TRPV1 Was Involved In Responses To Acupuncture And Moxibustion In Acupoint On Local Effective And Gastric Motility Related Cortex Responding Of Mice

Objective: 1.With different stimulation methods of acupuncture and moxibustion on different acupoints of mice,including ST36,DU14,RN4,to observe the difference starting substance in each acupoint area,such as mast cells,serotonin,histamine,calcitonin gene-related peptide,and substance P.2.To observe whether TRPV1 participates in the local initiation process of acupuncture and moxibustion,and how different stimulation methods affect local initiation process.3.To analyse whether TRPV1 is a key mediating substance in the central response of acupuncture to regulate gastric movement,and how different interventions of acupuncture and moxibustion affect the central response.Methods:1.Male C57BL/6 mice were randomly divided into normal group,acupuncture Zusanli group,acupuncture Dazhui group,acupuncture Guanyuan group,hot moxibustion Zusanli group,hot moxibustion Dazhui group,and hot moxibustion Guanyuan group.Male TRPV1 knockout mice were randomly divided into normal group,acupuncture Zusanli group,and hot moxibustion Zusanli group.Acupuncture needles were given to the C57BL/6 acupuncture group and the TRPV1-/-acupuncture group.The hand acupuncture stimulated the acupoints,and the C57BL/6 hot moxibustion group and the TRPV1-/-hot moxibustion group were treated with a hot moxibustion apparatus and stimulated warmly.2.Toluidine blue was used to detect the expression of mast cells in skin of acupoints.Immunofluorescence was used to detect the expression of chemical substances(HA,5-HT,CGRP,and SP)in skin areas.3.The contents of mast cells,HA,5-HT,CGRP,SP,VR1 in the local cryptal tissue and GSH-R and 5-HT1 A,5-HT2 C,5-HT3,5-HT4 in cerebral cortex were detected by ELISA and WB.5-HT7 protein expression.Results: 1.Acupuncture or moxibustion can increase the count of mast cells.Compared with the blank group,acupuncture at Zusanli,Dazhui and Guanyuan can increase the number of mast cells in the acupoints.(P<0.01,P<0.05,P<0.05),Moxibustion of Zusanli,Dazhui and Guanyuan can increase mast cells in acupoints(P<0.01,P<0.05,P<0.05),Hot moxibustion Zusanli group is better than acupuncture Zusanli group(P<0.05).After the acupuncture,HAand 5-HT mainly distributed in the subcutaneous tissue with local aggregation as the main form.After the heat moxibustion,HA and 5-HT were widely distributed in the epidermis,dermis and subcutaneous tissues.Compared with blanks,acupuncture at Zusanli,Dazhui and Guanyuan can lead to increased HA and 5-HT levels in cryptozones.(P<0.05),Moxibustion of Zusanli,Dazhui and Guanyuan can also increase the content of HA and 5-HT in cavernous areas(P<0.01,P<0.05,P<0.05);The expression of 5-HT in acupuncture at Zusanli was significantly better than acupuncture at Dazhui and Guanyuan Formation(P<0.01),Moxibustion Zusanli group is also superior to hot moxibustion Dazhui and Guanyuan group(P<0.01).CGRP and SP are located at the ends of nerve fibers in strips and distributed in the subdermal and subcutaneous tissues.Compared with the blank group,acupuncture at Zusanli,Dazhui,and Guanyuan groups can increase the expression of CGRP.(P<0.05,P<0.05,P<0.01),Heat moxibustion can also increase the expression of CGRP in Zusanli and Guanyuan(P<0.05);Acupuncture Zusanli is better than hot moxibustion Zusanli group(P<0.05).The difference was statistically significant.VR1 in the skin distributes mainly around the dermis and hair follicles,Compared with the blank Zusanli group,acupuncture and hot moxibustion Zusanli group can significantly increase the expression of VR1 positive nerve fibers in acupoints(P<0.05,P<0.001),Hot moxibustion group is better than acupuncture group(P<0.01);Compared with the blank Dazhui group,acupuncture and hot moxibustion Dazhui group significantly increased VR1 content(P<0.05,P<0.01)Hot moxibustion group is better than acupuncture group(P<0.05);The content of VR1 in Zusanli,which is better than Dazhui and Guanyuan Group(P<0.05),Zusanli is higher than that of acupuncture Guanyuan.2.Compared with the TRPV1-/-blank group,there was an increase in mast cell counts and degranulation in the TRPV1-/-acupuncture group.(P<0.05);The mast cell counts of acupuncture and hot moxibustion group in TRPV1 knockout mice were lower than those in C57BL/6 wild type mice.(P<0.05,P<0.01),The difference was statistically significant;The number of degranulation is basically the same as that of mast cell counts.Compared with the blank group,the expression of HA and 5-HT increased significantly in the acupuncture group and the hot moxibustion group,and accumulated in a large amount around the blood vessels and hair follicles Compared with the TRPV1-/-blank group,the expression of TRPV1-/-acupuncture groupincreased(P<0.05);HA expression in TRPV1-/-hot moxibustion group was significantly lower than that in C57BL/6 hot moxibustion group(P<0.01);5-HT expression in TRPV1-/-acupuncture group was lower than that of C57BL/6acupuncture group(P<0.05)The expression of 5-HT in TRPV1-/-moxibustion group was significantly lower than that in C57BL/6 group(P<0.01)Compared with C57BL/6acupuncture group,the expression of CGRP-positive nerve fibers was reduced in TRPV1-/-acupuncture group(P<0.05);Compared with C57BL/6 hot moxibustion group,CGRP and SP content in TRPV1-/moxibustion group decreased(P<0.05).3.Compared with C57BL/6 blank group,acupuncture can cause significant reduction of GHS-R content(P<0.05),5-HT1 A content is significantly reduced(P<0.05),5-HT2 Ccontent is significantly reduced(P<0.05),5-HT3 content is significantly reduced(P<0.05),The content of 5-HT7 in C57BL/6 acupuncture group is significantly reduced(p<0.05);Hot moxibustion can lead to a significant reduction in GHS-R content(P<0.05),5-HT3 content is significantly reduced(P<0.05)5-HT7 content is significantly reduced(P<0.05);The difference was statistically significant.Compared with TRPV1-/-blank group,TRPV1-/-acupuncture group caused a significant decrease in the content of 5-HT1A(P<0.05),TRPV1-/-Hot moxibustion group results in decreased 5-HT4 content(P<0.05),The difference was statistically significant.Conclusion:1.Acupuncture and moxibustion have different effect on mast cells and noxious neuropeptides,in detail,moxibustion affects mast cell degranulation more than acupuncture and acupuncture affects noxious neuropeptides more than moxibustion.2.The distribution of mast cells and nociceptive neuropeptides differes in different acupoints.Compared with DU14 and RN4,ST36 area has more biological active substances and is can be activated by both acupuncture and moxibustion more easily.3.TRPV1 receptors participate in the initiation process of acupuncture and moxibustion.The initiation effect of mediating moxibustion is stronger than that of needles,indicating that other receptors may participate in the initiation process of acupuncture.4.TRPV1 mediates the central response of acupuncture to regulate gastric movement,but there are differences in the neurotransmitters affected by both.