Trichosanthes Kirilowii Research Articles

Efficacy and safety of SKCPT in patients with knee osteoarthritis: A multicenter, randomized, double-blinded, active-controlled phase III clinical trial

Ethnopharmacological relevanceOsteoarthritis (OA) is the most prevalent type of arthritis worldwide and a leading cause of years lost to pain and disability. Among the current pharmacological treatments for OA, symptomatic slow-acting drugs for OA (SYSADOA) induce pain relief and aim to improve joint function by relieving inflammation while causing fewer gastrointestinal and cardiovascular adverse events than non-steroidal anti-inflammatory drugs (NSAIDs). SKCPT is a herbal SYSADOA formulated from Clematis mandshurica, Trichosanthes kirilowii, and Prunella vulgaris powdered extracts. This preparation has been shown to induce cartilage protection and anti-inflammatory effects in preclinical studies and inhibit glycosaminoglycan degradation and catabolic gene expression in human OA chondrocytes and cartilage. Aim of the studyWe aimed to evaluate the non-inferiority of SKCPT to celecoxib and safety for treating knee OA. Materials and methodsThis multicenter, randomized, double-blind, phase III clinical trial enrolled adults with primary knee OA who were randomized (1:1) to SKCPT 300 mg twice daily or celecoxib 200 mg once daily for 12 weeks. ResultsIn total, 278 patients were assigned to treatment (SKCPT, 136; celecoxib, 142) for approximately 12 weeks. The primary endpoint was the mean change of Korean Western Ontario and McMaster Universities Osteoarthritis Index (K-WOMAC) pain subscale scores from baseline to Day 84. The mean change (least squares [LS] mean ± standard error) from baseline to Day 84 was −23.74 ± 1.48 for SKCPT and −25.88 ± 1.44 for celecoxib. The two-sided 95% confidence interval of the difference (LS mean) between groups was [−1.94, 6.20], confirming that the upper limit was less than the non-inferiority margin of 10. Additionally, there were no significant differences in the secondary endpoints (mean changes of K-WOMAC pain, physical, stiffness subscale, and total score, and the frequency and number of doses of rescue medications) between groups at all time points. Differences between groups in adverse events and adverse drug reactions were not significant, and no serious adverse events occurred. ConclusionsSKCPT efficacy was non-inferior, and its safety profile was similar, to celecoxib. Building on previous results showing that SYSADOA reduce NSAID intake, the present results suggest that the SYSADOA SKCPT could effectively replace NSAIDs in knee OA treatment while avoiding long-term side effects.

The multi-herbal decoction SH003 alleviates LPS-induced acute lung injury by targeting inflammasome and extracellular traps in neutrophils

BackgroundAcute lung injury (ALI) is a devastating condition caused by sepsis, pneumonia, trauma, and more recently, COVID-19. SH003, an herbal formula consisted of Astragalus membranaceus, Angelica gigas and Trichosanthes kirilowii, is known for its effects on cancer and immunoregulation. Hypothesis/PurposePrevious studies show SH003 exerts a promising anti-inflammatory effect. This study investigates the effect of modified SH003 on ALI using in silico, in vivo, and in vitro models. Study Design and MethodsWe performed in silico-based analysis of SH003 on ALI-related pathways. C57BL/6 mice were intraperitoneally subjected to lipopolysaccharide (LPS) to induce septic ALI, followed by oral administration of SH003 for 2 weeks. Dexamethasone was used as the positive control. Human peripheral blood-derived polymorphonuclear neutrophils (PMN) were used to investigate the effect and mechanisms of SH003 on neutrophil extracellular trap (NET) formation. ResultsNetwork pharmacology analysis suggested SH003 regulates lung inflammation by modulating NET formation. SH003 significantly reduced mortality in sepsis in vivo by inhibiting local and systemic inflammation, likely via nuclear factor kappa B and mitogen-activated protein kinase pathways-mediated inflammasome suppression. SH003 also decreased NET-related markers in lung tissues and inhibited LPS- and phorbol myristate acetate-induced NET formation in PMN. Cytometry time-of-flight analysis confirmed regulation of NETosis-related pathways by SH003. ConclusionSH003 effectively inhibits excessive immune responses in the lung by suppressing inflammasome activation and NET formation. These findings suggest SH003 as a potential therapeutic agent for septic ALI.

Analgesic Effect of SKI306X on Chronic Postischemic Pain and Spinal Nerve Ligation-Induced Neuropathic Pain in Mice.

