Pinellia genus: A systematic review of active ingredients, pharmacological effects and action mechanism, toxicological evaluation, and multi-omics application.

The dried tuber of Pinellia ternata (Thunb.) Breit, Pinelliae Rhizoma (PR, also named 'Banxia' in Chinese), is widely used in traditional medicine. This review aims to provide detail summary of active ingredients, pharmacological effects, toxic ingredients, detoxification strategies, and omic researches, etc. Pharmacological ingredients from PR are mainly classified into six categories: alkaloids, amino acids, polysaccharides, phenylpropanoids, essential oils, and glucocerebrosides. Diversity of chemical composition determines the broad-spectrum efficacy and gives a foundation for the comprehensive utilization of P. ternata germplasm resources. The pharmacological compounds are involved in inhibition of cancer cells by targeting various pathways, including activation of immune system, inhibition of proliferation and cycle, induction of apoptosis, and inhibition of angiogenesis. The pharmacological components of PR act on nervous system by targeting neurotransmitters, activating immune system, decreasing apoptosis, and increasing redox system. Lectins, one major class of the toxic ingredients extracted from raw PR, possess significant toxic effects on human cells. Inflammatory factors, cytochrome P450 proteins (CYP) family enzymes, mammalian target of rapamycin (mTOR) signaling factors, transforming growth factor-ß (TGF-ß) signaling factors, and nervous system, are considered to be the target sites of lectins. Recently, omic analysis is widely applied in Pinellia genus studies. Plastome genome-based molecular markers are deeply used for identifying and resolving phylogeny of Pinellia genus plants. Various omic works revealed and functional identified a series of environmental stress responsive factors and active component biosynthesis-related genes. Our review summarizes the recent progress in active and toxic ingredient evaluation, pharmacological effects, detoxification strategies, and functional gene identification and accelerates efficient utilization of this traditional herb.

Regulating Th17/Treg Balance Contributes to the Therapeutic Effect of Ziyuglycoside I on Collagen-Induced Arthritis.

To investigate the therapeutic effect and primary pharmacological mechanism of Ziyuglycoside I (Ziyu I) on collagen-induced arthritis (CIA) mice. CIA mice were treated with 5, 10, or 20 mg/kg of Ziyu I or 2 mg/kg of methotrexate (MTX), and clinical manifestations, as well as pathological changes, were observed. T cell viability and subset type were determined, and serum levels of transforming growth factor-beta (TGF-ß) and interleukin-17 (IL-17) were detected. The mRNA expression of retinoid-related orphan receptor-γt (RORγt) and transcription factor forkhead box protein 3 (Foxp3) in mouse spleen lymphocytes was ascertained by the real-time reverse transcriptase-polymerase chain reaction (RT-qPCR). Molecular docking was used to detect whether there was a molecular interaction between Ziyu I and protein kinase B (Akt). The activation of mechanistic target of rapamycin (mTOR) in T cells was verified by Western blotting or immunofluorescence. Ziyu I treatment effectively alleviated arthritis symptoms of CIA mice, including body weight, global score, arthritis index, and a number of swollen joints. Similarly, pathological changes of joints and spleens in arthritic mice were improved. The thymic index, T cell activity, and RORγt production of Ziyu I-treated mice were significantly reduced. Notably, through molecular docking, western blotting, and immunofluorescence data analysis, it was found that Ziyu I could interact directly with Akt to reduce downstream mTOR activation and inhibit helper T cell 17 (Th17) differentiation, thereby regulating Th17/regulatory T cell (Treg) balance and improving arthritis symptoms. Ziyu I effectively improves arthritic symptoms in CIA mice by inhibiting mTOR activation, thereby affecting Th17 differentiation and regulating Th17/Treg balance.

Ziyuglycoside I attenuates collagen-induced arthritis through inhibiting plasma cell expansion.

