| Objective:To observe the Jichuanjian therapeutic effect on cathartic colon, and discussing the function of the SCF/c-kit signal channel, AQP3,AQP4and AQP9on Cathartic Colon mechanism.Methods:1. Clinical studyThe70"cathartic colon" patients were given Jichuanjian Decoction treatment (cistanche20grams, angelica30grams, Radix10grams, alisma10grams, cohosh10grams,20grams of Fructus Aurantii). A agent daily, early and late two times daily, ten days as a course of treatment, a total of two courses. The patients’ constipation symptom score was recorded (including frequency of defecation score, defecation time score, characteristics of feces score, abdominal distension score and the use of laxatives score) in Prior treatment and Post treatment for one month and Patient Assessment of Constipation Quality of Life (PAC-QOL). What’s more, the differences in Prior treatment and Post treatment were observed.2. Experimental studyThe70healthy adult SD rats in SPF level were randomly divided into control group (Group1:11rats; group2:12rats) and experimental group (model group1:11rats; model group2:12rats; Jichuanjian group:12rats; polyethylene glycol group:12rats).The control group rats were perfused with physiological saline:the control group1and model group1, as well as control group2and model group2were drew materials at the same time. The "cathartic colon" animal model was made on experimental group rats by "Rhein suspension gavage method". For the model group1drawing materials after the end of the molding; for model group2perfused with physiological saline for30days after the end of the molding; for Jichuanjian group:after the end of the molding, perfused with Jichuanjian Decoction for30days; polyethylene glycol group:after the end of the molding, perfused with polyethylene glycol aqueous solutions for30days. Then I tested the first black stool time and fecal dry and wet weight ratio to reflect intestinal transmission function and the nature of stool. The level expression of mRNA of c-kit, SCF and AQP3and AQP4, AQP9, TFF3in colon tissue was detected by the application of reverse transcription polymerase chain reaction (RT-PCR). C-kit and SCF were positioned in histology by immunohistochemical method. The level expression of protein of c-kit, SCF and AQP3, AQP4and AQP9, TFF3in colon tissue was detected by Western Blot method. Results:1. Clinical researchThe symptoms of constipation and quality of life for the "Cathartic colon" patients were significantly improved after the treatment by Jichuanjian. Defecation frequency was increased in the comparison of prior treatment and post treatment:2.37±1.18vs0.70±1.04, P<0.01; defecation time was shorter in the comparison of prior treatment and post treatment:1.66±0.56vs0.61±0.62, P<0.01; the stool was softer in the comparison of prior treatment and post treatment:2.22±0.84vs0.83±0.90, P<0.01; abdominal distension was eased in the comparison of prior treatment and post treatment:1.23±0.49vs0.49±0.50, P<0.01; the use of laxative was reduced in the comparison of prior treatment and post treatment:2.83±0.72vs0.83±0.61, P<0.01. The PAC-QOL score was decreased in the comparison of prior treatment and post treatment:58.39±15.2vs32.10±13.97, P<0.01.2. Experimental studyAfter the rhein suspension gavage for110days, the first black stool time of rats in model group1was prolonged significantly, and the fecal dry and wet weight ratio was increased significantly, so the differences were significant comparing with the control group1(P<0.01). After the treatment process for30days, the first black stool time of rats in Jichuanjian group was shortened significantly, and the fecal dry and wet weight ratio was decreased significantly, so the differences were significant comparing with the model group2(P<0.01). But compared with the control group2, the first black stool time was still extended, so it has significance in statistic (P<0.05), however it has no obvious difference in the fecal dry and wet weight ratio (P>0.05). Compared polyethylene glycol group with model group2, it has no difference statistically in the first black stool time and fecal dry and wet weight ratio (P>0.05).Through the immunohistochemical detection, It was found that the c-kit was distributed in the whole colon muscle layer, in which the circular muscle layer and the myenteric nerve plexus in the myenteric plexus regional is the most obvious. It was expressed in ICC cell membrane and neuritis. In the model group1, ICC is light brown staining and weak positive, and in contrast to the control group1, the expression of c-kit was weakened significantly. The SCF was expressed in the whole colon wall, and it has only slight expression in model group l,so in contrast to the control group1, the expression was decreased obviously.Through the RT-PCR and Western blot detection, It was found that c-kit and mRNA of SCF and protein expression in model group1are weaker, and in contrast to the control groupl, there was significant difference (P<0.05). In Jichuanjian group, c-kit and mRNA of SCF and protein expression in the colonic tissues were increased significantly, which was significant statistically (P<0.05) compared with model group2, but it has no significant difference (P>0.05) compared with the control group2. In polyethylene glycol group, compared with model group2, c-kit and mRNA of SCF and protein expression in the colonic tissues have no significant changes (P>0.05) but it has significant difference (P<0.05) compared with the control group2.Compared with the control group2, the mRNA of AQP3and protein expression in colon tissue in model group2were significantly increased, yet the mRNA of AQP9and TFF3and protein expression were significantly lower, which has significance statistically (P<0.05). However compared with the control group2, the mRNA of AQP4and protein expressions have no significant difference (P>0.05). In Jichuanjian group, the mRNA of AQP3and protein expression in colon tissue was lower significantly, yet the mRNA of AQP9and TFF3and protein expression were significantly increased. Compared with the model group2, it has significant difference (P<0.05), but compared with the control group2, it has no significant difference (P>0.05). In polyethylene glycol group, compared with the model group2the mRNA of AQP3, AQP9, TFF3and protein expression have no significance statistically (P>0.05).Conclusion:1. The satisfactory curative effect can be obtained on the treatment of "cathartic colon" by JiChuanJian therapy, as well as the constipation symptoms and quality of life of the patients were improved significantly.2. The intestinal transmission function and fecal dry and wet weight ratio can be improved significantly for the "cathartic colon" rats by JiChuanJian therapy, so as to take the therapeutic effect.3. The abnormal expression of c-kit, SCF, AQP3, AQP9and TFF3in colon tissue can be caused by long-term stimulation on the rats’intestinal tract from anthraquinone laxative, which was perhaps the one of the "mechanism of cathartic colon".4. The mechanism of the treatment on "cathartic colon" by Jichuanjian therapy may be related to the repair of SCF/c-kit signal channel and the correct of abnormal expression of AQP3, AQP9, and TFF3in colonic mucosa. |
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