| Background: Syndrome of deficiency of spleen-Qi (SDSQ) is a specific term inChinese traditional medicine, which is characterized with anorexia, abdominaldistension and loose watery stools. It commonly induces to the dysfunction ofgastrointestinal motility. Therefore the recovery of gastrointestinal motility function isthe first and critical step to remedy the damage of gastrointestinal dysfunction. ICC iswidely distributed in gastrointestinal tissues which show a highly branched morphologyand form unique networks in gastrointestinal tract which coordinate gastrointestinalmotility. ICC is the medium of the nerve signal conveying from the enteric nerve system(ENS) to the smooth muscle of the gastrointestinal tract. ENS, ICC and smooth musclecell (SMC) connect each other to form network structure and constitute the basicfunctional unit of gastrointestinal motility. ENS, ICC and SMC has importantrelationship with gastrointestinal motility dysfunction. The results of clinical and animalstudy showed that there were serious gastrointestinal motility dysfunction in the patientswith SDSQ, and acupuncture in ZU SAN LI (st36) and TIAN SHU (s25) can correct thegastrointestinal motility dysfunction and effectively treat the SDSQ. But the mechanismwas not clear. Objective: To make the model of SDSQ in rats and observe the morphologicalchanges of enteric nerve, ICC and smooth muscle cells in rats with SDSQ, and toinvestigate the therapeutic effects of acupuncture in st36and s25. The mechanism ofgastrointestinal motility disorders caused by SDSQ and the therapeutic effects ofacupuncture in st36and s25would be explored.Methods: Sixty healthy Wistar rats of both sexes weighing180to220g wererandomly divided into six groups:①control group (n=10),②SDSQ group (n=10),③natural recovery group (n=10),④acupuncture in st36treated group (n=10),⑤acupuncture in s25treated group (n=10),⑥acupuncture in st36and s25treated group(n=10). The model of SDSQ was established according to Chen’s method. According tothe SDSQ group, the normal saline was gavaged for the rats in control group. Afterestablishment of SDSQ was successful, the proximal10cm segment of jejunumbeginning2cm distal to the pylorus from each group was taked and was studied usingc-Kit, α-actin and protein gene product9.5(PGP9.5), vesicular acetylcholinetransporter (VAChT), substance P (SP), vasoactive intestinal polypeptide (VIP) andneuronal nitric oxide synthase (nNOS), and immunohistochemical simple ordouble-staining with whole-mount preparation technique and confocal laser scanningmicroscopy and transmission electron microscope.Results:1.Anatomical changes Compared with those in control group, the gastrointestinaltracts were distended and intestinal obstruction significantly in the rats in SDSQ group;Compared with the SDSQ group, the distention was markedly alleviated in the threeacupuncture group.2. immunofluorescence:(1) Enteric Nerve: Before the SDSQ model was established, the number andintegrated optical density (IOD) value of PGP9.5, Ach, SP, VIP and NO were nostatistically significance in each group. After the SDSQ model was successfullyestablished, the number and IOD value of PGP9.5, Ach, SP, VIP and NO weresignificantly decreased compared with the control group (P<0.05). The number and IOD value of PGP9.5, Ach, SP, VIP and NO were significantly decreased in naturalrecovery group compared with control group (P<0.05). After treatment of acupuncture,the number and IOD value of PGP9.5, Ach, SP, VIP and NO in the three acupuncturegroups were significantly increased compared with the group SDSQ and naturalrecovery group (P<0.05). There was no statistically significance between the controlgroup and three treated groups in the number and IOD value of PGP9.5, Ach, SP, VIPand NO.(2) Network of ICC: Before the SDSQ model was established, the number of ICCand IOD value of ICC were no statistically significance in each group. After the SDSQmodel was successfully established, the number and IOD value of ICC weresignificantly decreased compared with the control group(P<0.05). The number and IODvalue of ICC were significantly decreased in natural recovery group compared withcontrol group (P<0.05). After treatment of acupuncture, the number and IOD value ofICC in three acupuncture groups were significantly increased compared with the SDSQgroup and natural recovery group (P<0.05). There was no statistically significancebetween the control group and three treated groups in the number and IOD value ofICC.(2) SMC: Before the SDSQ model was established, the IOD value of SMC were nostatistically significance in each group. After the SDSQ model was successfullyestablished, the IOD value of SMC were significantly decreased compared with thecontrol group(P<0.05). The IOD value of SMC were significantly decreased in naturalrecovery group compared with control group (P<0.05). After treatment of acupuncture,the IOD value of SMC in the three acupuncture groups were significantly increasedcompared with the SDSQ group and natural recovery group (P<0.05). There was nostatistically significance between the control group and three treated groups in the IODvalue of SMC.3. Transmission electron microscopy:The ICC rendered as large and oval nucleus with less cytoplasm while chromatindistributed around the nucleus. Mitochondria, smooth endoplasmic reticulum and Golgi bodies were abundant. Compared the ultrastructural features of ICC of SDSQ group andnatural recovery group with the control group, ICC displayed severe ultrastructuralabnormalities. The ultrastructural features of ICC were severely damaged. Injury wasdisplayed in both the perinuclear cytoplasm and the ICC processes; the nuclei of ICCwere shrunken, and the plasma membrane bleb was observed. Mitochondria wasdecreasing and swelling. Compared the ultrastructural features of ICC of theacupuncture treated group with SDSQ group and natural recovery group, the shape ofultrastructural features of ICC were were significantly recovered, the nuclei was nearlynormal.Conclusions:(1) Gross morphological observation of the gastrointestinal tractsrevealed a marked intestinal distention of gastrointestinal tracts in the rats with SDSQ;acupuncture in st36and s25could alleviate the distention significantly.(2) The numberand IOD of PGP9.5, Ach, SP, VIP and NO were decreased in SDSQ. The enteric nervewas disrupted significantly in SDSQ. Acupuncture in st36and s25could protect theenteric nerve and repair the enteric nerve network.(3) The number and IOD of ICC wassignificantly decreased, ICC network was severely damaged and the ultrastructuralfeatures of ICC were injured in rats with SDSQ. Acupuncture in st36and s25couldenhance the number and IOD of the ICC and repair the ICC network.(4) The IOD ofSMC was significantly decreased, SMC was severely damaged and the ultrastructuralfeatures of SMC were injured in rats with SDSQ. Acupuncture in st36and s25couldenhance the IOD of the ICC and repair the ICC-SMC network.(5) The ultrastructuralfeatures of ICC were damaged significantly in SDSQ. Acupuncture in st36and s25could maintain the ultrastructural features of ICC. |
PDF Full Text Request