ETHNOBOTANICAL RELEVANCE: With most of the anti-rheumatic drugs having severe adverse drug reactions and poor tolerance, the active components from natural herbs provides a repository for novel, safe, and effective drug development. Sanguisorba officinalis L. exhibits definite anti-inflammatory capacity, however, whether it has anti-rheumatic effects has not been revealed. AIM OF THE STUDY: In the present study, the effect of Ziyuglycoside I (Ziyu I), one of the most important active components in Sanguisorba officinalis L., was investigated in treating collagen-induced arthritis (CIA), illuminating its potential pharmacological mechanisms. MATERIAL AND METHODS: CIA mice were treated with 5, 10, or 20 mg/kg of Ziyu I or 2 mg/kg of MTX, and clinical manifestations as well as pathological changes were observed. T and B cell viability was determined using cell counting kit-8, plasma autoantibodies and cytokines were tested with ELISA, T and B cell subsets were identified by flow cytometry, Blimp1 expression was detected by RT-qPCR and in situ immunofluorescence. The expression of activation-induced cytidine deaminase (AID) was detected by immunohistochemistry. ERK activation in B cells was verified through western blotting and immunofluorescence. Meanwhile, bioinformatics retrieval and molecular docking/molecular dynamics were used to predict the relationship between Blimp1, ERK and Ziyu I with the pharmacokinetics and toxicity of Ziyu I being evaluated in the ADMETlab Web platform. RESULTS: Ziyu I treatment effectively alleviated the joint inflammatory manifestation including arthritis index, global scores, swollen joint count and body weight of CIA mice. It improved the pathological changes of joint and spleen of arthritic mice, especially in germinal center formation. Ziyu I displayed a moderate regulatory effect on T cell activation, the percentage of total T and helper T cells, and tumor necrosis factor-α, but transforming growth factor-ß was not restored. Increased spleen index, B cell viability and plasma auto-antibody production in CIA mice were significantly reduced by Ziyu I therapy. Of note, we found that Ziyu I administration substantially inhibited the excessive expansion of plasma cells in spleen through preventing the expression of B lymphocyte induced maturation protein 1 (Blimp1) and AID in B cells. Ziyu I was predicted in silico to directly interact with ERK2, and reduce ERK2 activation, contributing to the depressed expression of Blimp1. Moreover, Ziyu I was predicted to have a favorable pharmacokinetic profile and low toxicity. CONCLUSION: Ziyu I effectively ameliorates CIA in mice by inhibiting plasma cell generation through prevention of ERK2-mediated Blimp1 expression in B cells. Therefore, Ziyu I is a promising candidate for anti-arthritic drug development.

Sodium houttuyfonate in vitro inhibits biofilm dispersion and expression of bdlA in Pseudomonas aeruginosa.

Biofilm dispersion is the last step in the development of biofilms, and allows bacteria to spawn novel biofilms in new locales. In the previous studies, we found that sodium houttuyfonate (SH) is effective at inhibiting biofilm formation and motility of Pseudomonas aeruginosa. Here, we investigated the effect of SH against the biofilm dispersion of P. aeruginosa by an in vitro model. The results show that the plant derivative, SH, could effectively inhibit both biofilm dispersion of P. aeruginosa, and gene and protein expression of the key biofilm regulator BdlA in a dose-dependent manner. Furthermore, our presented results suggest that SH can penetrate into the biofilm of P. aeruginosa to repress the biofilm life cycle. Therefore, these results indicate that the antimicrobial activity of SH may be partially due to its ability to disrupt biofilm dispersion in P. aeruginosa.

Effect of sodium houttuyfonate on symptom pattern of lung-Qi deficiency in rats induced by bacterialbiofilm infection.

OBJECTIVE: To investigate the in vivo inhibitory effects of sodium houttuyfonate (SH) on symptom pattern of Qi-deficiency in rats induced by infection of bacterial biofilm on rat respiratory tract. METHODS: Symptom pattern is a term used in Traditional Chinese Medicine (TCM) to define a cluster of symptoms in a medical condition. Based on the pattern, TCM therapies are administered. The symptom pattern used in this study was lung-Qi deficiency pattern identified in rats, which was induced by nasal intubation drip of Pseudomonas aeruginosa (P. aeruginosa) (two strains) to form bacterial biofilm on airway combined with stimulation of cold and fatigue. We measured the variations of the symptoms of the pattern, weight, spleen and thymus index, blood gas, lung bronchial tissue pathology and cytokine of rat in different treatments and control groups. RESULTS: The rats of SH-treatment groups had not showed typical symptoms comparing with model group in the early stage of infection. The weight, spleen and thymus index of the SH-treatment groups were significantly higher comparing with untreated model group. The SH-treatment groups also showed higher O(2) partial pressure and lower CO(2) partial pressure than model group. Furthermore, we found that the bronchopulmonary section of SH-treatment groups not showed typical pathogenic variation in model group. The comparison of cytokine concentration in different groups indicated that SH could prevent the over-production of cytokine to reduce the inflammation occurrence. CONCLUSION: In the early stage of airway infection by biofilm of P. aeruginosa, application of SH can prevent the occurrence of lung-Qi deficiency pattern.

