Tripchlorolide

T4 reduces cisplatin resistance by inhibiting AEG-1 gene expression in lung cancer cells.[Pubmed:35794136]

Sci Rep. 2022 Jul 6;12(1):11462.

Lung cancer is the most malignant form of cancer and has the highest morbidity and mortality worldwide. Due to drug resistance, the current chemotherapy for lung cancer is not effective and has poor therapeutic effects. Tripchlorolide (T4), a natural extract from the plant Tripterygium wilfordii, has powerful immunosuppressive and antitumour effects and may become a potential therapeutic agent for lung cancer. Therefore, this study aimed to investigate the effect of T4 on reducing chemoresistance in lung cancer cells and to explore the mechanism. 1. A549 and A549/DDP cells were separately transfected with AEG-1 overexpression and AEG-1 knockdown plasmids. A549/DDP cells were divided into the A549/DDP empty group, T4 group, and T4 + AEG-1 overexpression group. A CCK-8 assay was used to evaluate the proliferation of cells in each group. RT-qPCR and Western blotting were used to detect the expression of AEG-1 and MDR-1. Expression of AEG-1 in A549 and A549/DDP cells was positively correlated with cisplatin resistance. When the AEG-1 protein was overexpressed in A549 cells, the lethal effect of cisplatin on A549 cells was attenuated (all P < 0.05). After the AEG-1 protein was knocked down in A549/DDP cells, cisplatin was applied. The lethal effect was significantly increased compared to that in the corresponding control cells (all P < 0.05). AEG-1 protein expression gradually decreased with increasing T4 concentration in A549 and A549/DDP cells. Resistance to cisplatin was reduced after the addition of T4 to A549/DDP cells (P < 0.05), and this effect was enhanced after transfection with the AEG-1 knockdown plasmid. T4 plays an important role in increasing the sensitivity of lung cancer cells to cisplatin.

Tripchlorolide attenuates beta-amyloid generation by inducing NEP activity in N2a/APP695 cells.[Pubmed:34316383]

Transl Neurosci. 2021 Jul 20;12(1):301-308.

BACKGROUND AND PURPOSE: Alzheimer's disease (AD) is a neurodegeneration disease. The previous work from our research group demonstrated the neuroprotective effects of Tripchlorolide (T4) in AD animal models. MATERIALS AND METHODS: Neprilysin (NEP) is known as an important physiological amyloid-beta protein (Abeta) peptide-degrading enzyme in the brain due to its apparent rate-limiting function. In this study, we explored the effect of NEP on AD model N2a/APP695 cells. Western blots and enzyme-linked immunosorbent assays were performed to assess the expression of proteins, while quantitative real-time polymerase chain reaction assays were used to evaluate RNA levels. Cell vitality was detected by the MTT assay, and reactive oxygen species (ROS) levels were assessed using a ROS activity assay kit. RESULTS: We discovered that T4 was able to enhance the enzyme activity of NEP. T4 administration decreased the protein levels of the soluble amyloid precursor protein. In further experiments, we found that by using thiorphan the secretion of Abeta, oxidative stress, nitrosative stress, and inflammatory factors, which were suppressed by T4, were reversed. Due to its ability to attenuate Abeta generation and to protect neurons against the neurotoxicity of Abeta, T4 may be a potential therapy in the regulation of Abeta-related pathology in AD by affecting NEP activity. CONCLUSION: Tripchlorolide attenuates Abeta generation by inducing NEP activity in N2a/APP695 cells.

[Pharmacodynamic effect and virulent effect of Tripterygium wilfordii based on network pharmacology].[Pubmed:31602910]

Zhongguo Zhong Yao Za Zhi. 2019 Aug;44(16):3460-3467.

