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JKM > Volume 42(3); 2021 > Article
Kim, Yang, Lee, Kim, Lyu, and Park: Inhibitory Effects of GGX on Lung Injury of Chronic Obstructive Lung Disease (COPD) Mice Model

Abstract

Objectives

This study is aimed to evaluate the protective effects of GGX on lung injury of Chronic Obstructive Lung Disease (COPD) mice model.

Materials and Methods

C57BL/6 mice were challenged with lipopolysaccharide (LPS) and cigarette smoke extract (CSE) and then treated with vehicle only (Control group), dexamethasone 3 mg/kg (Dexa group), gam-gil-tang 200 mg/kg (GGT group), GGX 100, 200, and 400 mg/kg (GGX group). After sacrifice, its bronchoalveolar lavage fluid (BALF) or lung tissue was analyzed with cytospin, Enzyme-Linked Immunosorbent Assay (ELISA), real-time polymerase chain reaction (PCR) and hematoxylin & eosin (H&E), and Masson’s trichrome staining.

Results

In the COPD model, GGX significantly inhibited the increase of neutrophils, TNF-α, IL-17A, CXCL-1, MIP2 in BALF and TNF-α, IL-1β, IL-10 mRNA expression in lung tissue. It also decreased the severity of histological lung injury.

Conclusion

This study suggests the usability of GGX for COPD patients by controlling lung tissue injury.

