ABSTRACT
Natural Cordyceps sinensis as an insect-fungal complex, which is developed after Ophiocordyceps sinensis infects a larva of Hepialidae family. Seventeen genotypes of O. sinensis have been identified in natural C. sinensis. This paper summarized the literature reports and GenBank database regarding occurrence and transcription of the mating-type genes of MAT1-1 and MAT1-2 idiomorphs in natural C. sinensis, in Hirsutella sinensis(GC-biased Genotype #1 of O. sinensis), to infer the mating pattern of O. sinensis in the lifecycle of natural C. sinensis. The mating-type genes and transcripts of MAT1-1 and MAT1-2 idiomorphs were identified in the metagenomes and metatranscriptomes of natural C. sinensis. However, their fungal sources are unclear because of co-colonization of several genotypes of O. sinensis and multiple fungal species in natural C. sinensis. The mating-type genes of MAT1-1 and MAT1-2 idiomorphs were differentially present in 237 H. sinensis strains, constituting the genetic control of the O. sinensis reproduction. Transcriptional control of the O. sinensis reproduction includes: differential transcription or silencing of the mating-type genes of MAT1-1 and MAT1-2 idiomorphs, and the MAT1-2-1 transcript with unspliced intron I that contains 3 stop codons. Research on the H. sinensis transcriptome demonstrated differential and complementary transcriptions of the mating-type genes of MAT1-1 and MAT1-2 idiomorphs in Strains L0106 and 1229, which may become mating partners to accomplish physiological heterothallism. The differential occurrence and transcription of the mating-type genes in H. sinensis are inconsistent with the self-fertilization hypothesis under homothallism or pseudohomothallism, but instead indicate the need of mating partners of the same H. sinensis species, either monoecious or dioecious, for physiological heterothallism, or heterospecific species for hybridization. Multiple GC-and AT-biased genotypes of O. sinensis were identified in the stroma, stromal fertile portion(densely covered with numerous ascocarps) and ascospores of natural C. sinensis. It needs to be further explored if the genome-independent O. sinensis genotypes could become mating partners to accomplish sexual reproduction. S. hepiali Strain FENG experienced differential transcription of the mating-type genes with a pattern complementary to that of H. sinensis Strain L0106. Additional evidence is needed to explore a hybridization possibility between S. hepiali and H. sinensis, whether they are able to break the interspecific reproductive isolation. Genotypes #13～14 of O. sinensis feature large DNA segment reciprocal substitutions and genetic material recombination between 2 heterospecific parental fungi, H. sinensis and an AB067719-type fungus, indicating a possibility of hybridization or parasexuality. Our analysis provides important information at the genetic and transcriptional levels regarding the mating-type gene expression and reproduction physiology of O. sinensis in the sexual life of natural C. sinensis and offers crucial reproductive physiology evidence, to assist in the design of the artificial cultivation of C. sinensis to supplement the increasing scarcity of natural resource.
Subject(s)
Cordyceps/genetics , Genes, Mating Type, Fungal/genetics , Reproduction/geneticsABSTRACT
Objective To establish an HPLC method for simultaneous determination of uridine, vernine, adenosine, cordycepin, and N6-(2-hydroxyethyl)-adenosine in Cordyceps (Cordyceps militaris, Cordyceps Fungus Powder CS-4, Hirsutella sinensis, and C. sinensis), and determine the characteristic components of C. militaris to provide the scientific basis for quality control of C. militaris and its extract. Methods HPLC was performed on Inertsil ODS-3 column (250 mm × 4.6 mm, 5 μm) with mobile phase A (acetonitrile) and B (water) for gradient elution. The flow rate was 0.8 mL/min, detection wavelength was 260 nm, and column temperature was 30 ℃. Results All mentioned five nucleosides can be detected from C. militaris. However, cordycepin and N6-(2-hydroxyethyl)-adenosine were undetected in the other three Cordyceps species. The sample preparation method of C. militaris has a great effect on the content of nucleosides. The content of uridine, vernine and adenosine was the highest in samples prepared by ultrasonic extraction 180 min. Cordycepin was stable form six sample preparation methods, and N6-(2-hydroxyethyl)-adenosine was unbearable to heat and acid. The contents of cordycepin and N6-(2-hydroxyethyl)-adenosine were the same in four preparation methods. Conclusion This experiment provides a basis for the quality analysis of C. militaris and its extracts. Cordycepin and N6-(2-hydroxyethyl)-adenosine can be used as markers for the quality control of C. militaris and its extracts.
