Plant-derived phosphocholine facilitates cellular uptake of anti-pulmonary fibrotic HJT-sRNA-m7
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摘要
Pulmonary fibrosis, a progressive chronic disease with a high mortality rate, has limited treatment options. Currently, lung transplantation remains the only effective treatment. Here we report that a small RNA, HJT-sRNA-m7, from a Chinese herbal medicine Hong Jing Tian(HJT, RHODIOHAE CRENULATAE RADIX ET RHIZOMA, Rhodiola crenulata) can effectively reduce the expressions of fibrotic hallmark genes and proteins both in alveolar in vitro and in mouse lung tissues in vivo. We also discovered over one hundred oil-soluble chemicals from HJT decoctions, most of which are found in lipid extracts from other Chinese herbals decoctions, including Pu Gong Ying(PGY, TARAXACI HERBA, Taraxacum mongolicum), Chuan Xin Lian(CXL, changed to "ANDROGRAPHIS HERBA, Andrographis paniculata"), and Jin Yin Hua(JYH, lonicera japonica or Honeysuckle). We identified the active component in these decoctions as two forms of phosphocholines, PC(18:0/18:2) and PC(16:0/18:2). These PCs potentially could form liposomes with small RNAs to enter human alveolar and gastric cells. Our experimental results suggest an unprecendent lipid complex route through which botanic small RNA can enter human bodies.Our results provide an innovative treatment strategy for oral delivery of siRNAs as therapeutic medication.
Pulmonary fibrosis, a progressive chronic disease with a high mortality rate, has limited treatment options. Currently, lung transplantation remains the only effective treatment. Here we report that a small RNA, HJT-sRNA-m7, from a Chinese herbal medicine Hong Jing Tian(HJT, RHODIOHAE CRENULATAE RADIX ET RHIZOMA, Rhodiola crenulata) can effectively reduce the expressions of fibrotic hallmark genes and proteins both in alveolar in vitro and in mouse lung tissues in vivo. We also discovered over one hundred oil-soluble chemicals from HJT decoctions, most of which are found in lipid extracts from other Chinese herbals decoctions, including Pu Gong Ying(PGY, TARAXACI HERBA, Taraxacum mongolicum), Chuan Xin Lian(CXL, changed to "ANDROGRAPHIS HERBA, Andrographis paniculata"), and Jin Yin Hua(JYH, lonicera japonica or Honeysuckle). We identified the active component in these decoctions as two forms of phosphocholines, PC(18:0/18:2) and PC(16:0/18:2). These PCs potentially could form liposomes with small RNAs to enter human alveolar and gastric cells. Our experimental results suggest an unprecendent lipid complex route through which botanic small RNA can enter human bodies.Our results provide an innovative treatment strategy for oral delivery of siRNAs as therapeutic medication.
Pulmonary fibrosis, a progressive chronic disease with a high mortality rate, has limited treatment options. Currently, lung transplantation remains the only effective treatment. Here we report that a small RNA, HJT-sRNA-m7, from a Chinese herbal medicine Hong Jing Tian(HJT, RHODIOHAE CRENULATAE RADIX ET RHIZOMA, Rhodiola crenulata) can effectively reduce the expressions of fibrotic hallmark genes and proteins both in alveolar in vitro and in mouse lung tissues in vivo. We also discovered over one hundred oil-soluble chemicals from HJT decoctions, most of which are found in lipid extracts from other Chinese herbals decoctions, including Pu Gong Ying(PGY, TARAXACI HERBA, Taraxacum mongolicum), Chuan Xin Lian(CXL, changed to "ANDROGRAPHIS HERBA, Andrographis paniculata"), and Jin Yin Hua(JYH, lonicera japonica or Honeysuckle). We identified the active component in these decoctions as two forms of phosphocholines, PC(18:0/18:2) and PC(16:0/18:2). These PCs potentially could form liposomes with small RNAs to enter human alveolar and gastric cells. Our experimental results suggest an unprecendent lipid complex route through which botanic small RNA can enter human bodies.Our results provide an innovative treatment strategy for oral delivery of siRNAs as therapeutic medication.
引文
Beatty,M.,Guduric-Fuchs,J.,Brown,E.,Bridgett,S.,Chakravarthy,U.,Hogg,R.E.,and Simpson,D.A.(2014).Small RNAs from plants,bacteria and fungi within the order Hypocreales are ubiquitous in human plasma.BMC Genomics 15,933.
Bligh,E.G.,and Dyer,W.J.(1959).A rapid method of total lipid extraction and purification.Can J Biochem Physiol 37,911-917.
Cavalieri,D.,Rizzetto,L.,Tocci,N.,Rivero,D.,Asquini,E.,Si-Ammour,A.,Bonechi,E.,Ballerini,C.,and Viola,R.(2016).Plant microRNAs as novel immunomodulatory agents.Sci Rep 6,25761.