[Sodium houttuyfonate inhibits virulence related motility of Pseudomonas aeruginosa].

Sodium houttuyfonate (SH) is a derivative of effective component of a Chinese material medica, Houttuynia cordata, which is applied in anti-infection of microorganism. But, the antimicrobial mechanisms of SH still remain unclear. Here, we firstly discovered that SH effectively inhibits the three types of virulence related motility of.Pseudomonas aeruginosa, i.e., swimming, twitching and swarming. The plate assay results showed that the inhibitory action of SH against swimming and twitching in 24 h and swarming in 48 h is dose-dependent; and bacteria nearly lost all of the motile activities under the concentration of 1 x minimum inhibitory concentration (MIC) (512 mg x L(-1) same as azithromycin positive group (1 x MIC, 16 mg x L(-1)). Furthermore, we found that the expression of structural gene flgB and pilG is down-regulated by SH, which implies that inhibitory mechanism of SH against motility of P. aeruginosa may be due to the inhibition of flagella and pili bioformation of P. aeruginosa by SR Therefore, our presented results firstly demonstrate that SH effectively inhibits the motility activities of P. aeruginosa, and suggest that SH could be a promising antipseudomonas agents in clinic.

Sodium houttuyfonate affects production of N-acyl homoserine lactone and quorum sensing-regulated genes expression in Pseudomonas aeruginosa.

Quorum sensing (QS) is a means of cell-to-cell communication that uses diffusible signaling molecules that are sensed by the population to determine population density, thus allowing co-ordinate gene regulation in response to population density. In Pseudomonas aeruginosa, production of the QS signaling molecule, N-acyl homoserine lactone (AHL), co-ordinates expression of key factors of pathogenesis, including biofilm formation and toxin secretion. It is predicted that the inhibition of AHL sensing would provide an effective clinical treatment to reduce the expression of virulence factors and increase the effectiveness of antimicrobial agents. We previously demonstrated that sodium houttuyfonate (SH), commonly used in traditional Chinese medicine to treat infectious diseases, can effectively inhibit QS-regulated processes, including biofilm formation. Here, using a model system, we demonstrate that SH causes the dose-dependent inhibition of AHL production, through down-regulation of the AHL biosynthesis gene, lasI. Addition of SH also resulted in down-regulation of expression of the AHL sensor and transcriptional regulator, LasR, and inhibited the production of the QS-regulated virulence factors, pyocyanin and LasA. These results suggest that the antimicrobial activity of SH may be due to its ability to disrupt QS in P. aeruginosa.

[Effect of sodium houttuyfonate in enhancing imipenem's activity against Pseudomonas aeruginosa biofilms].

OBJECTIVE: To investigate the resistant effect of houttuyfonate sodium (SH) combined with imipenem (IMP) against Pseudomonas aeruginosa (Pa) biofilms. METHOD: The two-fold dilution method was used to examine the minimum inhibitory concentration (MIC) of the tested drug. The crystal violet staining was applied to detect the effect of the combination of 1/2MIC, 1MIC, 2MIC of SH, single IMP, 1/2MIC of SH and IMP of various concentrations on the clearance rate of adherent bacteria, growth of biofilms and alginate production. Fluorescein diacetate (FDA)-propidium iodide (PI) doubling staining assay was employed to observe the bacterial viability and morphological changes after membrane dispersion of each drug group. RESULT: Sodium houttuyfonate could enhance the effect of IMP against pseudomonas aeruginosa biofilms. Particularly, the combination group with the concentration of 2MIC showed the highest effect, with P < 0.001 compared with the negative control group. The above results were proved by the bacterial viability and biofilm morphology under fluorescence microscope. CONCLUSION: After being combined with imipenem, sodium houttuyfonate shows a higher effect against biofilms. It is expected that the combination of the two drugs could improve the clinical efficacy of associated infections.

