Silibinin inhibits ultraviolet B radiation‐induced mast cells recruitment and bone morphogenetic protein 2 expression in the skin at early stages in Ptch(+/−) mouse model of basal cell carcinoma

Around 80% of nonmelanoma skin cancers (NMSCs) are basal cell carcinoma (BCC), still studies evaluating the efficacy of chemopreventive agents during early stage/s of BCC development are lacking. Accordingly, utilizing the well‐established patched (Ptch)+/? mouse model of ultraviolet B (UVB) radiation‐induced BCC formation, we excised skin samples from UVB exposed Ptch+/? and Ptch+/+ mice before tumor formation to study the promotion/progression of BCC and to determine the efficacy and target/s of silibinin, a well‐known skin cancer chemopreventive agent. UVB exposure for 1 month increased the number of mast cells in Ptch+/? mice by ~48% (P < 0.05), which was completely inhibited by silibinin. Polymerase chain reaction profiler array analysis of skin samples showed strong molecular differences between Ptch+/+ and Ptch+/? mice which were either unexposed or UVB irradiated+/? silibinin treatment. Most notably, silibinin treatment significant decreased the expression of BMP‐2, Bbc3, PUMA, and Ccnd1 in Ptch+/? mice irradiated with silibinin + UVB. Additional studies showed that silibinin targets UVB‐induced expression of bone morphogenetic protein 2 (BMP‐2) in Ptch+/? mouse skin. Last, our studies found that silibinin strongly attenuates UVB‐induced BMP‐2 expression and DNA damage in Ptch+/? mouse skin ex vivo only after single UVB exposure. Together, our results suggest a possible role of mast cell recruitment and BMP‐2 activation in the early stages of BCC development; these are strongly inhibited by silibinin suggesting its possible chemopreventive efficacy against BCC formation in long‐term UVB exposure regimen.     

Activation of microRNA-494-targeting Bmi1 and ADAM10 by silibinin ablates cancer stemness and predicts favourable prognostic value in head and neck squamous cell carcinomas

Tumor initiating cells (TICs) possessing cancer stemness were shown to be enriched after therapy, resulting in the relapse and metastasis of head and neck squamous cell carcinomas (HNC). An effective therapeutic approach suppressing the HNC-TICs would be a potential method to improve the treatments for HNC. We observed that the treatment of silibinin (SB) dose dependently down-regulated the ALDH1 activity, CD133 positivity, stemness signatures expression, self-renewal property, and chemoresistance in ALDH1+CD44+ HNC-TICs. Using miRNA-microarray and mechanistic studies, SB increased the expression of microRNA-494 (miR-494) and both Bmi1 and ADAM10 were identified as the novel targets of miR-494. Moreover, overexpression of miR-494 results in a reduction in cancer stemness. However, knockdown of miR-494 in CD44⁻ALDH1⁻non-HNC-TICs enhanced cancer stemness and oncogenicity, while co-knockdown of Bmi1 and ADAM10 effectively reversed these phenomena. Mice model showed that SB treatment by oral gavage to xenograft tumors reduced tumor growth and prolonged the survival time of tumor-bearing mice by activation of miR-494-inhibiting Bmi1/ADAM10 expression. Survival analysis indicated that a miR494^highBmi1^lowADAM10^low phenotype predicted a favourable clinical outcome. We conclude that the inhibition of tumor aggressiveness in HNC-TICs by SB was mediated by up-regulation miR-494, suggesting that SB would be a valuable anti-cancer drug for treatment of HNC.     