Neuropathic pain (NP) results from lesions or diseases affecting the peripheral or central somatosensory system. However, there are currently no drugs that are particularly effective in treating this condition. SKI306X is a blend of purified extracts of three oriental herbs (Clematis mandshurica, Trichosanthes kirilowii, and Prunella vulgaris) commonly used to treat osteoarthritis for their chondroprotective effects. Chronic postischemic pain (CPIP) and spinal nerve ligation (SNL) models were created by binding the upper left ankle of mice with an O-ring for 3 h and ligating the L5 spinal nerve, respectively. Mice with allodynia were injected intraperitoneally with 0.9% normal saline (NS group) or different doses (25, 50, or 100 mg/kg) of SKI306X (SKI groups). We assessed allodynia using von Frey filaments before injection and 30, 60, 90, 120, 180, and 240 min and 24 h after injection to confirm the antiallodynic effect of SKI306X. We also measured glial fibrillary acidic protein (GFAP) levels in the spinal cord and dorsal root ganglia to confirm the change of SKI306X administration. Both models exhibited significant mechanical allodynia. The intraperitoneal injection of SKI306X significantly increased the paw withdrawal threshold in a dose-dependent manner, as the paw withdrawal threshold was significantly increased after SKI306X administration compared with at baseline or after NS administration. GFAP levels in the SKI group decreased significantly (p < 0.05). Intraperitoneal administration of SKI306X dose-dependently attenuated mechanical allodynia and decreased GFAP levels, suggesting that GFAP is involved in the antiallodynic effect of SKI306X in mice with CPIP and SNL-induced NP.

First Report of Root Rot Caused by Fusarium incarnatum on Mongolian Snake gourd in China.

First Report of Root Rot Caused by Fusarium incarnatum on Mongolian Snake gourd in China.

Effect of GHX02 on an Asthma-Rhinitis Mouse Model Induced by Ovalbumin and Diesel Particulate Matter.

Fine dust concentrations come in direct contact with the human respiratory system, thereby reducing lung function and causing respiratory diseases such as asthma and rhinitis. The aim of this study was to evaluate the efficacy of GHX02 (combination of four herbs [Trichosanthes kirilowii, Prunus armeniaca, Coptis japonica, and Scutellaria baicalensis]), a herbal extract with established efficacy against bronchitis and pulmonary disease, in the treatment of asthma accompanied by rhinitis aggravated by fine dust. Therefore, we constructed an asthma-rhinitis mouse model of Balb/c mice challenged with ovalbumin (OVA) and fine diesel particulate matter, which were administered with three concentrations of GHX02. GHX02 significantly inhibited the increase of total cells and immune cells in bronchoalveolar lavage fluid, lung tissue, and nasal ductal lymphoid tissue (NALT). GHX02 also reduced the severity of histological lung injury and the expression of interleukin (IL)-1α and nuclear factor kappa B (NF-κB), which regulate inflammatory responses. The results indicate that GHX02 inhibited the inflammatory immune response in mice. Therefore, this study highlights the potential of GHX02 as a treatment for patients with asthma accompanied by rhinitis. Balb/c mice were challenged with OVA and PM10D, and then treated with three concentration of GHX02. GHX02 significantly inhibited the increase of total cells, immune cells lymphocytes, neutrophils, and macrophages, as well as their expression in lung tissue. GHX02 significantly inhibited the increase of total cells and immune cells in NALT. GHX02 decreased the severity of histological lung injury, expression of IL-1α and NF-κB. This study suggests the probability that GHX02 is effective for asthma patients with rhinitis by inhibiting inflammatory immune response.

First report of root rot in Trichosanthes kirilowii (Chinese cucumber) caused by Fusarium sulawesiense in China.

Trichosanthes kirilowii (Chinese cucumber) is one of the important perennial herbaceous vines in China, with putative pharmacological activities including anti-tumor and lowering blood lipids. In July 2022, T. kirilowii plants with brownish roots and chlorotic leaves were observed in several orchards in Qianshan, Anhui province, China (30°34'N, 116° 30'E). The disease incidence reached approximately 10% within an area spanning 20 ha, and was higher in poorly drained orchards. To investigate this root rot disease, five symptomatic plants were collected from the diseased orchards in Qianshan. Subsequently, small sections of the diseased roots were surface sterilized using 1% sodium hypochlorite and 75% ethanol for 45 seconds each. Then, sterilized roots were placed onto PDA (20% diced potato, 2% glucose, and 1.5% agar, and distilled water) and incubated at 28℃ in the dark for 6 days. A total of eight isolates with similar morphology were obtained and purified by single spore culturing. Two representative isolates (QSJ4 and QSJ5) were chosen for further analysis. When grown on PDA, the surface of each colony was white with dense aerial mycelium and pale orange color in the center with a white edge on the reverse side. Macroconidia produced on carnation leaf agar plates were falcate, slightly curved, and 3 to 5 septate, with papillate apical cells and indistinct basal cells. Macroconidia were 17.4-42.3 × 2.4-5.8 μm (n = 100). Microconidia were ellipsoidal in shape, slightly curved or not curved, and most were 1-septate, 9.6-16.7 × 1.5-3.8 μm (n = 40). The identity was determined by sequencing four loci (i. e., ITS, CAL, EF1-α and RPB2) from two representative isolates (Liu et al. 1999; O'Donnell et al. 1998, 2000; Reeb et al. 2004; White et al. 1990). Sequences were deposited in GenBank [ITS (OR267397, OR267398), CAL (OR296634, OR296635), EF1-α (OR296637, OR296638) and RPB2 (OR296640, OR296641)]. A phylogenetic analysis was performed with three loci (CAL, EF1-α, RPB2) comprising a concatenated dataset of 68 strains in the Fusarium incarnatum-equiseti species complex (Han et al., 2023). The results showed that isolates QSJ4 and QSJ5 clustered closely together with reference strains of F. sulawesiense. Pathogenicity tests were conducted by inoculating three-week-old healthy T. kirilowii seedings (cv. Wanlou No. 9) cultivated in substrate soil in pots with a diameter of 17 cm and a height of 10.5 cm. A 20 mL aliquot of spore suspension (106 conidia/mL) of F. sulawesiense was inoculated to the roots of potted seedlings by irrigation. Each strain was inoculated onto three seedlings. The potted seedlings were inoculated with sterile water as the negative control. Inoculated seedlings were incubated in a growth chamber at 25℃ and 75% relative humidity. After one week, typical symptoms of root necrosis and leaf chlorosis were observed on the inoculated seedlings. Disease symptoms were not observed on the control seedlings. All seedlings showing root necrosis and leaf chlorosis caused by the inoculations were subjected to fungal isolation, and the results showed that the reisolated colonies matched the inoculated ones for morphologies and ITS sequences. Fusarium sulawesiense has been previously reported to cause disease on Cucumis melo L. in Brazil (Medeiros Araujo et al. 2021), Musa acuminata Colla in south Sulawesi (Maryani et al. 2019), Luffa aegyptiaca Miller and Musa nana Lour. in China (Wang et al. 2019). To our knowledge, this is the first report of F. sulawesiense causing Fusarium root rot of T. kirilowii in China.