Chin,A.R.,Fong,M.Y.,Somlo,G.,Wu,J.,Swiderski,P.,Wu,X.,and Wang,S.E.(2016).Cross-kingdom inhibition of breast cancer growth by plant miR159.Cell Res 26,217-228.
Cushing,L.,Kuang,P.P.,Qian,J.,Shao,F.,Wu,J.,Little,F.,Thannickal,V.J.,Cardoso,W.V.,and Lü,J.(2011).miR-29 is a major regulator of genes associated with pulmonary fibrosis.Am J Respir Cell Mol Biol45,287-294.
Dickinson,B.,Zhang,Y.,Petrick,J.S.,Heck,G.,Ivashuta,S.,and Marshall,W.S.(2013).Lack of detectable oral bioavailability of plant microRNAs after feeding in mice.Nat Biotechnol 31,965-967.
Gross,T.J.,and Hunninghake,G.W.(2001).Idiopathic pulmonary fibrosis.N Engl J Med 345,517-525.
Ji,X.,Wu,B.,Fan,J.,Han,R.,Luo,C.,Wang,T.,Yang,J.,Han,L.,Zhu,B.,Wei,D.,Chen,J.,and Ni,C.(2015).The anti-fibrotic effects and mechanisms of microRNA-486-5p in pulmonary fibrosis.Sci Rep 5,14131.
Kesharwani,P.,Gajbhiye,V.,and Jain,N.K.(2012).A review of nanocarriers for the delivery of small interfering RNA.Biomaterials33,7138-7150.
Kuhn,C.3rd.,Boldt,J.,King Jr.,T.E.,Crouch,E.,Vartio,T.,and McDonald,J.A.(1989).An immunohistochemical study of architectural remodeling and connective tissue synthesis in pulmonary fibrosis.Am Rev Respir Dis 140,1693-1703.
LaManna,C.M.,Lusic,H.,Camplo,M.,McIntosh,T.J.,Barthélémy,P.,and Grinstaff,M.W.(2012).Charge-reversal lipids,peptide-based lipids,and nucleoside-based lipids for gene delivery.Acc Chem Res 45,1026-1038.
Liang,G.F.,Zhu,Y.L.,Sun,B.,Shao,Y.H.,Jing,A.H.,Wang,J.H.,and Xiao,Z.D.(2014).Assessing the survival of exogenous plant microRNA in mice.Food Sci Nutr 2,380-388.
Liang,H.,Zhang,S.,Fu,Z.,Wang,Y.,Wang,N.,Liu,Y.,Zhao,C.,Wu,J.,Hu,Y.,Zhang,J.,Chen,X.,Zen,K.,and Zhang,C.Y.(2015).Effective detection and quantification of dietetically absorbed plant microRNAs in human plasma.J Nutr Biochem 26,505-512.
Mlotshwa,S.,Pruss,G.J.,MacArthur,J.L.,Endres,M.W.,Davis,C.,Hofseth,L.J.,Pe?a,M.M.,and Vance,V.(2015).A novel chemopreventive strategy based on therapeutic microRNAs produced in plants.Cell Res 25,521-524.
Pastrello,C.,Tsay,M.,McQuaid,R.,Abovsky,M.,Pasini,E.,Shirdel,E.,Angeli,M.,Tokar,T.,Jamnik,J.,Kotlyar,M.,Jurisicova,A.,Kotsopoulos,J.,El-Sohemy,A.,and Jurisica,I.(2016).Circulating plant miRNAs can regulate human gene expression in vitro.Sci Rep 6,32773.
Pierrat,P.,Laverny,G.,Creusat,G.,Wehrung,P.,Strub,J.M.,VanDorsselaer,A.,Pons,F.,Zuber,G.,and Lebeau,L.(2013).Phospholipid-detergent conjugates as novel tools for siRNA delivery.Chem Eur J 19,2344-2355.
Scotton,C.J.,and Chambers,R.C.(2007).Molecular targets in pulmonary fibrosis.Chest 132,1311-1321.
Selman,M.,King,T.E.,Pardo,A.,American Thoracic Society,,European Respiratory Society,,and American College of Chest Physicians,(2001).Idiopathic pulmonary fibrosis:prevailing and evolving hypotheses about its pathogenesis and implications for therapy.Ann Intern Med 134,136-151.
Silahtaroglu,A.N.,Nolting,D.,Dyrskj?t,L.,Berezikov,E.,M?ller,M.,Tommerup,N.,and Kauppinen,S.(2007).Detection of microRNAs in frozen tissue sections by fluorescence in situ hybridization using locked nucleic acid probes and tyramide signal amplification.Nat Protoc 2,2520-2528.
