查找使用阿拉丁产品的科研文献

Preparation of sensitive monoclonal antibodies against triptolide and establishment of a rapid and cost-effective icELISA method for authentication of Tripterygium wilfordii and related products

Yaping Duan,Congmin Liu,Xiaowen Dou,Liu Wu,Hao Liu,Linan Shan,Shihai Yang,Jiaoyang Luo,Meihua Yang

Triptolide is a characteristic chemical component and quality marker of Tripterygium wilfordii Hook. F., a widely used traditional Chinese medicine with various pharmacological activities. Triptolide-14-succinate is a derivative of triptolide that can be combined with bovine serum albumin and ovalbumin to form an immunogen and coating antigen. Monoclonal antibody specific to triptolide-14-succinate conjugated with ovalbumin was produced and named mAb 2D10 1H8. An indirect competitive enzyme-linked immunosorbent assay (icELISA) was established by using mAb 2D10 1H8. The 50% inhibition concentration and working range of triptolide for the icELISA were 0.28 and 0.09–0.87 μg mL −1 , (based on 20–80% binding inhibition of mAb 2D10 1H8 to triptolide, R 2  = 0.9997, y = -0.571Logx + 0.246), respectively. The icELISA displayed crossreactivity values of 48.6, 20.5, 3.27, and < 1.00% for triptonide, triptophenolide, tripterifordin, and other analogs of triptolide, respectively. The average recovery of triptolide from the Tripterygium wilfordii samples as determined by icELISA ranged from 90.05 to 106.6%. Triptolide was detected in 38 batches of Tripterygium wilfordii samples by icELISA, which was validated by ultra-fast liquid chromatography combined with electrospray ionization tandem-mass spectrometry. In conclusion, the icELISA method established in the present study is applicable for rapid authentication of Tripterygium wilfordii and related products.

HSP60-mediated UPRmt Overactivation drives triptolide-induced hepatocellular lipoapoptosis

Zhichao Yu,Lan Hu,Jing Ding,Xiang Li,Qi Zhang,Qiaoyu Zhang,Shuoyu Chen,Pei Zhang,Jiaqi Zhang,Baoping Jiang,Xueping Zhou,Lingling Zhou

Background The mitochondrial unfolded protein response (UPR mt ) maintains proteostasis, but its dysregulation dictates cell fate. This study aimed to elucidate the signaling mechanism by which triptolide (TP), a bioactive component of Tripterygium wilfordii Hook. f., triggers hepatocellular Lipoapoptosis, focusing on HSP60-mediated UPR mt overactivation. Methods A TP-induced mouse liver injury model was established. Serum biochemistry, histopathology, and liver proteomics were performed. Key proteins (HSP60, UPR mt components, SREBP-1c, FASN, cleaved caspase-3) were validated by Western blot and RT-qPCR both in vivo and in AML-12 hepatocytes, while co-immunoprecipitation (Co-IP) was conducted in AML-12 cells. Functional validation used siRNA knockdown. Results TP induced liver injury, lipid deposition, and apoptosis. Proteomic screening implicated UPR mt activation, with HSP60 subsequently validated as a central upregulated component. TP downregulated MFN2 and upregulated HSP60, ClpP, DRP1, SREBP-1c, FASN, and cleaved caspase-3. Co-IP confirmed a direct HSP60–SREBP-1c interaction. Moreover, HSP60 knockdown attenuated the entire TP-induced cascade, including UPR mt overactivation, mitochondrial dysfunction, lipid dysregulation, and apoptosis. Conclusion This study delineates a novel mitochondria-to-nucleus pathway wherein TP induces HSP60-mediated UPR mt overactivation, which promotes SREBP-1c/FASN-driven lipid accumulation, culminating in lipoapoptosis. The “HSP60–UPR mt –SREBP-1c/FASN–lipoapoptosis” axis links dysregulated organelle stress signaling to metabolic cell death. These findings establish HSP60 as a key signaling node and a potential therapeutic target for intercepting this pathological cascade.