To investigate the pharmacodynamic effect and virulent effect of the main components of the toxic Chinese medicine Tripterygium wilfordii,such as triptolide,Tripchlorolide,tripterine,demethylzeylasteral,wilfotrine and euonine,the admet SAR online assessment system was used to calculate the properties of the main components of T. wilfordii. The potential targets of the components were mined and collected through multiple databases,and the potential targets were enriched by the bioinformatics database DAVID.Cytoscape software was used to establish a " target-pathway" network and perform topology analysis on the network. The main chemical components of T. wilfordii were able to penetrate the blood-brain barrier and had intestinal permeability. A total of 65 targets were predicted,including pathways in cancer,hepatitis B,rheumatoid arthritis,and chagas disease( American trypanosomiasis),Toll-like receptor signaling pathway,apoptosis,colorectal cancer,NF-kappa B signaling pathway,etc. T. wilfordii mainly plays a role in the treatment of immune diseases and cancer by regulating inflammatory signaling pathways and cancer signaling pathways. Its action on apoptosis pathway and drug metabolism enzymes may be the mechanism of its toxicity.

[Overview of reproductive toxicity studies on Tripterygium wilfordii in recent 40 years].[Pubmed:31602902]

Zhongguo Zhong Yao Za Zhi. 2019 Aug;44(16):3406-3414.

This paper summarizes the research progress of reproductive toxicity of Tripterygium wilfordii from 1979,and the toxicity characterization,damage mechanism,and attenuated measures are summarized. It was found that,the reproductive toxicity caused by T. wilfordii is mainly distributed on components of Tripterygium glycosides,triptolide,Tripchlorolide,and clinically preparations,such as Leigongteng Tablets and Tripterygium Glycosides Tablets. Adverse reactions to male reproductive system caused by Tripterygium preparations mainly include decreased sperm motility,oligospermia or spermatozoa,decreased fertility or infertility,etc. Long-term drug use may also lead to testicular atrophy and decreased sexual desire. Adverse reactions to women are mainly manifested as menstrual disorders,decreased menstrual volume or even amenorrhea,decreased sexual desire,infertility,etc. The reproductive toxicity of T. wilfordii is related to apoptosis of reproductive cells,disturbance of spermatogenesis or oogenesis,damage of testis and ovary in reproductive target tissues,and changes of internal environment in gonad tissues( hormones,hormone synthesis rate-limiting enzymes and energy metabolism). Drug compatibility,hormone replacement,medication duration and dosage form changes can help reduce the damage of T. wilfordii to the reproductive system. In addition,in view of the existing problems in the current study,the author proposes new directions in clinical studies,pharmacological metabolism mechanism,preparation quality standards and new therapeutic effects,etc.,to provide a basis for the safe and reasonable clinical application of T. wilfordii.

Epigenetic mechanisms underlying the effects of triptolide and tripchlorolide on the expression of neuroligin-1 in the hippocampus of APP/PS1 transgenic mice.[Pubmed:31311385]

Pharm Biol. 2019 Dec;57(1):453-459.

Context: Neuroligin-1 (NLGN1) is a cell adhesion protein located on the excitatory postsynaptic membrane. beta-Amyloid (Abeta)-induced neuroinflammation decreases NLGN1 expression through epigenetic mechanisms. Triptolide (T10) and Tripchlorolide (T4) exert protective effects on synapses in Alzheimer's disease (AD) mice, but the mechanisms remain unclear. Objective: The effects of T10 and T4 on hippocampal NLGN1 expression in AD mice and the epigenetic mechanisms were assessed using chromatin immunoprecipitation and methylated DNA immunoprecipitation. Materials and methods: Sixty APP/PS1 transgenic mice were randomly divided into an AD model group, a T10-treated group and a T4-treated group (n = 20); 20 wild-type littermates served as the control group. APP/PS1 transgenic mice were intraperitoneally injected with T10 (0.1 mg/kg) and T4 (25 mug/kg) once per day for 60 days. NLGN1 expression was examined using western blotting and quantitative PCR. Results: T10 and T4 increased the levels of the NLGN1 protein and mRNA in hippocampus of AD mice. T10 and T4 inhibited the binding of HDAC2 (p< 0.01) and MeCP2 (p< 0.01 and p< 0.05, respectively) to the NLGN1 promoter, and cytosine methylation (1.2305 +/- 0.1482/1.2554 +/- 0.3570 vs. 1.6578 +/- 0.1818, p< 0.01) at the NLGN1 promoter in the hippocampus of AD mice. T10 and T4 increased the level of acetylated histone H3 (0.7733 +/- 0.1611/0.8241 +/- 0.0964 vs. 0.5587 +/- 0.0925, p< 0.01) at the NLGN1 promoter in the hippocampus of AD mice. Conclusions: T10 and T4 may increase hippocampal NLGN1 expression in AD mice through epigenetic mechanisms, providing a new explanation for the mechanism underlying the protective effects of T10 and T4 on synapses.

