| Background: Functional dyspepsia(FD) are very common in daily clinical practice. They are characterized by disturbances in motility patterns and/or visceral hypersensitivity. Increasing evidence is indicating that acupuncture(AP) at the Zusanli point(ST36) can alleviate FD. However, the potential mechanisms underlying this effects is not very clear.Objective: To investigate the pathogenesis mechanisms of FD and the potential mechanisms of theraputic effect of AP at the Zusanli point(ST36).Methods: In each group except for control group, FD rat models were established by repeated exposure to water avoidance stress. Daily food intake, weight and intestinal sensitivity in control group and model group were investigated. The proximal jejunum was cut, deep muscular plexus of interstitial cell of Cajal(ICC) was investigated using confocal laser scanning microscopy. Serum Ig G, Ig M, IL-2, and IL-6 levels were detected. Gene expression profile in small intestine was detected using gene chip technology. Differrential genes and pathways were analyzed. Next, rats in AP group underwent electroacupuncture(EA) at ST36 for 15 days. In sham-AP group, rats underwent sham AP. In model group, rats did not undergo AP. In control group, rats were bred naturally. Then, daily food intake, weight and intestinal sensitivity in control group, model group, AP group and sham-AP group were investigated. The proximal jejunum was cut, deep muscular plexus of ICC was investigated using confocal laser scanning microscopy. Serum Ig G, Ig M, IL-2, and IL-6 levels were detected. Gene expression profile in small intestine was detected using gene chip technology. Differrential genes and pathways were analyzed.Results:1. After modeling, daily food intake was lower in model group than in control group(T=-5.360, P<0.01), weight was lower in model group than in control group(T=-3.144, P<0.05)and intestinal sensitivity was higher(when CRD is 20,40,60 or 80 mm Hg, AWR was higher in model group than in control group, T=5.00, P<0.01; T=2.71, P<0.05; T=2.71, P<0.05; T=3.16, P<0.05 respectively). After EA at ST36, daily food intake was higher in AP group than in model group(P<0.05), and weight was higher in AP group than in model group(P<0.05). When CRD is 20 mm Hg, AWR was no significant difference between AP group and model group(P>0.05). When CRD is 40, 60 or 80 mm Hg, AWR was lower in AP group than in model group(P<0.05; P<0.05; P<0.05 respectively)2. After modeling, in model group, the morphology of deep muscular plexus of ICCs in the small intestine was abnormal. The number of ICCs was lower. The integrated optical density(IOD) was lower than in control group(T=-21.45, P<0.01). After EA at ST36, the morphology of ICCs in the small intestine was normal. The number of ICCs was higher in AP group than in model group. The IOD was higher in AP group than in model group(P<0.05).3. After modeling, the serum Ig G, Ig M and IL-2 levels were lower in model group than in control group(T=-11.67, P<0.01; T=-11.26, P<0.01; T=-4.44, P<0.01 respectively). Serum IL-6 level was higher in model group than in control group(T=5.23, P<0.01). After EA at ST36, serum Ig G, Ig M and IL-2 levels were higher in AP group than in model group(P<0.05; P<0.05; P<0.05 respectively) and serum IL-6 level was lower in AP group than in model group(P<0.05).4. Differential genes between model group rats and control group rats were screened using SAS software provided by gene chip company. The genes that no gene ID were ruled out. P<0.01 and foldchange(FC) >2 or <0.5. There were 255 differential genes between model group and control group, eleven genes were up-regulated and 244 genes were down-regulated. Top 10 genes that up-regulated were RGD1559622, Tsks, Gramd2, Colq, Kcnq4, Mecom, Tulp1, Thrb, Acot1, Lss. Top 10 genes that down-regulated were LOC100362150, Mcpt2, Igh-6, Mcpt1l4, Cpa3, Mcpt8l2, Fcer1 a, LOC363827, Gpr174, LOC100363695. There were 19 pathways related to differential genes :(1)Reninangiotensin system,(2)Primary