TP-CSO: A Triptolide Prodrug for Pancreatic Cancer Treatment

Xinlong Wang,Huahui Zeng,Xin Zhu,Duanjie Xu,Qikang Tian,Can Wang,Lingzhou Zhao,Junwei Zhao,Mingsan Miao,Xiangxiang Wu

Triptolide (TP) is a potential drug candidate for the treatment of cancer, but its use was hampered by its systemic toxicity and poor water solubility. Hence, a TP-CSO prodrug was synthesized by conjugating TP to chitosan oligosaccharide (CSO), and characterized by1H NMR, FTIR, DSC and XRD analyses. The TP-CSO containing about 4 wt% of TP exhibited excellent water solubility (15 mg/mL) compared to TP (0.017 mg/mL). Compared with TP, the pharmacokinetics of the conjugate after oral administration showed a three-fold increase in the half-life in the blood circulation and a 3.2-fold increase in AUC(0–∞). The orally administered TP-CSO could more effectively inhibit tumor progression but with much lower systemic toxicity compared with TP, indicating significant potential for further clinical trials. In conclusion, CSO-based conjugate systems may be useful as a platform for the oral delivery of other sparingly soluble drugs.Keywords:triptolide;toxicity;water solubility;pharmacokinetics;pancreatic cancer

Transdermal delivery of triptolide–phospholipid complex to treat rheumatoid arthritis

Xin-Yi Liu,Wen-Jun Pei,Ye-Zhen Wu,Fang-Li Ren,Si-Yu Yang,Xiu Wang

The aim of this study was to develop and evaluate a triptolide phospholipid complex (TPCX) for the treatment of rheumatoid arthritis (RA) by transdermal delivery. TPCX was prepared and characterized by differential scanning calorimetry (DSC), Fourier-transform infrared spectroscopy (FTIR) analysis, transmission electron microscope (TEM), and scanning electron microscope (SEM). The solubility of TPCX was determined. Then, a TPCX cream was prepared to evaluate its percutaneous permeability and the antiarthritis effect. The transdermal permeability was determined using the Franz method, and a microdialysis system was used for skin pharmacokinetic study. A rat model of RA was prepared to evaluate the pharmacological effects. TPCX increased the solubility of triptolide in water, and the percutaneous permeability of TPCX cream was greatly enhanced compared with triptolide cream. The skin pharmacokinetic study indicated that TPCX cream has a longer biological half-life (t1/2) and mean residence time (MRT), but it has a shorter Tmax than that of triptolide cream in vivo. The area under the curve (AUC0–t)/AUC0–∞) and the peak concentration (Cmax) of TPCX cream were obviously higher than those of triptolide cream. The TPCX-loaded cream alleviated paw swelling and slowed down the progression of arthritis by inhibiting the inflammatory response by down regulating the TNF-α, IL-1β, and IL-6 levels, thus exhibiting excellent antiarthritic effects. In summary, the prepared TPCX effectively increases the hydrophilicity of triptolide, which is good for its percutaneous absorption and enhances its effect on RA rats. TPCX can be a good candidate for the transdermal delivery to treat RA.

Targeting tryptophan metabolism reveals Clematichinenoside AR alleviates triptolide-induced hepatotoxicity

Xin-Nan Wang,Wen-Rui Xia,Jian-Qun Liu,Fang-Yuan Sun,Zhu-Jun Zhong,Li-Fang Liu,Gui-Zhong Xin