A comparative evaluation of the structure, functionality and volatile profiles of Trichosanthes kirilowii seed protein isolates based on different extraction methods

In the present study, we hypothesised that Trichosanthes kirilowii seed protein isolate (TPI) obtained by different extraction methods have distinct structure, functional attributes and volatile profiles. Alkaline-extracted isolate (AE-TPI) exhibited lower protein content and a darker colour than the other two isolates because more polyphenols and pigments were coextracted. Salt-extracted isolate (SE-TPI) and AE-TPI had higher in vitro protein digestibility than reverse micelle-extracted isolate (RME-TPI) due to higher degrees of denaturation, which enabled them to be more susceptible to proteolysis. The SE-TPI gel resulted in a stronger gel network and greater hardness than the other two isolate gels. In the volatile profile, SE-TPI (22) yielded the largest number of volatile compounds, followed by AE-TPI (20) and RME-TPI (15). The current results indicated that the structure, functional properties and volatile profiles of TPI are largely influenced by the extraction technique.

Integrated serum pharmacochemistry and investigation of the anti-influenza A virus pneumonia effect of Qingjin Huatan decoction

Ethnopharmacological relevanceQingjin Huatan Decoction (QJHTT) consists of 11 herbal medicines: Scutellaria baicalensis Georgi, Gardenia jasminoides J. Ellis, Platycodon grandiflorus (Jacq.) A. DC., Ophiopogon japonicus (Thunb.) Ker Gawl., Morus alba L., Fritillaria thunbergii Miq., Anemarrhena asphodeloides Bunge, Trichosanthes kirilowii Maxim., Citrus reticulata Blanco, Poria cocos (Schw.) Wolf, and Glycyrrhiza uralensis Fisch. As a traditional Chinese medicinal formula, QJHTT has been used for more than 400 years in China. It has shown promising results in treating influenza A virus (IAV) pneumonia. Aim of the studyTo elusive the specific pharmacological constituents and mechanisms underlying its anti-IAV pneumonia effects. Materials and methodsThe components in QJHTT were analyzed through the use of a serum pharmacology-based ultra high-performance liquid chromatography Q- Exactive Orbitrap mass spectrometry (UHPLC-Q Exactive Orbitrap-MS) method. Simultaneously, the dynamic changes in IAV-infected mouse lung viral load, lung index, and expression of lung inflammation factors were monitored by qRT-PCR. ResultsWe successfully identified 152 chemical components within QJHTT, along with 59 absorbed chemical prototype constituents found in the serum of mice treated with QJHTT. 43.45% of these chemical components and 43.10% of the prototype constituents were derived from the monarch drugs, namely Huangqin and Zhizi, aligning perfectly with traditional Chinese medicine theory. Notably, our analysis led to the discovery of 14 compounds within QJHTT for the first time, three of which were absorbed into the bloodstream. Simultaneously, we observed that QJHTT not only reduced the viral load but also modulated the expression of inflammation factors in the lung tissue including TNF-α, IL-1β, IL-4, IL-6, IFN-γ, and IL17A. A time-effect analysis further revealed that QJHTT intervention effectively suppressed the peak of inflammatory responses, demonstrating a robust anti-IAV pneumonia effect. ConclusionsWe comprehensively analyzed the pharmacological material basis of QJHTT by a highly sensitive and high-resolution UHPLC-Q Exactive Orbitrap-MS method, and demonstrated its efficacy in combating IAV pneumonia by reducing lung viral load and inflammatory factors. This study has significant importance for elucidating the pharmacological basis and pharmacological mechanism of QJHTT in combating IAV pneumonia.