immunodeficiency,(3)Staphylococcus aureus infection,(4)Intestinal immune network for Ig A production- Rattus norvegicus(rat),(5)Natural killer cell mediated cytotoxicity,(6)NOD-like receptor signaling pathway- Rattus norvegicus(rat),(7) Cell adhesion molecules(CAMs)- Rattus norvegicus(rat),(8) Ether lipid metabolism,( 9) Fc epsilon RI signaling pathway,( 10) Leukocyte transendothelial migration,( 11) Cytokine-cytokine receptor interaction- Rattus norvegicus(rat),(12)Toll-like receptor signaling pathway,(13)T cell receptor signaling pathway,(14)Chemokine signaling pathway,(15) Phagosome,(16) Fc gamma Rmediated phagocytosis- Rattus norvegicus(rat),( 17) Antigen processing and presentation,( 18) Lysosome- Rattus norvegicus(rat),( 19) Regulation of actin cytoskeleton. Fifteen pathways were related to immunoreaction. Three pathways were related to mast cells reaction. Five pathways were related to cell proliferation, differentiation and apoptosis. Two pathways were related to movement of cell and activities or metabolism in the cell.There were 169 differential genes between AP group and model group, 96 genes were up-regulated and 73 genes were down-regulated. Top 10 genes that up-regulated were Slc34a2, Trpv6, Gucy2 g, Hbb, Hbb-b1, LOC287167, RGD1564463, Akp3, Alas2, Gsg1. Top 10 genes that down-regulated were Acot1, LOC690226, Hmgcs2, Gbx1, Retsat, Klrb1 b, Ppp1r1 c, Slc27a2, LOC683474, Masp2. There were 25 pathways related to differential genes:(1)Biosynthesis of unsaturated fatty acids,(2)Steroid biosynthesis,(3)Synthesis and degradation of ketone bodies,(4)Glycosphingolipid biosynthesis- globo series,(5)PPAR signaling pathway,(6)Fatty acid metabolism,(7)Butanoate metabolism,(8)Mineral absorption,(9)Valine,leucine and isoleucine degradation,( 10) Steroid hormone biosynthesis,( 11) Ascorbate and aldarate metabolism,( 12) Porphyrin and chlorophyll metabolism,( 13) Glycosphingolipid biosynthesis- lacto and neolacto series,( 14) Retinol metabolism,( 15) ABC transporters,(16) Pentose and glucuronate interconversions,(17)Drug metabolism- cytochrome P450- Rattus norvegicus(rat),(18)Drug metabolism- other enzymes-Rattus norvegicus(rat),(19)Metabolism of xenobiotics by cytochrome P450- Rattus norvegicus(rat),(20)Carbohydrate digestion and absorption,(21) Peroxisome-Rattus norvegicus(rat),(22) Fat digestion and absorption,(23)Starch and sucrose metabolism,( 24) Arachidonic acid metabolism,( 25) Glycerophospholipid metabolism. Eight pathways were related to lipid metabolism. Seven pathways were related to carbohydrate metabolism. Three pathways were related to metabolism of hormone and enzyme. Two pathways were related to activities of the cell membrane and transport or catabolism in cell. One pathway was related to cell proliferation and differentiation.Conclusions: In stress-related FD, EA at ST36 could improve food intake and weight, and could improve the symptom of visceral hypersensitivity. It is a good therapeutic approach; in stress-related FD, in small intestine, the morphology of ICCs was abnormal and the number was decreased, which may be causes of gastrointestinal motility dysfunction. There was immune suppression and immune imbalance, which may be causes of visceral hypersensitivity; in stress-related FD, EA at ST36 showed good therapeutic effects. The mechanisms may be related to its repairing effects on ICCs damages in gastrointestinal tract and its regulation on immune suppression and immune imbalance; in molecular mechanism of FD, the results indicated the pathways were related to immunoreaction, mast cells reaction, cell proliferation, differentiation and apoptosis, et al. Further research could be done based on the differential genes and the pathways. In molecular mechanism of therapeutic effect of AP at ST36, the results indicated the pathways were related to lipid metabolism, carbohydrate metabolism, hormone and enzyme metabolism, et al. Further research could be done based on the differential genes and the pathways. |
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