| Objective:This study aims to explore the effects of JIAWEIJIAOTAIWAN(JWJTW)in the treatment of IBS-D patients with heart-kidney disinterchange combined with liver depression and spleen deficiency and analysis of changes in intestinal flora.Methods:In this study,60 IBS-D patients complicated with mood disorder(depression or anxiety)were recruited by the outpatient department of our hospital from October,2018 to October,2020.After TCM syndrome differentiation,they were divided into TCM treatment group(Group T)and western medicine control group(Group C)according to the syndrome of liver stagnation and spleen deficiency combined with heart-kidney imbalance.The patients of Group T were given JWJTW,and the patients of Group C were treatet by Bacillus licheniformis for 4 weeks.All the patients were evaluated every 2 weeks.Symptomatic index score,IBS symptom severity scale(IBS-SSS),mood disorder(HAMD/HAMA)score and intestinal flora analysis were analyzed in two groups at the beginning and end of treatment.Results:(1)The levels of three primary symptoms,including abdominal pain,defecate character,defecate number,were significantly alleviated after 4-week treatment in Group T(p<0.05),while only the levels of abdominal pain and defecate frequency could be found significantly improved in Group C(p<0.05).The degree of abdominal pain were significantly reduced on 4 weeks incomparison of those in 2 weeks in both of two groups(p<0.05).In addition,the improvement of stool frequency in Groups was better than that in Group C(p=0.013)after 2 weeks of treatment.However,there was no significant difference in the three main symptoms between the two groups after 4 weeks of treatment(p>0.05).(2)The improvements of depression or irritability and palpitation in Group T was better than those of Group C(p=0.04,p<0.05).The insomnia was improved after 2 weeks and 4 weeks treatment in Group T(p<0.05).The bowel singing and abdominal distension were relieved after 4 weeks compared with those before treatment(p<0.05).(3)IBS-SSS significantly decreased in both groups after 2 weeks and 4 weeks of treatment(p<0.001).There was no significant difference in IBS-SSS between the two groups before and after 2 weeks of treatment.However,levels of IBS-SSS in Group T were significantly lowercompared with those in Group Cafter after 4 weeks of treatment(p=0.0047).(4)Compared with those before treatment,the levels of depression were reduced in both of two groups after 4 weeks treatment(p<0.01,p=0.0052).The degrees of depression improvement in Group T was significantly higher than that in Group C after 4 weeks of treatment(p=0.0187).Similarly,compared with those before treatment,the levels of anxiety were reduced in both of two groups after 4 weeks treatment(p<0.01,p=0.008).The degrees of anxiety improvement in Group T was significantly higher than that in Group C after 4 weeks of treatment(p=0.0014).(5)Group T on yeast was stronger than that of Group C(p<0.01,p=0.0015)after 2 weeks and 4 weeks of treatment,while the Group T on Enterococcus was stronger than that of Group C(p<0.01).The promotion effect of Group T and Group C on Lactobacillus and Bifidobacterium after 4 weeks of treatment was stronger than that after 2 weeks of treatment(p=0.0209,p<0.05).Conclusions:JWJTW can improve the clinical symptoms in IBS-D patients with heart-kidney disinterchange combined with liver depression and spleen deficiency diarrhea,especially for depression or anxiety patients,which is better than Bacillus licheniformis treatment.Objective:To evaluate the effects and mechanism of JIAWEIJIAOTAIWAN(JWJTW)on IBS-D rats with heart-kidney disinterchange combined with liver depression and spleen deficiency via BDNF/TrkB pathway.Methods:A total of 50 rats were divided into five groups:normal group(Con,n=10),model group(Mo,n=10),low dose of JWJTW group(L,n=10),middle dose of JWJTW group(M,n=10)and high dose of JWJTW(H,n=10).The models of IBS-D and with heart-kidney disinterchange combined with liver depression and spleen deficiency were built by three factors and PCPA injection methods,respectively.Gastrointestinal function was evaluated by fecal water content and 1h sugar water intake.The abdominal withdrawal response(AWR)pressure threshold was used to evaluate the degree of intestinal sensitivity in rats.The pain pressure threshold and the maximum tolerance pressure threshold were analyzed.Two pro-inflammatory factors,including Tumor necrosis factor TNF-α and interleukin(IL)-6,and two anti-inflammatory factors,including IL-4 and transforming growth factor β(TGFβ),were selected to evaluate the tension of the pro-anti-inflammatory system in rats.RT-qPCR and Western Blot were used to detect the effects of different doses of JWJTW on the mRNA and protein levels of BDNF-TrkB pathway.16S rRNA was used to analyze the changes of intestinal flora of IBS-D feces before and after JWJTW intervention.Results:(1)Fecal water content:At T1,the water content of feces in Mo group(t=4.606,p<0.01),L group(t=4.301,p=0.001),M group(t=4.8205,p<0.01)and H group(t=2.494,p=0.031)was significantly higher than that in Con group.At T2,the water content of feces in Mo group(t=6.694,p<0.01),L group(t=3.337,p=0.007)and M group(t=3.018,p=0.012)was significantly higher than that in Con group.The water content of feces in the M and H groups increased significantly at T2,which showed no significant difference in comparison of those at T0(M group:t=1.983,p=0.075;H group:t=1.055,p=0.315).These results suggested that the fecal water content in the M group and H group was significantly corrected.