Effect of Huangqintang on Oxidative Stress and Ferroptosis-related Indexes GSH-Px4，P53，SLC7A11 in Ulcerative Colitis Mice / 中国实验方剂学杂志

Objective:To explore the possible mechanism of Huangqintang in treating ulcerative colitis （UC）. Method:The animal model of UC was induced by dextran sodium sulfate （DSS）．The experimental animals were divided into control group， model group，Huangqintang low dose （4.55 g·kg^-1）， medium dose （9.1 g·kg^-1）， and high dose（18.2 g·kg^-1） groups. Intragastric administration was also given in the modeling process for 7 consecutive days. At the end of the 8th day， colon tissues were collected to measure colon length and mass， and calculate the colon mass index. Pathological changes were observed by hematoxylin-eosin （HE） staining. Serum iron content， superoxide dismutase （SOD）， glutathione （GSH）， catalase （CAT） and myeloperoxidase （MPO） were determined by biochemical assay. Western blot was used to detect the protein expression of glutathione peroxidase 4 （GSH-Px4）， long-chain acyl-CoA synthetase 4 （ACSL4） and ferritin heavy chain 1（FTH1）. The mRNA expression levels of tumor trotein 53 （P53） and solute carrier family 7 member 11 （SLC7A11） in colon tissues were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Result:The experimental studies showed that compared with normal group， serum MPO and iron content， ACSL4 protein level and relative P53 mRNA expression in the model group significantly increased （<italic>P</italic><0.05）， while serum SOD， CAT， GSH content， GSH-Px4， FTH1 relative protein expression level and relative SLC7A11 mRNA expression in the model group significantly decreased （<italic>P</italic><0.01）. Compared with model group， serum MPO and iron content， ACSL4 protein level and relative P53 mRNA expression significantly decreased （<italic>P</italic><0.05）， while serum SOD， CAT， GSH content， GSH-Px4， FTH1 relative protein expression level and relative SLC7A11 mRNA expression significantly increased （<italic>P</italic><0.05） after the intervention of Huangqintang， and the effect was most significant in the high-dose group （<italic>P</italic><0.05）. The results of general condition， colon length， colon mass index and HE staining showed that Huangqintang could relieve clinical symptoms and histopathological changes in UC mice. Conclusion:These results indicated that Huangqintang had therapeutic effect on ulcerative colitis mice， and its mechanism might be related to inhibiting the oxidative stress and ferroptosis.

Effect of Huangqin Decoction on pyroptosis pathway of NLRP3/caspase-1 in mice with ulcerative colitis / 中国中药杂志

To explore the effect of Huangqin Decoction on ulcerative colitis(UC) pyroptosis, and to explain the mechanism of pyroptosis based on NOD-like receptor thermoprotein domain 3(NLRP3)/cysteine proteinase 1(caspase-1) pathway. The animal model of UC induced with 3% dextran sodium sulfate(DSS) was established. The experimental animals were divided into control group, model group, low-dose(4.55 g·kg~(-1)), medium-dose(9.1 g·kg~(-1)) and high-dose(18.2 g·kg~(-1)) Huangqin Decoction groups and salazosulfapyridine group(0.45 g·kg~(-1)). While modeling, intragastric administration was given for 7 consecutive days. On the 8 th day, the mice were euthanized, the colon length was collected, and the histopathological changes were observed by HE staining. The content of interleukin-18(IL-18) was observed by ELISA. The content of lactatedehydrogenase(LDH)was determined by microplate method. TUNEL assay kit was used to detect the cell death. The immunohistochemical staining was used to detect the expressions of NLRP3 and apoptosis-associated speck-like protein containing a CARD(ASC). Western blot was used to detect the expressions of interleukin-1β(IL-1β), caspase-1 and gasdermin D(GSDMD).The experimental study showed that compared with normal group, the LDH content, TUNEL positive staining, inflammatory factors(IL-18, IL-1β), and proteins associated with pyroptosis were significantly increased(P<0.05). Compared with model control group, the LDH content, TUNEL positive staining, inflammatory factors(IL-18, IL-1β), and proteins associated with pyroptosis were decreased, and these results were more significant in high-dose groups(P<0.05). The results of HE staining showed that Huangqin Decoction could improve the pathological changes of colon. Huangqin Decoction could inhibit UC cell pyroptosis, and the mechanism may be closely related to NLRP3/caspase-1 signaling pathway.

Bioinformatics analysis and systematic review of key genes and therapeutic traditional Chinese medicine of ulcerative colitis active stage / 中国中药杂志

Data GSE75214 and GSE48959 that contained ulcerative colitis(UC) in the active stage was download from GEO database. Differential genes of UC in the active phase were obtained by using adjusted P<0.05 and |log_2 FC|≥1.5, which was the screening criteria. PPI analysis was performed in the STRING database, and GO and KEGG pathway analysis was performed in DAVID database. Cytoscape was used to visualize differential genes, and calculate key genes in the active phase. Coremine Medical was used to analyze and systematically evaluate traditional Chinese medicines for treating key genes. Finally, 139 differentially expressed genes in the active phase were screened out, which included the 109 up-regulated genes and 30 down-regulated genes. DAVID analyzed that the biology and pathways of these differential genes were mainly concentrated in inflammatory response, immune response, chemokine activity, TNF pathway, NF-κB pathway, and Toll-like receptor pathway. Cytoscape software calculated that IL-6, CXCL8, IL-1β, MMP9, CXCL1, ICAM1, CXCL10, TIMP1, PTGS2 and CXCL9 were the key genes of UC in the active phase. According to Coremine Medical analysis, traditional Chinese medicines for UC in the active stage included Curcumae Longae Rhizoma, Scutellariae Radix, Curcumae Radix had clearing heat clearing damp, reducing fire and detoxifying effects, which was in line with the pathogenesis of UC active stage, and was often used in clinical treatment of dampness-heat diarrhea. Therefore, Huangqin Decoction, which Scutellariae Radix was the principal drug, was selected for systematic evaluation. The evaluation showed that Scutellariae Radix was superior to Western medicine in terms of improving clinical efficiency, reducing inflammatory factors and immunoglobulin levels, with statistically significant differences and fewer adverse reactions. This study provided a new idea for further research on the pathogenesis of UC in the active phase by analyzing the genes and their mechanism of action, and the systematic evaluation of Chinese medicine for the treatment of UC active stage provided a basis for the clinical prevention and treatment of UC by Chinese medicine.