Assembly and evolutionary analysis of the complete mitochondrial genome of Trichosanthes kirilowii, a traditional Chinese medicinal plant.

Trichosanthes kirilowii (T. kirilowii) is a valuable plant used for both medicinal and edible purposes. It belongs to the Cucurbitaceae family. However, its phylogenetic position and relatives have been difficult to accurately determine due to the lack of mitochondrial genomic information. This limitation has been an obstacle to the potential applications of T. kirilowii in various fields. To address this issue, Illumina and Nanopore HiFi sequencing were used to assemble the mitogenome of T. kirilowii into two circular molecules with sizes of 245,700 bp and 107,049 bp, forming a unique multi-branched structure. The mitogenome contains 61 genes, including 38 protein-coding genes (PCGs), 20 tRNAs, and three rRNAs. Within the 38 PCGs of the T. kirilowii mitochondrial genome, 518 potential RNA editing sites were identified. The study also revealed the presence of 15 homologous fragments that span both the chloroplast and mitochondrial genomes. The phylogenetic analysis strongly supports that T. kirilowii belongs to the Cucurbitaceae family and is closely related to Luffa. Collinearity analysis of five Cucurbitaceae mitogenomes shows a high degree of structural variability. Interestingly, four genes, namely atp1, ccmFC, ccmFN, and matR, played significant roles in the evolution of T. kirilowii through selection pressure analysis. The comparative analysis of the T. kirilowii mitogenome not only sheds light on its functional and structural features but also provides essential information for genetic studies of the genus of Cucurbitaceae.

Elucidating the pharmacodynamic mechanisms of Yuquan pill in T2DM rats through comprehensive multi-omics analyses.

Background: Yuquan Pill (YQW) is a modern concentrated pill preparation of six herbs, namely, Ge Gen (Pueraria lobata Ohwi), Di huang (Rehmannia glutinosa Libosch.), Tian Huafen (Trichosanthes kirilowii Maxim.), Mai Dong (Ophiopogon japonicus (L. f.) Ker Gawl.), Wu Weizi (Schisandra chinensis (Turcz.) Baill.) and Gan Cao (Glycyrrhiza uralensis Fisch.). It is extensively used to treat type 2 diabetes-related glucose and lipid metabolism disorders. But what's the pharmacodynamic substance and how it works in the treatment of Type 2 diabetes mellitus (T2DM) are still unclear. Purpose: The purpose of this study is to determine the likely pharmacological components and molecular mechanism of YQW's intervention on T2DM by combining serum pharmacochemistry, network analysis and transcriptomics. Methods: The efficacy and prototypical components of blood entry were determined after oral administration of YQW aqueous solution to T2DM rats induced by high-fat feed and low-dose streptozotocin (STZ), and the key targets and pathways for these compounds to intervene in T2DM rats were predicted and integrated using network analysis and transcriptomics techniques. Results: In diabetic rats, YQW can lower TG, CHO, NO, and MDA levels (p < 0.05) while increasing HDL-C levels (p < 0.01), and protecting the liver and kidney. 22 prototype components (including puerarin, daidzein, 3'-methoxypuerarin, and liquiritigenin, among others) were found in the serum of rats after oral administration of YQW for 90min, which might be used as a possible important ingredient for YQW to intervene in T2DM rats. 538 YQW pharmacodynamic components-related targets and 1,667 disease-related targets were projected through the PharmMapper database, with 217 common targets between the two, all of which were engaged in regulating PI3K-Akt, MAPK, Ras and FoxO signal pathway. Finally, the mRNA expression profiles of liver tissues from rats in the control, model, and YQW groups were investigated using high-throughput mRNA sequencing technology. YQW can regulate the abnormal expression of 89 differential genes in a disease state, including 28 genes with abnormally high expression and 61 genes with abnormally low expression. Five common genes (Kit, Ppard, Ppara, Fabp4, and Tymp) and two extensively used regulatory pathways (PI3K-Akt and MAPK signaling pathways) were revealed by the integrated transcriptomics and network analysis study. Conclusion: The mechanism of YQW's intervention in T2DM rats could be linked to 22 important components like puerarin, daidzein, and glycyrrhetinic acid further activating PI3K-Akt and MAPK signaling pathways by regulating key targets Kit, Ppard, Ppara, Fabp4, and Tymp, and thus improving lipid metabolism disorder, oxidative stress, and inflammation levels in T2DM rats. On the topic, more research into the pharmacological ingredient foundation and mechanism of YQW intervention in T2DM rats can be done.

