| Background: Syndrome of deficiency of spleen-Qi(SDSQ) is a clinical syndrome,which may be induced by eating and drinking without temperance, overstrain, eatingcold and uncooked food excessively and exhaustion of Qi due to protracted disease. Itis characterized with loss of appetite, epigastralgia, tiredness fatigue, deficiency of Qiwith disinclination to talk, loose watery stools, sallow complexion, pale tongue withfur, weak pulse. In traditional Chinese medicine, it is usually be seen in these diseases,such as:diarrhea, abdominal pain, edema, retention of phlegm and fluid, asthma,flaccidity syndrome, epigastric pain, infantile malnutrition. In western medicine, it isalso can be seen in these diseases, such as: chronic gastritis, peptic ulcer, chronicbronchitis, bronchial asthma, and so on. In recent years, it has been confirmed thatinterstitial cells of Cajal(ICC) is the pacemaker cells of the gastrointestinal tract. ICChave a close relationship between enteric nerve system(ENS) and smooth muscle cells(SMC), and it is widely distributed in gastrointestinal tissues. ICC, the nerve ending ofENS and SMC connect each other, they form net work struction, and constitute thebasic functional unit of gastrointestinal motility(BFUGM). It is believed that ICC havethree major functions. The first, ICC is the pacemaker cells of the gastrointestinal tract,which can produce and conduct the physiological slow wave, and control the smoothmuscle of the gastrointestinal tract. The second, ICC facilitates the propagation ofelectrical activity in gut. That is to say ICC can facilitate the electrical activitypropagate to near SMC. The third, ICC can regulate neurotransmitter and convey thenerve signal from the ENS to SMC. So far, at least four items of ICC have beenconfirmed. Respectively are myenteric ICC(ICC-MY), intromuscular ICC(ICC-IM), deep muscular plexus ICC(ICC-DMP), submusocal ICC(ICC-SM). ICC-MY andICC-SM are the pacemaker cells of the gastrointestinal slow wave activity.Disappearing of the gastrointestinal slow wave has a close relationship with ICC-MYand ICC-SM. ICC-IM and ICC-DMP have the following functions: convey theelectrical activity from the ENS to SMC, participate in the neurotransmitter signaltransduction of the gastrointestinal tract.Objective: To make the model of syndrome of deficiency of spleen-Qi in rats inducedwith the method of bitter cold diarrhoea and qi consumption, observing themorphological changes of ENS, expression changes of signal transduction pathway ofENS-ICC-DMP, and the therapeutic effects of Xiang Sha Liu Jun Zi Decoction(XSLJZD).Methods: Sixty Wistar rats of both sexes weighing180to220g were randomlydivided into three groups: control group(n=20), SDSQ group(n=20), treatmentgroup(n=20). The model of SDSQ group and treatment group were administrated byalternate gavage with prescription①and②. Prescription①was made up of rheumofficinale, magnoliae officinalis, immature bitter orange, which was according to(2:1:1) made of200%concentration decoction, Prescription②was made up ofsubprostrate sophora root, inula britannical and areca catechu, which was according to(3:1:1) made of250%concentration decoction. In the model procession, the first18days, the gavage was three times a day, the following12days, the gavage was fourtimes a day, and each time4ml. According to the mothod of SDSQ group, the normalsaline was gavaged for the rats in control group. After the model procession, treatmentgroup were administrated by gavage with XSLJZD(once a day)in two weeks. Themodel of SDSQ group and control group were administrated by gavage with nomalsaline in two weeks according to the treatment group. The proximal segment ofjejunum was taken and studied using the antibody of vesicular acetylcholine transpoter(VAChT), substance P(SP), vasoactive intestinal polypeptide(VIP), neuronal nitricoxide synthase (nNOS), protein gene product9.5(PGP9.5), c-kit, protein kinase A andprotein kinase C, and immunohistochemical simple or double staining withwhole-mount preparation technique. The confocal laser scanning microscopy andtransmission electron microscope were used.Results: Anatomical changes Compared with those in control group, thegastrointestinal tracts were distended significantly in the rats with SDSQ group. Comparaed with the SDSQ group, the changes in rats with XSLJZD was markedlyalleviated.Immunofluorescence Before the SDSQ model was established, the nervenetwork/the number of ICC/integrated optical density(IOD) value of ICC and proteinkinase A(PKA)/protein kinase C(PKC) were analyzed. There were no statisticallysignificance among the three groups. When the model of SDSQ was established,compared with the control group, the quantity and the integrity optical density(IOD) ofAch/SP/VIP/NO/PGP9.5in SDSQ group were significantly decreased(P<0.01), thenerve network was significantly disrupted. After treatment of XSLJZD, compared withSDSQ group, the quantity and the IOD of Ach/SP/VIP/NO/PGP9.5in XSLJZDtreatment group were significantly increased(P<0.01). The nerve network wassignificantly recovered. Compared with the control group, the quantity and the IOD ofICC of intestine in SDSQ group were significantly decreased(P<0.01), the ICCnetwork was significantly disrupted. Compared with SDSQ group, the quantity and theIOD of ICC of intestine in XSLJZD treatment group were significantlyincreased(P<0.01). After treated with XSLJZD, the ICC network was significantlyrecovered. Compared with control group,the IOD of protein kinase A(PKA)/proteinkinase C(PKC) in SDSQ group was significantly decreased(P<0.01). The signaltransduction pathway was destroyed. Compared with SDSQ, the IOD of protein kinaseA(PKA)/protein kinase C(PKC) in XSLJZD treatment group was significantlyincreased. The signal transduction pathway was significantly recovered.Conclusions: The gastrointestinal tracts of the rats with SDSQ which induced by bittercold diarrhoea and qi consumption were significantly distension, and the syndrome ofgastrointestinal obstruction and syndrome of enteroparalysis were significant.Thenumber of Ach/SP/VIP/NO/PGP9.5were decreased in SDSQ. The nerve network wasdisrupted in SDSQ significantly. XSLJZD could prevent the decreasing of the entericnerve and repair the enteric nerve network. The number of ICC was reduced in SDSQsignificantly. ICC network was severely destroyed in rats with SDSQ. The expressionof protein kinase A(PKA)/protein kinase C(PKC) around ICC-DMP was significantlyreduced in SDSQ. Signal transduction pathway of ENS-ICC was destroyed. XSLJZDcould repair the signal transduction pathway of ENS-ICC. |
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