Effects Of Tongxieercao Formula On The Exp-ression Of The Molecules Related To NF-κB P-athway On Ulcerative Colitis Disease Of Liver-depression And Spleen-deficiency Syndrome

Objective: This study was designed to established ulcerative colitis (UC) model ofliver-depression and spleen-deficiency syndrome by outside bondage and enema composed ofTNBS (2,4,6-three trinitrobenzene sulfonic acid) and ethanol, and to observe thehistopathological changing effects and to find the expressions of genes and proteins in NF-κBpathway in the UC rat with liver-depression with spleen-deficiency syndrome that treated byTongxieercao formula (TXECF). So this study aims at elucidate the mechanisms andtherapeutic targets of TXECF on UC model with liver-depression and spleen-deficiencysyndrome.Methods:90SPF Wistar rats were divided into six groups randomly and equally inweight and gender, which were as follows: blank group, model group, TXECF high-dosegroup, TXECF middle-dose group, TXECF low-dose group and positive group (SASP group).Rats in blank group was treated with distilled water. The rats in other groups were treated withenema composed of TNBS with ethanol and suffered from bondage stimulation toestablish liver-stagnation and spleen-deficiency syndrome as UC modal. Followed by that,The rats in traditional Chinese medicine treatment groups were given TXECF gavage ofcorresponding concentration. The rats in positive group were treated with SASP gavage, andthe rats in blank group and modal group were treated distilled water gavage in thesame volume. All rats were treated by oral gavage per day for21days. After completing thesetreatment steps, the rats in every group were fasted except water-deprivation for24h after lastgavage. Based on that preparation, all rats were administrated with chloral hydrate anesthesiafor sample collection. The methods of which were as follows: to dissect the entire colon tissueand observe each rats’ colonic mucosal hyperemia, edema, ulcers, erosion number and area ofpathological changes with eyes. Then a small amount of colon tissue were taken and fixed byparaffin embedding with HE staining for further diagnosis with pathological morphologicalchanges with microscope of each colon tissue, and the rest of colon tissue specimens werecollected for PCR-Array, RT-PCR and Western blot experiments.Results:1. Compared with the blank group, certain degree of congestion, edema, ulcer and erosion in colonic mucosa were found in model group and treatment groups with eyes, aswell as those different degrees of ulcer, inflammation, concurrent granulation hyperplasia andfibrosis with microscope. Compared with the model groups, the rats in each TXECF treatmentgroups showed lower grades in colonic mucosa injury and pathological histology, and whichdemonstrate significant difference (P<0.01or P<0.05).2. Compared with the blank group, the gene expressions of Nfkb1, Nfrkb, Ikbkb in colontissues of model group elevated which showed statistical significance (P<0.01). After thetreatment of TXECF, the gene expressions of Nfkb1, Nfrkb, Ikbkb in colon tissues oftreatment groups are lower than model group (P<0.01or P<0.05), and high-dose groupillustrated significant difference (P<0.05). The differences of Nfkb1and Nfrkb betweenhigh-dose group and SASP group have no statistical significance, but that of Ikbkb showedstatistical significance (P<0.05).3. Compared with the blank group, the protein expressions of Nfkb1, Nfrkb, Ikbkb incolon tissues of model group elevated which showed statistical significance (P<0.01). Afterthe treatment of TXECF, the protein expressions of Nfkb1, Nfrkb, Ikbkb in colon tissues oftreatment groups are lower than model group (P<0.01or P<0.05), and high-dose groupillustrated significant difference (P<0.05). The differences between high-dose group andSASP group showed no statistical significance (P<0.01or P<0.05).Conclusion:1. TXECF can inhibit inflammatory cells invasion in colon tissue of UCmodel with liver-depression and spleen-deficiency syndrome, reduce inflammation reaction,repair damaged intestinal mucosal, and it can help restore intestinal mucosa secretion. Itproves the model reconstruction of UC with liver-depression and spleen-deficiency syndromewere accomplished successfully, and TXECF can treat liver-depression and spleen-deficiencysyndrome UC by reducing inflammation reaction and repairing damaged intestinal mucosal.2. It shows that the expression levels of Nfkb1, Nfrkb, Ikbkb can reflect the damagedegree of colonic mucosa, and has potential implications in assessment and judgment ofdisease.3. The treatment effects of TXECF were associated with downregulation of theexpression levels of Nfkb1, Nfrkb and Ikbkb, and reduction of mucosal inflammation and repairment of mucous membrane. And as a result: Nfkb1, Nfrkb and Ikbkb may be the keytargeted of the TXECF when treating UC with liver-depression and spleen-deficiencysyndrome.4. TXECF could down regulate the colonic mucosa protein expressions of NF-κB-P65,IκB-α and IκB-β in UC rats model with liver-depression and spleen-deficiency syndrome, andit can suppress the development and progression of inflammatory and repair damaged mucousmembrane. And therefore, the therapeutic effects of TXECF on UC rats model withliver-depression and spleen-deficiency syndrome may be associated with downregulation ofthe expression levels of Nfkb1, Nfrkb and Ikbkb.