Home | Register | Login | Inquiries | Alerts | Sitemap |  


Advanced Search
JKM > Volume 34(3); 2013 > Article
Lee, Han, Kim, Namgung, Yeo, and Park: Effects of Inhalable Microparticles of Socheongryong-tang on Chronic Obstructive Pulmonary Disease in a Mouse Model

Abstract

Objectives:

This study aimed to evaluate the effects of microparticles of Socheongryong-tang (SCRT) on chronic obstructive pulmonary disease (COPD) in a mouse model.

Methods:

The inhalable microparticles containing SCRT were produced by spray-drying with leucine as an excipient, and evaluated with respect to the aerodynamic properties of the powder by Andersen cascade impactor (ACI). Its equivalence to SCRT extract was evaluated using lipopolysaccharide (LPS) and a cigarette-smoking (CS)-induced murine COPD model.

Results:

SCRT microparticles provided desirable aerodynamic properties (fine particle fraction of 49.6±5.5% and mass median aerodynamic diameter of 4.8±0.3 μm). SCRT microparticles did not show mortality or clinical signs over 14 days. Also there were no significant differences in body weight, organ weights or serum chemical parameters between SCRT microparticle-treated and non-treated groups. After 14 days the platelet count significantly increased compared with the non-treated group, but the values were within the normal range. Inhalation of SCRT microparticles decreased the rate of neutrophils in blood, granulocytes in peripheral blood mononuclear cells (PBMC) and bronchoalveolar lavage fluid (BALF) and level of TNF-α and IL-6 in BALF on COPD mouse model induced by LPS plus CS. This effect was verified by histological findings including immunofluorescence staining of elastin, collagen, and caspase 3 protein in lung tissue.

Conclusions:

These data demonstrate that SCRT microparticles are equivalent to SCRT extract in pharmaceutical properties for COPD. This study suggests that SCRT microparticles would be a potential agent of inhalation therapy for the treatment of COPD.

Fig. 1.
Experimental plan of repeated LPS+CS exposure.
jkm-34-3-54-5f1.tif
Fig. 2.
Effect of SCRT-MP on neutrophil % of blood in COPD mice. Mice were challenged by aspiration of LPS+CS, and then treated with dexamethasone (PC: positive control, 0.5 mg/kg), SCRT (160 mg/kg, p.o), SCRT microparticle (50, 100 mg/kg) for 21 days (n=4).
*: p<0.05 †: p<0.01 compared to COPD-CT by T-test.
‡: p<0.05 compared to Intact by T-test.
jkm-34-3-54-5f2.tif
Fig. 3.
Effect of SCRT-MP on CD11b/Gr-1 cell rate of PBMC (A) and BALF (B) in COPD mice. Mice were challenged by aspiration of LPS+CS, and then treated with dexamethasone (PC: positive control, 0.5 mg/kg), SCRT (160 mg/kg, p.o), SCRT microparticle (50, 100 mg/kg) for 21 days.
jkm-34-3-54-5f3.tif
Fig. 4.
Effects of SCRT-MP on TNF-α and IL-6 production in COPD mice. Mice were challenged by aspiration of LPS+CS, and then treated with dexamethasone (PC: positive control, 0.5 mg/kg), SCRT (160 mg/kg, p.o), SCRT microparticle(50, 100 mg/kg) for 21 days (n=4). (A) TNF-α, (B) IL-6.
*: p<0.05 †: p<0.01 ‡: p<0.001 compared to COPD_CT by T-test.
∮ : p<0.001 compared to Intact by T-test.
jkm-34-3-54-5f4.tif
Fig. 5.
Histological analysis(A) of lung tissues. Mice were challenged by aspiration of LPS+CS, and then treated with dexamethasone (PC: positive control, 0.5 mg/kg), SCRT (160 mg/kg, p.o), SCRT microparticle (50, 100 mg/kg) for 21 days. Lung tissues were examined under microscope (200 × magnification) after H&E staining. Asterisks denote alveoli. Immunofluorescence analysis(B) for caspase 3, elastin, collagen in lung tissue. Mice were challenged by aspiration of LPS+CS, and then treated with dexamethasone (PC: positive control, 0.5 mg/kg), SCRT (160 mg/kg, p.o), SCRT microparticle(50, 100 mg/kg) for 21 days. Lung tissues were examined under microscope (200 × magnification) after immunofluorescence staining against caspase 3, elastin, collagen. Asterisks denote alveoli.
jkm-34-3-54-5f5.tif
Table 1.
The Composition of Socheongryong-tang (SCRT)
Herbal name Pharmacognostic name Lot No. Country of origin Amount (g)
Mahwang Ephedrae Herba M011005 China 6.0
Baekjakyak Paeoniae Radix J031008 Gyeongbuk, Korea 6.0
Omija Schizandrae Fructus A151008 Gyeongbuk, Korea 6.0
Banha Pinelliae Tuber B031003 China 6.0
Sesin Asiasari Radix et Rhizoma S381004 China 4.0
Geongang Zingiberis Rhizoma K021008 Chungnam, Korea 4.0
Gyeji Cinnamomi Cortex K031006 Vietnam 4.0
Gamcho Glycyrrhizae Radix et Rhizoma K370910 China 4.0
Total amount 40.0
Table 2.
Aerodynamic Properties of Microparticle of SCRT
ED* (%) FPF of CD (%) FPF of ED (%) FPF of ND (%) MMAD (μm)
89.0±18.0 49.6±5.5 27.8±6.5 25.4±9.7 4.8±0.3

