| Objective:To study the acute toxicity of Jian Pi An Chang Pill.To measure LD50 of the rats and observe and record the situations of the acute toxicity,such as the appearances、activities、mental state、breathing change、toxic reactions and deaths of the rats.To study effects of Jian Pi An Chang Pill for Diarrhea type irritable bowel syndrome and Preliminary discuss on its mechanism of action,establishing the model of stagnation of liver qi and spleen deficiency(the model of Diarrhea type irritable bowel syndrome),observing Each dose group of animal liver 、 kidney function index,measuring the contents of VIP、CGRP in serum and Colon tissue,observing and recording Colonic pathological alterant situations and protein expressions of VIP、CGRP.It provide scientific gist for clinical treatment of irritable bowel syndrome further.Methods: 1、Gavage of Jian Pi An Chang Pill mixed suspension to rats is by the largest drug concentration and volume.At the same time,the control group rats were given the same volume normal saline.Observe and record the food intakes 、 the weight changes 、 deaths and activities of the rats.2 、 With reference to Bristol stool Scale,establish the model of stagnation of liver qi and spleen deficiency,observe and record Defecate classification and grading value of rats.3、After 1 hours of the last dosing,rats were narcotized and taken blood from abdominal aortic vein.Then to measure liver function(ALT、AST)、renal function(Cr、BUN)and the content of VIP、CGRP.4、Colon tissues which is 1cm、2cm、3cm far from ileocecal junction were taken、washed 、conserved and standby used.Colon tissue which is 1cm far from ileocecal junction was blotted up by filter paper、weighed、tissue homogenizedcentrifuged、taken the supernatant fluids,and then was measured its’ contents of VIP、CGRP.After 24 hours kept in 10% neutral buffer paraformaldehyde solution,the other were taken、cut along the longitudinal、unfolded、embedded and sliced.Observe the morphology change of colon tissues by HE staining method.Measure the protein expression situations of VIP、CGRP by immunohistochemical method.Results: 1 、 Part of the rats who were given JianPi AnChang Pill reduced their activities after 20 minutes and recovered 1 hour later.The food intakes of the Dosage groups of rats are less than the control group in 1 day,but there is no significant difference.The rats given drugs all survived in Continuous 14 days,had no obvious toxic effects and their food intakes、weight changes had no obvious difference than the normal saline group.Their appearances、activities、mental state and so on were all normal.After 14 days,the organic colors、volume、qualities of the rats killed and checked alive were all normal.Compared with normal saline group,those were no changes.2、Establishing model after 10 days,with reference to Bristol stool Scale,all rats’ models were made and loose stools rate was 99%.Giving drugs after 3weeks,Compared with model group,the rats’ loose situation of Jian Pi An Chang Pill large,medium dose group and pinaverium bromide group obviously improved(P<0.01 or P<0.05);small dose group rats’ loose stools scores are lower than model group,but it don’t have statistical differences.3、Compared with normal group,the model group、Jian Pi An Chang Pill group and pinaverium bromide group rats’ ALT、AST、Cr、BUN don’t have statistical differences.The results show that rhubarb water decoction、Jian Pi An Chang Pill and pinaverium bromide are no harmful for rats’ liver and renal.4 、 The results of VIP:Compared with model group,Jian Pi An Chang Pill each does group rats’ serum contents of VIP obviously reduce(P<0.01 or P<0.05),pinaverium bromide group’ serum levels of VIP decreased but it don’t have statistical differences.The contents of VIP in colon tissues of all does group obviously reduce(P<0.05).The results show that each does Jian Pi An Chang Pill can effectively improve diarrhoea situation by decreasing the contents of VIP in serum and colontissue.The results of CGRP:Compared with model group,all does groups rats’ serum contents of CGRP don’t have very large difference,while the levels of CGRP in colon tissues remarkable rise,which have statistical differences(P<0.01 or P<0.05).The results show that each does Jian Pi An Chang Pill can increase secretion of CGRP in colon tissues and have role for Diarrhea type irritable bowel syndrome.5 、 The results of HE staining method:The normal control group rats’ structures of the villi and gland in colon tissues are clear and complete,forms on the mucosa and submucosa are basically normal,the boundaries of the cells are clear,goblet cells are complete.The model group rats’ part of goblet cells rupture.There are focal inflammatory cells invading on the mucosa and submucosa.The model group rats’ Inflammatory conditions are obvious,while each dosage groups’ conditions significantly reduce.Compared with normal group,large dose group is almost no obvious difference.6、The protein expression results of VIP、CGRP:Compared with model group,all dosage groups’ positive expressions of VIP proteins in colon tissues significantly decrease,while expressions of CGRP proteins obviously increase.The best effects are Jian Pi An Chang Pil large dose group and pinaverium bromide group(P<0.01).Compared with pinaverium bromide group,Jian Pi An Chang Pil large dose group has a significant increase(P<0.05).Conclusion:There are no rats death through JianPi AnChang Pill’ largest drug concentration and volume lavage and no LD50.Within 21 continuous dosing days,the rats’ livers and kidneys function without damage.These explain that the drug is safety and low-toxic.Jian Pi An Chang Pill can decrease the model of stagnation of liver qi and spleen deficiency rats ’ loose quantities and scores,who were through bound and rhubarb lavage,significantly decrease the contents and protein expressions of VIP in serum and colon tissues,increase the levels and protein expressions of CGRP in colon tissues.Jian Pi An Chang Pill can ease diarrhea type irritable bowel syndrome. |
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