Root Extract of Trichosanthes kirilowii Suppresses Metastatic Activity of EGFR TKI-Resistant Human Lung Cancer Cells by Inhibiting Src-Mediated EMT

The roots of Trichosanthes kirilowii (TK) have been used in traditional oriental medicine for the treatment of respiratory diseases. In this study, we investigated whether an ethanolic root extract of TK (ETK) can regulate the metastatic potency of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI)-resistant human lung cancer cells. The relative migration and invasion abilities of erlotinib-resistant PC9 (PC9/ER) and gefitinib-resistant PC9 (PC9/GR) cells were higher than those of parental PC9 cells. Mesenchymal markers were overexpressed, whereas epithelial markers were downregulated in resistant cells, suggesting that resistant cells acquired the EMT phenotype. ETK reduced migration and invasion of resistant cells. The expression levels of N-cadherin and Twist were downregulated, whereas Claudin-1 was upregulated by ETK, demonstrating that ETK suppresses EMT. As a molecular mechanism, Src was dephosphorylated by ETK. The anti-metastatic effect of ETK was reduced by transfecting PC9/ER cells with a constitutively active form of c-Src. Dasatinib downregulated N-cadherin, Twist, and vimentin, suggesting that Src regulates EMT in resistant cells. Notably, CuB played a key role in mediating the anti-metastatic activity of ETK. Collectively, our results demonstrate that ETK can attenuate the metastatic ability of EGFR-TKI-resistant lung cancer cells by inhibiting Src-mediated EMT.

Molecular identification and development of an infectious cDNA clone of Trichosanthes kirilowii-infecting cucurbit mild mosaic virus

Trichosanthes kirilowii has been mainly grown for use in traditional Chinese medicine. In this study, cucurbit mild mosaic virus (CuMMV) belonging to the genus Fabavirus was identified from T. kirilowii plants. CuMMV possesses a segmented, bipartite linear single-stranded RNA genome composed of RNA1 and RNA2. Sequence analysis showed that each genomic segment shares the highest sequence similarity with those of CuMMV isolated from pumpkin. A full-length infectious cDNA clone of CuMMV was further constructed and was found to induce typical symptoms in T. kirilowii, Cucumis sativus, C. melo, Citrullus lanatus, and Cucurbita pepo. The sap inoculum derived from the infectious cDNA clone of CuMMV could be mechanically transmitted and reproduce similar symptoms in the tested plants. This is the first report on the construction of a biologically active, full-length infectious cDNA clone of CuMMV, which will provide a useful tool in understanding CuMMV-encoded proteins and plant-CuMMV interactions.

First Report of Tomato mottle mosaic virus infecting Chinese snake gourd (Trichosanthes kirilowii) in China.

Tomato mottled mosaic virus (ToMMV) was first identified in tomato in Mexico (Li et al. 2013). It belongs to the genus Tobamovirus and family Virgaviridae, and is a positive-sense single-stranded RNA virus. The viral genome contains about 6400 nucleotides, encoding four proteins, including the 126 K protein, 183 K protein, movement protein (MP) and coat protein (CP) (Tu et al. 2021). ToMMV mainly poses a serious risk to solanaceous crops. The virus-infected plants appear stunted growth and top necrosis, and the disease leaves show mottled, shrinkage and necrosis symptoms, resulting in a significant decline in tomato fruit yield and quality (Li et al. 2017; Tu et al. 2021). Chinese snake gourd (Trichosanthes kirilowii Maxim) is a perennial climbing herb in the family Cucurbitaceae, and the fruit, seed, peel and root can all be used as traditional Chinese medicine. In May of 2021, twenty-seven symptomless seedlings (developed from tissue culture plantlets) were randomly collected from nursery in Fengyang, Anhui Province. Total RNA of each sample was extracted, and RT-PCR was performed using degenerate tobamovirus primers Tob-Uni1 (5'-ATTTAAGTGGASGGAAAAVCACT-3') and Tob-Uni2 (5'-GTYGTT GATGAGTTCRTGGA-3') (Letschert et al. 2002). Amplicons with expected size were obtained from 6 of 27 samples and sequenced. Alignment results showed that the nucleotide sequence identities ranged from 98.7 to 100% with all ToMMV isolates deposited in NCBI GenBank. Then, ToMMV coat protein (CP) gene was amplified using specific primers CP-F (5'-ATGTCTTACGCTATTACTT CTCCG-3') and CP-R (5'-TTAGGACGCTGGCGCAGAAG-3'). The CP fragment was obtained and sequenced. Sequence alignment indicated that CP sequence of isolate FY (GenBank accession no. ON924176) exhibited a 100% identity with ToMMV isolate LN (MN853592.1). The anti-ToMMV polyclonal antibody (PAb) was prepared by the author (S.L.) by immunizing rabbit with purified virus from Nicotiana benthamiana, and serological tests (dot-enzyme linked immunosorbent assay, Dot-ELISA) of RNA-positive T. kirilowii leaf samples using anti-ToMMV PAb were also positive. To fulfill a Koch's postulate, a pure culture of ToMMV was obtained from N. benthamiana using infectious cDNA clone of ToMMV (Tu et al. 2021), and then healthy T. kirilowii plants were mechanically inoculated with a prepared inoculum from ToMMV-infected N. benthamiana, as described previously (Sui et al. 2017). T. kirilowii seedlings showed chlorosis and leaf tip necrosis symptoms at 10 and 20 day post-inoculation respectively, and ToMMV infection on symptomatic plants was also verified by RT-PCR detection using primers CP-F and CP-R. These results demonstrated that T. kirilowii is a host of ToMMV under natural conditions, which might threaten the production of this medicinal plant. The seedlings from nursery appeared to be asymptomatic, but the plants showed chlorosis and necrosis symptoms after indoor inoculation. In qRT-PCR analysis, viral accumulation level in greenhouse-inoculated plants was a 25.6-fold of that in field-collected samples, which may be the reason of different symptom expression between field samples and inoculated samples. ToMMV has now been detected from the solanaceous (tomato, pepper and eggplant) and leguminous (pea) crops in the field (Li et al. 2014; Ambrós et al. 2017; Zhang et al. 2022). To our knowledge, this is the first report of natural infection of ToMMV in T. kirilowii as well as its natural infection on Cucurbitaceae plants.