水飞蓟宾抑制人乳头状甲状腺癌TPC-1细胞增殖及诱导凋亡作用的实验研究

目的:研究水飞蓟宾(Silibinin,SB)在体外对人乳头状甲状腺癌细胞株TPC-1增殖的抑制作用。方法:采用CCK-8法观察不同浓度水飞蓟宾对TPC-1细胞株增殖的抑制作用;流式细胞仪分析细胞周期及凋亡情况;Western blot检测细胞周期分布及凋亡相关蛋白的表达变化。结果:CCK-8检测发现,水飞蓟宾呈时间-剂量依赖性地抑制TPC-1细胞株的增殖,诱导细胞凋亡及G0/G1期阻滞,并伴有细胞周期调节蛋白CDK2和CDK6的表达水平降低和凋亡相关蛋白的升高。结论:水飞蓟宾具有抑制人乳头状甲状腺癌细胞株TPC-1的增殖和诱导其凋亡的作用。     

Silibinin inhibits myofibroblast transdifferentiation in human tenon fibroblasts and reduces fibrosis in a rabbit trabeculectomy model

Purpose: To investigate the effect of silibinin in myofibroblast transdifferentiation and in animal trabeculectomy models. Methods: The effect of silibinin on the expression of α‐smooth muscle actin (α‐SMA) and vimentin in response to transforming growth factor‐β1 (TGF‐β1) was determined in human tenon fibroblasts (HTFs). Cell migration and collagen contraction arrays were used to demonstrate the functionality of silibinin‐modulated HTFs. ELISA analysis was used to determine the effect of silibinin on the release of type 1 collagen and connective tissue growth factor (CTGF). The effect of silibinin on the activation of the TGF‐β receptor–related pathway was evaluated by Western blotting. A rabbit model of trabeculectomy was established to assess the effect of silibinin in vivo. Results: TGF‐β1 elevated the expression of α‐SMA and vimentin in HTFs; this elevation was inhibited by silibinin. TGF‐β1 increased cell migration and collagen contraction of HTFs, which were also suppressed by silibinin. The production of both CTGF and type 1 collagen in TGF‐β1‐treated HTFs was inhibited by silibinin. The effects of silibinin on TGF‐β1‐stimulated HTFs were mediated via the down‐regulation of TGF‐β receptor–related SMAD signalling pathways. In the rabbit model of trabeculectomy, silibinin increased the period of decreasing intraocular pressure after trabeculectomy and reduced the production of collagen and α‐SMA at the site of blebs in vivo. Conclusion: Silibinin inhibited the TGF‐β receptor–related signalling pathway in TGF‐β‐treated HTFs and several of the downstream events associated with myofibroblast transdifferentiation. Silibinin also improved the outcome of trabeculectomies by reducing the fibrotic response in the bleb tissue in vivo.     

Inhibiting NF-κB activation and ROS production are involved in the mechanism of silibinin's protection against D-galactose-induced senescence

Aging is featured by intelligence decline, behavioral disorders and cognitive disability. Autophagy is related to senescent development. In this study, we investigated the roles of NF-κB and autophagy in hippocampal neurons of d-galactose-induced senescent mice, and examined the protective roles of silibinin. Senescence was induced in 6-month-old mice by subcutaneous injection of d-galactose (150 mg/kg/d, for 6 weeks). Silibinin (50 mg/kg/d, intramuscular injection, for 6 weeks) or inhibitors (PDTC, 3-MA or rapamycin, 50 mg/kg/d, subcutaneous injection, for 6 weeks) were given 1 h before d-galactose exposure. Senescent control animals received vehicle for the same time. Ethological analysis, immunofluorescence staining, flow cytometric analysis, western blot and enzyme activity assays were used. Compared with senescent controls, silibinin, PDTC or rapamycin-treated mice showed upregulations of spatial recognition memory (P < 0.05), cellular oxidoreductase activities (P < 0.05) and autophagy (P < 0.05) as well as downregulations of MDA (P < 0.05) and ROS (P < 0.05) levels. We propose in d-galactose-induced murine senescence, autophagy is inhibited by NF-κB, inducing the deactivations of celluar oxidoreductases and upregulation of ROS level. The protection by autophagy and the promotion of cellular oxidoreductase activities via inhibiting NF-κB activation and ROS production are involved in the mechanism of silibinin's protection against d-galactose-induced senescence.     

