Sanhuang decoction inhibits autophagy of periodontal ligament fibroblasts during orthodontic tooth movement by activating PI3K-Akt-mTOR pathway.

BACKGROUND: Orthodontic tooth movement (OTM) is a typical treatment that corrects malaligned teeth by applying mechanical forces. However, mechanical overload often leads to damage of PDL fibroblasts. Sanhuang decoction (SHD) is commonly used to inhibit inflammation and oxidative stress. However, the mechanism of SHD for OTM treatment is still unclear. Therefore, this study attempts to explore the underlying mechanism through relevant experiments. METHODS: In the present paper, we established a OTM rat model and further explored the effects of SHD on the PDL of OTM rats. The OTM model and effects of SHD were determined by micro-CT, and the PDL pathological changes, PDL width and capillaries in PDL were observed by H&E staining. Subsequently, the ROS levels in PDL was determined using flow cytometry analysis with DCFH-DA staining, MDA contents and antioxidative enzymes activities were also measured using commercial kits. Furthermore, the autophagy of PDL fibroblasts and proteins in the PI3K/Akt/mTOR pathway were detected using immunoluminescence, qPCR and western blotting assays. RESULTS: The results showed SHD treatment can alleviate the decrease of PDL cells and capillaries induced by OTM, and improve the MDA and ROS levels in PDL, as well as enhance the activities of SOD and GSH-Px. Further experiments indicated SHD decreased the autophagy levels of PDL fibroblasts via promoting the phosphorylation levels of mTOR, PI3K and Akt proteins. CONCLUSION: SHD inhibited autophagy of periodontal ligament fibroblasts during orthodontic tooth movement by inhibiting oxidative stress via activating PI3K-Akt-mTOR pathway. Our present findings suggested SHD treatment would be useful for management of the possible disorders occurs in orthodontic tooth movement therapy.

Chinese herbal compound SanHuang decoction reverses axitinib resistance in ccRCC through regulating immune cell infiltration by affecting ADAMTS18 expression.

This investigation aims to study the reversal effect of the Chinese herbal compound SanHuang decoction on axitinib resistance in clear cell renal cell carcinoma (ccRCC) cells and its mechanistic role by employing cellular and mouse models. Axitinib-resistant ccRCC cell lines (A498-DR and 786-O-DR) were cultured and treated with SanHuang decoction. The apoptosis and migration of tumor cells were observed by flow cytometry and wound healing assays, respectively, and the expression of a disintegrin-like and metalloprotease with thrombospondin type 1 motif 18 (ADAMTS18) was evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting (WB). In addition, A498-DR cells were inoculated into mice to establish tumorigenic models, and the models were treated with normal saline, axitinib, or different concentrations of SanHuang decoction plus axitinib. Then, the tumor diameter in each group was measured, and the expression of ADAMTS18 was evaluated by RT-PCR, WB and immunohistochemistry. In addition, the distribution of T cells (CD45+, CD4+, CD8+) and PD-L1 expression was analyzed by flow cytometry to evaluate the level of immune cell infiltration. SanHuang decoction significantly reduced the proliferative activity of axitinib-resistant tumor cells and enhanced the sensitivity of tumors to axitinib in vitro (cell lines) and in mice. In the SanHuang decoction group, the expression level of ADAMTS18 was increased to some extent, and several phenomena were observed, including (1) subcutaneous transplanted tumors grew slower, (2) the CD45+/PD-L1 ratio was decreased and (3) the proportions of CD8+ and CD4+ T cells were increased. Overexpression of ADAMTS18 was synergistic with SanHuang decoction treatment to jointly improve tumor immune infiltration and inhibit immune escape. Pearson correlation analysis of sample data showed that there was a negative correlation between the expression of ADAMTS18 and PD-L1 in tumor tissues. In conclusion, the Chinese herbal compound SanHuang decoction can reverse axitinib resistance in ccRCC cells by regulating immune cell infiltration and affecting ADAMTS18 expression.

Sanhuang Decoction Controls Tumor Microenvironment by Ameliorating Chronic Stress in Breast Cancer: A Report of Ninety Cases.

Long-term endocrine treatment which results in estrogen deprivation causes chronic stress associated with a series of uncomfortable symptoms leading not only to a decrease in quality of life but also to cancer recurrence, which may be mediated primarily through the enhanced expression of angiogenic factors, as well as a series of inflammatory microenvironmental changes that favor tumor progression. In this study, we designed a clinical trial and aimed to explore the effects of Sanhuang Decoction (SHD) treatment on chronic stress, inflammatory factors, and breast cancer recovery. A total of 90 patients with breast cancer who met the inclusion/exclusion criteria were randomly allocated to a treatment or control group. The treatment group received the standard endocrine treatment and the traditional Chinese medicine decoction known as SHD. The control group received the standard endocrine treatment only. The treatment period was 6 months. The modified Kupperman Menopausal Index, the self-rating anxiety scale, and the self-rating depression scale were evaluated once per month. The body microenvironment plasma indices related to chronic stress, such as oxidative and antioxidative stress markers, inflammatory factors, hemorheology, coagulation, lipid and D-dimer, immunologic functions, tumor biomarkers, and angiogenic factors of the vascular endothelial growth factor (VEGF) were measured before and after 6 months of treatment. After treatment for 5 months, the scores in the treatment group decreased to nearly normal levels and the control group showed no significant improvement. After treatment for 6 months, all indices related to the body microenvironment, as well as the tumor biomarkers and carcinoembryonic antigen, carbohydrate antigen 153, and angiogenic factor VEGF levels improved significantly to normal levels in the treatment group. Our primary research showed that treatment with SHD effectively improved the quality of life of breast cancer patients by facilitating a change in the body microenvironment that controlled tumor growth and prevented drug resistance. CLINICAL TRIAL REGISTRATION: Chinese Clinical Trial Registry, identifier ChiCTR-IIR-2000041413. Date of registration: 2017-06-07 (retrospective registration).

