Chemical constituents from Hypericum curvisepalum / 中国中药杂志

This study explored the chemical constituents of the aerial part of Hypericum curvisepalum. Sixteen compounds were isolated from the 95% ethanol extract of H. curvisepalum with various chromatographic techniques, including a new prenylated phenyl polyketide, mysorenone D(1). Other compounds were mysorenone-A(2), mysorenone-C(3), mysorenone-B(4), peplidiforone A(5), 4-methoxy-3-(2-methylbut-3-en-2-yl)-6-phenyl-2H-pyran-2-one(6), hyperenone-A(7), 4-(3,3-dimethylallyl)oxy-6-phenyl-α-pyrone(8), peplidiforone B(9), elegaphenone(10), hypercohin A(11), hyperisampsin G(12), spathulenol(13), quercetin(14), β-sitosterol(15), and β-amyrin(16).

Polyphyllin VI Induces Caspase-1-Mediated Pyroptosis via the Induction of ROS/NF-κB/NLRP3/GSDMD Signal Axis in Non-Small Cell Lung Cancer.

Trillium tschonoskii Maxim (TTM), a traditional Chinese medicine, has been demonstrated to have a potent anti-tumor effect. Recently, polyphyllin VI (PPVI), a main saponin isolated from TTM, was reported by us to significantly suppress the proliferation of non-small cell lung cancer (NSCLC) via the induction of apoptosis and autophagy in vitro and in vivo. In this study, we further found that the NLRP3 inflammasome was activated in PPVI administrated A549-bearing athymic nude mice. As is known to us, pyroptosis is an inflammatory form of caspase-1-dependent programmed cell death that plays an important role in cancer. By using A549 and H1299 cells, the in vitro effect and action mechanism by which PPVI induces activation of the NLRP3 inflammasome in NSCLC were investigated. The anti-proliferative effect of PPVI in A549 and H1299 cells was firstly measured and validated by MTT assay. The activation of the NLRP3 inflammasome was detected by using Hoechst33324/PI staining, flow cytometry analysis and real-time live cell imaging methods. We found that PPVI significantly increased the percentage of cells with PI signal in A549 and H1299, and the dynamic change in cell morphology and the process of cell death of A549 cells indicated that PPVI induced an apoptosis-to-pyroptosis switch, and, ultimately, lytic cell death. In addition, belnacasan (VX-765), an inhibitor of caspase-1, could remarkably decrease the pyroptotic cell death of PPVI-treated A549 and H1299 cells. Moreover, by detecting the expression of NLRP3, ASC, caspase-1, IL-1ß, IL-18 and GSDMD in A549 and h1299 cells using Western blotting, immunofluorescence imaging and flow cytometric analysis, measuring the caspase-1 activity using colorimetric assay, and quantifying the cytokines level of IL-1ß and IL-18 using ELISA, the NLRP3 inflammasome was found to be activated in a dose manner, while VX-765 and necrosulfonamide (NSA), an inhibitor of GSDMD, could inhibit PPVI-induced activation of the NLRP3 inflammasome. Furthermore, the mechanism study found that PPVI could activate the NF-κB signaling pathway via increasing reactive oxygen species (ROS) levels in A549 and H1299 cells, and N-acetyl-L-cysteine (NAC), a scavenger of ROS, remarkably inhibited the cell death, and the activation of NF-κB and the NLRP3 inflammasome in PPVI-treated A549 and H1299 cells. Taken together, these data suggested that PPVI-induced, caspase-1-mediated pyroptosis via the induction of the ROS/NF-κB/NLRP3/GSDMD signal axis in NSCLC, which further clarified the mechanism of PPVI in the inhibition of NSCLC, and thereby provided a possibility for PPVI to serve as a novel therapeutic agent for NSCLC in the future.

Astragaloside IV Downregulates ß-Catenin in Rat Keratinocytes to Counter LiCl-Induced Inhibition of Proliferation and Migration.

Re-epithelialization is a crucial step towards wound healing. The traditional Chinese medicine, Astragalus membranaceus (Fisch) Bge, has been used for hundreds of years for many kinds of ulcerated wounds. Recent research has identified the active compound in this drug as astragaloside IV (AS-IV), but the underlying molecular mechanisms of its therapeutic action on keratinocytes remain poorly understood. In this study, we used an in vitro model of ulcer-like wound processes, lithium chloride (LiCl)-induced cultured mouse keratinocytes, to investigate the effects of AS-IV treatment. The effects on cell proliferation were evaluated by the MTS/PMS colorimetric assay, effects on cell migration were determined by a wound-healing scratch experiment, effects on the cell cycle were analyzed by flow cytometry, and effects on protein expression were analyzed by immunoblotting and immunofluorescence. LiCl strongly inhibited cell proliferation and migration, up-regulated ß-catenin expression, and down-regulated proliferating cell nuclear antigen (PCNA) expression. AS-IV treatment attenuat the inhibition of proliferation and migration, significantly reducing the enhanced ß-catenin expression, and recovering PCNA and ß-tubulin expression. Thus, AS-IV mediates mouse keratinocyte proliferation and migration via regulation of the Wnt signaling pathway. Down-regulating ß-catenin to increase keratinocyte migration and proliferation is one mechanism by which AS-IV can promote ulcerated wound healing.

