Blocking of Cell Proliferation, Cytokines Production and Genes Expression Following Administration of Cordyceps Sinensis in the Bronchoalveolar Lavage Fluids Cells

다국어 초록 (Multilingual Abstract)

Yuh-Chi Kuo¹,Chia-Lin Wn¹,Wei-jern Tasi¹,Jir-Yenn Wang²,Shi-Chung Chang₃,Ching-Yung Lin⁴,and Ming-Shi Shiao⁴¹National Research Institute of Chinese Medicine, ²Department of Biology,Fu Jen University, ³Department of Chest, ⁴Department of Pediatrics, □De[art,amt of Medical Research and Education. Taipei Veterans General Hospital, Taipei, Taiwan Yuh-Chi Kuo¹,Chia-Lin Wn¹,Wei-jern Tasi¹,Jir-Yenn Wang²,Shi-Chung Chang²,Ching-Yung Lin³,and Ming-Shi Shiao⁴.¹National Research Institute of Chinese Medicine, ²Department of Biology,Fu Jen University, ³Department of Chest, ⁴Department of Pediatrics, □De[art,amt of Medical Research and Education. Taipei Veterans General Hospital, Taipei, Taiwan. Blocking of Cell Proliferation, Cytokines Production and Genes Expression Following Administration of Cordyceps sinensis in the Bronchoalveolar Lavage Fluids Cells. Proceedings of international Symposium on East-West Medicine, Seoul. 44-64, 1999.-Cordyceps sinensis is a major parasitic fungus on larva of Lepidoptera. It is one of the well know fungi used in traditional Chinese medicine for treatment asthma, lung and kideney diseases. Many fungi belonging to the genus Cordyceps have been demonstrated to produce natural products with various biological activities. Reported biological activities of genus cordyceps include:(a)inhibition of DNA and RNA synthesis;(b)enhancement of cell differentiation;(c)restruction of cytoskeleton;(d)inhibition of protein kinase activity;(e)antitumor activity on bladder, colon, lung carcinoma as well as fibroblastoma;(f)inhibition of the infection and revers transciptase activity of human immunodeficiency virus type 1; and(g) inhibition of methylation of nucleic acid. Thus, it is suggested that the extracts of Cordyceps sp. Contain antitumor, antiviral, and many other biological activities. In our previous studies, we proved that C. sinensis contains the antitumor and immunomodulatory agents. In the present studies, effects of C. sinensis on bronchoalveolar lavage fluids(BAL)cells were demonstrated. The crude methanolic extracts of C. sinensis were fractionated into 15 fractions by silica gel column chromatography. Effects of various fraction on BAL cells proliferation and lipopolysaccharide(LPS) stimulated interleukin-1β(IL-8),platelet activating factor(PAF), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α),and interleukin-8(IL-8) production on BAL cells were determined. The result indicated that the CS-19-22 fraction suppressed BAL cells proliferation activated by LPS. The median inhibitory concentration(IC50) was 6㎍/ml. The CS-19-22 fraction also decreased IL-1β,PAF,IL-6, TNF-αand IL-8 production.The results of revers transcription-polymerase chain reaction(RT-PCR) demonstrated that the CS-19-22 fraction did not affect IL-1β,PAF,IL-6, TNF-α,and IL-8 mRNAs expression in BAL cells activated by LPS. By contrast, it inhibited the IL-10 mRNA expression but enhanced IFN-γand IL-12 mRNAs expression in activated BAL cells. Moreover, the CS-19-22 fraction blocked PAF-induced platelet aggregation. It is unlikely that cytotoxicity was involved. Because no cell deths were observable. We hypothesize that inhibitory mechanisms of CS-19-22 on BAL cells proliferation may be related to the impairments of gene expression and production of cytokines in BAL cells. The explanation of C. sinensis antiasthma function may involved the following(1)C.sinenesis contained immunomodulatory agents that stimulated IFN-γand IL-12 expression in TH1cells.(2)Enhanced secretion of IFN-γand IL-12 will inhibit the Tγcells immune responses Ex. Decreasing of IL-6 and IL-10 production.(3) The impairments of cytokines production in TH2 cells suppressed th BAL cells proliferation then more cytokines production Ex.IL-1β,PAF,IL-6,TNF-αand IL-8 in activated BAL cells were decreased. (4)The suppression of TH2cells immune responses will inhibit the growth and differentiation of B cells then IgE production will be decreased. (5)Reduced production of IgE will decreased asthma attack occurring in individuals. However, the results of the present study suggested that C. sinensis may also have acted to treat asthma in part by inhibiting BAL cells proliferation and cytokine gene expression and production. Future, experiments with treatment of animals with asthma with C. sinensis will be necessary to defined whether C. sinensis can reduce asthma injury. Plans are underway for the isolation of pure principal growth inhibitors and their mechanisms of action are subjected for further study.

목차 (Table of Contents)

• Introduction
• Materials and Methods Extraction and fractionation of the fruiting bodies of C. sinensis
• Human subjects
• Lavage Fluid Processing
• Determination of BALF cells proliferation
• Determination of cytokines production in BALF cells
• Extraction of total cellular RNA
• Synthesis of first strand complementary deoxyribonuclei acid
• RCR
• Preparation of washed platelet suspension
• Platelet aggregation test
• Determination of cell viability
• Statistical analysis
• Results
• Effects of 15 subfractions of C. sinensis on BALF cells proliferation
• Effects of the CS-19-22 and CS-103-111 fractions on eytokines production in BALF
• cells
• Effect of the CS-19-22 fraction on cytokine mRNAs expression in BALF cells
• Effect of the CS-19-22 fraction on platelets aggregation
• The viability of BALF cells treated with the CS-19-22 and CS-103-111 fractions
• Disussion
• Acknowledgements