| Objective:The mechanism study of acupoint catgut embedding in the fat layer on simple obesity through leptin-mediated JAK2/STAT3 pathway in hypothalamic.Methods:Sprague-Dawley male rats of the same batch were acclimated for a week.They were randomly divided into control group and high-fat diet model group(HFD).Rats in the control group were fed with normal diets,while the HFD group were fed with high fat diets to induced obesity.After 20 weeks,the HFD group was selected as an obesity model rat with a body weight above the average weight 20%of the control group.Then,it was randomly divided into model group,fat layer-embedded group(FLE),and fat layer-embedded+AG490 group(FLE+AG490),and the control group was used as normal group,10 rats in each group.The catgut was embedded into the fat layer of the acupoints of bilateral Tianshu(ST25),Shuidao(ST28),Pishu(BL20)and Sanjiaoshu(BL22),Zhongwan(CV12)in the FLE group,and treated once a week for 4 times.The FLE+AG490 group rats were treated with the same embedded method as in the FLE group,and intraperitoneal injection of AG490(1mg/kg)daily during the treatment period.The other two groups of rats were only fixed.After 4 weeks,observe the change of weight,use enzyme linked immunosorbent assay to analyze the serum leptin.Real-time polymerase chain reaction and Western Blot were used to analyze the expression of JAK2,STAT3,SOCS3,leptin receptors mRNA and protein in the hypothalamus.Results:1.Body weight1.1 Before and after modeling:There was no significant difference in weight between the control group and the HFD group before modeling(P>0.05),and the weight of the HFD group increased significantly after 20 weeks(P<0.Ol).1.2 After intervention:Comparison within the group:After the intervention,the weight of the FLE group was significantly lower than that before the intervention(P<0.01),while the FLE+AG490 group had no significant change in weight before and after intervention(P>0.05).Comparison between groups:Compared with the model group after the intervention,the FLE group was significantly reduced(P<0.01),while the FLE+AG490 group had no difference(P>0.05).The weight of the FLE group was significantly lower than that of the FLE+AG490 group.2.Serum leptin level:Compared with the normal group,the level of leptin in the model group significantly increased(P<0.01).Compared with the model group,the FLE group was significantly reduced(P<0.01),but there was no significant difference in the FLE+AG490 group(P>0.05).The level of leptin in the FLE group was significantly lower than that of the FLE+AG490 group(P<0.05).3.Expression of JAK2/STAT3 pathway-related mRNA in each group of rats after intervention(1)Leption receptor mRNA expression:Compared with the normal group,the expression level in the model group was significantly reduced(P<0.01).Compared with the model group,the FLE group and the FLE+AG490 group were significantly increased(P<0.01,P<0.05).The expression level of leption receptor in the FLE group was significantly higher than that of the FLE+AG490 group(P<0.01).(2)JAK2 mRNA expression:Compared with the normal group,the expression level in the model group was significantly reduced(P<0.01).Compared with the model group,the FLE group and the FLE+AG490 group were significantly increased(P<0.01).The expression level of JAK2 mRNA in the FLE group was significantly higher than that of the FLE+AG490 group(P<0.01).(3)STAT3 mRNA expression:Compared with the normal group,the expression level in the model group was significantly reduced(P<0.01).Compared with the model group,the FLE group was significantly increased(P<0.01),while the FLE+AG490 had no significant difference(P>0.05).The expression level of STAT3 mRNA in the FLE group was significantly higher than that of the FLE+AG490 group(P<0.01).(4)SOCS3 mRNA expression:Compared with the normal group,the expression level in the model group was significantly increased(P<0.01).Compared with the model group,the FLE group was significantly reduced(P<0.01),while the FLE+AG490 had no significant difference(P>0.05).The expression level of SOCS3 mRNA in the FLE group was significantly lower than that of the FLE+AG490 group(P<0.01).4.Expression of JAK2/STAT3 pathway related proteins in rats in each group after intervention(1)Leption receptor protein expression:Compared with the normal group,the expression level in the model group was significantly reduced(P<0.01).Compared with the model group,the FLE group was significantly increased(P<0.01),while the FLE+AG490 had no significant difference(P>0.05).The expression level of leption receptor protein in the FLE group was significantly higher than that of the FLE+AG490 group(P<0.01).(2)JAK2 protein expression:Compared with the normal group,the expression level in the model group was significantly reduced(P<0.01).Compared with the model group,the FLE group was significantly increased(P<0.01),while the FLE+AG490 had no significant difference(P>0.05).The expression level of JAK2 protein in the FLE group was significantly higher than that of the FLE+AG490 group(P<0.01).(3)STAT3 protein expression:Compared with the normal group,the expression level in the model group was significantly reduced(P<0.01).Compared with the model group,the FLE group was significantly increased(P<0.01),while the FLE+AG490 had no significant difference(P>0.05).The expression level of STAT3 protein in the FLE group was significantly higher than that of the FLE+AG490 group(P<0.01).(4)SOCS3 protein expression:Compared with the normal group,the expression level in the model group was significantly increased(P<0.01).Compared with the model group,the FLE group and the FLE+AG490 group were significantly reduced(P<0.01),while the expression level of SOCS3 protein in the FLE group was significantly lower than that of the FLE+AG490 group(P<0.01).Conclusion:1.Acupoint catgut embedding in the fat layer can down-regulate serum leptin levels,hypothalamic SOCS3 mRNA and protein expression,up-regulate hypothalamic leption receptor,JAK2,and STAT3 mRNA and protein expression,activate JAK2/STAT3 signaling pathway,and achieve the regulation of simple obesity.2.The JAK2/STAT3 signaling pathway in hypothalamic may be involved in the regulatory process of acupoint catgut embedding in the fat layer to reduce the weight of simple obese rats. |
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