| Objective By observing the clinical curative effect of Changjining decoction on patients with IBS-D due to liver-qi stagnation and spleen deficiency syndrome,the clinical advantage of developing Traditional Chinese Medicine in the treatment of IBS-D was evaluated through symptom scale assessment of patients’ symptom improvement and its’ overall effect.At the same time,we explore its mechanism by observing the effect of Changjining decoction on aquaporin and its related signal pathway in model rats with IBS-D of liver-qi stagnation and spleen deficiency syndrome.Methods Clinical research:From January 2016 to December 2017,96 IBS-D patients with liver-qi stagnation and spleen deficiency syndrome were selected from outpatient or inpatient department of the Second Chinese Medicine Hospital of Guangdong Province,and were randomly divided into experimental group and control group by SPSS statistical software.There were 48 cases in each group,the experimental group was treated with Changjining decoction,and the control group was treated with Tongxining granule for 4 weeks.Before and after treatment,the clinical symptom scale were respectively scored,and the data of symptom improvement and recurrence rate were analyzed to explore the mechanism of Chang Jining prescription on how it works.Animal experiment:Eighty SPF rats were randomly divided into model group(n=60)and blank control group(n=20).Rats in the model group followed an irregular diet and were fed with Senna leaf gavage combined with swimming induced fatigue test.The model of IBS-D rats with live qi stagnation and spleen deficiency syndrome was simulated for 14 days.After successful modeling,the rats were randomly divided into model group(n = 20),positive control group(n = 20)and model group(n = 20),each group of rats was half male and half female.The experimental group was given the Chinese herbal medicine the Changjining granule solution gavage,the positive control group was given the Tongxiening granule solution gavage,and the model group and the blank group were given the same volume distilled water gavage once a day for a total of 14 days.After14 days,the general condition(body weight,smoothness of fur etc.)and the activity of the rats was observed.The fecal water content,the degree of dilute stool,the rate of dilute stool and the sensitivity of colon were measured and the sugar water preference experiment was analyzed.The sugar preference experiment and the activity degree of rats were censused and analyzed.Guts from pylorus to the ileocecum was collected,and the carbon propelling rate was calculated.The 3cm colons was collected 5 cm from anus to grind into tissue homogenate.The expression levels of protein of protein kinase A(PKA),cyclic adenosine monophosphate(c AMP)and aquaporin 4(AQP4)in rat colon tissues were detected and analyzed statistically to explore the mechanism of Changjining prescription works.Results Clinical study:The clinical symptom scale: After treatment,the score of Changjining group was 6.75±3.32,and that of Tongxiening group was 8.58±4.02.The improvement degree of Changjining group was better than that of Tongxiening group(P<0.05).The effective rate of symptom improvement: The effective rate of symptom improvement was 93.75% in Changjining group,which was significantly higher than that of Tongxiening group(77.08%)(P<0.05).Symptoms recurrence rate: The symptom recurrence rate of the Changjining group was 4.17%,which was significantly lower than that of the group of Tongxiening(20.83%)(P<0.05).Animal Experiment:The body mass of rats: the body weight of model group increased 13.36 ±3.45 g,22.96±5.07 g in Changjining group,18.44±5.78 g in Tongxiening group,28.79±5.79 g in blank group.The body weight of Changjining group,Tongxieninggroup and blank control group were significantly higher than that of model group(P<0.01),and the weight gain in the group of Changjining was higher than the group of Tongxiening and the difference was significant(P<0.05).IBS-D model of moisture of the rat feces: model group rats stool water content(unit:%)was 44.32 ± 2.99,Changjining group was 33.47 ± 3.10,Tongxiening group was 39.32±2.62,the blank group was 32.71±2.42.Compared with the model group,the water content of feces(P <0.05)in the blank group,the Changjining group and the Tongxiening group decreased significantly.Compared with Tongxiening,the fecal water content in Changjining group decreased significantly(P<0.05).IBS-D model rat stool rate results: model group rat stool rate(unit:%)was 65.22±9.38,Changjining group was 37.92±7.21,Tongxiening group was56.95±7.67,the blank group was 36.13±6.07.Compared with the model group,the stool ratio of the blank group,Changjining group and the Tongxiening group decreased significantly(P<0.05).Compared with Tongxiening,the stool rate of Changjinning group was significantly lower(P<0.05).IBS-D model rats sugary water preference results: the sugar water preference rate of IBS-D model rats(unit:%)was 55.87±3.20,Changjining group was 70.56±2.24,the Tongxiening group was 61.95±2.95,blank The group was 73.95±2.45.The preference level of sugar water in the blank group,the