* ED: Emitted dose,

† FPF: fine particle fraction,

‡ CD: collection dose,

∮ ND: nominal dose (11.1±1.7 mg),

‖ MMAD: mass median aerodynamic diameter.

Table 3.
Body Weight Changes of Male Mice on 14 Day Single Dose Toxicity Study of SCRT Microparticle
Body weight (g)
Intact* Mock SCRT-MP
0 day 22.54±0.27 21.69±0.28 22.14±0.20
5 day 23.29±0.46 22.00±0.31 22.57±0.30
10 day 24.43±0.37 23.36±0.39 23.64±0.34
14 day 24.58±0.49 23.64±0.53 24.00±0.61

Values are expressed as mean±S.D. (n=7).

* Intact: negative, not inhaled;

† Mock: inhaled air;

‡ SCRT-MP: inhaled SCRT microparticle (240 mg/kg).

Table 4.
Organ Weight of Male Mice Inhaled SCRT Microparticle on 14 Day Single Dose Toxicity Study
Intact* Mock SCRT-MP
Liver (g) 1.35±0.06 1.26±0.11 1.12±0.06
Kidney (g) 0.35±0.02 0.32±0.01 0.31±0.01

Values are expressed as mean±S.D. (n=7). Intact: negative, not inhaled;

† Mock: inhaled air;

‡ SCRT-MP: inhaled SCRT microparticle (240 mg/kg).

Table 5.
Hematological Values of Male Mice Inhaled SCRT Microparticle on 1 and 14 Day in Single Dose Toxicity Study
Intact* Mock SCRT-MP
1 Day 14 Day 1 Day 14 Day 1 Day 14 Day
WBC (103/mm3) 3.93±2.23 5.99±2.14 4.06±2.33 4.55±1.55 6.02±1.03 6.49±1.84
RBC (106/mm3) 7.96±1.52 9.02±0.84 8.75±0.06 8.03±2.46 7.19±4.20 9.02±1.30
HGB (g/dl) 9.83±2.11 10.80±0.93 10.47±0.86 9.66±3.11 8.40±5.29 10.28±2.46
HCT (%) 41.50±7.62 45.49±4.11 44.23±0.68 40.52±12.39 35.37±21.10 44.28±7.44
PLT (103/mm3) 467.00±139.86 598.14±83.23 524.00±128.64 604.20±204.95 411.67±285.63 1,015.20±142.25*

Values are expressed as mean±S.D. (n=7).

* Intact: negative, not inhaled;

† Mock: inhaled air;

‡ SCRT-MP: inhaled SCRT microparticle (240 mg/kg).

*: p<0.01 compared to Intact by T-test.

Table 6.
Clinical Chemistry Values of Male Mice Inhaled SCRT Microparticle on 1 Day Single Dose Toxicity Study
Intact* Mock SCRT-MP
1 Day 14 Day 1 Day 14 Day 1 Day 14 Day
GOT (IU/L) 10.00±3.43 19.29±6.66 7.57±1.78 14.86±2.48 11.29±2.78 10.00±5.08
GPT (IU/L) 0.33±0.56 8.86±3.93 0.00±0.00 2.00±0.38 0.14±0.14 1.17±0.48
ALP (IU/L) 9.17±1.05 7.29±1.29 10.29±1.04 6.71±0.89 8.57±1.15 6.17±0.60
Creatinine (IU/L) 0.05±0.02 0.00±0.00 0.04±0.02 0.00±0.00 0.04±0.02 0.00±0.00

Values are expressed as mean±S.D. (n=7).

* Intact: negative, not inhaled;

† Mock: inhaled air;

‡ SCRT-MP: inhaled SCRT microparticle (240 mg/kg).

