Observation of Inflammatory Marker Levels in Sprague-Dawley Rats with Youngyopaedoc-san-related Anti-atherogenic Effect

Da-rae Yoon; Sung-in Hong; Hyun-in Noh; Seo-ra Yi; In-hee Lee; Jae-hwan Lew; Yoon, Da-rae; Hong, Sung-in; Noh, Hyun-in; Yi, Seo-ra; Lee, In-hee; Lew, Jae-hwan

doi:10.13048/jkm.13014

JKM > Volume 34(3); 2013 > Article

Yoon, Hong, Noh, Yi, Lee, and Lew: Observation of Inflammatory Marker Levels in Sprague-Dawley Rats with Youngyopaedoc-san-related Anti-atherogenic Effect

Original Article

Journal of Korean Medicine 2013; 34(3): 86-95.

Published online: September 30, 2013

DOI: https://doi.org/10.13048/jkm.13014

Observation of Inflammatory Marker Levels in Sprague-Dawley Rats with Youngyopaedoc-san-related Anti-atherogenic Effect

Da-rae Yoon, Sung-in Hong, Hyun-in Noh, Seo-ra Yi, In-hee Lee, Jae-hwan Lew

Dept. of Oriental Internal Medicine, College of Oriental Medicine, Kyunhee University Dept. of East-west integrated medicine, Kyung-Hee Oriental Medical Hospital

Correspondence to: 류재환(Jae-hwan Lew), 서울시 동대문구 회기동 1번지 경희의료원 한방병원 3313호 동서협진실 의국, Tel: +82-2-958-1812, E-mail: intmed@khu.ac.kr

Received May 30, 2013 Revised July 17, 2013 Accepted July 17, 2013

Abstract

Objectives:

Sprague-Dawley rats were fed with high fat diet, and atherogenic changes were seen in the aorta. However, when Sprague-Dawley rats were fed with a high fat diet and administered Youngyopaedoc-san together, these atherogenic changes were rarely seen. This study was aimed to find the inflammatory marker level changes in Sprague-Dawley rats with Youngyopaedoc-san-related anti-atherogenic effect.

Methods:

The extract from Youngyopaedoc-san was made by the pharmacy department of Kyung-hee Oriental Medical Hospital. The animals were divided into five groups: normal diet, high fat diet, high fat diet with Youngyopaedoc-san, high fat diet with Vytorin, and high fat diet with Youngyopaedoc-san and Vytorin. A light microscopic image of a cross section taken from the aorta of the Sprague-Dawley rat was analyzed. We compared inflammatory marker levels among the five groups.

Results:

The complex of Youngyopaedoc-san and Vytorin has more anti-atherogenic effects in the aorta of Sprague-Dawley rats fed with high fat diet than Vytorin alone. Youngyopaedoc-san has inhibitory effect on the increase of IFN-ɣ and IL-2 levels. The difference on eosinophil levels of each group was statistically significant, but the eosinophil level of each group was within normal limits, so the difference on eosinophil levels was not clinically significant.

Conclusions:

Youngyopaedoc-san-related anti-atherogenic effect could be a result of inhibitory mechanism on IFN-ɣ and IL-2.

Keywords: Youngyopaedoc-san, high fat diet, atherosclerosis, histology, inflammatory marker

Fig. 1.

The light microscopic images of a cross section taken through the wall of the aorta of Sprague Dawley rats.

As shown in (A) and (B), the innermost layer, the endothelial cells are supported by a thin bed of subendothelial connective tissue that rest on thick sheet of elastic fiber. In (C) and (D), The arrow shows subendothelial foam cell deposits. In (E), The arrow shows the tunica intima composed of endothelial cells and elastic lamina tightly. The foamy vacuolization and fat deposits in the tunica media mean degenerative changes of wall. In (F), The arrow shows degenerative chages in the tunica media at the bifurcation of aorta. In (G), The arrow shows the foamy vacuolization in the tunica intima apparently. In (H), The arrow shows the vacuolization and the fat infiltration in the tunica media. In (I), The arrow shows minor fat deposits in the subendothelial layer. In (J), The arrow slightly shows minor vacuolizaion in the endothelial layer. It is almost normal. Animals were divided into five groups: Normal, fed with normal diet; Control, fed with high fat diet; YG, fed with high fat diet with Youngyopaedoc-san; VY, fed with high fat diet with Vytorin; and YG+VY, fed with high fat diet with Youngyopaedoc-san and Vytorin.