Antimicrobial effect of sodium houttuyfonate on Staphylococcus epidermidis and Candida albicans biofilms.

OBJECTIVE: To study antimicrobial effect of Sodium houttuyfonate (SH) on Staphylococcus epidermidis (SE) and Candida albicans (CA). METHODS: The prepared strain broths (OD,o=0.05) containing SE and CA were firstly used to test the minimal inhibitory concentrations (MICs) of SH, azithromycin (AZM) and fluconazole (FLU) by micro-dilution method. Then the biofilms of SE and CA were matured in 96-well plates, and co-cultured with SH, AZM and FLU for 1, 2 and 3 days to assess the antibiofilm efficacies of the agents with different concentrations by crystal violet staining method. At last, the treated biofilms of SE and CA by 2x MIC agents were observed by scanning electronic microscope. RESULTS: The MICs of SE and CA were 256 and 1024 microg/ml, respectively. After the 1st, 2nd and 3rd day of medications, the suppressions of biofilm were about 60% (P < 0.01), 76% (P = 0.000) and 75% (P = 0.000) by 2 x MIC SH, the suppressions of biofilm were about 90% (P = 0.000), 88% (P = 0.000) and 90% (P = 0.000) by 2 x MIC SH, which could be testified by scanning electron microscope results. However, the inhibitions of biofilm attachment had no significant difference for SE by SH and azithromycin and CA by SH and fluconazole. CONCLUSION: SH had widely anti-pathogenic effect on pathogenic biofilm formation of either bacteria or fungus, had more influence on enclosed cells of SE and CA than the traditional antibiotics, revealing its target might be the extracellular polymeric substances, and was more active to inhibit the growth of CA than SE.

[Study on andrographolide-induced apoptosis of Candida albicans biofilm dispersion cells].

OBJECTIVE: To detect the effect of andrographolide on apoptosis of Candida albicans biofilm dispersion cells. METHOD: The morphological changes of apoptotic C. albicans biofilm cells were observed by using Hoechst 33258 staining Fluorescence microscope; changes of mitochondrial membrane potential (MMP) of C. albicans biofilm cells were detected by rhodamine 123 staining flow cytometry; and reactive oxygen species (ROS) was detected by DHR staining flow cytometry. RESULT: 1 000, 100 micromol x L(-1) of andrographolide could cause pyknosis and dense staining of C. albicans biofilm cells, 1 000, 100, 10 micromol x L(-1) of andrographolide could decrease MMP and increase ROS of C. albicans biofilm cells. CONCLUSION: Andrographolide of appropriate concentrations could induce apoptosis of dispersion cells of C. albicans biofilms.

[Effects of houttuyfonate sodium on eliminating adhesion of Psedomonas aeruginosa and forming biofilms].

OBJECTIVE: To investigate the effect of houttuyfonate sodium (HS) on eliminating adhesion of Psedomonas aeruginosa (Pa) and forming biofilms. METHOD: Pa biofilms were established in 96-hold plates. MTT assay was used to evaluate the changes in metabolism of biofilms and assess the minimum eliminating concentration and minimum biofilm inhibitory concentration for adherent Pa. The colony counting method was used to observe the effect of HS on Pa adhesion and biomass in biofilms. SEM was employed to examine the effect of HS on adhesion of tested Pa and morphology of biofilms. RESULT: MEC80 and MEC50 of HS for adherent Pa was 500 mg x L(-1) and 125 mg x L(-1), respectively. Meanwhile, its SMIC80 for either early or mature biofilms of Pa was 500 mg x L(-1), and SMIC50 for early and mature biofilms of Pa were 31.25, 1.95 mg x L(-1), respectively. At the concentration of 250 mg x L(-1), the number of viable bacteria in the state of adhesion and in initial and mature biofilms decreased significantly, compared with the control group (P < 0.05). The number of bacteria on adherent carriers notably reduced under SEM. Following the continuous administration, there were no visible biofilms on carriers in the mature biofilm phase, with the biomass remarkably shrinking and the bacterial morphology changing from bacillus into coccobacillus. CONCLUSION: HS displayed powerful effect on eliminating adherent Pa, and can inhibit Pa biofilm from being formed through continuous administration.