Liver toxicity induced by Triptolide (TP) has limited its clinical application on rheumatoid arthritis (RA). Saponins have been proved as an efficacious remedy to mitigate hepatotoxicity. However, the mechanism of reducing hepatotoxicity by saponins intervention remains incompletely characterized. Tryptophan (Trp) metabolites activate transcriptional regulators to mediate host detoxification responses. Our study aimed to investigate whether Clematichinenoside AR (C-AR) could attenuate TP-induced liver damage by regulating Trp metabolism. We used targeted metabolomics to quantify Trp metabolites in the serum and liver samples of collagen-induced arthritis rats treated by TP. Multiple comparison analyses helped the evaluation of promising biomarkers. The pronounced changed levels of Trp, indole acetic acid, and indole-3-carboxaldehyde in the serum and indole acetic acid , indole , and tryptamine in the liver are relevant to TP-induced liver injury. Intervention with C-AR could relieve TP-induced hepatotoxicity evidenced by ameliorative serum parameters and hepatic histology. In addition, C-AR regulated the levels of these indoles biomarker candidates to normal. Therapeutic modulation with natural compounds might be a useful clinical strategy to ameliorate toxicity induced by TP. Deciphering Trp metabolism will facilitate a better understanding of the pathogenesis of diseases and drug responding.

Folic Acid-Targeted Mixed Pluronic Micelles for Delivery of Triptolide

Meizhen Yin,Xinying Zhang,Tongguang Zhang,Zhiqiang Bao,Zhihui He

The present study aimed to explore an ideal delivery system for triptolide (TPL) by utilizing the thin-film hydration method to prepare drug-loaded, folate-modified mixed pluronic micelles (FA–F-127/F-68–TPL). Scanning electron microscopy and atomic force microscopy showed that the drug-loaded micelles had a spherical shape with a small particle size, with an average of 30.7 nm. Cell viability experiments showed that FA–F-127/F-68–TPL significantly reduced HepG2 cell viability, exhibiting strong cytotoxicity. Its cytotoxicity was markedly enhanced compared with bare TPL. Nile red (Nr) was used as a model drug to prepare FA–F-127/F-68–Nr to further validate its tumor-targeting and cellular uptake capability. After coincubation with HepG2 cells, a multifunctional microplate reader showed that intracellular fluorescence intensity significantly increased, indicating that FA–F-127/F-68–Nr could more effectively enter the cells. A nude mouse model of subcutaneous hepatocellular carcinoma was constructed. Following tail vein injection of FA–F-127/F-68–Nr, the fluorescence imaging system showed that FA–F127/F-68–Nr could significantly target tumor tissue, and even if entering the small-sized tumor was challenging, it could be excreted through urine. Nude mice with subcutaneous hepatocellular carcinoma were treated with tail vein injections of FA–F-127/F-68–TPL (45 µg/kg) every other day for 21 days. The results showed that the growth of the transplanted tumors was significantly slowed, with no significant difference compared with bare TPL. In summary, the FA–F-127/F-68–TPL exhibits the advantages of low cost, excellent biological properties, active/passive targeting capabilities, notable cytotoxicity against liver cancer cells, and significant inhibition of transplanted hepatocellular carcinoma growth. Significantly, the FA–F-127/F-68–TPL, despite challenges in targeting tumors with an insignificant EPR effect, can be efficiently excreted via the kidneys, thereby preventing the release of the drug during prolonged circulation and potential damage to normal tissues. Therefore, FA–F-127/F-68–TPL represents a promising antitumor drug delivery system.

Glycyrrhizic Acid-Lipid Framework Nanovehicle Loading Triptolide for Combined Immunochemotherapy

Ziyi Xu,Yu Huang,Yihan Wu,Jiamei Chen,Sai-Wang Seto,George Pak-Heng Leung,Yin Cai,Jingjing Li,Jinming Zhang