Tripchlorolide May Improve Spatial Cognition Dysfunction and Synaptic Plasticity after Chronic Cerebral Hypoperfusion.[Pubmed:30923551]

Neural Plast. 2019 Feb 24;2019:2158285.

Chronic cerebral hypoperfusion (CCH) is a common pathophysiological mechanism that underlies cognitive decline and degenerative processes in dementia and other neurodegenerative diseases. Low cerebral blood flow (CBF) during CCH leads to disturbances in the homeostasis of hemodynamics and energy metabolism, which in turn results in oxidative stress, astroglia overactivation, and synaptic protein downregulation. These events contribute to synaptic plasticity and cognitive dysfunction after CCH. Tripchlorolide (TRC) is an herbal compound with potent neuroprotective effects. The potential of TRC to improve CCH-induced cognitive impairment has not yet been determined. In the current study, we employed behavioral techniques, electrophysiology, Western blotting, immunofluorescence, and Golgi staining to investigate the effect of TRC on spatial learning and memory impairment and on synaptic plasticity changes in rats after CCH. Our findings showed that TRC could rescue CCH-induced spatial learning and memory dysfunction and improve long-term potentiation (LTP) disorders. We also found that TRC could prevent CCH-induced reductions in N-methyl-D-aspartic acid receptor 2B, synapsin I, and postsynaptic density protein 95 levels. Moreover, TRC upregulated cAMP-response element binding protein, which is an important transcription factor for synaptic proteins. TRC also prevented the reduction in dendritic spine density that is caused by CCH. However, sham rats treated with TRC did not show any improvement in cognition. Because CCH causes disturbances in brain energy homeostasis, TRC therapy may resolve this instability by correcting a variety of cognitive-related signaling pathways. However, for the normal brain, TRC treatment led to neither disturbance nor improvement in neural plasticity. Additionally, this treatment neither impaired nor further improved cognition. In conclusion, we found that TRC can improve spatial learning and memory, enhance synaptic plasticity, upregulate the expression of some synaptic proteins, and increase the density of dendritic spines. Our findings suggest that TRC may be beneficial in the treatment of cognitive impairment induced by CCH.

Tripchlorolide induces autophagy in lung cancer cells by inhibiting the PI3K/AKT/mTOR pathway and improves cisplatin sensitivity in A549/DDP cells.[Pubmed:28969040]

Oncotarget. 2017 Jul 12;8(38):63911-63922.