References

1.. Statistics Korea. Cause of death statistics 2011. 2012. [2screens]. Available at: URL:http://kosis.kr/ups/ups_01List01.jsp?pubcode=YD. Accessed Sep 20, 2012.


2.. Mannino DM, Kiriz VA. Changing the burden of COPD mortality. Int J Chron Obstruct Pulmon Dis. 2006; 1:3. 219–33.
crossref pmid pmc

3.. Mannino DM. COPD: epidemiology, prevalence, morbidity and mortality, and disease heterogeneity. Chest. 2002; 121:121S–26S.
crossref pmid

4.. Celli BR, MacNee W. Standards for the diagnosis and treatment of patients with COPD: a summary of the ATS/ERS position paper. Eur Respir J. 2004; 23:6. 932–46.
crossref pmid

5.. Barnes PJ, Hansel TT. Prospects for new drugs for chronic obstructive pulmonary disease. Lancet. 2004; 364:9438. 985–96.
crossref pmid

6.. Jung SK, Jung HJ, Kim JD, Choi HY, Park MY, Park YC, et al. Pye-gye-nae-gwa-hak. Seoul: Nado;2011. p. 510–1.


7.. Lee H, Kim Y, Kim HJ, Park S, Jang YP, Jung S, et al. Herbal Formula, PM014, Attenuates Lung Inflammation in a Murine Model of Chronic Obstructive Pulmonary Disease. Evid Based Complement Alternat Med. 2012; 2012:769830Epub 2012 Jun 12.
crossref pmid pmc

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


9.. Yoon JM, Park YC. Protective effects of Seonpyejeongcheon-tang on Elastase-induced Lung Injury. J Korean Oriental Med. 2010; 31:1. 84–101.


10.. Choi HJ, Bang NY, Song BW, Kim NJ, Rhyu BH. Survey on the preference for the dosage forms of oriental herbal medicine. Kyunghee Medicine. 2004; 20:1. 46–57.


11.. Derendorf H, Nave R, Drollmann A, Cerasoli F, Wurst W. Relevance of pharmacokinetics and pharmacodynamics of inhaled corticosteroids to asthma. Eur Respir J. 2006; 28:5. 1042–50.
crossref pmid

12.. Courrier HM, Butz N, Vandamme TF. Pulmonary drug delivery systems: recent developments and prospects. Crit Rev Ther Drug Carrier Syst. 2002; 19:4–5. 425–98.
crossref pmid

13.. Zhang J. Shang-han-za-bing-lun. Shijiazhuang. Hebei Kezue Jishu Chubanshe. 1994; 27


14.. Jung S, Cho SJ, Moon KI, Kim HW, Kim BY, Cho SI. Effects of Socheongryong-Tang on Immunoglobulin Production in Asthmatic Mice. Kor. J. Herbology. 2008; 23:1. 23–8.


15.. Kim KY, Lee JH, Kim YJ, Choi SY, Kim TH, Lyu YS, et al. Anti-allergic effects of Socheongyoung-tang on RBL-2H3 mast cell and mice-mediated allergy model. Korean J. Oriental Physiology & Pathology. 2007; 21:5. 1260–70.


16.. Hwang WS, Lee JS, Choi JY, Jung HJ, Rhee HK, Jung SK. Two Cases of Chronic Sinusitis with Asthma Improved by Socheongryong-tang. J Korean Oriental Med. 2003; 24:1. 207–12.


17.. Jung SK, Heo TS, Hwang WS, Ju CY, Kim YW, Jung HJ. The Effects of Sochongryong-tang on Serum IL-4, IL-5, and IFN-ɣ in asthmatic Patients. J Korean Oriental Med. 2002; 23:2. 70–7.


18.. Ibrahim BM, Jun SW, Lee MY, Kang SH, Yeo Y. Development of inhalable dry powder formulation of basic fibroblast growth factor. Int J Pharm. 2010; 385:1–2. 66–72.
crossref pmid

19.. Thiel CG. Cascade impactor data and the log-normal distribution: nonlinear regression for a better fit. J Aerosol Med. 2002; 15:4. 369–78.
crossref pmid

20.. Nemzek JA, Bolgos GL, Williams BA, Remick DG. Differences in normal values for murine white blood cell counts and other hematological parameters based on sampling site. Inflamm Res. 2001; 50:10. 523–7.
crossref pmid

21.. Hillery AM, Lloyd AW, Swarbrick J. Drug delivery and targeting; for pharmacists and pharmaceutical scientists. London: Taylor and Francis;2001. p. 2


22.. Tayab ZR, Hochhaus G. Pharmacokinetic/pharmacodynamic evaluation of inhalation drugs: application to targeted pulmonary delivery systems. Expert Opin Drug Deliv. 2005; 2:3. 519–32.
crossref pmid