Despite the acknowledged advantages of combined immunochemotherapy for tumor treatment, the high efficiency of co-delivery of these combined agents into the targeted tumor tissue is still challenging. Herein, based on a “three-birds-with-one-stone” strategy, a facile glycyrrhizic acid (GL)-lipid hybrid nanoplatform loading triptolide (TP/GLLNP) is designed to better address the dilemma. Differing from the traditional liposomes with dual-drug co-delivery NPs, GL with a cholesterol-like structure is primarily employed to construct the lipid membrane skeleton of the GL-based lipid nanoparticle (GLLNP), and then triptolide (TP) is readily loaded in the lipid bilayer of GLLNP. The fabricated GLLNP possessed similar drug loading efficacy, particle size, and storage stability; none of the hemolysis; even higher membrane fluidity; and lower absorbed opsonin proteins compared with the conventional liposomes. Compared to TP-loaded traditional liposomes (TP/Lipo), TP/GLLNP exhibits significantly enhanced cellular uptake, cytotoxicity, and apoptosis of HepG2 cells. In addition, GLLNP could ameliorate tumor immunosuppression by promoting tumor-associated macrophage polarization from M2 to M1 phenotype. Furthermore, enhanced retention and accumulation in the tumor area of GLLNP could be found. As expected, TP/GLLNP displayed synergistic anti-hepatocellular carcinoma efficacy in vivo. In conclusion, this study provides an inspirational strategy to combine the anti-HCC benefits of GL and TP using a novel dual-drug co-delivery nanosystem.

Triptolide improves spinal cord injury by promoting autophagy and inhibiting apoptosis

Ning Zhu,Jianwei Ruan,Xinming Yang,Yang Huang,Yuting Jiang,Yucheng Wang,Danyang Cai,Yu Geng,Marong Fang

We investigated the effect of triptolide (TP) on spinal cord injury (SCI), and its underlying mechanism. Following the establishment of the SCI model using YFP H-line transgenic mice, TP was intraperitoneally injected at a dose of 0.2 mg/kg once daily for 7 days. Behavioral tests, Nissl staining, and hematoxylin–eosin staining were employed to assess motor function recovery and neuronal cell death. Western blot and immunofluorescence staining were used to assess autophagy-associated proteins (LC3B, p62, Beclin-1) and the apoptosis-associated proteins (Bcl-2, caspase-3, Bax). The TP-treated group showed improved motor functions, and reduced neuronal cell death. Also, significant upregulation of Bcl-2 and LC3B expressions, with the downregulation of p62, Bax and caspase-3 expressions were found in the TP-treated group. Additionally, phosphorylation of extracellular signal-regulated protein kinases 1 and 2 (ERK1 and ERK2) was decreased in the TP-treated group. TP mediates its protective effect in SCI by promoting the autophagic pathway while inhibiting the MAPK/ERK1/2 signaling pathway. These results demonstrate the therapeutic potential of TP in SCI.

Triptolide alleviates psoriasis through inhibiting the Wnt5a/β-Catenin signaling pathway

Eryang Chen,Lei Wang,Qu Wang,Yan Cai,Yaning Dou,Hongyan Qu,Junyi Zhu,Haiyang Zhao,Suqing Zheng,Chengguang Zhao,Bin Chen

Background: Psoriasis, an immune-mediated chronic inflammatory skin disease, is characterized by keratinocyte proliferation and inflammatory cell infiltration. T ripterygium wilfordii is a potential treatment option for psoriasis, and triptolide (TP) is one of its active components. TP may possess the potential to treat psoriasis; however, its mechanism of action remains unknown.Objective: The research aims to explore the therapeutic effect of TP on psoriasis and elucidate its potential targets.Methods: The imiquimod-induced psoriasis-like lesion mouse model was used to identify the mechanism underlying the therapeutic effect of TP.RNA-seq strategy was utilized to forecast the targets and mechanisms of TP in the context of psoriasis.Finally, we verify the effect of TP in the IL-17A-induced keratinocyte hyperproliferation and inflammation model.Results: TP reduced epidermal hyperplasia as well as psoriasis area and severity index scoring. Moreover, treatment with TP inhibited IMQ-induced splenomegaly and T-helper 17 cell differentiation in the psoriatic mice. Additionally, the treatment reduced the serum levels of pro-inflammatory cytokines such as interleukin (IL)-17A, IL-22, IL-23, IL-6, and tumor necrosis factor-α in the mice. The sequencing of RNA obtained from skin lesions of the psoriatic mice indicated that treatment with TP significantly downregulated Wnt5a RNA levels. Moreover, the Wnt5a/β-catenin pathway upregulated by IMQ was downregulated by treatment with TP. Additionally, IL-17A induced and upregulated Wnt5A and β-catenin mRNA expression, and TP inhibited this upregulated expression in HaCaT cells. Furthermore, TP inhibited proliferation, promoted apoptosis, and arrested the cell cycle in the IL-17A-induced keratinocyte hyperproliferation and inflammation model, thereby exhibiting its anti-inflammatory properties.Conclusion: TP alleviated psoriasis in mice by exerting anti-inflammatory effects and inhibited keratinocyte proliferation, which was partly achieved by regulating the Wnt5a/β-catenin signaling pathway.