Urban,M.L.,Manenti,L.,and Vaglio,A.(2015).Fibrosis-A common pathway to organ injury and failure.N Engl J Med 373,95-96.
Yang,J.(2014).A clinical study on treating idiopathic pulmonary fibrosis with Dazhu Hongjingtian injection.Clin J Chinese Med 6,8-9.
Yang,J.,Farmer,L.M.,Agyekum,A.A.A.,and Hirschi,K.D.(2015).Detection of dietary plant-based small RNAs in animals.Cell Res 25,517-520.
Yang,J.,Hotz,T.,Broadnax,L.C.,Yarmarkovich,M.,Elbaz-Younes,I.,and Hirschi,K.D.(2016).Anomalous uptake and circulatory characteristics of the plant-based small RNA MIR2911.Sci Rep 6,26834.
Yu,B.,Yang,Z.,Li,J.,Minakhina,S.,Yang,M.,Padgett,R.W.,Steward,R.,and Chen,X.(2005).Methylation as a crucial step in plant microRNA biogenesis.Science 307,932-935.
Zhang,L.,Hou,D.,Chen,X.,Li,D.,Zhu,L.,Zhang,Y.,Li,J.,Bian,Z.,Liang,X.,Cai,X.,Yin,Y.,Wang,C.,Zhang,T.,Zhu,D.,Zhang,D.,Xu,J.,Chen,Q.,Ba,Y.,Liu,J.,Wang,Q.,Chen,J.,Wang,J.,Wang,M.,Zhang,Q.,Zhang,J.,Zen,K.,and Zhang,C.Y.(2012).Exogenous plant MIR168a specifically targets mammalian LDLRAP1:evidence of cross-kingdom regulation by microRNA.Cell Res 22,107-126.
Zhou,Z.,Li,X.,Liu,J.,Dong,L.,Chen,Q.,Liu,J.,Kong,H.,Zhang,Q.,Qi,X.,Hou,D.,Zhang,L.,Zhang,G.,Liu,Y.,Zhang,Y.,Li,J.,Wang,J.,Chen,X.,Wang,H.,Zhang,J.,Chen,H.,Zen,K.,and Zhang,C.Y.(2015).Honeysuckle-encoded atypical microRNA2911 directly targets influenza A viruses.Cell Res 25,39-49.
Bligh,E.G.,and Dyer,W.J.(1959).A rapid method of total lipid extraction and purification.Can J Biochem Physiol 37,911-917.
Cavalieri,D.,Rizzetto,L.,Tocci,N.,Rivero,D.,Asquini,E.,Si-Ammour,A.,Bonechi,E.,Ballerini,C.,and Viola,R.(2016).Plant microRNAs as novel immunomodulatory agents.Sci Rep 6,25761.
Chin,A.R.,Fong,M.Y.,Somlo,G.,Wu,J.,Swiderski,P.,Wu,X.,and Wang,S.E.(2016).Cross-kingdom inhibition of breast cancer growth by plant miR159.Cell Res 26,217-228.
Cushing,L.,Kuang,P.P.,Qian,J.,Shao,F.,Wu,J.,Little,F.,Thannickal,V.J.,Cardoso,W.V.,and Lü,J.(2011).miR-29 is a major regulator of genes associated with pulmonary fibrosis.Am J Respir Cell Mol Biol45,287-294.
Dickinson,B.,Zhang,Y.,Petrick,J.S.,Heck,G.,Ivashuta,S.,and Marshall,W.S.(2013).Lack of detectable oral bioavailability of plant microRNAs after feeding in mice.Nat Biotechnol 31,965-967.
Gross,T.J.,and Hunninghake,G.W.(2001).Idiopathic pulmonary fibrosis.N Engl J Med 345,517-525.
Ji,X.,Wu,B.,Fan,J.,Han,R.,Luo,C.,Wang,T.,Yang,J.,Han,L.,Zhu,B.,Wei,D.,Chen,J.,and Ni,C.(2015).The anti-fibrotic effects and mechanisms of microRNA-486-5p in pulmonary fibrosis.Sci Rep 5,14131.
Kesharwani,P.,Gajbhiye,V.,and Jain,N.K.(2012).A review of nanocarriers for the delivery of small interfering RNA.Biomaterials33,7138-7150.
Kuhn,C.3rd.,Boldt,J.,King Jr.,T.E.,Crouch,E.,Vartio,T.,and McDonald,J.A.(1989).An immunohistochemical study of architectural remodeling and connective tissue synthesis in pulmonary fibrosis.Am Rev Respir Dis 140,1693-1703.
LaManna,C.M.,Lusic,H.,Camplo,M.,McIntosh,T.J.,Barthélémy,P.,and Grinstaff,M.W.(2012).Charge-reversal lipids,peptide-based lipids,and nucleoside-based lipids for gene delivery.Acc Chem Res 45,1026-1038.
