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1.甘肃中医药大学,兰州 730000
2.深圳市宝安纯中医治疗医院,广州 深圳 518101
曹思齐,在读硕士,从事中医四诊及病证合参临床应用研究,E-mail:382983532@qq.com
王凤仪,博士,教授,硕士生导师,从事中医四诊理论研究与分析工作,E-mail:wfy@gszy.edu.cn
收稿日期:2020-10-13,
网络出版日期:2020-11-19,
纸质出版日期:2021-03-05
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曹思齐,王凤仪,汤胜男等.芍药汤对大肠湿热型溃疡性结肠炎大鼠结肠组织中CD14,FADD,Caspase-8表达的影响[J].中国实验方剂学杂志,2021,27(05):1-7.
CAO Si-qi,WANG Feng-yi,TANG Sheng-nan,et al.Effect of Shaoyaotang on Expressions of CD14, FADD and Caspase-8 in Colonic Tissues of Rats with Large Intestinal Damp-heat Syndrome of Ulcerative Colitis[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(05):1-7.
曹思齐,王凤仪,汤胜男等.芍药汤对大肠湿热型溃疡性结肠炎大鼠结肠组织中CD14,FADD,Caspase-8表达的影响[J].中国实验方剂学杂志,2021,27(05):1-7. DOI: 10.13422/j.cnki.syfjx.20202401.
CAO Si-qi,WANG Feng-yi,TANG Sheng-nan,et al.Effect of Shaoyaotang on Expressions of CD14, FADD and Caspase-8 in Colonic Tissues of Rats with Large Intestinal Damp-heat Syndrome of Ulcerative Colitis[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(05):1-7. DOI: 10.13422/j.cnki.syfjx.20202401.
目的
2
观察芍药汤对大肠湿热型溃疡性结肠炎(UC)模型大鼠血清中细胞黏附分子-1(ICAM-1),转化生长因子-
β
1
(TGF-
β
1
)含量及病灶结肠组织中白细胞分化抗原14(CD14),Fas相关死亡结构域蛋白(FADD)与天冬半胱氨酸蛋白酶-8(Caspase-8) mRNA及蛋白表达的影响。
方法
2
将SPF级Wistar大鼠80只随机分为正常组10只、造模组70只,采用病证结合的方式复制大肠湿热型UC大鼠,即高脂高糖辛辣饮食+2,4-二硝基苯磺酸(DNBS)结合乙醇复合法造模,造模成功后,按随机数字表法将造模组分为模型组、柳氮磺砒啶(SASP)组及芍药汤低、中、高剂量组,每组14只,给予柳氮磺嘧啶0.2 g·kg
-1
·d
-1
,芍药汤低、中、高剂量(6,12,24 g·kg
-1
·d
-1
)灌胃,正常组及模型组给予等体积生理盐水灌胃,连续21 d。采用酶联免疫吸附测定(ELISA)法检测血清ICAM-1,TGF-
β
1
含量,实时荧光定量聚合酶链式反应(Real-time PCR)检测结肠组织CD14,FADD,Caspase-8 mRNA表达,蛋白免疫印迹法(Western blot)检测结肠组织CD14,FADD,Caspase-8蛋白表达。
结果
2
与正常组比较,模型组大鼠血清中ICAM-1含量明显升高,TGF-
β
1
含量明显降低(
P
<
0.05);CD14,FADD,Caspase-8 mRNA及蛋白相对表达量明显升高(
P
<
0.05)。与模型组比较,芍药汤中、高剂量组及SASP组大鼠血清中ICAM-1含量明显降低,而芍药汤低、中、高剂量组及SASP组大鼠血清中TGF-
β
1
含量明显升高(
P
<
0.05);各干预组CD14,FADD,Caspase-8 mRNA及蛋白表达量明显降低(
P
<
0.05),并以芍药汤高剂量组及SASP组更为明显。
结论
2
芍药汤对大肠湿热型UC大鼠具有一定干预作用,其机制可能与抑制CD14,FADD及Caspase-8 mRNA及蛋白的表达有关。
Objective
2
To observe the effect of Shaoyaotang on the contents of cell adhesion molecule-1 (ICAM-1) and transforming growth factor-
β
1
(TGF-
β
1
) in serum of large intestine damp-heat syndrome of ulcerative colitis (UC) in rats, and the gene and protein expressions of leukocyte differentiation antigen14 (CD14), Fas-related death domain protein (FADD) and cysteinyl aspartate specific protease-8 (Caspase-8) in the focal colon tissue.
Method
2
A total of 80 SPF Wistar rats were randomly divided into the blank group (
n
=10) and modeling group (
n
=70). The large intestine damp-heat syndrome of UC rats was replicated by the combination of disease and syndrome, which was high-fat, high-sugar and spicy diets combined with 2, 4-dinitrobenzene sulfonic acid (DNBS) and ethanol. After successful modeling, the modeled groups were divided into model group, sulfasalazine (SASP)control group, and low, medium and high-dose Shaoyaotang groups by the method of random number table, with14 rats in each group. Low, medium and high doses of Sulfasalazine 0.2 g·kg
-1
·d
-1
and Shaoyaotang (6, 12, 24 g·kg
-1
·d
-1
)were given by gavage. The blank group and the model group were given equal volume of normal saline for 21 days. The contents of serum ICAM-1 and TGF-
β
1
were detected by enzyme-linked immunosorbent assay (ELISA), the expressions of CD14, FADD and Caspase-8 mRNA in colon tissues were detected by Real-time quantitative polymerase chain reaction (Real-time PCR), and the expressions of CD14, FADD and Caspase-8 protein in colon tissues were detected by Western blot.
Result
2
Compared with the blank group, the serum ICAM-1 level in the model group were significantly increased, whereas the content of TGF-
β
1
were significantly decreased (
P
<
0.05). The relative expression levels of CD14, FADD, Caspase-8 mRNA and protein were significantly increased (
P
<
0.05). Compared with the model group, the content of ICAM-1 in the serum of the rats in the medium, high-dose Shaoyaotang groups and the SASP group were significantly decreased, while the content of TGF-
β
1
in the serum of the rats in the low, medium, high-dose Shaoyaotang groups and the SASP group were significantly increased (
P
<
0.05). The expression levels of CD14, FADD, Caspase-8 mRNA and protein in each intervention group were significantly decreased (
P
<
0.05), especially in the high-dose Shaoyaotang group and the SASP group.
Conclusion
2
Shaoyaotang has a certain intervention effect on UC rats with large intestine damp-heat syndrome, and its mechanism may be related to the inhibition of CD14, FADD and Caspase-8 genes and proteins expression.
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