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1.山东中医药大学,济南 250355;
2.山东省中医经典名方协同创新中心,济南 250355;
3.山东中医药大学 附属医院,济南 250011;
4.济宁医学院,山东 济宁 272067
罗光芝,在读博士,从事中医医史文献及临床文献研究,E-mail:13606414698@163.com
张成博,博士,教授,从事中医医史文献及临床文献研究,Tel:0537-3616779,E-mail:doctorzcb@sina.com
收稿日期:2019-01-16,
网络出版日期:2019-05-16,
纸质出版日期:2019-09-20
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罗光芝, 韩成恩, 韩晓春, 等. 基于和法探讨黄连汤治疗慢性非萎缩性胃炎的机制[J]. 中国实验方剂学杂志, 2019,25(18):36-42.
Guang-zhi LUO, Cheng-en HAN, Xiao-chun HAN, et al. Mechanism of Huangliantang in Treatment of Chronic Non-atrophic Gastritis Based on Harmonization Method[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(18): 36-42.
罗光芝, 韩成恩, 韩晓春, 等. 基于和法探讨黄连汤治疗慢性非萎缩性胃炎的机制[J]. 中国实验方剂学杂志, 2019,25(18):36-42. DOI: 10.13422/j.cnki.syfjx.20191702.
Guang-zhi LUO, Cheng-en HAN, Xiao-chun HAN, et al. Mechanism of Huangliantang in Treatment of Chronic Non-atrophic Gastritis Based on Harmonization Method[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(18): 36-42. DOI: 10.13422/j.cnki.syfjx.20191702.
目的:
2
基于和法调节寒热,以慢性非萎缩性胃炎(CNAG)模型大鼠为研究对象,探讨黄连汤治疗CNAG的作用机制。
方法:
2
将60只健康SD大鼠随机分成2组,正常组10只和CNAG造模组50只。造模组采用化学刺激结合饥饱失常诱导CNAG大鼠模型,造模成功后将其随机分为5个组,分别为模型组、荆花胃康丸治疗组、黄连汤高、中、低剂量组,每组8只。荆花胃康丸治疗组(0.04 g·kg
-1
),黄连汤高、中、低剂量组(11.00,5.48,2.74 g·kg
-1
),空白组及模型组给予同剂量生理盐水连续灌胃4周后收集样本。苏木素-伊红(HE)染色观察胃黏膜组织形态改变,酶联免疫吸附测定(ELISA)检测大鼠血清白细胞介素-6(IL-6),白细胞介素-1
β
(IL-1
β
),白细胞介素-8(IL-8)含量,免疫组化(IHC)检测核转录因子-
κ
B(NF-
κ
B)及其抑制蛋白受体(I
κ
B
α
)蛋白表达,实时荧光定量聚合酶链式反应(Real-time PCR)检测I
κ
B
α
,NF-
κ
B mRNA表达。
结果:
2
与正常组比较,模型组大鼠胃组织黏膜受损,可见大量炎细胞浸润,血清炎性因子显著升高,胃组织I
κ
B
α
mRNA及蛋白表达降低,NF-
κ
B mRNA,蛋白表达升高(
P
<
0.01);与模型组比较,各治疗组大鼠胃黏膜组织炎症均有不同程度的改善,血清炎性因子下降,胃组织I
κ
B
α
mRNA表达上调,I
κ
B
α
蛋白增高,NF-
κ
B mRNA表达下调,NF-
κ
B蛋白降低,其中以荆花胃康丸治疗组及黄连汤高剂量组改善最明显(
P
<
0.05,
P
<
0.01)。
结论:
2
基于和法调节寒热黄连汤能够有效改善CNAG大鼠胃黏膜损伤程度,降低血清炎性因子,其作用机制与上调I
κ
B
α
mRNA,增加胃黏膜I
κ
B
α
蛋白表达,下调NF-
κ
B mRNA,降低NF-
κ
B蛋白表达有关。
Objective:
2
Harmonization method is one of the eight unique methods of traditional Chinese medicine with important application value in clinic. Based on the effect of harmonization method in regulating cold and heat
the mechanism of Huangliantang in treating chronic non-atrophic gastritis(CNAG) on rats were studied.
Method:
2
Rats were divided into normal group (
n
=10) and CNAG model group (
n
=50). The model of CNAG rats was induced by chemical stimulation combined with hunger and satiety. The model group was randomly divided into 5 groups
namely the model group
the Jinghua Weikang pill treatment group
and the high
middle and low-dose Huangliantang groups
with 8 rats in each group. After the model was successfully established
the Jinghua Weikang pill treatment group (0.04 g·kg
-1
)
the high
middle
low dose Huangliantang group (11.00
5.48
2.74 g·kg
-1
)
the blank group and the model group were given the same dose of saline for 4 weeks
and then the samples were collected. The histological changes of gastric mucosa were observed by hematoxylin-eosin (HE) staining. The serum levels of interleukin-6 (IL-6)
interleukin-1
β
(IL-1
β
) and interleukin-8 (IL-8) were measured by enzyme-linked immunosorbent assay (ELISA). immunohistochemistry(IHC) was used to detect nuclear factor-
κ
B (NF-
κ
B) and its inhibitory protein receptor (I
κ
B
α
)
protein expression. Real-time quantitative fluorescence polymerase chain reaction (Real-time PCR) was used to detect I
κ
B
α
NF-
κ
B mRNA expressions.
Result:
2
In the model group
the gastric mucosa was damaged
a large number of inflammatory cells were infiltrated
the serum inflammatory factors increased significantly
mRNA and protein expressions of I
κ
B
α
decreased
and mRNA and protein expressions of NF-
κ
B increased in the gastric tissue (
P
<
0.01). In each treatment group
the inflammation of gastric mucosa was improved to some extent
the serum inflammatory factor was decreased
the mRNA expression of I
κ
B
α
was up-regulated
I
κ
B
α
protein was increased
while the expression of NF-
κ
B mRNA was down-regulated
and NF-
κ
B protein was decreased. The Jinghua Weikang pill treatment group and the high-dose Huangliantang group had the most obvious improvement (
P
<
0.05
P
<
0.01).
Conclusion:
2
Huangliantang for regulating cold and heat based on the harmonization method can effectively alleviate the degree of gastric mucosal injury
and reduce serum inflammatory factors in CNAG rats. The mechanism is related to the up-regulation of I
κ
B
α
mRNA expression
and the down-regulation of NF-
κ
B mRNA expression and NF-
κ
B protein expression in gastric mucosa.
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