ZHANG Man¹, YANG Jun¹, WANG Ping², WANG Pin¹

(1 ZheJiang University of Traditional Chinese Medicine Second Hospital,Hangzhou 310005;2Anhui University of Traditional Chinese Medicine First Hospital, Hefei 230038)

Abstract Objective To investigate the in vitro intervening effect of rat serum effector substances on  vascular  endothelial  cell  proliferation  and  migration  after  stasis-eliminating  and  meridianunblocking moxibustion and explore the possible proangiogenic mechanism of stasis-eliminating and  meridian-unblocking  moxibustion  treatment  for  vascular  dementia.  Method  Male  adult Wistar rats were used. A rat model of vascular dementia was made by four-vessel occlusion (4-VO). The rats were randomized to moxibustion, Western medication and model groups. A sham operation group was established as a control. There was a  total of four groups. The moxibustion group received aconite cake moxibustion on point Baihui and suspended moxibustion on points Dazhui(GV14) and Shenting(GV24) and the Western medication, an oral gavage of aniracetam tablets. Treatment was given 15 days  as one course for a total of two courses. Blood was taken from the Orbital cavity in the moxibustion, sham operation and model groups after each course of treatment.  After  stasis-eliminating  and  meridian-unblocking  moxibustion,  animal  serum  was prepared  to  intervene  in  human  umbilical  vein  endothelial  cell  (HUVEC-12)  culture. Another 

parallel liquid group was set up. RPMI-1640 culture medium containing 10% fetal bovine serum (FBS) was prepared. Endothelial cell proliferation was examined by methyl thiazolyl tetrazolium (MTT)  colorimetric  assay.  Endothelial  cell  migration  was  examined  by  cell  scratch  labeling. Result   After  two  courses  of  treatment,  the  examination  of  endothelial  cell  proliferation  by methyl  thiazolyl  tetrazolium  (MTT)  colorimetric  assay  showed  that  rat  serum  intervened significantly  in  human  umbilical  vein  endothelial  cell  (HUVEC-12)  proliferation  in  the moxibustion serum group compared with the model serum, sham operation serum and parallel liquid groups (P＜0.01). At 24 hrs after two courses of treatment, the examination of endothelial cell migration by cell scratch labeling showed that rat serum intervention significantly increased the HUVEC-12 migration rate in the moxibustion serum group compared with the  model serum, sham operation serum and parallel liquid groups (P＜0.01). Conclusion  Stasis-eliminating and meridian-unblocking  moxibustion  has  a  marked  improving  effect  on  vascular  endothelial  cell proliferation and migration in vascular dementia rats, which provides strong evidence for its in vitro proangiogenic effect.

Key  words ： Dementia,  vascular,  Moxibustion,  Endothelial  cell,  Cell  proliferation,Cell migration,Angiogenesis,Rats