Table 1.

Youngyopaedoc-san Prescription of Kyung-hee Oriental Medical Hospital

Constitute Herbs	Weight
Ostericii Radix	3.0g
Angelicae Pubescentis Radix	3.0g
Bupleuri Radix	3.0g
Peucedani Radix	3.0g
Platycodi Radix	3.0g
Cnidii Rhizoma	3.0g
Poria	3.0g
Lonicerae Flos	3.0g
Aurantii Fructus	3.0g
Forsythiae Fructus	3.0g
Saposhnikovia Radix	3.0g
Schizonepetae Spica	3.0g
Menthae Herba	3.0g
Glycyrrhizae Radix	3.0g

Total amount	42g

Table 2.

Components of Normal Diet

Ingredient	Proportion (%)
Moisture	11.03
Protein	20.89
Fat	4.75
Fiber	5.77
Ash	6.04
Calcium	1.24
Phosphorus	0.55
etc.	49.73

Total amount	100

Table 3.

Components of High Fat Diet

Ingredient	Proportion (%) by weight
Protein	23.5
Cardohydrate	27.3
Fat	34.3
etc.	14.9

Total amount	100

Ingredient	Proportion (%) kcal from
Protein	18.4
Cardohydrate	21.3
Fat	60.3

Total amount	100

Casein	26.50
L-Cystine	0.40
Maltodextrin	16.00
Sucrose	9.00
Lard	31.00
Soybean Oil	3.00
Cellulose	6.55
Mineral Mix AIN-93G-MX (94046)	4.80
Calcium Phosphate, dibasic	0.34
Vitamin Mix V10001, AIN-93-VX (94047)	2.10
Choline Bitartrate	0.30
Blue Food Color	0.01

Total amount	100

Table 4.

Inflammatory Marker Level of Each Group

	Normal	Control	YG	VY	YG+VY
WBC (10³/μl)	8.73±2.16	8.07±2.55	7.11±3.05	7.02±2.41	7.35±2.92
S-Neutrophil (%)	12.89±4.99	13.65±2.79	15.25±4.56	15.14±2.11	13.43±3.71
Lymphocyte (%)	83.70±5.88	82.49±2.77	80.59±4.92	79.39±3.18	83.40±4.08
Monocyte (%)	1.45±0.67	1.50±0.46	1.49±0.50	1.45±0.52	1.54±0.40
Eosinophil (%)	1.75±0.90	2.14±0.50	2.50±1.67	3.83±2.51* (P=0.008)	1.45±0.42* (P=0.035)
hs-CRP (mg/L)	1.40±0.20	1.43±0.74	1.43±0.07	1.41±0.83	1.49±1.25
IFN-γ (ρg/μl)	74.15±45.76	137.96±67.52	46.34±15.37*	151.03±117.38	61.56±35.15*
IL-1β (ρg/μl)	300.56±118.97	339.00±102.63	485.28±248.43	341.33±110.23	294.84±71.95
IL-2 (ρg/μl)	315.58±98.38	749.08±417.33	305.60±134.92*	655.06±268.36	495.38±237.24
IL-4 (ρg/μl)	40.44±32.87	55.69±38.73	26.00±15.49	38.24±16.34	13.42±7.08
IL-12p70 (ρg/μl)	1.68±0.11	2.24±0.48	2.00±0.00	2.13±0.51	4.08±3.27
TNF-α (ρg/μl)	464.82±154.05	940.92±427.56	1533.76±991.65	1240.23±587.81	1553.17±989.89
IL-6 (ρg/μl)	385.04±147.93	498.14±252.64	958.64±903.15	452.69±67.08	460.67±208.90

The results are expressed as mean±S.D (n=8). The significant differences with the control group were statistically calculated by Mann-Whitney test. YG: The group administered Youngyopaedoc-san, VY: The group administered Vytorin, YG+VY: The group administered Youngyopaedoc-san and Vytorin.

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