Tongxiening group and the Changjining group were significantly higher than those in the model group(P<0.050).Compared with the preference rate of the sugar water in the Tonxiening group,that in the Changjinning group was significantly increased,with significant difference(P<0.050).Evaluation of open field experiment in IBS-D model rats: the scores of open field test in model group were 101.74±5.25,in Changjining group was115.41±8.50,in Tongxiening group was 107.67±7.89,and in blank group was124.78±5.77.The vertical score of open-field test in model group rats was12.92±3.06,that in Changjining was 17.96±3.17,and in Tongxiening group group was 14.86±2.41,and 18.02±3.15 in blank group.The vertical score(P<0.050)and horizontal score(P<0.050)in the blank group,Tongxiening group and Changjining group were significantly higher than those in the model group(P<0.050).Among them,Changjining group had higher level scores and higher vertical scores than those in the Tongxiening group,and the difference was significant(P<0.050). Results of colonic sensitivity of IBS-D model rats:the threshold of abdominal lift pressure(unit: mm Hg)was 29.92±4.01 in model group,36.16±3.82 in Changjining group,33.06±5.02 in Tongxiening group and 40.87±4.86 in blank group.The threshold value of abdominal lift pressure of the blank group,the Changjining group and the Tongxiening group was higher than that of the model group,with significant difference(P<0.050),and the pressure threshold in the Changjining group was higher than that in the Tongxiening group,with significant difference(P<0.050).The threshold of arch back pressure was 32.54±3.04 in model group,38.00±4.50 in Changjining group,34.78±4.06 in Tongxiening group and 39.98±4.03 in blank group.The threshold value of the blank group,the Changjining group,the Tongxiening group and the model group were significantly different from those of the model group.(P<0.050)and the Changjining group was higher than that of the model group,and the difference was significant(P<0.050).The results of carbon powder propelling rate in IBS-D model rats: model group rats charcoal propulsion rate(unit: mm Hg)was 55.06 ± 5.95,Changjining group was 49.28 ± 7.41,Tongxiening group was 54.30 ± 5.81 and blank group was 49.79 ± 6.68.The propulsive rate in blank group,Changjining group and Tongxiening group was lower than that in small intestine in model group(P<0.050).The rate of pressure propulsion in the Changjining group was lower than Tongxiening,and was statistically significant(P<0.050).IBS-D model rat PKA protein expression levels: Model group rats PKA protein expression(unit: ng/ml)was 0.558± 0.107,Changjining group was 0.743±0.077,Tongxiening group was 0.654±0.078 and the blank group was 0.828±0.073.Compared with the PKA protein content of the model group,the blank group,the Changjining group and the Tongxiening group was higher,and the difference was significant(P<0.050).The content of PKA in Changjining group was higher than that in Tongxiening group(P<0.050),and had statistical significance(P<0.050).The expression level of c AMP protein in IBS-D model rats:Model group rats was 0.319 ± 0.086(ng/ml),0.465 ± 0.073 in Changjining group,0.380 ± 0.087 in Tongxiening group,and the blank group was 0.533 ± 0.065.Comparing with the c AMP protein content in the model group,the content in the blank group,Changjining group and the Tong Xie Ning group was higher,and the difference was significant(P<0.050).The content of c AMP in Changjining group was higher thanthat in Tongxiening group(P<0.050),and had statistical significance(P<0.050).The expression of AQP4 protein in IBS-D model rats: the content of AQP4protein(unit: ng/ml)in model group was 0.352 ± 0.079,0.478 ± 0.079 in Changjining group,0.423 ± 0.082 in Tong Xie Ning group,the blank group was0.507 ± 0.081.The content of AQP4 protein in blank group,Changjinning group and Tongxianning group was higher than that in model group(P <0.050).The expression of AQP4 protein in Changjining group was higher than that in Tongxiening group(P < 0.050),and had statistical significance(P <0.050).Conclusion Clinical trial results1.Changjining decoction has the effect of improving the clinical symptoms of IBS-D patients with liver depression and spleen deficiency,and has good clinical efficacy.2.The patients in this clinical study did not show any significant adverse reactions,indicating that Changjining prescription has good safety.Animal experiment results1.Changjining Prescription may interfere with the colonic mucosa PKA/c AMP signaling pathway,by affecting the membrane vesicle shuttling mechanism of AQP4;to up-regulate the expression of AQP4 and promote colon water transport,so as to improve the diarrhea of IBS-D and achieve the treatment of IBS-D with liver-qi stagnation and spleen deficiency.2.Changjining Prescription has an good improvement on rat model with IBS-D in regard to the irritant symptoms and loose stools.In conclusion,Chang Jining Prescription has a good clinical effect on IBS-D with liver depression and spleen deficiency,which worth widely disseminated. |
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