Liang,G.F.,Zhu,Y.L.,Sun,B.,Shao,Y.H.,Jing,A.H.,Wang,J.H.,and Xiao,Z.D.(2014).Assessing the survival of exogenous plant microRNA in mice.Food Sci Nutr 2,380-388.
Liang,H.,Zhang,S.,Fu,Z.,Wang,Y.,Wang,N.,Liu,Y.,Zhao,C.,Wu,J.,Hu,Y.,Zhang,J.,Chen,X.,Zen,K.,and Zhang,C.Y.(2015).Effective detection and quantification of dietetically absorbed plant microRNAs in human plasma.J Nutr Biochem 26,505-512.
Mlotshwa,S.,Pruss,G.J.,MacArthur,J.L.,Endres,M.W.,Davis,C.,Hofseth,L.J.,Pe?a,M.M.,and Vance,V.(2015).A novel chemopreventive strategy based on therapeutic microRNAs produced in plants.Cell Res 25,521-524.
Pastrello,C.,Tsay,M.,McQuaid,R.,Abovsky,M.,Pasini,E.,Shirdel,E.,Angeli,M.,Tokar,T.,Jamnik,J.,Kotlyar,M.,Jurisicova,A.,Kotsopoulos,J.,El-Sohemy,A.,and Jurisica,I.(2016).Circulating plant miRNAs can regulate human gene expression in vitro.Sci Rep 6,32773.
Pierrat,P.,Laverny,G.,Creusat,G.,Wehrung,P.,Strub,J.M.,VanDorsselaer,A.,Pons,F.,Zuber,G.,and Lebeau,L.(2013).Phospholipid-detergent conjugates as novel tools for siRNA delivery.Chem Eur J 19,2344-2355.
Scotton,C.J.,and Chambers,R.C.(2007).Molecular targets in pulmonary fibrosis.Chest 132,1311-1321.
Selman,M.,King,T.E.,Pardo,A.,American Thoracic Society,,European Respiratory Society,,and American College of Chest Physicians,(2001).Idiopathic pulmonary fibrosis:prevailing and evolving hypotheses about its pathogenesis and implications for therapy.Ann Intern Med 134,136-151.
Silahtaroglu,A.N.,Nolting,D.,Dyrskj?t,L.,Berezikov,E.,M?ller,M.,Tommerup,N.,and Kauppinen,S.(2007).Detection of microRNAs in frozen tissue sections by fluorescence in situ hybridization using locked nucleic acid probes and tyramide signal amplification.Nat Protoc 2,2520-2528.
Urban,M.L.,Manenti,L.,and Vaglio,A.(2015).Fibrosis-A common pathway to organ injury and failure.N Engl J Med 373,95-96.
Yang,J.(2014).A clinical study on treating idiopathic pulmonary fibrosis with Dazhu Hongjingtian injection.Clin J Chinese Med 6,8-9.
Yang,J.,Farmer,L.M.,Agyekum,A.A.A.,and Hirschi,K.D.(2015).Detection of dietary plant-based small RNAs in animals.Cell Res 25,517-520.
Yang,J.,Hotz,T.,Broadnax,L.C.,Yarmarkovich,M.,Elbaz-Younes,I.,and Hirschi,K.D.(2016).Anomalous uptake and circulatory characteristics of the plant-based small RNA MIR2911.Sci Rep 6,26834.
Yu,B.,Yang,Z.,Li,J.,Minakhina,S.,Yang,M.,Padgett,R.W.,Steward,R.,and Chen,X.(2005).Methylation as a crucial step in plant microRNA biogenesis.Science 307,932-935.
Zhang,L.,Hou,D.,Chen,X.,Li,D.,Zhu,L.,Zhang,Y.,Li,J.,Bian,Z.,Liang,X.,Cai,X.,Yin,Y.,Wang,C.,Zhang,T.,Zhu,D.,Zhang,D.,Xu,J.,Chen,Q.,Ba,Y.,Liu,J.,Wang,Q.,Chen,J.,Wang,J.,Wang,M.,Zhang,Q.,Zhang,J.,Zen,K.,and Zhang,C.Y.(2012).Exogenous plant MIR168a specifically targets mammalian LDLRAP1:evidence of cross-kingdom regulation by microRNA.Cell Res 22,107-126.
Zhou,Z.,Li,X.,Liu,J.,Dong,L.,Chen,Q.,Liu,J.,Kong,H.,Zhang,Q.,Qi,X.,Hou,D.,Zhang,L.,Zhang,G.,Liu,Y.,Zhang,Y.,Li,J.,Wang,J.,Chen,X.,Wang,H.,Zhang,J.,Chen,H.,Zen,K.,and Zhang,C.Y.(2015).Honeysuckle-encoded atypical microRNA2911 directly targets influenza A viruses.Cell Res 25,39-49.