Establishment and comparison of combining disease and syndrome model of asthma with "kidney yang deficiency" and "abnormal savda"

Abstract

THE STUDY WAS THE FIRST TIME TO ESTABLISH AND COMPARE TWO RAT MODELS OF TWO COMMON SYNDROMES: Kidney Yang Deficiency syndrome (KYDS) in traditional Chinese medicine (TCM) and abnormal savda syndrome (ASS) in traditional Uighur medicine (TUM). Then, we also established and evaluated rat models of combining disease and syndrome models of asthma with KYDS or ASS. Results showed that usage of the high dose of corticosterone (CORT) injection or external factors could successfully establish the KYDS or ASS rat models, and the two models had similar changes in biological characterization, abnormal behaviors, dysfunction of hypothalamic-pituitary-target organ axes (HPTOA), and sympathetic/parasympathetic (S/P) nerve system but varied in different degrees. The rat models of combining disease and syndrome of asthma with KYDS or ASS had either pathological characteristics of asthma such as airway hyperresponsiveness (AHR), airway inflammation, airway remodeling, which were more serious than allergy exposure alone, or the syndrome performance of Kidney Yang Deficiency in TCM and abnormal savda in TUM. These findings provide a biological rationale for further investigation of combining disease and syndrome model of asthma as an effective animal model for exploring asthma based on the theory of traditional medicine.

Figure 1

Rat models of combined KYDS or ASS with OVA sensitization and challenge.

Figure 2

Body weight was measured and recorded before and after experiment. There was no significant difference among six groups before the experiment. However, when rats were sacrificed, the body weight gain was statistically different. Data are expressed as mean ± SD, n = 10 rats per group. (a): **P < 0.01, *P < 0.05, versus the NC group; ^# P < 0.05, versus the KYDS group. (b): **P < 0.01, *P < 0.05, versus the NC group; ^# P < 0.05, versus the AC group. (c): *P < 0.05, versus the KYDS group or the ASS group.

Figure 3

Food and water intake were calculated during the three point time of the whole experiment process. Before the experiment, there was no significant difference in food and water intake among six groups. Data are expressed as mean ± SD, n = 10 rats per group. (a) and (d): **P < 0.01, *P < 0.05, versus the NC group. (b) and (e): **P < 0.01, *P < 0.05, versus the NC group. (c) and (f): *P < 0.05, versus the KYDS group or the ASS group.

Figure 4

In the open field test, spontaneous activity was evaluated when rats were response to a new environment. Data are expressed as mean ± SD, n = 10 rats per group. (a): **P < 0.01, *P < 0.05, versus the NC group; ^# P < 0.05, versus the KYDS group. (b): **P < 0.01, *P < 0.05, versus the NC group; ^# P < 0.05, versus the KYDSA group. (c): **P < 0.01, *P < 0.05, versus the KYDS group or the ASS group.

Figure 5

In the sucrose preference test, total water intake, sucrose water intake and pure water intake were recorded, and then the index of sucrose preference was calculated. Data are expressed as mean ± SD, n = 10 rats per group. (a): *P < 0.05, versus the NC group. (b): *P < 0.05, versus the NC group; ^# P < 0.05, versus the AC group. (c): *P < 0.05, versus the KYDS group or the ASS group.

Figure 6

In order to evaluate possible atrophic and/or hyperplasia effects on immune organs and HPTOA glands due to modeling, thymus, spleen, adrenal, thyroid and testicular gland were removed, carefully trimmed, and weighed after rats were sacrificed. Data are expressed as mean ± SD, n = 10 rats per group. (a) and (b): **P < 0.01, *P < 0.05, versus the NC group. (c): **P < 0.01, versus the KYDS group or the ASS group. (d): **P < 0.01, *P < 0.05, versus the NC group; ^# P < 0.05, versus the KYDS group. (e): **P < 0.01, *P < 0.05, versus the AC group. (f): *P < 0.05, versus the KYDS group or the KYDSA group or the ASSA group.

Figure 7

Function of HPTOA was evaluated by the analysis of plasma hormones. Data are expressed as mean ± SD, n = 10 rats per group. (a1): **P < 0.01, *P < 0.05, versus the NC group. (a2): **P < 0.01, *P < 0.05, versus the NC group or the AC group. (a3): *P < 0.05, versus the KYDS group or the ASS group. (b1): **P < 0.01, *P < 0.05, versus the NC group; ^# P < 0.05, versus the KYDS group. (b2): **P < 0.01, *P < 0.05, versus the NC group. (b3): *P < 0.05, versus the ASS group. (c1): **P < 0.01, versus the NC group. (c2): **P < 0.01, *P < 0.05, versus the NC group. (c3): *P < 0.05, versus the ASS group.

Figure 8

Dysfunction of S/P nerve system was induced in the five modeling groups compared to the NC group. Data are expressed as mean ± SD, n = 10 rats per group. **P < 0.01, *P < 0.05, versus the NC group.

Figure 9

Airway hyperresponsiveness (AHR) was assessed more directly by measuring changes in airway resistance in response to increasing concentrations of inhaled methacholine (Mch). Data are expressed as mean ± SD, n = 10 rats per group. (b): **P < 0.01, *P < 0.05, versus the NC group; ^# P < 0.05, versus the AC group. (c): **P < 0.01, *P < 0.05, versus the KYDS group or the KYDSA group or the ASSA group; ^# P < 0.05, versus the KYDSA group.

Figure 10

Analysis of pro-inflammatory and anti-inflammatory cytokines in bronchoalveolar lavage fluid (BALF). BALF was harvested during the 24 h after last OVA challenge. Data are expressed as mean ± SD, n = 10 rats per group. (a1): *P < 0.05, versus the NC group; (a2) and (b2): **P < 0.01, *P < 0.05, versus the NC group; (a3) and (b3): **P < 0.01, *P < 0.05, versus the KYDS group or the ASS group. (c2) and (d2): **P < 0.01, *P < 0.05, versus the NC group; ^# P < 0.05, versus the KYDSA group. (c3) and (d3): **P < 0.01, *P < 0.05, versus the KYDS group or the ASS group.

Figure 11

Detection of pulmonary inflammation according to the lung pathology. Lung tissues were obtained on the day after the last OVA challenge. Tissues were stained with hematoxylin and eosin (H&E, 200x). (a): NC group; (b): AC group; (c): KYDS group; (d): ASS group; (e): KYDSA group; (f): ASSA group. Note: Lung tissues were stained by hematoxylin and eosin (H&E), periodic acid-schiff (PAS) and masson, and scores mirrored to airway inflammation and airway remodeling were performed blindly by two researchers. Data are expressed as mean ± SD, n = 10 rats per group. **P < 0.01, *P < 0.05, versus the NC group; ^# P < 0.05, vs the AC group.