| Objectives To investigate the effects of Jianpi Huashi granule in colon of Na+-K+-ATPase,Occludin,AQP3 and AQP4 expression levels of rats with diarrhea-predominant irritable bowel syndrome(D-IBS),to further explore the mechanism of Jianpi Huashi granule improving diarrhea symptoms and enrich the theory of traditional Chinese medicine.Methods 1 SD rats were randomly divided into normal group(N),model group(M),Jianpi Huashi granule low(LD),middle(MD)and high dose(HD)groups,and positive drug group(pinaverium bromide,0.03g/kg),n=10 each of group.Except the normal group of rats,the other groups received Senna for gastric administration combined with restraint stress to establish D-IBS models.After the model was successfully established,the normal groups were always given a gavage of normal saline at the volume.the model groups were given a gavage of normal saline after giving Senna for gastric and restraint stress 60min.The positive drug group were given pinaverium bromide(1.5mg/ml)after giving Senna for gastric and restraint stress 60min.Jianpi Huashi granule low,middle and high dose groups were given low,middle,high dose(6.25g/kg,12.5g/kg,25g/kg)after giving Senna for gastric and restraint stress 60min,administered at 10ml/kg two weeks.2 Index 1)Enzyme-linked immunosorbent assay(ELISA)was used to assay Na+-K+-ATPase activity in D-IBS Colon.2)Immunohistochemical(IHC)method was used to detect the expression levels of Occludin,AQP3 and AQP4 in D-IBS Colon.3)Western blot was used to detect the protein expression levels of Occludin,AQP3 and AQP4 in D-IBS Colon.4)Reverse transcription-PCR(RT-PCR)method was used to detect the expression levels of mRNA in Occludin AQP3 and AQP4.Results 1 Body weight change(n=10 rats/group):There was no significant difference in body weights between rats in each group before the establishment of the D-IBS model(P>0.05).After the establishment of the D-IBS model,Compared to N group,body weights of M,LD,MD,HD and PC groups(250.50±8.67,256.50±14.4,257.10±14.48,252.50±17.16,257.30±13.55 vs 294.30±21.50)were decreased significantly(P ?0.05).2 The diarrhea grade change(n=10 rats/group):There was no significant difference in body weights between rats in each group before the establishment of the D-IBS model(P?0.05).The successfully established on day 14,compared to N group,The diarrhea grade of M,LD,MD,HD,PCgroups(3.36±0.72,3.36±0.68,3.38±0.68,3.36±0.34,3.40±0.68 vs 0.10±0.22)were significantly increased(P?0.05).3 Colonic tissue pathology observation:Observation of colon and microscopic tissue staining.Each group showed no edema,ulcer,congestion,inflammatory cell infiltration and so on,Compared with NC group,there were no significant difference in other groups.4 The results of ELISA test of the Na+-K+-ATPase activity in D-IBS Colon:Compared to M group,the expression levels of Na+-K+-ATPase activity in colon were significantly increased in LD,MD,HD and PC group(4.47±0.69,6.15±0.15,7.03±0.70,4.67±0.66 vs 1.66±0.20)(P?0.05).5 The results of IHC test of the Occludin,AQP3 and AQP4 in D-IBS Colon:Compared to M group,the expression levels of Occludin(0.33±0.02,0.43±0.01,0.46±0.01,0.23±0.01 vs 0.16±0.01),AQP3(0.35±0.01,0.41±0.01,0.44±0.01,0.30±0.01 vs 0.18±0.01)and AQP4(0.27±0.01,0.32±0.02,0.39±0.01,0.28±0.02 vs 0.17±0.01)in colon were significantly increased in LD,MD,HD and PC group(P?0.05).6 The results of WB test of the Occludin,AQP3 and AQP4 in D-IBS Colon:Compared to M group,the protein expression levels of Occludin(0.99±0.01,1.08±0.02,1.16±0.03,0.70±0.02 vs 0.47±0.03),AQP3(0.84±0.05,0.94±0.02,1.12±0.04,0.77±0.02 vs 0.59±0.04)and AQP4(1.12±0.02,1.24±0.03,1.37±0.02,0.42±0.01 vs 0.10±0.01)in colon were significantly increased in LD,MD,HD and PC group(P?0.05).7 The results of RT-PCR test of the mRNA in Occludin,AQP3 and AQP4 in D-IBS Colon:Compared to M group,the protein expression levels of mRNA of O ccludin(0.99±0.01,1.09±0.01,1.19±0.03,0.61±0.01 vs 0.53±0.01),AQP3(0.95±0.01,1.01±0.02,1.10±0.03,0.82±0.01 vs 0.74±0.02)and AQP4(0.95±0.02,1.07±0.02,1.18±0.01,0.82±0.01 vs 0.66±0.01)in colon were significantly increased in LD,MD,H D and PC group(P?0.05).Conclusions 1.Jianpi Huashi Granules can effectively improve the symptoms of D-IBS diarrhea.2.Jianpi Huashi granule can increase Na+-K+-ATPase activity,the expression levels of Occludin,AQP3,AQP4 and mRNA of Occludin,AQP3,AQP4 in colon,so as to influense the effect of the water absorption ability in colon.This is probably one of the therapeutic mechanisms of Jianpi Huashi granule of D-IBS.Figure 4;Table 12;Reference 55... |
PDF Full Text Request