Fig. 1
Total particulate matter of cigarette smoke solution.
TPM: total particulate matter, WFHA: Weight of filter holder after smoke, WFHB: Weight of filter holder before smoke, N: Cigarette number of each trap.
jkm-42-3-56f1.gif
Fig. 2
Experimental plan of repeated CSE+LPS exposure. CSE+LPS: Intranasal instillation of LPS 100 μg/μl and cigarette smoke extract 1 mg/ml.
jkm-42-3-56f2.gif
Fig. 3
Effect of GGX on cytospin image (A) and neutrophils count (B) in BALF of COPD mice.
Mice were challenged by aspiration of LPS+CSE (Control), and then treated with Dexa (dexamethasone 3 mg/kg), GGT (200 mg /kg) and GGX (100, 200, 400 mg /kg) for 21 days (n=8). All values are presented as mean±SE. †: Significant difference with the non-treated group (††† p<0.001), *: Significant difference with the Control (*** p<0.001).
jkm-42-3-56f3.gif
Fig. 4
Effect of GGX on TNF-α production in BALF of COPD mice.
Mice were challenged by an aspiration of LPS+CSE (Control), and treated with Dexa (dexamethasone 3 mg /kg), GGT (200 mg /kg) and GGX (100, 200, 400 mg /kg) for 21 days (n=8). Level of TNF-α was determined with ELISA. All values are presented as mean±SE. †: Significant difference with the non-treated group (††† p<0.001), *: Significant difference with the Control (** p<0.01, *** p<0.001).
jkm-42-3-56f4.gif
Fig. 5
Effect of GGX on IL-17A production of BALF in COPD mice.
Mice were challenged by an aspiration of LPS+CSE (Control), and treated with Dexa (dexamethasone 3 mg /kg), GGT (200 mg /kg) and GGX (100, 200, 400 mg /kg) for 21 days (n=8). Level of TNF-α was determined with ELISA. All values are presented as mean±SE. †: Significant difference with the non-treated group (††† p<0.001), *: Significant difference with the Control (* p<0.05, ** p<0.01).
jkm-42-3-56f5.gif
Fig. 6
Effect of GGX on MIP2 production in BALF of COPD mice.
Mice were challenged by an aspiration of LPS+CSE (Control), and treated with Dexa (dexamethasone 3 mg /kg), GGT (200 mg /kg) and GGX (100, 200, 400 mg /kg) for 21 days (n=8). Level of TNF-α was determined with ELISA. All values are presented as mean±SE. †: Significant difference with the non-treated group (††† p<0.001), *: Significant difference with the Control (** p<0.01, *** p<0.001).
jkm-42-3-56f6.gif
Fig. 7
Effect of GGX on CXCL-1 production in BALF of COPD mice.
Mice were challenged by an aspiration of LPS+CSE (Control), and treated with Dexa (dexamethasone 3 mg /kg), GGT (200 mg /kg) and GGX (100, 200, 400 mg /kg) for 21 days (n=8). Level of TNF-α was determined with ELISA. All values are presented as mean±SE. †: Significant difference with the non-treated group (††† p<0.001), *: Significant difference with the Control (*** p<0.001).
jkm-42-3-56f7.gif
Fig. 8
Effect of GGX on TNF-α mRNA expression in lung tissue of COPD mice.
Mice were challenged by an aspiration of LPS+CSE (Control), and treated with Dexa (dexamethasone 3 mg /kg), GGT (200 mg /kg) and GGX (100, 200, 400 mg /kg) for 21 days (n=4). Level of TNF-α was determined with real time PCR. All values are presented as mean±SE. †: Significant difference with the non-treated group († p<0.05), *: Significant difference with the Control (* p<0.05).
jkm-42-3-56f8.gif
Fig. 9
Effect of GGX on IL-1β mRNA expression in lung tissue of COPD mice.
Mice were challenged by an aspiration of LPS+CSE (Control), and treated with Dexa (dexamethasone 3 mg /kg), GGT (200 mg /kg) and GGX (100, 200, 400 mg /kg) for 21 days (n=4). Level of IL-1β was determined with real time PCR. All values are presented as mean±SE. †: Significant difference with the non-treated group (†† p<0.01), *: Significant difference with the Control (** p<0.01).
jkm-42-3-56f9.gif
Fig. 10
Effect of GGX on IL-6 mRNA expression in lung tissue of COPD mice.
Mice were challenged by an aspiration of LPS+CSE (Control), and treated with Dexa (dexamethasone 3 mg /kg), GGT (200 mg /kg) and GGX (100, 200, 400 mg /kg) for 21 days (n=4). Level of IL-6 was determined with real time PCR. All values are presented as mean±SE.
jkm-42-3-56f10.gif
Fig. 10
Effect of GGX on IL-10 mRNA expression in lung tissue of COPD mice.
Mice were challenged by an aspiration of LPS+CSE (Control), and treated with Dexa (dexamethasone 3 mg /kg), GGT (200 mg /kg) and GGX (100, 200, 400 mg /kg) for 21 days (n=4). Level of IL-10 was determined with real time PCR. All values are presented as mean±SE. †: Significant difference with the non-treated group († p<0.05), *: Significant difference with the Control (* p<0.05, ** p<0.01).
jkm-42-3-56f11.gif
Fig. 12
Effect of GGX on histopathological changes and histology scores in the lung of COPD mice.
Mice were challenged by an aspiration of LPS+CSE (Control), and treated with Dexa (dexamethasone 3 mg /kg), GGT (200 mg /kg) and GGX (100, 200, 400 mg /kg) for 21 days (n=4). (A) Representative sections from each treatment group are shown (Light microscope at 100×magnification). (B) Quantitative analysis of the degree of lung tissue damage in the sections. All values are presented as mean±SE. †: Significant difference with the non-treated group (††† p<0.001), *: Significant difference with the Control (** p<0.01, *** p<0.001).
jkm-42-3-56f12.gif
Table 1
The Composition of GGX
Herb Pharmacognostic name Amount (g)
Gilgyeong Platycodi Radix 14.00
Gamcho Glycyrrhizae Radix 6.00
Geumeunhwa Lonicerae Flos 14.00
Sangbaekpi Mori Radicis Cortex 6.00
Total amount 40.00
Table 2
Oligonucleotide Sequence Used for Mouse Real-time PCR
Gene Primer Sequence
TNF-α FAM 5′-CACGTCGTAGCAAACCACCAAGTGGA-3′
Forward 5′-CACCTTCTTTTCCTTCATCTT-3′
IL-1β Reverse 5′-GTCGTTGCTTGTCTCTCCTTGTA-3′
Forward 5′-TACCCCCAGGAGAAGATTCC-3′
IL-6 Reverse 5′-TTTTCTGCCAGTGCC TCTTT-3′
Forward 5′-GATGCCTTCAGCAGAGTGAAGA-3′
IL-10 Reverse 5′-CATGGCTTTGTAGATGCCTTTC-3′
G3PDH VIC 5′-TGCATCCTGCACCACCAACTGCTTAG-3′

참고문헌

1. Park YB, Rhee CK, Yoon HK, Oh YM, Lim SY, Lee JH, et al. 2018; COPD clinical practice guideline of the Korean Academy of Tuberculosis and Respiratory Disease: a summary. Tuberc Respir Dis (Seoul). 81:261–73.
crossref pmid pmc

2. Barnes PJ, Shapiro SD, Pauwels RA. 2003; Chronic obstructive pulmonary disease: molecular and cellular mechanisms. Eur Respir J. 22:4. 672–88.
crossref pmid

3. Kim IA, Park YB, Yoo KH. 2004; Pharmacotherapy for chronic obstructive pulmonary disease. J Korean Med Assoc. Sep. 61:9. 545–551.
crossref

4. Anthonisen NR, Connett JE, Kiley JP, Altose MD, Bailey WC, Buist AS, et al. 1994; Effects of smoking intervention and the use of an inhaled anticholinergic bronchodilator on the rate of decline of FEV1: the Lung Health Study. JAMA. 272:1497–505.
crossref pmid