ABSTRACT
Cordyceps powder is the mycelium obtained by fermentation, and the mycelium is separated from fresh Chinese Cordyceps. Due to its pharmacological activities, it has come to the foreground in recent years. It is found that there were significant differences in the chemical constituents and pharmacological activities of Hirsutella sinensis and Paeceilomyces hepiali from Cordyceps powder. Based on a detailed and objective analysis for the published literatures on the research progress of the two types of mycelium, it could provide the reference for the further studies and development of Cordyceps powder.
ABSTRACT
Objective:To investigate the effect of HSM on the inflammation and pulmonary fibrosis in mice with cecal ligation and puncture ( CLP) . Methods: Both 5 day and 10 day timepoint pulmonary fibrosis were induced by CLP;mice were randomly divided into three groups,including sham group (sham,n=5),model group (CLP,n=11),therapy group (HSM,n=11). HSM extract (200 mg/kg,less than human dose of 0. 27 g/kg) was orally administered to HSM group 2 hour before surgery and repeated everyday, while sham group and model group were given the same dose of normal saline. We used Q-PCR assay to analyze the expression of in-flammatory molecules ( IL-1β and TNF-α) and fibrogenic cytokines ( TGF-β, MMP9 and TIMP1 ) from lung tissues , we used FACS assay to analyze the amount of Th1 cells in PBMC and spleen sections,as well as Treg in spleen sections. Besides,lung sections were subjected to HE stain and immunohistochemical staining withα-SMA and fibronectin. Results:The amount of model group's Th1 cells in PBMC and spleen sections,as well as Treg in spleen sections were significantly lower than sham group's,whereas HSM succeeded in raising them. By day 5, the expression of inflammatory molecules IL-1β and fibrogenic cytokines (TGF-β,MMP9 and TIMP1) from CLP group's lung sections were remarkably improved,by day 10,the expression had decreased but was higher than sham group's;HSM inhibited these cytokines' expression. The expression of TNF-α of CLP group and HSM group had no significant changes. The CLP group's HE results showed obvious pulmonary inflammatory damage,by day 10,the situation had improved,whereas HSM reduced the histopathologic alterations;contrast to sham group,model group's expression of α-SMA and fibronectin was remarkably improved,while HSM group showed lower expression. Conclusion: HSM extract can suppress inflammation, balances immune system and relieves pulmonary fibrosis.
ABSTRACT
Objective:To investigate the effect of HSM on the immunoreaction and renal fibrosis in mice with cecal ligation and puncture (CLP).Methods: Renal fibrosis was induced by CLP;mice were divided into three groups,including sham group (sham,n =5),model group (CLP,n =11),therapy group (HSM,n =11).HSM extract [200 mg/(kg?d)] was orally administered to HSM group2 hour before surgery and repeated everyday throughout 10 days,while sham group and model group were given the same dose of normalsaline.FACS assay was used to analyze the amount of macrophages ,neutrophils and Treg in PBMC,as well as macrophages in peritonealfluid;we used Q-PCR assay to analyze the expression of inflammatory molecules (IL-1βand TNF-α) and fibrogenic cytokines (TGF-β,MMP9 and TIMP1) from renal sections.Besides,renal sections were subjected to HE stain and immunohistochemical staining with α-SMA and fibronectin.Results: The amount of model group′s macrophages and neutrophils in PBMC,as well as macrophages in theperitoneal fluid were significantly higher than sham group ′s,whereas HSM succeeded in lowering them;contrast to sham group,Tregs′amount of CLP group and HSM group in PBMC had no significant changes .The expression of inflammatory molecules (IL-1βand TNF-α) and fibrogenic cytokines (TGF-β,MMP9 and TIMP1) from CLP group′s renal sections were remarkably improved ,whereas HSM inhibitedthat.The CLP group′s HE results showed obvious renal inflammatory damage ,whereas HSM reduced the histopathologicalterations;contrast to sham group,model group′s expression of α-SMA and fibronectin was remarkably improved,while HSM groupshowed lower expression.Conclusion: HSM extract could regulate immunity response and had effect in improving renal fibrosis .
ABSTRACT
A HPLC-QTOF MS method was established for analysis of components in Ophiocordyceps sinensis . The HPLC analysis was performed on an Agilent Zorbax SB Aq (150 mmí4.6 mm, 5 μm) with gr adient elution (5 mmol·L-1 ammonium acetate aqueous solution-acetonitrile), flow rate was 0.8 mL·min-1 and detection wave-length was 260 nm. The developed method was successfully applied in analysis of three different samples in-cluding O. sinensis, Hirsutella sinensis ( anamorph of O.sinensis) and Cordyceps militaris. Nine compounds were i-dentified in both O.sinensis and H.sinensis, which eight compounds were identified in C.militaris.
