| Irritable bowel syndrome is a chronic functional gastrointestinal disorder that is divided into four subtypes depending on clinical symptoms such as bowel habits and stool properties,of which those with chronic abdominal pain and diarrhoea symptoms are classified as diarrhoeal.The biological clock is an endogenous regulatory system that synchronises the physiological and behavioural activities of the body with the external environment.There is growing evidence that biological clock disorders is involved in the development of IBS.Abnormal expression of the biological clock genes BMAL1 and CLOCK,which are core components of the circadian pacemaker,disrupts normal biological rhythms and may be an important factor contributing to the development of IBS.Sishen pill has remarkable efficacy in treating IBS-D of the spleen and kidney Yang deficiency type,but whether its therapeutic effect is related to the regulation of biological clock gene expression is not yet clear.Objective: This experiment was conducted to explore the effect of Sishen pill intervention on the m RNA of CLOCK and BMAL1 proteins in the colonic mucosa of rats,to explore the mechanism of IBS-D from the perspective of biological clock genes,to provide new ideas for the treatment of IBS-D,and also to provide new scientific basis for the effect of Sishen pill on the treatment of IBS-D with Yang deficiency of spleen and kidney.It also provides a new scientific basis for the mechanism of action of Sishen pill in treating IBS-D with spleen-kidney Yang deficiency.Methods:A total of 24 SD rats at the age of 8 weeks were randomly divided into 6 rats in the normal group,6 rats in the model group,6 rats in the Sishen pill group and 6 rats in the Desutex group.Except for the normal group,all rats were given senna leaf granules(1g/ml,1ml/kg)by gavage for 28 days and then 25mg/(kg-d)of hydrocortisone was injected intramuscularly at the right and left buttocks daily for 10 days to establish a rat model of IBS-D with spleen and kidney yang deficiency.After the modeling was completed,the normal group and the model group were given equal volume of saline gavage once a day.The rats in the Sishen pill group were given 1.83g/kg of Sishen pill suspension by gavage once daily,while the rats in the Desutex group were given15.23mg/kg of Desutex aqueous solution once daily.14 days later,the rats’ diarrhea index and abdominal wall withdrawal reflex were observed and recorded to assess the degree of diarrhea and visceral sensitivity.The expression of BMAL1 and CLOCK m RNA in rat colon tissues was measured by q-PCR,and the expression of BMAL1 and CLOCK protein in rat colon tissues was measured by Western blot.Results:1.Compared with the control group,the diarrhea index of rats in the model group and each administration group increased(P<0.05);the diarrhea index of the Sishen pill and Desutex groups was lower than that of the model group,and the difference was not statistically significant(P>0.05);2.Compared with the control group,the AWR score of the model group increased significantly(P<0.05),and compared with the model group,the AWR score of the Sishen pill group decreased significantly(P<0.01)Compared with the model group,CLOCK m RNA expression in the colon tissues of rats in the Desutex group increased(P<0.01),while no significant difference was found between the model group and the Sishen pill group(P>0.05);BMAL1 m RNA expression in the colon tissues of rats in all groups was not significantly different(P < 0.05).4.compared with the model group,BMAL1 protein expression was not significantly changed and CLOCK protein expression was significantly up-regulated in the Sishen pill group(P<0.05),while BMAL1 protein expression was significantly down-regulated(P<0.001)and CLOCK protein expression was significantly up-regulated in the Desutex group(P<0.001)compared with the model group.Conclusion: 1.Sishen pill can significantly reduce the sensitivity of the internal organs of rats with irritable bowel syndrome of the spleen-kidney-yang deficiency type,and the efficacy is better than that of Desutex.2.Sishen pill up-regulates the expression of CLOCK gene protein level in the colon of IBS-D rats with spleen and kidney yang deficiency. |
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