5. Singh S, Amin AV, Loke YK. 2009; Long-term use of inhaled corticosteroids and the risk of pneumonia in chronic obstructive pulmonary disease: a meta-analysis. Arch Intern Med. 169:219–29.
crossref pmid

6. Barnes PJ. 2013; New anti-inflammatory targets for chronic obstructive pulmonary disease. Nat Rev Drug Discov. 12:7. 543–59.
crossref pmid

7. Lee ES, Han JM, Kim MH, Namgung U, Yeo Y, Park YC. 2013; Effects of inhalable microparticles of Socheongryong-tang on chronic obstructive pulmonary disease in a mouse model. J Korean Med. 34:3. 54–68.
crossref

8. Lee JG, Yang SY, Kim MH, Namgung U, Park YC. 2011; Protective effects of Socheongryong-tang on elastase-induced lung injury. J Korean Oriental Med. 32:4. 83–99.


9. Kim Y, Yang SY, Kim MH, Namgung U, Park YC. 2011; Effects of Saengmaekcheongpye-eum on LPS-induced COPD model. Korean J Oriental Int Med. 2011; 32:2. 217–31.


10. Kim HW, Yang SY, Kim MH, Namgung U, Park YC. 2011; Protective effects of Maekmundong-tang on elastase-induced lung injury. J Korean Oriental Med. 32:2. 63–78.


11. Lee H, Kim Y, Kim HJ, Park S, Jang YP, Jung S, et al. 2012; Herbal Formula, PM014, Attenuates Lung Inflammation in a Murine Model of Chronic Obstructive Pulmonary Disease. Evid Based Complement Alternat Med. 2012; 769830
crossref

12. Han JM, Yang WK, Kim SH, Park YC. 2015; Effects of Sagan-tang and individual herbs on COPD mice model. J Korean Med Soc Herb Formula Study. 23:2. 171–87.


13. Park JJ, Yang WK, Lyu YR, Kim SH, Park YC. 2019; Inhibitory effects of SGX01 on lung injury of COPD mice model. Korean J Int Korean Med. 40:4. 567–81.
crossref

14. Yang WK, Lyu YR, Kim SH, Park YC. 2018; Effects of GHX02 on Chronic Obstructive Pulmonary Disease Mouse Model. J Korean Med. 39:4. 126–35.
crossref

15. Kim SH, Hong JH, Yang WK, Geum , et al. 2020; Herbal combinational medication of Glycyrrhiza glabra, Agastache rugosa containing Glycyrrhizic acid, Tilianin inhibits Neutrophilic lung inflammation by affecting CXCL2, Interleukin-17/STAT3 signal pathways in a murine model of COPD. Nutrients. 12:4. 926
crossref pmc

16. Yang L, Li J, Li Y, Tian Y, Li S, Jiang , et al. 2015; Identification of metabolites and metabolic pathways related to treatment with Bufei Yishen formula in a rat COPD model using HPLC Q-TOF/MS. Evidence-Based Complementary and Alternative Medicine, 2015.
crossref

17. Hwang DY. 1986. Bang-yak-hap-pyeon. Seoul: Namsandang;p. 240


18. Lyu YR. 2020. Inhibitory effects of GGX in a particulate matter-induced lung injury mouse model. doctoral dissertation. Daejeon university.


19. Hong HW. 2011. The Effects of Kamgiltang on Passive Smoking in Rats. doctoral dissertation. Dong-eui University.


20. Mizutani N, Fuchikami J, Takahashi M, Nabe T, Yoshino S, Kohno S. 2009; Pulmonary emphysema induced by cigarette smoke solution and lipopolysaccharide in guinea pigs. Biol Pharm Bull. 32:9. 1559–64.
crossref pmid

21. Lopez AD, Shibuya K, Rao C, Mathers CD, Hansell AL, Held LS, et al. 2006; Chronic obstructive pulmonary disease: current burden and future projections. Eur Rspir J. 27:2. 397–412.
crossref

22. Mathers CD, Loncar D. 2006; Projections of global mortality and burden of disease from 2002 to 2030. PLoS Med. 3:11. e442
crossref pmid pmc

23. An TJ, Yoon HK. 2018; Prevalence and socioeconomic burden of chronic obstructive pulmonary disease. J Korean Med Assoc. 61:9. 533–8.
crossref

24. GBD 2015 Chronic Respiratory Disease Collaborators. 2017; Global, regional, and national deaths, prevalence, disability adjusted life years, and years lived with disability for chronic obstructive pulmonary disease and asthma, 1990–2015 a systematic analysis for the Global Burden of Disease Study 2015. Lancet Respir Med. 5:9. 691–706.
pmid pmc

25. Jung YM, Lee H. 2011; Chronic obstructive pulmonary disease in Korea: Prevalence, risk factors, and quality of life. J Korean Acad Nurs. 41:2. 149–56.
crossref pmid

26. Ministry of Gender Equality & Family (MOGEF). 2014. 2014 Comprehensive survey on the contact with the harmful environment for youth. Available at: URL: http://www.mogef.go.kr Accessed April 1.