[Effects of Fuhuang Shengji Yuchuang Ointment on expressions of types I and III collagens in granulation tissue of wound in rats with diabetes].

OBJECTIVE: To observe the effects of Fuhuang Shengji Yuchuang (FHSJYC) Ointment, a compound traditional Chinese herbal medicine, on the expressions of types I and III collagens in granulation tissue of wound in rats with diabetes. METHODS: Fifty-four male Wistar rats were randomly divided into three groups: wound control group, normal saline (NS) group and FHSJYC Ointment group. Diabetes was induced by injection of 1.5% alloxan and oral gavage of 5% glucose, and skin wound was made in rats of the NS group and the FHSJYC Ointment group. Skin wounds of the rats in the FHSJYC Ointment group were treated with FHSJYC Ointment gauze dressing, while those in the NS group were treated with NS gauze dressing once daily. The rats were executed in turn on the third day and the eleventh day of the treatment, and the changes of the content of types I and III collagens in the wound granulation tissue were observed by immunohistochemical technology. RESULTS: Compared with the NS group, the wound closure index in the FHSJYC Ointment group was increased (P<0.05). After 3-day treatment, the expression of type I collagen showed no significant differences among the three groups, while the expression of type III collagen in the FHSJYC Ointment group was higher than that in the NS group (P<0.05), similar with that in the wound control group. After 11-day treatment, the expressions of both types I and type III collagens in the FHSJYC Ointment group were higher than those in the NS group (P<0.05), similar with those in the wound control group. CONCLUSION: FHSJYC Ointment can affect the process of wound healing by promoting and regulating the expressions of types I and III collagens.

[Influence of stasis-removing and muscle-regenerating Chinese hebal medicine on the expression of collagen type I and matrix metalloproteinase-1 in granulation tissue of ordinary wound].

OBJECTIVE: To understand the effects of Chinese herbal medicine for stasis -removing (SR) and muscle-regenerating (MR), in different constitutions, on granulation tissue of ordinary wound. METHODS: Shengji Huayu recipe, a common used Chinese recipe for SR & MR (SR-MR), and its disassembled recipe, Shengji recipe (MR) and Huayu recipe (SR), were applied resectively on the full-layer skin injured wound in 120 rats to observe their effects on collagen type I (C I ) and matrix metalloproteinase-1 (MMP-1) in granulation tissue of wound at various stages of healing with immunohistochemistry technique and image-analysis system. RESULTS: The expression of C I of the normal group was obviously delayed and the level of MMP-1 increased in a ladder type from day 3 to day 11. Shengji group could promote the secretion of C I and MMP-1 at the early stage and keep a high level from day 3 to day 11. The expression of MMP-1 of Huayu group maintained a high level on the first 7 days with significant difference as compared with the Shengji group (P < 0.05), and reduced from day 11 showing significant difference when compared with the normal group and Shengji group (all P < 0.05). Shengji Huayu low dose group has two periods of peak time in promoting the secretion of C I on day 7 and 15, and showed significant difference as compared with the normal group (P < 0.05); Shengji Huayu high dose group could reduce the MMP-1 on day 11 and obviously lower when compared with the normal group (P<0.05). CONCLUSION: The mechanism of Chinese herbal medicine for SR-MR in promoting wound healing was probably through inhibiting the secretion of MMP-1 to increase the C I content in granulation tissue of wound.

[Effects of Shengji Huayu Recipe and its decomposed formulas on synthesis of collagen types I and III in granulation tissue of rats in early wound healing].

OBJECTIVE: To study the effects of Shengji Huayu Recipe (a traditional Chinese medicine compound recipe for resolving stagnation and promoting granulation) and its decomposed formulas (Huayu Recipe for resolving stagnation and Shengji Recipe for promoting granulation) on the synthesis of collagen types I and III in granulation tissue of rats in early wound healing. METHODS: Twenty-four male Sprague-Dawley (SD) rats with full-thickness skin lesion were randomized into 4 groups: Shengji Huayu Recipe-treated group, Shengji Recipe-treated group, Huayu Recipe-treated group and untreated group. Collagen types I and III in granulation tissue of the rats were tested with immunohistochemical methods and image analysis. RESULTS: On the third day of wound healing, collagen I of the rats in both Shengji Huayu Recipe-treated group and Shengji Recipe-treated group was higher than that in the untreated group, and collagen I of the rats in Huayu Recipe-treated group was lower than that in the untreated group (P<0.05). Collagen III of the rats in the three treated groups were lower than that in the untreated group (P<0.05). On the seventh day of wound healing, Collagen I of the rats in both Shengji Huayu Recipe-treated group and Shengji Recipe-treated group was higher than that in the untreated group (P<0.05), and collagen III of the rats in both Shengji Recipe-treated group and Huayu Recipe-treated group was higher than that in the untreated group (P<0.05). CONCLUSION: Resolving stagnation and promoting granulation therapy can promote the wound healing in rats.