[Mechanism of Sanhuang Decoction in alleviating dextran sulfate sodium induced ulcerative colitis in mice with Candida albicans colonization:based on high-throughput transcriptome sequencing].

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 ß-1,3-glucan in serum, and TNF-α, IL-1ß, 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-1ß 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 ß-1,3-glucan in serum and TNF-α, IL-1ß, 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-1ß. 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.

Systems pharmacology and transcriptomics reveal the mechanisms of Sanhuang decoction enema in the treatment of ulcerative colitis with additional Candida albicans infection.

BACKGROUND: Ulcerative colitis (UC) is an important inflammatory phenotype in bowel disease (IBD), which is caused by multiple potential factors, including fungal dysbiosis. Candida albicans (C. albicans) was confirmed to be an important factor promoting the occurrence and development of UC. Sanhuang decoction (SHD) has been used for UC therapy in China for thousand of years, although its core active constituents and pharmacological mechanism remain undefined. METHODS: In this work, a murine model of UC with C. albicans colonization was established with dextran sodium sulfate (DSS) and C. albicans intragastric administration. The major bioactive constituents and potential mechanism of SHD against UC with fungal dysbiosis were comprehensively examined by combining systems pharmacology and in vivo transcriptomics. RESULTS: SHD attenuated C. albicans burden, reduced DAI, increased mucosal integrity and relived systemic inflammation in UC mice. Systems pharmacology analysis identified 9 core bioactive ingredients and 45 hub targets of SHD against UC. Transcriptomics analysis confirmed 370 differentially expressed genes (DEGs) after SHD treatment, which were mainly enriched in inflammatory and immune response related signaling pathways. Toll-like receptor and PI3K-Akt signaling pathway were screened out as the candidate targets involved in the action of SHD on fungal dysbiosis-associated UC, which were consistent with the findings in systems pharmacology. The expression of TLR4, IL-1ß, NF-κB, PI3K and Akt proteins were stimulated by C. albicans, and partially reversed by SHD in UC mice. CONCLUSION: These findings suggested SHD could be a candidate for the treatment of fungal dysbiosis-associated UC via TLR4-NF-κB and PI3K-Akt signaling pathways.

Mechanism of Sanhuang Decoction in alleviating dextran sulfate sodium induced ulcerative colitis in mice with Candida albicans colonization:based on high-throughput transcriptome sequencing / 中国中药杂志

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.

Effect of Shenteng Sanhuang Decoction and Gegen Qinlian Tang on Diabetic Kidney Disease Rats / 中国实验方剂学杂志

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.

Effects of Astragaloside IV on treatment of breast cancer cells execute possibly through regulation of Nrf2 via PI3K/AKT/mTOR signaling pathway.

Traditional Chinese medicine (TCM) has from ancient times been applied in China for the treatment of breast cancer with its own unique theoretical system. Sanhuang decoction composed of astragalus membranaceus, prepared rhubarb, and rhizoma curcumae longae has traditionally been used for antioxidant stress, inflammatory reaction, and angiogenesis. However, the role and mechanism of Sanhuang decoction in breast cancer remains unknown. The present study demonstrated the antitumor activity of Sanhuang decoction against breast cancer xenografts in nude mice. Notably, Sanhuang decoction promoted severe necrosis and induced cell death. In addition, Sanhuang decoction obviously regulated the inflammation and oxidative stress. Despite these, Sanhuang decoction could increase the expression of Nrf2. Moreover, si-Nrf2 exhibited the opposite effects compared with the Sanhuang decoction treatment group and reversed the antibreast cancer role of Sanhuang decoction. Further, Sanhuang decoction remarkably suppressed the expression of PI3K/AKT/mTOR signaling pathway. Taken together, Sanhuang decoction was firstly evaluated to possess potent antibreast cancer effect in vivo through regulation of inflammation and oxidative stress accomplished by up-regulation of Nrf2 via PI3K/AKT/mTOR signaling pathway and Sanhuang decoction might be a powerful candidate formula for antibreast cancer.

Effect of Sanhuang Decoction on glucose and lipid metabolism in 3 T3-L1 adipocytes / 中国药理学通报

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.

Pharmacokinetics of Baicalin of Sanhuang Decoction in Rats / 中国药房

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.