| Purpose：To observe the change of the liver-stagnation and spleen-deficiency irritable bowelsyndrome (IBS) rat model hippocampus, hypothalamus, colon tissue5-hydroxytryptamine(5-HT), vasoactive intestinal peptide (VIP) and vasoactive intestinal peptide receptor (VIP-R),the expression of aquaporin8(AQP8) in colon, and the influence of the eye acupoint (EA). Toexplore the pathophysiology basis of the liver-stagnation and spleen-deficiency IBS diseasecombine model from the central and peripheral two aspects, and the biological effect of the eyeacupoint. To provide the experimental basis for illustrating the happening of theliver-stagnation and spleen-deficiency IBS and the mechanisms of the eye acupuncture.Material and method：1.28Wistar SPF male rats were randomly chose from40to establish the liver-stagnation andspleen-deficiency IBS animal model with the combination methods of “the mentalstimulation and improper diet and overstrain”. To use six kinds of stimulation as follows: coldwater swimming to exhaustive, fasting for24h, water deprivation for24h,45℃hyperthermyoven for15min, clip tail for5min, electric shock (30V, shock for5s, pause for5s, process for3min in all), randomly selected a kind of stimulation everyday, combining with homemadecages bound for1h, process for3round,18d in all. Observed the change of the rats’ generalstate, body weight change and growth rate of body mass, and evaluated the model. Afterpassing the evaluation, selected24successfully molding rats, randomly divided into threegroups as Normal, Model, Eye acupoint (EA),12per group. After the EA group molded, used31size (diameter13mm) acupuncture needle, needled the rats’ lower energizer, large intestine,hepatic region, spleen intervention for7d.2. Enzyme-linked immuno sorbent assay (ELISA) was used to detect the content of5-HT, SPand VIP in serum and colon tissue.3. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNAof VIP, VIP-R and AQP8in colon tissue.4. Strept actividin-biotin complex (SABC) immunohistochemistry was used to detect and analyze the expressions of5-HT, VIP and VIP-R in hippocampus, hypothalamus and colon.Results：1. The effects of eye acupuncture on general state and the content of5-HT and SP in serum ofthe liver-stagnation and spleen-deficiency IBS rat1.1The effects of eye acupuncture on general state of the liver-stagnation and spleen-deficiency IBS ratThe molded rats appeared depression, agitated, lose flesh, stool soft or deformed, anddefecate times increased gradually and stool turned thin. Expressed as liver-stagnation andspleen-deficiency, after acupuncture treatment for7d, the eye acupuncture rats looked likebetter and the stool improved to be normal, while model group had no obvious change.Compared to the Normal, in the18th day, the growth rate decreased significantly (P<0.01)in Model and EA; Compared to the Normal in the25th day, the growth rate of the EA setincreased dramatically (P<0.01). Compared to the Normal in the18th day, the rate of fecesmoisture content increased significantly (P<0.01) in Model and EA; Compared to the Normalin the25th day, the rate of feces moisture content decreased dramatically (P<0.01) in EA.1.2The effects of eye acupuncture on the content of5-HT and SP in serum of theliver-stagnation and spleen-deficiency IBS ratCompared to the Normal, the content of5-HT and SP in serum increased dramatically inModel and EA (P<0.01). Compared to the Model, the content of5-HT and SP in serumdecreased dramatically in eye acupuncture (P<0.01).2. The effects of eye acupuncture on the colon tissue5-HT, VIP, VIP-R and AQP8of theliver-stagnation and spleen-deficiency IBS rat2.1The effects of eye acupuncture on the content and expression of5-HT in colon tissue ofthe liver-stagnation and spleen-deficiency IBS ratCompared to the Normal, the content of5-HT in colon tissue increased significantly inModel and EA (P<0.01). Compared to the Model, the expression of5-HT dramaticallydecreased (P<0.01) in eye acupuncture rat colon tissue.2.2The effects of eye acupuncture on serum, the content and expression of VIP