(2)1h sugar water intake:At T2,the intake of 1h sugar water in H group(t=2.256,p=0.047)was significantly higher than that in Mo group.The 1h sugar water intake of rats in Con group(t=2.937,p=0.014)and H group(t=2.370,p=0.039)at T2 was significantly higher than that at T0,indicating that the 1h sugar water intake of rats in H group was significantly rectified.(3)Intestinal sensitivity:At T1,the pain pressure threshold(Mo group:t=8.54 7,p<0.01;L group:t=5.736,p<0.01;M dose group:t=6.355,p<0.01;H group:t=5.950,p<0.01)and maximum tolerable pressure threshold(Mo group:t=6.424,p<0.01;L group:t=5.256,p<0.01;M group:t=5.238,p<0.01;H group:t=4.094,p<0.01)were significantly lower than the Con group.At T2,the pain pressure threshold(Mo group:t=6.518,p<0.01;L group:t=3.737,p<0.01;M group:t=3.013,p=0.003)and maximum tolerable pressure threshold(Mo group:t=7.951,p<0.01;L group:t=4.578,p<0.01;M group:t=3.045,p=0.003)were significantly lower than the Con group.At T2,the pain pressure threshold and the maximum tolerable pressure threshold were significantly increased in M and H groups,which showed no significance difference in comparison of those at T0.These results suggested that the pain pressure threshold and the maximum tolerance pressure threshold were significantly improved in M and H groups.(4)Pro-and anti-inflammatory system:At T2,pro-inflammatory factor TNFa in treatment group(L group:t=4.886,p<0.01;M group:t=5.244,p<0.01;H group:t=7.936,p<0.01)and IL-6(L group:t=4.211,p<0.01;M group:t=5.483,p<0.01;H group:t=7.501,p<0.01)was significantly lower than that in Mo group.At T2,the levels of pro-inflammatory factor TNFα(t=2.752,p=0.007)and IL-6(t=3.973,p<0.01)in the H group were significantly lower than those in the L group,indicating that the high-dose JTW had the strongest ability to improve the level of pro-inflammatory factor in rats.At T1,IL-4(M group:t=2.439,p=0.017;H group:t=3.404,p=0.001)and TGFβ(M group:t=2.625,p=0.011;H group:t=3.628,p<0.01)was significantly higher than that in Mo group.At T2,the level of IL-4 in H group was significantly higher than that in Mo group(t=3.393,p=0.001),indicating that high-dose JTW had the strongest ability to improve the level of anti-inflammatory factor in rats.(5)Activity of BDNF-TrkB pathway:At T2,BDNF mRNA in colon tissue of rats in treatment group(L group:t=4.254,p<0.01;M group:t=4.919,p<0.01;H group:t=5.504,p<0.01)and TrkB mRNA(L group:t=4.561,p<0.01;M group:t=4.827,p<0.01;H group:t=5.661,p<0.01),BDNF protein(L group:5.232,p<0.01;M group:t=4.889,p<0.01;H group:t=4.677,p<0.01)and TrkB protein(L group:5.535,p<0.01;M group:t=4.827,p<0.01;H group:t=4.262,p<0.01)was significantly lower than that in Mo group.(6)The results showed that the diversity of intestinal microbiota in IBS-D feces was changed.Firmicutes,Bacteroides and Actinomycetes were the main microflora.There was no significant difference in the composition of the microflora in each group at the phylum level(p>0.05).At the genus level,the richness of intestinal microbiota in the middle dose group was the most similar to that in the normal control group.Eubacterium,Faecalibacterium,Lactobacillus,Ruminococcaceae,Staphylococcus,Jeotgalicoccus and Enterococcus were significantly enriched in IBS-D rats compared to the normal control group(p<0.05),while Lachnospiraceae,Ruminococcus_torques,Mucispirillum and Anaeroplasma were significantly depleted in IBS-D rats.When compared the treatment group with the disease group,we found that Eubacterium,Faecalibacterium,Lactobacillus,Ruminococcaceae and Jeotgalicoccus that increased in disease modeling were significantly depleted after treatment of medium doses of TCM while Lachnospiraceae and Anaeroplasma were enriched in treatment group(p<0.05).Several bacteria with high relative abundance such as Lactobacillus,Lachnospiraceae and Ruminococcaceae had more dense networks in middle dose group than that in the disease group.In addition,it was found that the bacteria in the treatment group varied greatly such as,Lachnospiraceae and Ruminococcus(correlation index:0.69),Ruminococcaceae and Faecalibacterium(correlation index:0.58).Conclusion:(1)The intestinal function is obviously abnormal in IBS-D rats with heart-kidney disinterchange combined with liver depression and spleen deficiency,which is improved by JWJTW treatment in a dose-depentant manner.(2)The intestinal sensitivity is obviously abnormal in IBS-D rats with heart-kidney disinterchange combined with liver depression and spleen deficiency,which is improved by JWJTW treatment.High-dose JWJTW has the strongest ability to improve the maximum pressure tolerance threshold of rats(3)The pro-and anti-inflammation system is obviously abnormal in IBS-D rats with heart-kidney disinterchange combined with liver depression and spleen deficiency,which is improved by JWJTW treatment.High dose JWJTW shows the strongest ability to improve TNFα,IL-6 and IL-4 in rats(4)The BDNF/TrkB pathway is obviously abnormal in IBS rats with heart-kidney disinterchange combined with liver depression and spleen deficiency,which is improved by JWJTW treatment,especially high dose JWJTW.(5)The structure,composition and function of intestinal microbiota in feces of IBS-D rats with heart-kidney disinterchange combined with liver depression and spleen deficiency were out of balance.JWJTW can improve the intestinal microbiota and increase the production of beneficial bacteria to a certain extent after treatment,and form a relevant intestinal microbiota network for IBS-D to further elaborate the role of intestinal microbiota in the occurrence,development and treatment of IBS-D. |
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