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Objective To explore the effects of Sishen Pill and Gegen Qinlian Tablet on cytokines of acute and chronic colitis induced by dextran sulfate sodium (DSS) in mice. Methods Mice freely drank 4%DSS dissolved in drinking water for continuous 5 days to establish acute colitis model. Mice circularly drank 3% DSS for 4 times for the establishment of chronic colitis model. In the acute or chronic colitis experiment, mice were randomly divided into control group, acute and chronic model groups, Sishen Pill group, and Gegen Qinlian Tablet group. Acute model group was administrated one day after DSS drinking for 8 days. Chronic model group was administrated after the second time of circular drinking for 16 days. ELISA was used to detect the contents of IFN-γ, IL-17 and IL-22 in cultural supernatant of mouse colon. Results Contents of IFN-γ, IL-17A and IL-22 in cultural supernatant increased significantly in acute models (P<0.01). Gegen Qinlian Tablet can significantly decrease the contents of IFN-γ and IL-17A of acute models (P<0.05). Sishen Pill can significantly decrease the content of IL-17A (P<0.05). IFN-γand IL-17A in cultural supernatant in chronic model mice significantly increased compared with control group (P<0.01). Sishen Pill and Gegen Qinlian Tablet inhibited the contents of IFN-γ and IL-17A. Compared with control group, Sishen Pill significantly increased the content of IL-22 (P<0.05). Conclusion Sishen Pill and Gegen Qinlian Tablet can treat colitis by decreasing the contents of IFN-γ and IL-17A in acute colitis model mice;Sishen Pill can treat chronic colitis by promoting IL-22 to increase.
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Objective To observe the influence of Sishen Pill on the NF-κB p65 mRNA and protein expressions of colonic mucosa in rats with experimental ulcerative colitis (UC), and identify its underlying mechanism of action. Methods The experimental rats were divided into blank group, model group, Sishen Pill group and SASP group. The models were prepared by TNBS/ethanol enema. Sishen Pill group was intragastrically administrated by Sishen Pill extract 5 g/kg, SASP group by SASP 0.3 g/kg, and blank group and model group by equal volume of normal saline. The morphological injury of colonic mucosa was observed and scored with the naked eyes, and NF-κB p65 gene and protein expression were detected by RT-PCR and immunohistochemical method. Results Inflammation and ulceration on the colonic mucous membrane were found in the model group by naked eyes, and had significant difference with the blank group (P<0.05). The relative expression amount of NF-κB p65 gene and protein of colonic tissues were increased in the model group compared with the blank group (P<0.01). Compared with the model group, the relative expression amount of NF-κB p65 gene and protein in Sishen Pill group were decreased (P<0.05, P<0.01). Conclusion Sishen Pill has effect for treating UC, which is probably related to the activation of NF-κB signal transduction pathway.
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Objective To observe the effect of Sishen Pill on ICAM-1 mRNA and protein expression of colonic mucosa in rats with ulcerative colitis (UC), and identify its mechanism. Methods Taolly 40 SPF Wistar rats were randomly divided into blank group, model group, Pill group and SASP group. Except the blank group, UC model was prepared with TNBS/ethanol enema. Pill group was given Sishen Pill 5 g/kg, and SASP group was given SASP 0.3 g/kg by gavage, blank group and model group was given the same volume physiological saline for three weeks. Morphological injury of colonic mucosa was observed and scored. ICAM-1 gene and protein expression were detected by RT-PCR and immunohistochemical method. Results Inflammation and ulceration were found on the colonic mucous membrane of rats in the model group. The expression of ICAM-1 gene and protein of colonic tissues of rats in the model group increased compared with that of the blank group (P<0.01). Compared with the model group, the expression of ICAM-1 in Pill group decreased (P<0.05, P<0.01). Conclusion Sishen Pill can decrease the expression of ICAM-1 mRNA and ICAM-1 protein, inhibit the infiltration of inflammation cells, prevent and reduce colon tissue damage, and play a vital role in the treatment of UC.
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AIM: To establish HPLC fingerprint of Sishen Pill and a determination of main component of Sishen Pills(Fructus Evodiae, Semen Myristicae, Fructus Psoraleae, etc.) by HPLC. METHODS: The samples were extracted with the solvent methanol. The separation was performed on YWG-C 18(4.6mm?250mm, 10?m) column with water- methanol-acetonitrile as a mobile phase gradient elution. The flow rate was 1.0mL?min -1, the column temperature was at 30℃ and the wavelength of UV detector was at 225nm. RESULTS: There are 21 higher peaks and 12 marker peaks in the fingerprint of eight batches of samples. For evodiamine, the regression equation was Y=147.37X+1186.32, r=0.9999, the liner range was from 5 to 1000?g?mL -1 and the recovery was 99.83%. For rutaecarpin, the regression equation was Y=101.19X+279.81, r= 0.9999, the liner range was from 4.5 to 900?g?mL -1. The recovery was 98.72%. CONCLUSION: The quality standard of Sishen Pill was established firstly. It can be used for quality cantrol of Sishen Pill.