Study On The Effect And Mechanism Of "Key Herb" Compatibility In Buzhong Yiqi Decoction On Constipation Induced By Loperamide In Rats

OBJECTIVE:To observe the effect of"Key Herb"of Shengma Chaihu compatibility in Buzhong Yiqi Decoction and different dosage of Buzhong Yiqi Decoction on loperamide-induced constipation model in rats,and to explore the mechanism of synergy effect of"Key Herb"compatibility in Buzhong Yiqi DecoctionMethod:1.SD rats were injected subcutaneously with loperamide twice every day at a dose of 0 mg/kg/d,2 mg/kg/d,4 mg/kg/d,and 6 mg/kg/d respectively.Intestinal propulsion rate was measured at the time of 2^nd and 7^th day during treatment.2.SD rats were injected subcutaneously with loperamide at a dose of 4 mg/kg/d twice every day to replicate the model of delayed intestinal propulsion.After successful modeling,rats were administrate nomal dose of Buzhong Yiqi Decoction,dose of 0.5 times Buzhong Yiqi Decoction,dose of 0.25 times of Buzhong Yiqi Decoction and 5 g/kg/d of Rhubarb orally.Rats were sacrificed after 5 days treatment,and tests were studied as follows:?1?Small intestine propulsion rate was measured in each group;?2?Serum contents of MTL,Gas and VIP were measured in each group;?3?Changes of morphology of small intestine in each group were observed.3.SD rats are injected subcutaneously with loperamide at a dose of 4 mg/kg/d twice every day to replicate the model of constipation with spleen deficiency.After successful modeling,drugs of mosapride,0.25 times dose of Shengma are administered.0.25 times the dose of Chaihu,0.25 times the dose of Shengma and Chaihu,0.25 times the dose of Buzhong Yiqi Decoction without“key herbs of Shengma and Chaihu,0.25 times the dose of Buzhong Yiqi Decoction were admininstrated orally.Rats were sacrificed after 5 days treatment,and tests were studied as follows:?1?Different administration group rat small intestine Changes in propulsive rate;?2?Changes in serum MTL,Gas,VIP,and SS contents in each group;?3?The changes of sc F/c-kit,PKC?,Drd2,and CHR-M3 m RNA expression in the small intestine of each group are determined.Result:1.The effect of different doses of loperamide on the intestinal propulsion rate in rats:Compared with the normal group,the small intestine propulsion rate in the model group 1 was significantly decreased?P<0.05?,and the small intestine propulsion rate in the model group 2 was significantly decreased?P<0.01?.In the model group 3,the small intestine propulsion rate decreased significantly?P<0.01?.Model 2?4 mg/kg/d?and model 3?6 mg/kg?were more significant.2.The effect of different doses of Buzhong Yiqi Decoction on small intestine propulsion rate in rats with constipation induced by loperamide:Compared with the normal group,the small intestine propulsion rate of the model group was significantly decreased?p<0.05?,compared with the model group.Compared with the reduction of the therapeutic dose,the small intestine propulsion rate showed a gradual increase trend.When the dose reached 0.25 times,the small intestine propulsion rate increased significantly?p<0.05?.Compared with the model group,the rhubarb treatment group decreased slightly,no significance different.?p>0.05?.3.Effects of different doses of Buzhong Yiqi Decoction on serum gastrointestinal hormones MTL,GAS and VIP in loperamide-induced constipation rats:Compared with the normal group,MTL?GAS?VIP in serum of each gradient dose group?Compared with the model group,the MTL and GAS levels in the serum of the model group were decreased?p<0.01?,and the VIP content was increased?P<0.01?.Compared with the model group,the 1 times dose of the original group and the0.5-fold dose of the original group The MTL content of rats in the 0.25 times dose of the Buzhong Yiqi decoction group gradually increased?P<0.01,P<0.01,P<0.01?,1time dose of the Buzhong Yiqi decoction group,0.5 times the dose of the Buzhong Yiqi decoction group,0.25 times the dose of the Buzhong Yiqi decoction group of rats.The serum GAS content of the rats increased gradually?P<0.05,P<0.01,P<0.01?,and the serum VIP content of the rats in the 1 times dose of the Buzhong Yiqi decoction prescription group,the 0.5 times dose of the Buzhong Yiqi decoction group,and the 0.25 times dose of the Buzhong Yiqi decoction prescription group gradually decreased?P<0.05,P<0.01,P<0.01?,the effect of rhubarb group on MTL,GAS and VIP was not statistically significant?P>0.05?,and it also showed the opposite trend to the treatment group.4.Different doses of Buzhong Yiqi Decoction on the gastrointestinal morphology of rats with constipation induced by loperamide:The gastrointestinal abdomen of the model group rats,the 1 times dose of Buzhong Yiqi Decoction group had slight