Research On The Mechanism Of Different Acupoints In Reducing Weight Based On Neuro-immune Interaction

ObjectiveThis study was based on a High fat diet-induced obese rat model,the effects of different acupoints in reducing weight were observed.To explore the neurobiological mechanism of acupuncture about Nerve-macrophage-adipose cross-talk,the nerves,and immune activity were investigated in inguinal white adipose tissue(iWAT).This research provides a certain scientific basis for acupuncture in weight loss.Methods:1.Weaning Sprague-Dawley(SD)rats were randomly divided into normal group,high-fat diet(HFD)group,HFD+Tianshu(ST25)group,HFD+Zusanli(ST36)group,HFD+Quchi(LI11)group,the electroacupuncture(EA)parameters are 2/15 Hz,2 mA,20min/times,6 times a week,total 4 weeks.The weight loss effect of electroacupuncture was measured by weight,length,and fat mass,and the improvement of blood fat was evaluated by serum cholesterol,triglycerides,low/high-density cholesterol,glutamic-pyruvic transaminase,and free fatty acids.2.Using neuroelectrophysiological technique to measure iWAT sympathetic nerve activity,the double-hook electrode was hook by the nerve,and an amplifier was connected.After the nerve discharge signal stabilized,the Spike2 software was used to measure the activity of the adipose sympathetic nerve during EA.The effects of EA at ST25,ST3 6,LI11(2/15 Hz,2 mA,120s)on the frequency of sympathetic nerve discharge in rats at different periods were recorded.3.Elisa was used to measuring the serum inflammatory factors(IL-6,TNF?,TGF?,IL-4,IL-10)in the normal group,high-fat diet(HFD)group,HFD+ST25 group,HFD+ST36 group,HFD+LI11 group;real-time quantitative PCR was used to measure the levels of iWAT inflammatory factors(IL-1?,TNF?)and anti-inflammatory factors(IL-10,Argl)in normal group,high-fat diet(HFD)group,HFD+ST25 group.4.The whole-mount immunostaining was used to measure the co-expression of sympathetic nerve-associated macrophages(SAMs)in the normal group,high-fat diet(HFD)group,HFD+ST25 group.and the relative protein levels of norepinephrine transporter(Slc6a2),?3 receptor,tyrosine hydroxylase(TH)and uncoupling protein 1(UCP1)in iWAT was measured by Western-blot.Results:1.All acupoints can significantly resist weight gain from the high-fat diet.ST25 has the most significant improvements in body weight,Lee's index,fat mass,and liver weight.ST36 has the most significant effect on improving total cholesterol.LI11 has the most significant effect on improving high/low-density cholesterol,ST25 and ST36 have the most significant effect on improving serum free fatty acids.ST25 and LI11 can significantly improve glutamic-pyruvic transaminase.2.All acupoints(ST25,ST36,LI11)have different effects on nerve activity in iWAT.Under normal conditions,ST36 had the best excitatory effect on neural activity;in the obese state,the ST25 had the best excitatory effect on neural activity;after EA for four weeks,the ST36 had the best excitatory effect on neural activity.3.Under normal conditions,the peak discharge frequency of ST36 is 60-90s,and ST25 is 30-60s;Under the obesity condition,the peak discharge frequency of ST36 is 90-120s,and ST25 is 60-90s.After the EA intervention,the peak discharge frequency of ST36 is 60-90s,and ST25 is 0-30s.The time of the peak discharge frequency of ST25 is earlier than that of ST36.EA can improve the neural signal delay in the obese state.4.All acupoints(ST25,ST36,LI11)can improve the inflammatory status of obese rats.ST25 and LI11 significantly decrease the inflammatory factors(IL-6,TNF?,TGF?),and the ST26 significantly increases the anti-inflammatory factors(IL-4,IL-10).ST25 decreases Inflammation factor(IL-1?,TNF?)in iWAT,and increases anti-inflammatory factor(IL-10,Argl).5.ST25 reduced sympathetic nerve-associated macrophages(SAMs),reduced the protein norepinephrine transporter(Slc6a2)expression,increased protein expression of ?3 receptor and tyrosine hydroxylase(TH),increased adaptive thermogenic-related protein uncoupling protein 1(UCP1).Conclusion:1.EA at ST25,ST36,LI11 can effectively reduce weight,and there are differences in weight loss effects.Overall,ST25 has the most effective in losing weight.2.EA at ST25 can significantly excite the sympathetic nerve in iWAT and improve the passivation of the sympathetic nerve signal in high fat diet SD rats,and EA at ST36 can significantly excite the sympathetic nerve in normal and loss weight-loss SD rats.3.EA at ST25,LI11,and ST36 can effectively improve the systemic inflammatory state.EA at ST25 can significantly improve the inflammatory state of iWAT.4.EA at ST25 reduces the sympathetic nerve associated macrophages(SAMs)through nerve-immune crosstalk,thereby reducing the transport of norepinephrine,improving sympathetic nerve passivation;increasing the UCP1,?3 receptor and TH protein expression.It has been shown that neural-immune interactions play an important role in electroacupuncture for weight loss.So,EA at ST25 plays the best weight loss effect which improves the passivation of the sympathetic nerve signal and the state of inflammation under the state of obesity through neuro-immunity interaction.The mechanism may be to activate the adipose sympathetic nerve,reduce the number of sympathetic nerve-associated macrophages(SAMs),reduce the transport of norepinephrine,and thereby increase the energy consumption of fat production.