23.. Rabe KF, Hurd S, Anzueto A, Barnes PJ, Buist SA, Calverley P, et al. Global strategy for the diagnosis, management, and prevention of chronic obstructive pulmonary disease: GOLD executive summary. Am J Respir Crit Care Med. 2007; 176:6. 532–55.
crossref pmid

24.. Geller DE. Comparing clinical features of the nebulizer, metered-dose inhaler, and dry powder inhaler. Respir Care. 2005; 50:10. 1313–21.
pmid

25.. Borgstrom L. On the use of dry powder inhalers in situations perceived as constrained. J Aerosol Med. 2001; 14:3. 281–7.
crossref pmid

26.. Yang Y, Tsifansky MD, Shin S, Lin Q, Yeo Y. Mannitol-guided delivery of Ciprofloxacin in artificial cystic fibrosis mucus model. Biotechnol Bioeng. 2011; 108:6. 1441–9.
crossref pmid

27.. Yang Y, Tsifansky MD, Wu CJ, Yang HI, Schmidt G, Yeo Y. Inhalable antibiotic delivery using a dry powder co-delivering recombinant deoxyribonuclease and ciprofloxacin for treatment of cystic fibrosis. Pharm Res. 2010; 27:1. 151–60.
crossref pmid

28.. Lipworth BJ. Targets for inhaled treatment. Respir Med. 2000; 94:S13–6.
crossref pmid

29.. Azarmi S, Lobenberg R, Roa WH, Tai S, Finlay WH. Formulation and in vivo evaluation of effervescent inhalable carrier particles for pulmonary delivery of nanoparticles. Drug Dev Ind Pharm. 2008; 34:9. 943–7.
crossref pmid

30.. Molfino NA, Jeffery PK. Chronic obstructive pulmonary disease: Histopathology, inflammation and potential therapies. Pulm Pharmacol Ther. 2007; 20:5. 462–72.
crossref pmid

31.. Yoo CG. Pathogenesis and pathophysiology of COPD. The Korean Journal of Medicine. 2009; 77:383–400.


32.. Niewoehner DE, Kleinerman J, Rice DB. Pathologic changes in the peripheral airways of young cigarette smokers. N Engl J Med. 1974; 291:15. 755–8.
crossref pmid

33.. Schaberg T, Haller H, Rau M, Kaiser D, Fassbender M, Lode H. Superoxide anion release induced by platelet-activating factor is increased in human alveolar macrophages from smokers. Eur Respir J. 1992; 5:4. 387–93.
pmid

34.. Wright JL, Cosio M, Churg A. Animal models of chronic obstructive pulmonary disease. Am J Physiol Lung Cell Mol Physiol. 2008; 295:1. L1–15.
crossref pmid pmc

35.. Wright JL, Sun JP. Effect of smoking cessation on pulmonary and cardiovascular function and structure: analysis of guinea pig model. J Appl Physiol. 1994; 76:5. 2163–8.
pmid

36.. Yang IA, Clarke MS, Sim EH, Fong KM. Inhaled corticosteroids for stable chronic obstructive pulmonary disease. Cochrane Database Syst Rev. 2012; 7:CD002991
crossref

37.. Tanino M, Betsuyaku T, Takeyabu K, Tanino Y, Yamaguchi E, Miyamoto K, et al. Increased levels of interleukin-8 in BAL fluid from smokers susceptible to pulmonary emphysema. Thorax. 2002; 57:5. 405–11.
crossref pmid pmc

38.. Keatings VM, Collins PD, Scott DM, Barnes PJ. Differences in interleukin-8 and tumor necrosis factor-alpha in induced sputum from patients with chronic obstructive pulmonary disease or asthma. Am J Respir Crit Care Med. 1996; 153:2. 530–4.
crossref pmid

39.. Demedts IK, Demoor T, Bracke KR, Joos GF, Brusselle GG. Role of apoptosis in the pathogenesis of COPD and pulmonary emphysema. Respir Res. 2006; 7:53
crossref pmid pmc

40.. Foronjy R, D’Armiento J. The role of collagenase in emphysema. Respir Res. 2001; 2:6. 348–52.
crossref pmid pmc

Editorial office contact information
3F, #26-27 Gayang-dong, Gangseo-gu Seoul, 157-200 Seoul, Korea
The Society of Korean Medicine
Tel : +82-2-2658-3627   Fax : +82-2-2658-3631   E-mail : skom1953@daum.net
About |  Browse Articles |  Current Issue |  For Authors and Reviewers
Developed in M2PI