Transcript-wide identification and expression pattern analysis to comprehend the roles of AP2/ERF genes under development and abiotic stress in Trichosanthes kirilowii

BackgroundThe APETALA 2/ ethylene-responsive element binding factors (AP2/ERF), are thought to be associated with plant abiotic stress response, and involved in some plant hormone signaling pathways. Trichosanthes kirilowii is an important edible and medicinal crop, so far no research has been conducted on the TkAP2/ERF genes.ResultIn this study, a total of 135 TkERFs were identified, these genes were divided into 4 subfamilies and clustered into 13 groups. Moreover, 37 paralogous pairs were identified, with only two having Ka/Ks values greater than 1, proving that most TkERF genes underwent purifying selection during evolution. Co-expression networks constructed using transcriptome data at various flowering stages revealed that 50, 64, and 67 AP2/ERF genes correlated with members of the ethylene, gibberellin, and abscisic acid signaling pathways, respectively. When tissue cultured seedlings were treated with ETH, GA3 and ABA, 11, 12 and 17 genes were found to be up-regulated, respectively, suggesting that some members of the TkERF gene family may be involved in plant hormone signaling pathways. And under 4 ℃, PEG and NaCl treatment, 15, 20 and 19 genes were up-regulated, respectively, this suggested that these selected genes might be involved in plant abiotic stresses.ConclusionsOverall, we identified 135 AP2/ERF family members, a comprehensive analysis of AP2/ERF gene expression patterns by RNA-seq and qRT-PCR showed that they played important roles in flower development and abiotic stress. This study provided a theoretical basis for the functional study of TkAP2/ERF genes and the genetic improvement of T. kirilowii.

Qingjin Huatan decoction protects mice against influenza a virus pneumonia via the chemokine signaling pathways

Ethnopharmacological relevanceQingjin Huatan Decoction (QJHTT) consists of 11 herbal medicines: Scutellaria baicalensis Georgi, Gardenia jasminoides J.Ellis, Platycodon grandiflorus (Jacq.) A.DC., Ophiopogon japonicus (Thunb.) Ker Gawl., Morus alba L., Fritillaria thunbergii Miq., Anemarrhena asphodeloides Bunge, Trichosanthes kirilowii Maxim., Citrus reticulata Blanco, Poria cocos (Schw.) Wolf, and Glycyrrhiza uralensis Fisch. As a traditional compound Chinese medicinal formula, QJHTT has been used for more than 400 years in China. Historically, it was used to treat respiratory diseases and had shown beneficial clinical results for diseases related to lung inflammation. Aim of the studyTo investigate the therapeutic effect of QJHTT on influenza A virus (IAV) pneumonia in mice and explore its possible mechanism of action. Materials and methodsThe components in QJHTT were analyzed by UPLC-Q-TOF-MS and some antiviral active components reported in the literature were determined and quantified by HPLC. The protective effects of QJHTT were investigated using lethal and sublethal doses (2 LD50 or 0.8 LD50 viral suspension, separately) of H1N1-infected mice. Mortality and lung lesions in H1N1-infected mice were used to evaluate the efficacy of QJHTT. The potential mechanism of QJHTT in the treatment of viral pneumonia was determined at the gene level by RNA sequencing and validated by qRT-PCR. Following this, the changes in protein levels of JAK2/STAT3 were analyzed since it is a key downstream target of the chemokine signaling pathways. Preliminary elucidation of the mechanism of QJHTT to protect mice against IAV pneumonia through this pathway was conducted. ResultsIn this study, 12 antiviral active constituents including baicalin, geniposide, and mangiferin were identified from QJHTT. In vivo treatment of QJHTT reduced the virus titers of lung tissue significantly and improved the survival rate, lung index, and pulmonary histopathological changes; additionally, a reduction in the serum levels of TNF-α, IL-1β, IL-6, and IFN-γ inflammatory factors in H1N1-infected mice was observed. RNA-seq analysis and qRT-PCR showed that QJHTT primarily reversed the activities CCL2, CCL7, CCR1, and other chemokines and their reception-related genes, suggesting that QJHTT may produce disease-resistant pneumonia by inhibiting the downstream JAK2/STAT3 pathway. Western blot analysis confirmed that QJHTT effectively reduced the protein levels of JAK2, STAT3, and related phosphorylated products in the lung tissue of H1N1-infected mice. ConclusionsOur results indicated that QJHTT alleviated IAV pneumonia in mice by regulating related chemokines and their receptor-related genes in lung tissue, thereby inhibiting JAK2/STAT3 pathway. This could pave way for the design of novel therapeutic strategies to treat viral pneumonia.