flatulence,0.5 times dose Buzhong Yiqi Decoction group,0.25 times dose Buzhong Yiqi Decoction group,normal group gastrointestinal color,normal shape,no congestion,edema and so on.Under the light microscope,the mucosal damage of the small intestine tissue in the model group and the rhubarb group was severe,a large number of necrotic cells were shed,a large number of inflammatory cells infiltrated,and the mucosa and muscle layers were narrowed.In each treatment group,there was a small amount of inflammatory cell infiltration,and occasionally,the edema of the lamina propria of individual samples was found,and the degree of damage was slight.There were no obvious abnormalities such as obvious damage in the normal group.5.The effect of Buzhong Yiqi Decoction on the small intestine propulsion rate of constipation rats with spleen deficiency:Compared with the normal group,the small intestine propulsion rate of the model group was significantly decreased?p<0.05?;Compared with the model group,the positive group,0.25 times Shengma Chaihu group?Sheng Chai group?,0.25 times Buzhong Yiqi Decoction without“key herbs”of Shengma and Chaihu group,0.25 times Buzhong Yiqi Decoction group,the small intestine propulsion rate was significantly increased?p<0.05,p<0.01,p<0.01,p<0.05,?.6.Changes of the contents of gastrointestinal hormones Ghrelin,VIP,MTL,GAS,SS in constipation rats with spleen deficiency in Buzhong Yiqi Decoction:In the serum of constipation model rats,the vasoactive intestinal peptide that inhibits gastrointestinal motility and the motilin and gastrin that promote gastrointestinal motility show opposite trends,namely,the inhibition of sex hormone VIP content,hormone MTL promoting gastrointestinal motility,GAS decline;and ghrelin which promotes gastrointestinal motility and SS which inhibits gastrointestinal motility were no significant change.Both Buzhong Yiqi Decoction group and the drug“Shengma Chaihu”group increased the content of excitatory gastrointestinal hormones GAS and MTL.The change of GAS and MTL on Shengma or Chaihu group were not significant.The Buzhong Yiqi Decoction group can effectively reduce the content of VIP.Shengma or Chaihu group can also reduce the content of VIP.The Buzhong Yiqi Decoction had no effect on the content of VIP.The mosapride had a certain effect on VIP,and had little effect on the contents of MTL,GAS,SS and Ghrelin.7.The expression of SCF/c-kit,Drd2,CHR-M3 and PKC?m RNA in the small intestine of rats with spleen deficiency and constipation in Buzhong Yiqi Decoction group:Compared with the normal group,the constipation model group rats There was no significant change in the expression of SCF/c-kit,Drd2,CHR-M3 and PKC?m RNA in the small intestine.Compared with the model group,Buzhong Yiqi Decoction treatment group can significantly increase the expression of PKC?m RNA in the small intestine of model rats,and Buzhong Yiqi Decoction can slightly increase c-kit in the small intestine of model rats.The expression level of c-kit?m RNA,while the combination of whole prescription and other“Key Herb”can not significantly affect other indicators,indicating the mechanism of action of Buzhong Yiqi Decoction on intestinal motility and SCF/c-kit,Drd2,CHR-M3.There is no change in the expression of PKC?m RNA,and its specific possible mechanism needs further investigation.Conclusion:1.The 4 mg/kg/d dosage of loperamide is the preferred dose to induce constipation model in rats.2.The original dosage of Buzhong Yiqi Decoction in"Spleen and Stomach theory"has perfect effect on promoting intestinal motility.3.Loperamide-induced constipation model in rats is a constipation model of spleen deficiency.4."Key Herb"of Shengma and Chaihu has a good promoting effect on small intestine motility.The effect of Buzhong Yiqi Decoction with"Key Herb"of Shengma and Chaihu on promoting small intestine motility is better Compared with Buzhong Yiqi Decoction without"Key Herb"of Shengma and Chaihu.Indicated that“Key Herb”plays a important role in the fomular of Buzhong Yiqi Decoction.5.The mechanism of promoting intestinal motility effect of"Key Herb"compatibility in Buzhong Yiqi Decoction on the constipation model in rats may be related to the regulation effects on changing serum content of MTL,GAS,VIP and other gut hormones and repairing small intestinal mucosa but with few relationship on regulating Drd2,CHR-M3,PKC?,SCF/c-kit m RNA expression.6.Buzhong Yiqi Decoction and“Key Herb”have equivalent effects on promoting intestinal motility in our research.We take a hypothesis that the advantages of Buzhong Yiqi Decoction may effect on other aspects,such as promoting intestinal absorption function and so on,and more researches should be done in future.