Objective： To observe the effects of repeated electroacupuncture （EA） of Zusanli （ST36）Yanglingquan （GB34） on hypothalamic acetylcholinesterase （AchE） and vesicular acetylcholine （ACh） transporter （VAChT） activities and choline acetyltransferase （CHAT） mRNA and muscarinic M1 receptor （MIR） mRNA expression in chronic constrictive injury （CCI） and/or ovariectomy （OVX） rats so as to reveal its underlying mechanism in cumulative analgesia. Methods： A total of 103 female Wistar rats were randomly divided into normal control （n=15）, CCI （n=15）, CCI＋EA2d （n=15）, CCI＋EA2W （n=15）, OVX＋CCI （n=13）, OVX＋CCI＋EA2d （n=15）, and OVX＋CCI＋EA2W groups （n=15）. CCI model was established by ligature of the unilateral sciatic nerve with surgical suture. Memory impairment model was established by removal of the bilateral ovaries. Morris water test was conducted to evaluate the OVX rats＇ memorylearning ability, and the thermal pain threshold （PT） of the bilateral paws was detected the next morning after EA. EA （2/15 Hz, 1 mA） was applied to bilateral ST36-GB34 for 30 min, once daily for 2 days or 2 weeks, respectively. Hypothalamic AChE activity was detected by histochemistry, VAChT immunoactivity was determined by immunohistochemistry, and ChAT mRNA and MIR mRNA expressions were assayed by reverse transcription-polymerase chain reaction （RT-PCR）. Results： In comparison with the normal control group, the AChE activity in hypothalamic arcuate nucleus （ARC） and supraoptic nucleus （SON） regions of CCI group, AChE activity in paraventricular nucleus （PVN）, ARC, and SON regions of OVX＋CCI group, and hypothalamic muscarinic M1R mRNA expression levels in both CCI and OVX＋CCI groups were down-regulated significantly （P〈0.05）. Compared with the CCI group, the AChE activities in hypothalamic ARC and SON regions of CCI＋EA2d and CCI＋EA2W groups and PVN region of CCI＋EA2W group and hypothalamic ChAT mRNA and M1R mRNA expres
