| Objective1 To observe the effects of Bufei Yishen granule combined with "Shu Fei Tie" acupoint sticking therapy on pulmonary function and morphormetry based on chronic obstructive pulmonary disease (COPD) rat model.2 To explore the mechanisms of Bufei Yishen granule combined with "Shu Fei Tie" acupoint sticking therapy on inflammation, including inflammatory cytokines, oxidative indexes, alveolar surface active protein, as well as JNK/p38MAPK, PPARy pathway.Methods120 Rats were randomized into Control group (CG), Model group (MG), Bufei Yishen group (BYG), Acupoint sticking group (ASG), Bufei Yishen+Acupoint sticking group (BY+ASG), aminophyline group (AG). COPD rats were duplicated by repeated cigarette smoke and bacterial exposures. The rats were treated with normal saline, Bufei Yishen granule, acupoint sticking, Bufei Yishen+Acupoint sticking and aminophyline respectively from the 9th through 20th week of the experiment. Pulmonary function was detected by using whole body plethysmograph every 4 weeks. The rats were sacrificed at the end of the 20th week, and lung tissue histology as well as ultrastructure was observed under light and electron microscope; interleukine (IL)-2, IL-6, IL-10, monocyte chemoattractant protein(MCP)-l as well as surfactant protein(SP)-A, SP-B, SP-C, SP-D in serum and bronchoalveolar lavage fluid(BALF) were detected by enzyme-linked immunosorbent assay (ELISA); IL-2, IL-6, IL-10, MCP-1, superoxide dismutase (SOD), Malondialdehyde (MDA) protein expression were detected by Immunohistochemistry; .INK, p38 MAPK, PPARy and SOD mRNA were detected by real-time quantitative polymerase chain reaction (QPCR); JNK, p38 MAPK, PPARy protein expressions and phorspholation were detected by western blotting.Results1 Pulmonary functionFrom week 8, VT, PEF and EF50 in MG decreased compared with CG (P<0.01); At week 16, VT in BYG, ASG, BY+ASG and AG was higher than MG (P<0.01, P<0.05); EF50 in BYG and BY+ASG was higher than MG (P<0.01, P<0.05). At week 20, VT, PEF, EF50 in BYG and BY+ASG increased compared with MG (P<0.01, P<0.05), while EF50 in ASG was higher than MG (P<0.05) and VT in AG higher than MG (P <0.05).2 Pulmonary histopathologyMLI in MG increased significantly compared with CG, while MAN decreased significantly (P<0.01). MLI in ASG, BYG, BY+ASG, AG decreased compared with MG (P<0.01), and that in BYG was lower than ASG and AG, in BY+ASG were lower thanASG, BYG and AG (P<0.01).MLI in ASG was lower than AG (P<0.01). MAN in ASG, BYG, BY+ASG, AG increased compared with MG (P<0.01), and that in BYG was higher than ASG and AG, in BY+ASG were higher than ASG, BYG and AG (P<0.01).Under the electron microscope, the basal lamina of respiratory membrane are homogeny in CG and the thickness was relatively homogeneous, while it in MG was illegibility and rough, and it was significantly thicker than CG (P<0.01). The thickness of respiratory membrane in BYG, BY+ASG and AG decreased compared with MG (P <0.01), while it in BY+ASG and AG decreased compared with ASG (P<0.01, P O.05).In MG, the swollen mitochondria, shortened cell ridges and the thinned microvilli were observed in type Ⅱ alveoli epithelial cells, and the alveolar wall was thickened. In ASG, some of type Ⅱ alveoli epithelial cells had shed and detached from the basement membrane, the lamellar bodies were reduced, and the microvilli were thinned and increased in number. The changes in BYG, AG were milder than those of MG in different degrees. In BY+ASG, type Ⅱ alveoli epithelial cells had shed and detached from the basement membrane, the lamellar bodies were reduced, and the microvilli were thinned and increased in number.3 Inflammation3.1 Inflammatory cytokinesIL-2, IL-6, MCP-1 in lung, serum and BALF in MG increased compared with CG (P＜0.01), IL-10 decreased compared with CG (P＜0.01); IL-2, IL-6, MCP-1 in lung, serum and BALF in ASG, BYG, BY+ASG and AG decreased compared with MG (P＜0.01), while IL-10 increased (P＜0.01).3.2 JNK/p38MAPKJNK, p38 MAPK mRNA and phorspholation of JNK (p-JNK) and p38 (p-p38) in MG were higher than in CG (P＜0.01). while they in ASG, BYG and BY+ASG were lower than in MG (P＜0.05,P＜0.01); JNK mRNA in BY+ASG was lower than in ASG (P＜0.05), p38MAPK mRNA in BY+ASG was lower than AG (P<0.05). p-p38 in BYG, BY+ASG and AG was lower than ASG (P<0.05, P＜0.01), p-p38 in BY+ASG was lower than AG (P<0.05).3.3 SOD and MDASOD in lung in MG was lower than in CG (P＜0.01), MDA was higher than CG (P＜0.01); SOD in each treatment group was higher than in MG(P＜0.01), MDA was lower than in MG (P＜0.01); SOD in BYG, BY+ASG and AG was higher than in ASG (P＜0.01), SOD in BYG, BY+ASG was higher than in AG (P＜0.05, PP＜0.01), MDA in BYG, BY+ASG was lower than in AG (P＜0.05, PP＜0.01).3.4 PPARyPPARy mRNA, PPARy and p-PPARy protein expression decreased in MG compared with that in CG(P＜0.01), while that in BYG, ASG, BY+ASG and AG increased compared with in MG(P<0.01); PPARy mRNA in BY+ASG was higher than that inASG(P<0.05), PPARy and p-PPARy protein expression in BY+ASG were higher than that in ASG and AG(P<0.05, P＜0.01); PPARy protein expression in BYG was higher than that in AG(P＜0.05).3.5 surfactant proteinSerum SP-A, SP-B, SP-C, SP-D in MG were higher than that in CG(P<0.01), while that in all treatment groups were lower than in MG(P<0.01). Serum SP-A, SP-B, SP-C, SP-D in BYG and BY+ASG were lower than that in ASG (P＜0.01). Serum SP-A, SP-C, SP-D in BY+ASG were lower than that in BYG and AG (PP＜0.05, P<0.01). Serum SP-D in BYG and SP-B in BY+ASG were lower than in AG (PP＜0.01).Conclusions1 Bufei Yishen granule combined with acupoint sticking plays an important role in improving pulmonary function and lung pathological impairment in COPD rat.2 Bufei Yishen granule combined with acupoint sticking can decrease IL-2,IL-6 and MCP-1 in COPD rats, the anti-inflammatory effect may be involved in regulating JNK/p38MAPK pathway.3 Bufei Yishen granule combined with acupoint sticking can improve mRNA and protein expression of PPARy as well as SOD activity, the beneficial effect may be involved in regulating PPARy pathway increase the antioxidant activity and alleviate inflammation.4 The abnormal synthesis and secretion of pulmonary surfactant protein is observed in COPD rats, and closely related to the degree of inflammation and restricted airflow,Bufei Yishen granule combined with acupoint sticking can regulate the level of pulmonary surfactant and inhibit the inflammatory response.
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