ABSTRACT
This study explored the mechanism of Sanhuang Decoction(SHD) in treating dextran sulfate sodium(DSS)-induced ulcerative colitis(UC) in mice with Candida albicans(Ca) colonization via high-throughput transcriptome sequencing. Specifically, the animal model was established by oral administration of 3.0% DSS for 7 days followed by intragastrical administration of Ca suspension at 1.0 × 10~8 cells for 4 days and then the mice were treated with SHD enema for 7 days. Afterwards, the general signs were observed and the disease activity index(DAI) was recorded every day. After mice were sacrificed, colon length and colon mucosa damage index(CMDI) were determined and the histomorphology was observed with the HE staining method. The fungal loads of feces were detected with the plate method. Anti-saccharomyces cerevisiae antibody(ASCA) and β-1,3-glucan in serum, and TNF-α, IL-1β, and IL-6 in serum and colon were detected by ELISA. High-throughput RNA sequencing method was adopted to identify transcriptome of colon tissues from the control, model and SHD(15.0 g·kg~(-1)) groups. Differentially expressed genes(DEGs) among groups were screened and the GO and KEGG pathway enrichment analysis of the DEGs was performed. The expression levels of NLRP3, ASC, caspase-1, and IL-1β genes related to the NOD-like receptor signaling pathway which involved 9 DEGs, were examined by qRT-PCR and Western blot. The results demonstrated that SHD improved the general signs, decreased DAI and Ca loads of feaces, alleviated colon edema, erosion, and shortening, and lowered the content of β-1,3-glucan in serum and TNF-α, IL-1β, and IL-6 in serum and colon tissues of mice. Transcriptome sequencing revealed 383 DEGs between SHD and model groups, which were mainly involved in the biological processes of immune system, response to bacterium, and innate immune response. They were mainly enriched in the NOD-like signaling pathway, cytokine-cytokine interaction pathway, and retinol metabolism pathway. Moreover, SHD down-regulated the mRNA and protein levels of NLRP3, caspase-1, and IL-1β. In a word, SHD ameliorates DSS-induced UC in mice colonized with Ca, which probably relates to its regulation of NOD-like receptor signaling pathway.
Subject(s)
Animals , Mice , Candida albicans/genetics , Colitis, Ulcerative/genetics , Colon , Dextran Sulfate/toxicity , Disease Models, Animal , Drugs, Chinese Herbal , High-Throughput Nucleotide Sequencing , TranscriptomeABSTRACT
Objective::To explore the protective mechanism of Shenteng Sanhuang decoction and Gegen Qinlian Tang on diabetic nephropathy (DN). Method::Rat DN models were duplicated with unilateral nephrectomy combined with streptozotocin (STZ). The rats were randomly divided into blank group, model group, irbesartan group and traditional Chinese medicine group. After 8 weeks of administration of corresponding drugs, the body weight, blood sugar, blood urea nitrogen (BUN), 24-hour urine volume (24 h U-vol), 24-hour urinary protein (24 h U-pro), serum creatinine (SCr), kidney weight/body weight (KW/BW) mass index of rats in each group were measured. The kidney tissues of rats in each group were homogenized, and supernatant was taken. Expressions of Smad ubiquitination regulatory factor 2 (Smurf2), matrix metalloprotein-9 (MMP-9), TGF-β1 and malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) were detected by Western blot or special test kit. Result::Compared with the blank group, the biochemical indicators, body weight, KW/BW, blood sugar, BUN, 24 h U-vol, 24 h U-pro, SCr and MDA were significantly higher or increased(P<0.05), while SOD and CAT were significantly decreased in the model group(P<0.05). Western blot showed that the expression of Smurf2 and TGF-β1 was high, while the expression of MMP-9 was low(P<0.05). Compared with the model group, the biochemical indicators of irbesartan group and traditional Chinese medicine group improved significantly(P<0.05), KW/BW were reduced, and blood sugar, BUN, 24 h U-vol, 24 h U-pro, SCr and MDA were significantly decreased (P<0.05), SOD and CAT was obviously increased (P<0.05), expressions of Smurf2 and TGF-β1 were decreased significantly(P<0.05), and expression of MMP-9 was increased significantly(P<0.05). Conclusion::Shenteng Sanhuang decoction and Gegen Qinlian Tang can effectively improve many biochemical indexes of rat DN models, and improve renal function. Its mechanism is closely related to reducing the expressions of Smurf2 and TGF-β1, and enhancing the expression of MMP-9.
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Aim ToobservetheeffectofSanHuang Decoction (SHD )on glucose and lipid metabolism in insulin resistance(IR)3T3-L1 adipocytes.Methods TheIRmodelof3T3-L1adipocyteswasinducedby high glucose and hyperinsulinism cultivation(also con-taining dexamethasone ).The adipocytes were treated with rosiglitazone(Ros)and different concentrations of SHD(2. 5,5,10,20,40 g·L-1 )for 24 h.The content of glucose disappeared from the culture medium was determined as glucose consumption of the cells. The transport of glucose was observed by 2-deoxidation-[3 H]-glucose uptake method.The efflux of nonesteri-fied fatty acids(NEFA)from adipocytes was observed by the concentration of NEFA in the culture medium. The mRNA expression of glucose transporter-4 (GLUT-4)was measured by real-time polymerase chain reac-tion (real-time PCR).The protein expression of GLUT-4wasdetectedbyWesternblot.Results Compared with the Con group,SHD(5,10,20,40 g·L-1 ) could significantly induce the glucose consumption and transportion(P0.05).Conclusion SHDcanin-crease insulin sensitivity by increasing glucose trans-portation and consumption in the 3T3-L1 adipocytes as well as decreasing the NEFA efflux from the cells.
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OBJECTIVE:To investigate the pharmacokinetic profiles of baicalin(a constituent in Sanhuang decoction) in rats.METHODS:Blood samples were collected as schedule from rats after intragastrically administration of Sanhuang decoction, then the content of baicalin was determined by HPLC and the chief pharmacokinetic parameters were computed.RE SU1TS:The chief pharmacokinetic parameters of baicalin in rats were as follows:t_(max1)=(15?4.23) min;t_(max2)=(6.8?0.54) h;C_(max1) =(4.49?1.56)?g?mL~(-1);C_(max2)=(3.25?1.23)?g?mL~(-1);AUC =(35.52?12.35)?g?h?mL ~(-1);t_(12)=(5.12?0.23)h;CL=(3.86?0.91) 1?h~(-1).CONCLUSION:The method is simple in sample treatment,rapid and accurate,specific and sensitive,and it can be used as a means for the monitoring of the serum concentration of baicalin containing Chinese patent medicines and so as to provide reference for clinical rational drug use.