in colontissue of the liver-stagnation and spleen-deficiency IBS ratCompared to Normal set serum, the content of serum VIP increased significantly in Model (P<0.01). Compared to the Model, the content of serum VIP decreased significantly ineye acupuncture (P<0.01), the expression of VIP mRNA in colon tissue increaseddramatically in Model (P<0.05or P≤0.01).2.3The effects of eye acupuncture on the expression of VIP receptor in colon tissue of theliver-stagnation and spleen-deficiency IBS ratCompared to Normal, the expression of VIP receptor mRNA in colon tissue increasedsignificantly in Model (P<0.01). Compared to the Model, the expression of VIP receptormRNA in colon tissue decreased dramatically in eye acupuncture (P<0.01).2.4The effects of eye acupuncture on the expression of AQP8in colon tissue of theliver-stagnation and spleen-deficiency IBS ratCompared to Normal, the expression of AQP8mRNA in colon tissue decreasedsignificantly in Model (P<0.01). Compared to Model, the expression of AQP8mRNA incolon tissue increased dramatically in eye acupuncture (P<0.01).3. The effects of eye acupuncture on the expression of CA3area of hippocampus and5-HT,VIP, VIP receptor in hypothalamus tissue of the liver-stagnation and spleen-deficiency IBSrat3.1The effects of eye acupuncture on the expression of CA3area of hippocampus and5-HTin hypothalamus tissue of the liver-stagnation and spleen-deficiency IBS ratCompared to Normal, the expression of CA3area of hippocampus and5-HT inhippocampus decreased significantly in Model (P<0.01). Compared to Model, the expressionof CA3area of hippocampus and5-HT in hypothalamus tissue increased dramatically in eyeacupuncture (P<0.01).3.2The effects of eye acupuncture on the expression of CA3area of hippocampus and VIP inhypothalamus tissue of the liver-stagnation and spleen-deficiency IBS ratCompared to Normal, the expression of CA3area of hippocampus and VIP inhypothalamus tissue increased significantly in Model (P<0.01). Compared to Model, theexpression of CA3area of hippocampus and VIP in hypothalamus tissue decreaseddramatically in eye acupuncture (P<0.05or P<0.01).3.3The effects of eye acupuncture on the expression of CA3area of hippocampus and VIPreceptor in hypothalamus tissue of the liver-stagnation and spleen-deficiency IBS rat Compared to the Normal, the expression of CA3area of hippocampus and VIP receptorin hypothalamus tissue increased significantly in Model and eye acupuncture (P<0.01orP<0.05). Compared to Model, the expression of VIP receptor in hypothalamus tissuedecreased dramatically in eye acupuncture (P<0.01).Conclusion：1. The liver-stagnation and spleen-deficiency IBS rat model was established successfully withthe combination methods of “mental stimulation and improper diet and overstrain”.2. The content of5-HT and SP in serum increased significantly in Model. Eye acupuncture canimprove the syndrome and decrease the content of5-HT and SP in serum.3. The expression of5-HT, VIP and VIP receptor in colon tissue increased significantly and theexpression of AQP8decreased markedly in Model. Eye acupuncture can changeover them.4. One of the possible mechanisms of soothing liver and strengthening spleen with eye acupu-ncture is down-regulation the expression of5-HT, VIP and VIP receptor and up-regulation theexpression of AQP8in colon tissue.5. The expression of5-HT in hippocampus and hypothalamus tissue decreased significantly inModel, and the expression of VIP and VIP receptor increased significantly. Eye acupuncturecan changeover them.6. There are two possible central zones of hippocampus and hypothalamus with theliver-stagnation and spleen-deficiency IBS rat, and there are two neurotransmitter of5-HT andVIP involved in the formation of the liver-stagnation and spleen-deficiency IBS.7. One of the possible mechanisms of soothing liver and strengthening spleen with eye acupu-ncture is up-regulation the expression of5-HT in hippocampus and hypothalamus tissue anddown-regulation the expression of VIP and VIP receptor. |
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