ABSTRACT
Objective Hirsutella sinensis (HS) is the anmorph of Ophiocordyceps sinensis (Cordyceps sinensis). O.sinensis and Panax notoginseng are two popular Chinese herbs, commonly used in traditional Chinese prescriptions for the treatment of various diseases. A combination of HS extract with P. notoginseng saponin (PNS) extract demonstrated more prominent lung-protective activity than the two herbs individually used in our preliminary studies. This study further investigated the action of their combination (HSPNS) on anti pulmonary fibrosis using a Bleomycin (BLM)induced mouse model. Methods BLM-treated Kunming mice was given HSPNS daily for 7, 14 or 28 d via ig administration. After treatment, following parameters were monitored using proper methods, respectively. Lung index, serum and lung malondialdehyde (MDA) and hydroxyproline (HYP) contents, lung superoxide dismutase (SOD) activity, transforming growth factor β1 (TGF-β1), and expression levels of collagen Ⅰ (Col- Ⅰ) and collagen Ⅲ (Col-Ⅲ). The lung biopsies were also dissected for semiquantitative histological analysis. Results The results indicated that HSPNS significantly reduced lung index, MDA and HYP contents, and expression levels of TGF-β1,Col- Ⅰ, and Col-Ⅲ. The combination also remarkably enhanced SOD activity compared with BLM-induced group.Moreover, the severe pulmonary fibrosis histopathological changes induced by BLM could be attenuated by HSPNS treatment. Conclusion These results suggest that HSPNS could significantly inhibit the progression of pulmonary fibrosis induced by BLM and its inhibitory effect might associate with its ability to scavenge free radicals, decrease TGF-β1 level, and inhibit collagen synthesis.
ABSTRACT
The particularities of oil suspension formulation can raise the invasive rate of Hirsutella sinensis to host of Hepialidae for the commercialization of artificial cultivation of Cordyceps sinensis. So it is important to develop a high cell viability oil suspension formulation of C. sinensis spawn. According to the characteristics of the oil suspension formulation, MTT assay is adapted and optimized. The result is as follows: reaction time 120 min, reaction temperature 37?C, methylbenzene as extracting agent, and a positive linear correlation established between active cell weights and cell viability. Varieties and concentrations of assistance agents in oil suspension formulation have been selected with the refined MTT assay, and further optimized together with cell concentrations through orthogonal experiment. The optimal combination project was obtained, namely, cell concentration 0.15 g/mL, aluminium stearate 60 mg/mL, and SPAN-80 50 ?L/mL. Results of stability test on the oil suspension formulation indicate that cell viability can maintain above 90% at 4?C after one month.
ABSTRACT
[Objective] To observe the proliferation of T and B Lymphocyte in mouse after treated by Hirsutella sinensis strain fermentation.[Methods] Mice were treated with 1.0g?kg-1,0.1g?kg-1 of Hirsutella sinensis strain fermentation by intragastric administration.The proliferation of T and B Lymphocyte from the spleen of mouse is detected by the MTT assay .[Results] Hirsutella sinensis strain fermentation and Cordyceps Sinensis can distinctly increasse the stimulation index (SI) of T and B lymphocyte transformation.[Conclusion] Hirsutella sinensis strain fermentation and Cordyceps Sinensis are able to enhance immune function of normal mouse.
ABSTRACT
Objective To study whether Hirsutella sinensis (HS) can antagonize aristolochic acid (AA) induced fibrogenesis on human proximal tubular epithelial cells (HKC). Methods HKC were incubated with medium alone, medium with HS 10 mg/L, medium with AA-Na 40 mg/L or medium with AA-Na 40 mg/L and HS 10 mg/L, respectively. After 12 h (for mRNA) or 36 h (for protein) ,cells were lysed,and the mRNA and protein expression level of transforming growth factor-?1 (TGF-?1), connective tissue growth factor (CTGF), tissue inhibitor of metalloproteinase-1 (TIMP-1)and plasminogen activator inhibitor-1 (PAI-1) of HKC were measured by RT-PCR, ELISA (for TGF-?1, TIMP-1 and PAI-1) and Western blotting (for CTGF), respectively. Results The mRNA and protein expression of TGF-?1, PAI-1, CTGF and TIMP-1 were significandy up-regulated by AA-Na 40 mg/L. Compared with the control group, the mRNA expression of TGF-?1,.CTGF, TIMP-1 and PAI-1 was up-regulated to 1.24,1.31,1.27 and 1.36 times,respectively (P