栀子提取物对脂肪肝大鼠增强胰岛素敏感性及抗脂肪肝作用

目的:研究栀子提取物对高脂诱导胰岛素抵抗-脂肪肝大鼠血糖、血脂、胰岛素抵抗及肝功能和肝细胞肪变性病理改变的作用。方法:用高脂、高糖饲料喂养大鼠,饲养56d,第57天测定大鼠空腹血糖(FBG)和口服耐量试验2h血糖(OGTT-2hBG),并依血糖和体质量将大鼠均衡随机分为脂肪肝模型对照组,水飞蓟宾组:0.154g.kg-1,栀子高剂量组:3.6g.kg-1,栀子低剂量组:1.8g.kg-1;连续灌胃给药28d后,测定大鼠FBG,OGTT-2hBG和血清胰岛素(Fins)水平,并计算胰岛素抵抗指数(HOMA-IRI)和胰岛素敏感指数(ISI);测定血浆三酰甘油(TC)、总胆固醇(TG)、低密度脂蛋白(LDL-C)、高密度脂蛋白(HDL-C)、游离脂肪酸(FFA)水平;测定天门冬氨氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、胆碱脂酶(ChE)、超氧化物歧化酶(SOD)、一氧化氮合酶(NOS)活性以及丙二醛(MDA)和一氧化氮(NO)含量等的变化,并评估肝组织脂肪变性。结果:与正常大鼠比较,模型组大鼠胰岛素含量和IRI明显升高(P<0.01),而ISI降低(P<0.01),水飞蓟宾及栀子3.6g.kg-1和1....     

水飞蓟宾胶囊治疗非酒精性脂肪肝50例的疗效

目的:探讨水飞蓟宾胶囊(水林佳)治疗非酒精性脂肪肝(nonalcoholic fatty liver disease,NAFLD)的临床疗效及不良反应。方法:选择NAFLD患者96例,随机分为试验组和对照组,其中,对照组46例,采用静脉点滴甘利欣注射液100 mg+5%葡萄糖注射液250 mL,qd,疗程15 d,复查肝功能,如正常即停药,不正常则继续用药,总观察期为3个月;试验组50例,在静脉点滴甘利欣注射液(qd)同时服用水飞蓟宾胶囊70 mg,tid,疗程15 d,复查肝功能,如正常即停药,不正常则继续用药,观察期为3个月。治疗前后检测转氨酶(TA)、总胆固醇(TC)、三酰甘油(TG)等及超声影像变化,同时观察两组患者用药期间的不良反应。结果:治疗结束时,试验组谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆固醇(TC)、三酰甘油(TG)等均明显下降,肝脏影像学也有明显改善。对照组虽然ALT和AST有所下降,但肝脏影像学无明显改善。治疗组总有效率为90%,对照组有效率为54.3%,有显著性差异(P<0.01),观察期间两组均未出现明显不良反应。结论:水飞蓟宾胶囊具有保肝、降血脂、改善肝纤维化的作用,对N...     

Amatoxin Poisoning: Case Reports and Review of Current Therapies

Background

Diagnosis and management of Amanita mushroom poisoning is a challenging problem for physicians across the United States. With 5902 mushroom exposures and two resultant deaths directly linked to Amanita ingestion in 2009, it is difficult for physicians to determine which patients are at risk for lethal toxicity. Identification of amatoxin poisoning can prove to be difficult due to delay in onset of symptoms and difficulty with identification of mushrooms. Consequently, it is difficult for the Emergency Physician to determine proper disposition. Further, treatment options are controversial.

Objectives

To review current data to help health care providers effectively identify and treat potentially deadly Amanita mushroom ingestions.

Case Reports

We present two cases of Amanita mushroom ingestion in the northeastern United States treated with N-acetylcysteine, high-dose penicillin, cimetidine, and silibinin, a semi-purified fraction of milk thistle-derived silymarin, as part of their treatment regimen. The mushroom species was identified by a consultant as Amanita Ocreata.

Conclusions

We present the successful treatment of 2 patients who ingested what we believe to be an Amanita species never before identified in the northeastern United States.