First report of the root-knot nematode Meloidogyne incognita on Salvia miltiorrhiza Bunge in Henan Province, China.

First report of the root-knot nematode Meloidogyne incognita on Salvia miltiorrhiza Bunge in Henan Province, China.

Identification and analysis of miRNAs differentially expressed in male and female Trichosanthes kirilowii maxim

Trichosanthes kirilowii Maxim. (TK) is a dioecious plant in the Cucurbitaceae family of which different sexes have separate medicinal uses. We used Illumina high-throughput sequencing technology to sequence miRNAs from male and female flower buds of TK. We performed bioinformatics analysis, miRNA identification, and target gene prediction on the data obtained from sequencing, and association analysis was performed in combination with the results of a previous transcriptome sequencing study. As a result, there were 80 differentially expressed miRNAs (DESs) between the female and male plants (48 upregulated and 32 downregulated in female plants). Moreover, 27 novel miRNAs in DESs were predicted to have 282 target genes, and 51 known miRNAs were predicted to have 3418 target genes. By establishing a regulatory network between miRNAs and target genes, 12 core genes were screened, including 7 miRNAs and 5 target genes. Among them, tkmiR157a-5p, tkmiR156c, tkmiR156_2, and tkmiR156k_2 jointly target the regulation of tkSPL18 and tkSPL13B. These two target genes are specifically expressed in male and female plants, respectively, and are involved in the biosynthesis process of BR, which is closely related to the sex differentiation process of TK. The identification of these miRNAs will provide a reference for the analysis of the sex differentiation mechanism of TK.

The Protective Effect of a Functional Food Consisting of Astragalus membranaceus, Trichosanthes kirilowii, and Angelica gigas or Its Active Component Formononetin against Inflammatory Skin Disorders through Suppression of TSLP via MDM2/HIF1α Signaling Pathways.

An herbal mixture (SH003) of Astragalus membranaceus, Trichosanthes kirilowii, and Angelica gigas exhibits therapeutic effects on carcinomas and immunosuppression. However, the role of JRP-SNF102, which is an advanced mixture of SH003, in regulating inflammatory responses is unexplored. We aim to substantiate the therapeutic potential of JRP-SNF102 and its active component, formononetin (FMN), as a functional food that moderates inflammatory responses. The inhibitory effects of JRP-SNF102 or FMN on thymic stromal lymphopoietin (TSLP) levels were evaluated in phorbol 12-myristate 13-acetate (PMA) plus A23187-activated human mast cell line-1 (HMC-1) cells and a mouse model of PMA-induced ear edema. The JRP-SNF102 or FMN inhibited the secretion and mRNA expression of TSLP and vascular endothelial growth factor (VEGF) in the activated HMC-1 cells. The expression levels of murine double minute 2 (MDM2), hypoxia-inducible factor 1α (HIF1α), and NF-κB were also suppressed by JRP-SNF102 or FMN in the activated HMC-1 cells. The JRP-SNF102 or FMN inhibited TSLP and VEGF levels, attenuating redness and ear thickness in mice with acute ear edema; JRP-SNF102 or FMN reduced the expression levels of MDM2, HIF1α, and NF-κB in the ear tissues. These findings suggest the potential for JRP-SNF102 as a functional food in the treatment of inflammatory skin disorders through suppression of TSLP and VEGF.