Tripchlorolide (T4) has been shown to induce A549 lung cancer cell death predominantly by activating an autophagy pathway. However, the underlying mechanism remains unclear. Herein, we demonstrated that compared with T4 treatment alone, pretreatment with wortmannin (an inhibitor of phosphatidylinositol 3-kinase), perifosine (an inhibitor of AKT) or rapamycin (an inhibitor of mTOR) combined with a subsequent T4 treatment significantly impaired the cell viability of A549 and A549/DDP lung cancer cells. We found that either treatment scheme markedly reduced the activity of P13K and AKT. Expression of LC3II increased in parallel to the increase of the T4 concentration in both A549 and A549/DDP cells and was repressed by overexpression of AKT. The expression levels of PI3-K, PI3-P, AKT, TSC2, mTOR, p70S6K and 4E-BP1 were minimally affected by the wortmannin, perifosine, or rapamycin plus T4 treatments, but their phosphorylated products were greatly affected in A549 lung cancer cells and slightly affected in A549/DDP lung cancer cells. These results indicate that T4 induces autophagy in lung cancer cells by inhibiting the PI3K/AKT/mTOR signaling pathway. We further found that T4 decreased expression of MDR1 and improved cisplatin sensitivity of A549/DDP cells. Altogether, these results have meaningful implications for tumor therapy in the future.

Tripchlorolide Attenuates beta-amyloid Generation via Suppressing PPARgamma-Regulated BACE1 Activity in N2a/APP695 Cells.[Pubmed:26582466]

Mol Neurobiol. 2016 Nov;53(9):6397-6406.

Due to its apparent rate-limiting function, BACE1 (beta-secretase) appears to be a prime target for prevention of amyloid-beta (Abeta) generation in brains with Alzheimer's disease (AD). The activity of BACE1 is regulated by peroxisome proliferator-activated receptor-gamma (PPARgamma), a transcription factor binding site of the BACE1 promoter, indicating that PPARgamma may be a potential target for AD treatment. Several studies have demonstrated that PPARgamma activation is involved in the immunostimulation of amyloid-beta precursor protein processing by nonsteroidal anti-inflammatory drugs (NSAIDs). The present study found that Tripchlorolide (T(4)), with a similar chemical structure to that of NSAIDs, decreased the levels of Abeta secreted in N2a-APP695 cells. T(4) treatment reduced the mRNA and protein levels of BACE1 and the protein level of sAPPbeta, a cleaved N-terminal fragment of APP by BACE1. The treatment also translocated PPARgamma from cytoplasm to nuclear. Intriguingly, T(4), like pioglitazone (a PPARgamma agonist), suppressed the BACE1 activity in N2a-APP695 cells, which was attenuated by GW9662 (a PPARgamma antagonist). These results indicate that T(4) may be a PPARgamma agonist to enhance the binding of nuclear PPARgamma to the BACE1 promoter, which may in turn inhibit the transcription and translation of BACE1, suppress the activity of BACE1, and ultimately attenuate the generation of Abeta. Due to its capability to alter Abeta generation and to protect central neural system against the neurotoxicity of Abeta, T(4) may serve as a promising agent in modulating Abeta-related pathology in Alzheimer's disease.

The Pharmacological Effects and Mechanism of Tripterygium wilfordii Hook F in Central Nervous System Autoimmunity.[Pubmed:27224044]

J Altern Complement Med. 2016 Jul;22(7):496-502.

OBJECTIVE: Extracts of the Chinese herb Tripterygium wilfordii Hook F (TwHF) have potent anti-inflammatory functions and are widely used to treat rheumatoid arthritis and Crohn's disease. They have also been considered as potential drugs in the treatment of multiple sclerosis and its animal model, experimental autoimmune encephalomyelitis. METHODS: A systematic search of MEDLINE, EMBASE, PubMed, and the China National Knowledge Infrastructure (CNKI) was performed. We reviewed many Chinese- and English-language articles. RESULTS: Recent studies have indicated that TwHF extracts, such as triptolide and Tripchlorolide, are able to attenuate progression of this neuroimmunologic disorder because of their immunoregulatory, neurotrophic, and neuroprotective effects, but use of these extracts is often accompanied by acute and chronic toxicity. CONCLUSIONS: This review systematically summarizes the effects, safety consideration, and molecular mechanisms of action of TwHF extracts with regard to their inhibition of microglia activation, T cell functions, and transcriptional activation of nuclear factor (NF)-kappaB signaling.

