Regulatory Mechanism Of Gastrointestinal Motility Disorder In FD Rats By Electro-acupuncture On Zusanli Based On ICC’s Ultrastructure And Ca²⁺Inside/outside ICC

Object:1.Verify the regulating effect of gastrointestinal motility disorder in FD rats by Electro-acupuncture on Zusanli,2.Observe the change of ICC’s ultrastructure and Cx43in FD rats by Electro-acupuncture on Zusanli,3.Discuss the regulatory Mechanism of Gastrointestinal Motility Disorder in FD rats by Electro-acupuncture on Zusanli based on Ca2+inside/outside ICC.Methods:1.FD rats model was made by stimulated using sponge forceps on their tails, using Electro-acupuncture on Zusanli as intervention means, using domperidone as positive control, obsive and record the appetite, weight, gastric emptying rate and small intestine advance rate of experimental rats.2.After the rats received the stimulation by sponge forceps on their tails and received intervention means using the methods in first part experiment, obsive the ultrastructure of ICC in stomach and intestine of FD rats by transmission electron microscope, detect the expression of Cx43in stomach and intestine of FD rats by immunohistochemistry.3.Separate and culture ICC from intestine of infantile SD rats, after the ICC received intervention by serum from FD rats, using serum from FD rats by Electro-acupuncture on Zusanli to intervene the ICC. Then, detect the concentration of Ca2+inside/outside ICC by microplate test, detect the expression of IP3R-. RyR in ICC by Western-Blot, detect the concentration of mRNA of IP3R、RyR in ICC by RT-PCR.Result:1. The model rats’spirits were drooping, sedentary, fur color was dim, baldness, faeces was watery, appetite and weight were lower than the normal group rats (P<0.01), the gastric emptying rate and small intestine advance rate were slower. The appetite and weight were higher than the model group rats in Electro-acupuncture group (P<0.01), the gastric emptying rate and small intestine advance rate recover to normal level in Electro-acupuncture group.2. ICC in normal group were present in triangular or fusifrom shapes. They had a higher electron-density cytoplasm compared with that of surrounding smooth muscle cells and contained abundant cell organelles, such as mitochondria, endoplasmic reticulum and caveolae et al. Along the length of overlapping processes, ICC predominantly fromed large gap junctions between adjoining cells, suggesting a functional relationship between these cell types. However, altered ICC ultrastructure (swollen or vacuolated mitochondria, lack of mitochondrial cristae, dilatated and vesiculated rER et al.) was observed frequently in model group. Damage of gap junctions between ICC and smooth muscle cells were also seen. In contrast, the ultrastructural abnormalities in electroacupuncture group exhibited some improvement compared to model group. But the ultrastructural morphology had not returned to normal as observed in normal group and minor injury could be seen occasionally. Immunohistochemistry showed that compared with the normal group, the expression of Cx43in the small intestine and antrum of the model group significantly decreased, with P<0.01. After electroacupuncture treatment, compared with the model group, the expression of Cx43in the small intestine and antrum significantly increased, with P<0.013. Cultured ICC show oval or fusifromis shape, with big cell nucleus and little cytoplasm. Under fluorescent microscope, the cytoplasm of ICC show green fluorescence, we say it c-kit positive, and the cell nucleus of ICC show blue fluorescence.4. Compared with normal ICC, the concentration of Ca2+inside ICC intervened by serum from FD rats was lower (F<0.01), and the concentration of Ca2+outside ICC was higher(P<0.05). Compared with ICC intervened by serum from FD rats, the concentration of Ca2+inside ICC intervened by serum from FD rats by Electro-acupuncture on Zusanli was higher, and the concentration of Ca2+outside ICC was lower (P<0.05)5. Compared with normal ICC, the expression of IP3R、RyR inside ICC intervened by serum from FD rats were lower(P<0.01). Compared with ICC intervened by serum from FD rats, the expression of IP3R、RyR inside ICC intervened by serum from FD rats by Electro-acupuncture on Zusanli was higher (P<0.01)6. Compared with normal ICC, the concentration of mRNA of IP3R%RyR inside ICC intervened by serum from FD rats were lower (P<0.01) Compared with ICC intervened by serum from FD rats, the concentration of mRNA of IP3R、RyR inside ICC intervened by serum from FD rats by Electro-acupuncture on Zusanli was higher (P<0.01)Conclusion:1. The method using sponge forceps to stimulate SD rats’tails could make them FD.2. Electro-acupuncture on Zusanli had regulating effect of gastrointestinal motility disorder in FD rats.3. The regulating effect of gastrointestinal motility disorder in FD rats by Electro-acupuncture on Zusanli maybe relate to the ICC and Cx43of the small intestine and antrum in FD rats.4. The concentration of Ca2+inside/outside ICC maybe the regulatory Mechanism of gastrointestinal motility disorder in FD rats by Electro-acupuncture on Zusanli.