硼添加对铝胁迫栝楼生长和生理的影响

PDF HTML阅读 XML下载 导出引用 引用提醒 硼添加对铝胁迫栝楼生长和生理的影响 DOI: 10.5846/stxb202202220419 作者: 作者单位: 作者简介: 通讯作者: 中图分类号: 基金项目: 国家自然科学基金（32001224，41702181）；2021年国家级大学生创新创业训练计划（202110345017） Effects of exogenous boron on growth and physiology of Trichosanthes kirilowii Maxim. under aluminum stress Author: Affiliation: Fund Project: The National Natural Science Foundation of China (General Program, Key Program, Major Research Plan)，the 2021 National Undergraduate Training Program for Innovation and Entrepreneurship 摘要 | 图/表 | 访问统计 | 参考文献 | 相似文献 | 引证文献 | 资源附件 | 文章评论 摘要:为揭示硼对缓解栝楼（Trichosanthes kirilowii Maxim.）铝（Aluminum）毒害的生理机制，以耐铝性强的安国栝楼和耐铝性弱的浦江栝楼为材料，研究了50 μmol/L硼酸（H3BO3）对300 μmol/L Al3+胁迫下栝楼幼苗生长、铝积累、抗氧化能力和细胞壁组分的影响。结果表明：铝胁迫下，植株的根长、株高、鲜重和干重降低，超氧化物歧化酶（SOD）、过氧化物酶（POD）、过氧化氢酶（CAT）活性受到显著抑制，安国栝楼、浦江栝楼根尖丙二醛（MDA）含量分别增加256.1%、278.2%，细胞壁多糖含量、果胶甲酯酶（PME）活性、根尖铝积累量显著提高，果胶甲基酯化度（DM）和3-脱氧-D-甘露-辛酮糖酸（KDO）含量降低。外源硼可以缓解铝胁迫对栝楼幼苗生长的抑制作用，安国与浦江栝楼的抗氧化酶活性显著提升，MDA含量、细胞壁多糖含量、PME酶活性均降低，安国栝楼果胶DM值恢复至正常水平的91.5%，浦江栝楼KDO含量较单铝处理组上升52.0%，活性铝结合位点减少，有效降低了根尖铝含量并维持其形态结构。因此，硼信号能通过调节细胞壁多糖组分且稳定其网格结构，减少根尖对铝的吸收，同时增强抗氧化酶活性，缓解铝胁迫导致栝楼幼苗生长中的次生氧化胁迫，提高栝楼幼苗对铝毒环境的耐受性，且对耐铝性强的安国栝楼作用效果较好。研究结果可为改良酸铝胁迫下作物的生长环境提供理论指导。 Abstract:Aluminum toxicity, a major abiotic stress that inhibits plant growth and development, has become one of the main factors reducing agricultural yield. To reveal the physiological mechanism of boron on alleviating aluminum toxicity of Trichosanthes kirilowii Maxim., seedlings from Anguo (with strong Al-tolerance) and Pujiang (with weak Al-tolerance) varieties of T. kirilowii were selected for water culture experiments and the effects of boron treatment (50 μmol/L) on varieties' growth, aluminum accumulation, antioxidant capacity, and cell wall components under aluminum stress (300 μmol/L) were studied. The results showed that 300 μmol/L Al3+ stress inhibited the root length, plant height, fresh weight, and dry weight of the plant. The activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were significantly decreased, and the content of malondialdehyde (MDA) in root tip of T. kirilowii in Anguo and Pujiang varieties increased by 256.1% and 278.2%, respectively. Aluminum stress also significantly enhanced the content of cell wall polysaccharide, pectin methylesterase (PME) activity, and root tip aluminum accumulation. However, the degree of pectin methyl-esterification (DM) and the content of 3-deoxy-d-mannose-octanoic acid (KDO) were strongly inhibited. Exogenous boron could alleviate the inhibitory effect of aluminum stress on the growth of T. kirilowii seedlings. The antioxidant enzyme activities of T. kirilowii in Anguo and Pujiang varieties were increased significantly. The MDA content, cell wall polysaccharide content, and PME enzyme activity were decreased. In addition, the pectin DM value of Trichosanthes in Anguo variety returned to 91.5% of the normal level, and the KDO content of T. kirilowii in Pujiang variety increased by 52.0% compared with that in the single aluminum treatment group. As a result, the active aluminum binding sites decreased, effectively reducing the aluminum content in the root tip and maintaining its morphological structure. Therefore, boron improved the tolerance of T. kirilowii seedlings to aluminum toxic environment by regulating the polysaccharide components of cell wall and stabilizing its grid structure to reduce the absorption of aluminum by root tips, and enhancing the activity of antioxidant enzymes to alleviate the secondary oxidative stress in the growth of T. kirilowii seedlings, which has a better effect on Anguo variety with stronger tolerance to aluminum toxicity. The research in this paper can provide theoretical guidance for improving the environment for crop growth under acid-aluminum stress. 参考文献 相似文献 引证文献

Influence of different extraction methods on the chemical composition, antioxidant activity, and overall quality attributes of oils from Trichosanthes kirilowii Maxim seed

Trichosanthes kirilowii Maxim seed oil is full of nutrition and exerts positive effects on the health of humans. The study detected and compared the quality characteristics, the lipid composition and active constituents of Trichosanthes kirilowii Maxim seed kernel and shell oil. This study used four extraction techniques: cold pressing, ultrasonic-assisted solvent extraction, supercritical fluid extraction, and subcritical butane extraction. The results indicated that the highest lipid yield (36.75%) was obtained by ultrasonic-assisted solvent extraction, while the lowest (32.24%) was obtained by supercritical fluid extraction. Meanwhile, Trichosanthes kirilowii Maxim seed oil is rich in linoleic acid (25.64–31.24%) and α-tocopherol (247.76–702.28 mg/kg). The extraction method and shells significantly affected beneficial phytochemicals (tocopherols, phytosterols and total phenols) and antioxidant activity (DPPH) (p < 0.05). Moreover, the tocopherols of supercritical fluid extraction oil (1020.15–1052.55 mg/kg) and the phytosterols of cold pressing oil (1033.84–1748.20 mg/kg) were higher than that of other methods under the same conditions. The content of sterols and tocopherol in the seed kernel oil is higher than that of the seed shell oil. Meanwhile, cold pressing oil had more abundant volatile compounds. The above information is valuable for manufacturers to select the appropriate method to produce Trichosanthes kirilowii Maxim seed oil with the desired quality attributes and chemical compositions for the target end-user.