[Lymphomatoid papulosis: a clinicopathologic study of 22 cases].[Pubmed:26850015]

Zhonghua Yi Xue Za Zhi. 2015 Dec 8;95(46):3750-2.

OBJECTIVE: To investigate the clinical presentation, histopathological features, progression, and treatment of lymphomatoid papulosis (LyP). METHODS: A retrospective review was performed on clinicopathological data of 22 patients diagnosed with LyP from June 2010 to March 2015 in Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College. RESULT: The mean age of the 22 LyP patients was 39 years (range: 7-83 years). The male-to-female ratio was 1:1. The areas predominantly affected were the trunks, followed by limbs and face. Most lesions presented as recurrent eruption of erythema, papule, nodules, ulcer, necrosis and crusting. Some of them leaved hyperpigmentation or atrophic scars on healing. Histopathologically, LyP were devided into types A, B, C, D and E, with 12, 1, 1, 3, and 4 cases in each type, respectively, and one case of mixed type B and C. One of the patients was also diagnosed with primary cutaneous anaplastic large-cell lymphoma besides the diagnosis of LyP. Among the 15 patients with follow-up information available, 11 patients were treated with regimens including oral corticosteroids, methotrexate, Tripchlorolide, intramuscular injection of interferon, phototherapy, and topical corticosteroids. The mean follow-up time was 22 (1-54) months.All the patients were alive at the end of the follow-up period. CONCLUSIONS: LyP is a low-grade malignant T-cell lymphoma with a benign clinical course but histologically malignant features. Multi-agent chemotherapy is unnecessary. Patients with LyP are likely to have an favorable prognosis.

Tripchlorolide ameliorates experimental autoimmune encephalomyelitis by down-regulating ERK1/2-NF-kappaB and JAK/STAT signaling pathways.[Pubmed:25662403]

J Neurochem. 2015 Apr;133(1):104-12.

Tripchlorolide (T4), an extract of the natural herb Tripterygium wilfordii Hook F, has been found to possess anti-inflammatory and immunosuppressive actions. In the current study, these actions were evaluated in experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis by scoring the clinical signs, observing the infiltration of inflammatory cells and myelin sheath in the lumbar spinal cord of EAE mice. The results demonstrated that T4 (at a dose of 40 mug/kg) significantly reduced the severity of EAE and slowed down the ongoing EAE. Further analysis showed that T4 suppressed the mRNA and protein levels of the transcription factors T-bet and RoRrt and mRNA levels of IFN-gamma and IL-17 in the spinal cords. Furthermore, T4 down-regulated the ERK1/2-NF-kappaB and JAK/STAT signaling pathways. At 40 mug/kg, T4 did not induce side effects on hematological parameters. These findings suggest that T4 ameliorates EAE by immunosuppression, providing a new insight into T4 application in multiple sclerosis treatment.

Tripchlorolide improves cognitive deficits by reducing amyloid beta and upregulating synapse-related proteins in a transgenic model of Alzheimer's Disease.[Pubmed:25661995]

J Neurochem. 2015 Apr;133(1):38-52.