27. Yoon J, Seo H, Oh IH, Yoon SJ. 2016; The Non-Communicable Disease Burden in Korea: Findings from the 2012 Korean Burden of Disease Study. J Korean Med Sci. Nov. 31:Suppl 2. S158–S167.
crossref pmid pmc

28. Park SK. 2002; Chronic obstructive pulmonary disease - definition, severity, risk factors, etiology, pathology, diagnosis. J Korean Med. 63:2. 389–99.


29. Lurwidya F, Damayanti T, Yunus F. 2016; The Role of Innate and Adaptive Immune Cells in the Immunopathogenesis of Chronic Obstructive Pulmonary Disease. Tuberc Respir Dis(Seoul). 79:1. 5–13.
crossref pmid

30. Lee S, et al. 2007; Antielastin autoimmunity in tobacco smokinginduced emphysema. Nature Med. 13:567–569.
crossref pmid

31. Yoo CG. 2009; Pathogenesis and pathophysiology of COPD. Korean J Med. 77:4. 383–400.


32. Barnes PJ. 2004; Macrophages as orchestrators of COPD. COPD. 1:59–70.
crossref pmid

33. Majo J, Ghezzo H, Cosio MG. 2001; Lymphocyte population and apoptosis in the lungs of smokers and their relation to emphysema. Eur Respir J. 17:946–53.
crossref pmid

34. Barnes PJ. 2008; The cytokine network in asthma and chronic obstructive pulmonary disease. J Clin Invest. 118:11. 3546–56.
crossref pmid pmc

35. Rovina N, Koutsoukou A, Koulouris NG. 2013; Inflammation and immune response in COPD: where do we stand? Mediators Inflammation. 2013; 413735
crossref

36. Herbology Editorial Committee of Korean Medicine schools. 1991. Boncho-hak. Seoul: Younglimsa;p. 124–5. p. 136–7. p. 214–5. p. 448–9. p. 534–5. p. 580–1. p. 588–9.


37. Park YC, Jin M, Kim SH, Kim MH, et al. 2014; Effects of inhalable microparticle of flower of Lonicera japonica in a mouse model of COPD. Journal of Ethnopharmacology. 151:1. 123–130.
crossref pmid

38. Di Stefano A, Capelli A, Lusuardi M, Balbo P, Vecchio C, Maestrelli P, et al. 1998; Severity of airflow limitation is associated with severity of airway inflammation in smokers. Am J Respir Crit Care Med. 158:4. 1277–85.
crossref pmid

39. Aaron SD, Angel JB, Lunau M, Wright K, Fex C, Le Saux N, et al. 2001; Granulocyte inflammatory markers and airway infection during acute exacerbation of chronic obstructive pulmonary disease. Am J Respir Crit Care Med. 163:2. 349–55.
crossref pmid

40. Deveci Y, Deveci F, Ilhan N, Karaca I, Turgut T, Muz MH. 2010; Serum ghrelin, IL-6 and TNF-α levels in patients with chronic obstructive pulmonary disease. Tuberk Toraks. 58:2. 162–72.
pmid

41. Traves SL, Donnelly LE. 2008; Th17 cells in airway diseases. Curr Mol Med. 8:5. 416–26.
crossref pmid

42. Levänen B, Glader P, Dahlén B, Billing B, Qvarfordt I, Palmberg L, et al. 2016; Impact of tobacco smoking on cytokine signaling via interleukin-17A in the peripheral airways. Int J Chron Obstruct Pulmon Dis. 11:2109–16.
crossref pmid pmc

43. Lukacs NW, Hogaboam CM, Kunkel SL. 2005; Chemokines and their receptors in chronic pulmonary disease. Curr Drug Targets Inflamm Allergy. 4:3. 313–7.
crossref pmid

44. Traves SL, Culpitt SV, Russell RE, Barnes PJ, Donnelly LE. 2002; Increased levels of the chemokines GROα and MCP-1 in sputum samples from patients with COPD. Thorax. 57:7. 590–5.
crossref pmid pmc

45. Culpitt SV, Rogers DF, Shah P, De Matos C, Russell RE, Donnelly LE, et al. 2003; Impaired inhibition by dexamethasone of cytokine release by alveolar macrophages from patients with chronic obstructive pulmonary disease. Am J Respir Crit Care Med. 167:1. 24–31.
crossref pmid

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