / Q000166:cytology / Q000276:immunology*; D050378:T-Lymphocytes, Regulatory / Q000166:cytology* / Q000276:immunology
D015704:CD4 Antigens; D053645:Interleukin-2 Receptor alpha Subunit