Alzheimer's disease (AD) is characterized by early impairments in memory and progressive neurodegeneration. Disruption of synaptic plasticity processes that underlie learning and memory contribute partly to this pathophysiology. Tripchlorolide (T4 ), an extract from a traditional Chinese herbal Tripterygium wilfordii Hook F, has been shown to be neuroprotective in animal models of Parkinson's disease and to improve cognitive deficits in senescence-accelerated mouse P8. In this study, we investigated the effect of T4 on cognitive decline and synaptic plasticity in five times familial AD (5XFAD) mice co-expressing mutated amyloid precursor protein and presenilin-1. Five-month-old 5XFAD mice and wild type littermates were intraperitoneally injected with T4 , 5 mug/kg or 25 mug/kg, every other day for 60 days. T4 treatment significantly improved spatial learning and memory, alleviated synaptic ultrastructure degradation, up-regulated expression of synapse-related proteins, including synaptophysin, post-synaptic density-95, N-methyl-D-aspartate receptor subunit 1, phosphorylation of calcium/calmodulin dependent protein kinase II alpha, and phosphorylation of cyclic AMP-response element binding protein, and promoted activation of the phophoinositide-3-kinase-Akt-mammalian target of rapamycin signaling pathway in 5XFAD mice. Accumulation of amyloid beta (Abeta) may contribute to synapse dysfunction and memory impairment in AD. We found that T4 treatment significantly reduced cerebral Abeta deposits and lowered Abeta levels in brain homogenates. These effects coincided with a reduction in cleavage of beta-carboxyl-terminal amyloid precursor protein (APP) fragment, levels of soluble APPbeta, and protein expression of beta-site APP cleaving enzyme 1. Taken together, our findings identify T4 as a potent negative regulator of brain Abeta levels and show that it significantly ameliorates synaptic degeneration and cognitive deficits in a mouse model of AD.

Tripchlorolide improves age-associated cognitive deficits by reversing hippocampal synaptic plasticity impairment and NMDA receptor dysfunction in SAMP8 mice.[Pubmed:24140565]

Behav Brain Res. 2014 Jan 1;258:8-18.

Deficits in cognition and performance accompanying age-related neurodegenerative diseases such as Alzheimer's disease (AD) are closely associated with the impairment of synaptic plasticity. Here, using a mouse model of senescence-accelerated P8 (SAMP8), we reported the role of Tripchlorolide (T4), an extract of the natural herb Tripterygium wilfordii Hook F, in improving cognitive deficits and promoting the long-term potentiation (LTP) of hippocampal slices via the N-methyl-D-aspartate receptor (NMDAR)-dependent signaling pathway. Our results demonstrated that chronic administration of T4 at low doses (0.25, 1.0, or 4.0 mug/kg per day, injected intraperitoneally for 75 days) significantly improved learning and memory function in aged SAMP8 mice, as indicated by a chain of behavioral tests including the Y-maze and Morris water maze. Additionally, T4 reversed the impaired LTP in hippocampal CA1 regions of SAMP8 mice in a dose-dependent manner. Moreover, it upregulated the levels of phospho-NMDAR1, postsynaptic density-95 (PSD-95), phospho-calcium-calmodulin dependent kinase II (CaMKII), phospho-CREB and brain derived neurotrophic factor (BDNF) in the hippocampus. This indicates that T4 prevents the impairment of NMDAR-mediated synaptic plasticity-related signal molecules. At optimal doses, T4 did not show significant side-effects on blood counts, blood biochemical measures, or survival of the mice. This novel mechanism in reversing age-related synaptic dysfunction and NMDAR functional deficits suggests that T4 can halt the manifestation of a key early-stage event in AD. With the consideration of SAMP8 mice as a model to develop therapeutic interventions for AD, our findings provide new insight into the clinical application of Tripchlorolide in AD treatment.

Tripchlorolide induces cell death in lung cancer cells by autophagy.[Pubmed:22139090]

Int J Oncol. 2012 Apr;40(4):1066-70.

It has been demonstrated that triptolide inhibits the growth of several types of cancer cells in vitro and prevents tumor growth in vivo by inducing apoptosis and autophagy. Here we showed that Tripchlorolide (T4) significantly suppressed the proliferation of A549 cells in a dose- and time-dependent manner. This suppressive effect was diminished when cells were pretreated with 3-Methylamphetamine (3-MA). After the cells were treated with T4, the LC3 II protein expression was significantly increased, and autophagosomes were observed by TEM. However, almost no apoptosis was observed in A549 treated with T4. These results suggest that T4 induces A549 cell death predominantly through the activation of the autophagy pathway instead of the apoptosis pathway.