ABSTRACT
This study aims to investigate the regulatory effect of Sishen Pills(SSP) and its split prescriptions Ershen Pills(EP) and Wuweizi Powder(WP) on T follicular helper(Tfh) cell subset in the dextran sodium sulfate(DSS)-induced colitis mice and the mechanism. A total of 60 male SPF BALB/c mice were used, 10 of which were randomly selected as the normal group. The rest 50 were induced with 3% DSS solution for colitis modeling. After modeling, they were randomized into 5 groups: model group, SSP group, EP group, WP group, and mesalazine group. Body mass, colon mass, colon mass index, colon length, and unit colon mass index in each group were observed. After hematoxylin-eosin(HE) staining, the pathological injury of colon tissue was scored. The expression levels of molecules related to the STAT/SOCS signaling pathway in colon tissues were analyzed by Western blot. Differentiation levels of Tfh cells such as CD4~+CXCR5~+IL-9~+(Tfh9), CD4~+CXCR5~+IL-17~+(Tfh17), and CD4~+CXCR5~+Foxp3~+(Tfr) in peripheral blood of mice were detected by flow cytometry. The results showed each treatment group demonstrated significant increase in body mass and colon length, decrease in colon mass, colon mass index, unit colon mass index, and histopathological score(P<0.05, P<0.01), reduction of the expression of p-STAT3, STAT3, p-STAT6, and STAT6(P<0.05, P<0.01), rise of the expression of SOCS1 and SOCS3(P<0.05, P<0.01), decrease of Tfh9 and Tfh17 cells, and increase of Tfr cells(P<0.05, P<0.01) compared with the model group. These results indicated that SSP and the split EP and WP may alleviate ulcerative colitis by inhibiting the activation of STAT/SOCS signaling pathway and regulating the balance of Tfr/Tfh9/Tfh17 cells.
Subject(s)
Animals , Male , Mice , Colitis/genetics , Colitis, Ulcerative/metabolism , Mice, Inbred BALB C , Prescriptions , T-Lymphocytes, RegulatoryABSTRACT
The present study aimed to explore the effect of nuclear factor erythroid 2 related factor 2(Nrf2)/heme oxygenase-1(HO-1) signaling pathway in intestinal protection by Sishen Pills against ulcerative colitis(UC). After the UC model was induced by 3% dextran sodium sulfate(DSS), experimental animals were randomly divided into control group, model group, salazosulfapyridine(SASP) group, and low-and high-dose Sishen Pills groups. Drug intervention(ig) was performed for seven consecutive days during modeling. On the 7 th day, the mice were euthanized. The body weight and colon length were recorded, and the histopathological changes of the colon were observed by HE staining. Serum interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), total antioxidant capacity(T-AOC), malondialdehyde(MDA), and reactive oxygen species(ROS) were detected by ELISA. The protein and mRNA expression of Nrf2, HO-1, and NADPH quinine oxidoreductase-1(NQO-1) was determined by Western blot and reverse transcription-polymerase chain reaction(RT-PCR). Compared with the normal group, the model group exhibited reduced body weight, colon length, and T-AOC, increased IL-6, TNF-α, MDA, and ROS, and diminished protein and mRNA expression of Nrf2, HO-1, and NQO-1 in the colon tissues. Compared with the model group, the SASP group and high-dose Sishen Pills group showed elevated body weight, colon length, and T-AOC, lowered IL-6, TNF-α, MDA, and ROS levels, and increased protein and mRNA expression of Nrf2, HO-1, and NQO-1 in the colon tissues. As assessed by HE staining, Sishen Pills could improve the pathological changes of the colon. The findings suggested that Sishen Pills could protect the colon against UC induced by 3% DSS. The specific mechanism of action may be related to the anti-inflammatory and anti-oxidative stress effects by the activation of the Nrf2/HO-1 signaling pathway.
Subject(s)
Animals , Mice , Colitis, Ulcerative/genetics , Dextran Sulfate , Heme Oxygenase-1/metabolism , NF-E2-Related Factor 2/metabolism , Signal TransductionABSTRACT
Objective To explore the effects of Sishen Pills on Ghrelin-CaM-MLCK signaling pathway with sinuses ventriculi in diarrhea rats of yang deficieney of spleen and kidney; To discuss its mechanism of action. Methods A model rats with diarrhea model of yang deficieney of spleen and kidney was established by treated with intragastric administration of adenine and Senna water decoction to replicate. 72 SD rats were randomly divided into normal group, model group, positive medicine group, Sishen Pills high-, medium- and low-dose groups, with 12 rats in each group. Each medication group was given relevant medicine for gavage, for 14 days. Immunohistochemistry and Western blot were used to detect the expressions of Ghrelin and GHSR, CaM, and MLCK protein in the sinuses ventriculi. Results In the model group, Ghrelin, GHSR, CaM and MLCK protein expressions of the sinuses ventriculi significantly increased (P<0.01). In each medication group, protein expression of the indexes was lower than the model group (P<0.05, P<0.01). The Sishen Pills high-dose group was particularly obvious. Conclusion The effect of Sishen Pills on diarrhea model rats of yang deficieney of spleen and kidney may be associated with regulation of index protein expression in Ghrelin-CaM-MLCK signaling pathway.
ABSTRACT
Objective To observe the effects of Sishen Pills on gene and protein expressions of toll-like receptor 2 (TLR-2) and toll-like receptor 4 (TLR-4) of colonic tissue in spleen-kidney yang deficiency rats with ulcerative colitis (UC); To discuss its mechanism of action for spleen-kidney yang deficiency UC. Methods Lavage of senna +intraperitoneal injection of hydrocortisone injection + enema of 2,4,6-trinitrobenzene sulfonic acid/ethanol were used to establish the model of UC of spleen-kidney yang deficiency type. 90 Wister rats were randomly divided into blank group, model group, Salazosulfadimidine group and the Sishen Pills high-, medium- and low-group. Each medication group was intervened by relevant medicine. RT-qPCR was used to detect the expressions of TLR-2 and TLR-4, and Western blot was used to detect the protein expressions of TLR-2 and TLR-4. SOD and MDA in serum were detected. Results Compared with the blank group, SOD activity decreased and MDA content increased in the model group (P<0.01); gene and protein expressions of TLR-2 and TLR-4 increased significantly (P<0.05). Compared with model group, SOD increased and MDA content decreased in all the medication groups (P<0.05, P<0.01); gene and protein expressions of TLR-2 and TLR-4 decreased (P<0.05, P<0.01). Conclusion Sishen Pills can achieve the treatment of UC by improving the metabolism of oxygen radicals in colonic tissue and regulating the immune system of the intestinal epithelium.
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Objective To observe the clinical efficacy of sishen pills plus trimebutine maleic acid tablets and bifid triple viable bacteria on treating ulcerative colitis.Methods 100 cases were randomly divided into two groups,with 50 cases for each group,the treatment group was given sishen pills plus trimebutine maleic tablets and bifid triple viable bacteria,the control group was treated with sishen pills.The clinical effect of the two groups were observed.Results The efficiency of the treatment group was 80%,which of the control group was 36%,there was statistically significant difference between the two groups (χ2 =8.16,P <0.05).Conclusion The efficacy of sishen pills plus trimebutine maleic tablets and bifid triple viable bacteria was better than that of sishen pills only.