[In vitro activity of baicalin against non-albicans Candida biofilms].

OBJECTIVE: To investigate the effects of baicalin against Candida glabrata, C. parapsilosis, C. krusei and C. guilliermondii biofilms. METHOD: 96-well microtitre plates were used to set up the biofilms; microdilution method was applied to detect minimal inhibitory concentration (MIC) of baicalin for the four non-albians Candida, and XTT reduction assay was adopted to determine sessile minimal inhibitory concentration (SMIC) of baicalin against the four isolates and to detect the effects on adhesion of the fungal cells. RESULT: MICs of baicalin for the four non-albians Candida cells were 125, 250, 125, 62.5 mg L(-1), respectively. The four non-albians Candida could form mature biofilms on 96-well microtitre plates. SMIC50 of baicalin for the four isolates were > 1000, 500, 125, 250 mg x L(-1), respectively. SMIC80 for the four isolates were greater than or equal to 1000 mg x L(-1). Baicalin showed potent inhibitory effects on adhesion of the four non-albians Candida cells. CONCLUSION: Baicalin displays substantial inhibitory effects on C. parapsilosis, C. krusei and C. guilliermondii biofilm.

[Inhibitory effect of baicalin on germ tube formation and adhesion of Candida albicans].

OBJECTIVE: To investigate the effects of baicalin against Candida albicans germ tube formation, and adherence to buccal epitherial and vaginal epitherial cells. METHOD: Various concentrations of baicalin (100, 50, 10 mg x L(-1)) were incubated with C. albicans suspension, the mixed suspension of C. albicans and human buccal epitherial cells, the mixed suspension of C. albicans and vaginal epitherial cells, respectively. The effects of baicalin on C. albicans germ tube formation, and adherence to buccal epitherial and vaginal epitherial cells were then assessed microscopically. RESULT: All concentrations of baicalin could inhibit C. albicans germ tube formation, and adherent to buccal epitherial and vaginal epitherial cells,while there was no significant difference between standard and clinical strains. CONCLUSION: Baicalin could inhibit C. albicans germ tube formation, and adherence to buccal epitherial and vaginal epitherial cells.

[In vitro activity of gallic acid against Candida albicans biofilms].

OBJECTIVE: To investigate the effects of gallic acid against Candida albicans biofilms in vitro. METHOD: XTT reduction assay was performed to determine the effect of gallic acid on C. albicans biofilms and its adherence, and microscopic examination was conducted to assess the effect of gallic acid on morphogenesis of C. albicans biofilms; and cytotoxic assay was used to measure the adverse effects of gallic acid. RESULT: SMIC50, SMIC50 of gallic acid against C. albicans biofilms were 500, 1000 mg x L(-1), respectively; 100 mg x L(-1) and 1000 mg x L(-1) of gallic acid could inhibit the initial adherence and filamentous growth, and the agent showed poor cytotoxic activity. CONCLUSION: gallic acid displayed potent activity against C. albicans biofilm.

[Preventive and therapeutic effects of extract from Rheum palmatum on hepatic encephalopathy in rats with acute liver failure].

OBJECTIVE: To probe the preventive, therapeutic effect and the possible mechanism of extract from Rheum palmatum (ERP) on hepatic encephalopathy(HE) in rats with acute liver failure. METHODS: HE was induced by the administration of 300 mg thioracetamide per kg body weight by gavage on two consecutive days. The effects of ERP were observed on neurology test, serum ammonium, serum endotoxin and liver impairment. RESULTS: ERP could improve rat neuro-reflexes, decrease the staging of HE and rat serum ammonium, endotoxin concentrations, and reduce the liver impairment. CONCLUSION: ERP significantly prevents and treats HE in rats with thioacetamide-induced acute liver failure.