Alteration of Androgen Receptor Protein Stability by Triptolide in LNCaP Cells

Wei Li,Bi-De Liu,Kai Liao,Yong Liu,Zi-Jin Wan,Yu-Fen Dong,Qian-Qian Cao,Qian Zhu,Xiao Gu

Background and Objective:Although triptolide was effective for prostate cancer (PCa), the mechanism is still unclear. Androgen receptor (AR) plays a large role in the development and progression of PCa, even after castration. The present study aimed at investigating the effects of triptolide on AR protein stability and the possible mechanism.Materials and Methods:By blocking protein synthesis with cycloheximide (CHX), the effect of triptolide on AR protein stability was investigated with western blot assay. The potential role of calpains in triptolide reduced AR protein stability was investigated with calpain inhibitor and Ca2+chelator.Results:Triptolide down-regulated AR protein level when protein synthesis was blocked by CHX, demonstrating the decrease of AR protein stability. The AR protein level was restored when the cells were co-treated with triptolide and calpain inhibitor or Ca2+chelator, indicating the important role of calpains.Conclusions:The results indicate that triptolide can activate calpain via promoting intracellular Ca2+accumulation, and thus decrease the stability of AR protein, subsequently resulting in the breakdown of the AR protein in LNCaP cells. This work provides an experimental basis and evidence to elucidate the anti-PCa mechanisms of triptolide.Keywords:triptolide;androgen receptor;calpain;protein stability

In vivo antitumor effects of carboxymethyl chitosan-conjugated triptolide after oral administration

Huahui Zeng,Xin Zhu,Qikang Tian,Yinyin Yan,Lan Zhang,Min Yan,Ruiqin Li,Xiaofang Li,Guoqiang Wang,Jinlian Ma,Yufang Su,Xiangbo Zhang,Linyu Ma,Zhenqiang Zhang,Xiangxiang Wu

Novel Carboxylated Chitosan-Based Triptolide Conjugate for the Treatment of Rheumatoid Arthritis

Lan Zhang,Min Yan,Kun Chen,Qikang Tian,Junying Song,Zijuan Zhang,Zhishen Xie,Yong Yuan,Yaquan Jia,Xin Zhu,Zhenqiang Zhang,Xiangxiang Wu,Huahui Zeng

A new platform for triptolide (TP) delivery was prepared by conjugating TP to a carboxylmethyl chitosan (CMCS). Compared with the natural TP, the TP-conjugate (TP-CMCS) containing TP of ~5 wt% exhibited excellent aqueous solubility (>5 mg/mL). Results of in vitro experiments showed that TP-CMCS could relieve TP-induced inhibition on RAW264.7 cells and apoptosis, respectively. Compared with the TP group, TP-CMCS could effectively alleviate the toxicity injury of TP and decreased the mortality rate of the mice (p< 0.05). TP-CMCS did not cause much damage to the liver (AST and ALT) and kidney (BUN and CRE) (p< 0.05). After administration, the levels of IL-6, IL-1β, and TNF-α decreased, and the arthritis detumescence percentages increased significantly, and the bony erosion degree was distinctly decreased in the TP-CMCS groups and TP group. Our results suggested that TP-CMCS was a useful carrier for the treatment of RA, which enhanced aqueous solubility of free TP and reduced drug toxicity in vitro and in vivo.Keywords:triptolide;toxicity;water solubility;carboxylmethyl chitosan;drug carrier system