针刺研究2024，Vol.49Issue(4)：341-348,375,9.DOI:10.13702/j.1000-0607.20221296
小鼠初级体感皮层后肢区神经元对"足三里"不同温度热刺激的响应规律
Responsive regularity of neurons in the hindlimb region of primary somatosensory cortex to different temperature thermal needle stimulation of"Zusanli"(ST36)in mice
摘要
Abstract
Objective To study the regularity of central response to thermal needle stimulation of"Zusanli"(ST36)at different temperature,and to analyze the temperature difference of central responses.Methods Six male C57BL/6j adult mice were used in the present study.For observing activities of neurons in the hindlimb region of left primary somatosensory cortex(S1HL,A/P=0.46 mm,M/L=1.32 mm,D/V=-0.14 mm)by using a fast high-resolution miniature two-photon microscopy(FHIRM-TPM),the mice were anesthetized with 3％isoflurane(inhalation),with its head fixed in a stereotaxic apparatus,then,adeno-associated virus(AAV-hSyn-GCaMP6f-WPRE-hGHpA,for showing intracellular calcium transients in neurons transfected)was injected into the left S1HL region using a micro-syringe after scalp surgical operation.The mice's right ST36 were stimulated using internal thermal needles with the temperature being 43℃,or 45℃,or 47℃,separately.Image J software and MATLAB 2020b software were used to process the image data of neuronal calcium activity(Ca2+ signaling)in the left S1HL region,including the instant maximum calcium peak value(ΔF/F)in 2 s,instant calcium spike frequency in 2 s,short-term calcium peak value(ΔF/F)in 3.5 min,short-term calcium spike frequency in 3.5 min,calcium peak duration in 3.5 min,maximum calcium peak value(ΔF/F)at the 1st,2nd and 3rd min,and calcium spike frequency at the 1st,2nd and 3rd min after thermal needle stimulation.Results In comparison with the normal temperature needle stimulation,the instant intracellular maximum calcium peak value,instant calcium spike frequency,short-term maximum calcium peak value,short-term calcium spike frequency,and calcium peak duration of S1HL neurons in response to 43℃,45℃ and 47℃ internal thermal needle stimulation of ST36 were significantly increased(P<0.001,P<0.01).Comparison among the 43℃,45℃ and 47℃ thermal needle stimulation showed that the 45℃ thermal needle stimulation was obviously superior to 43℃ and 47℃ thermal needle stimulation in increasing instant calcium spike frequency,short-term calcium spike frequency and calcium peak duration of S1HL neurons(P<0.001,P<0.01).The 47℃ thermal needle stimulation was stronger than 43℃ and 45℃ thermal needle stimulation in increasing the instant maximum calcium peak value(P<0.001).The maximum calcium peak value was apparently higher(P<0.001)at the 2nd min than that at the 1st and 3rd min after 43℃,45℃ and 47℃ thermal needle stimulation.No significant differences were found in the short-term maximum calcium peak value among the 3 thermal needle stimulation and in the calcium spike frequency among the 3 time points after 43℃,45℃ and 47℃ thermal needle stimulation.Conclusion S1HL neurons respond to all 43℃,45℃ and 47℃ thermal needle stimulation of ST36 in mice,while more actively to 45℃thermal needle stimulation.关键词
初级体感皮层/热刺激/足三里/神经元响应/双光子技术Key words
Primary somatosensory cortex/Thermal needle stimulation/Zusanli(ST36)/Neuron response/Two-photon technology引用本文复制引用
朱海滨,金传阳,周航,孙征,许骞,陆成轩,周子秋,熊嘉玮,张建斌..小鼠初级体感皮层后肢区神经元对"足三里"不同温度热刺激的响应规律[J].针刺研究,2024,49(4):341-348,375,9.基金项目
国家自然科学基金项目(No.81973947) （No.81973947）
南京中医药大学自然科学基金项目(No.XZR2020053) （No.XZR2020053）
2021年南京江北新区新型研发机构青年研究员项目 （）
