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Effect of Medicated Serum Prepared with Chang'an Ⅰ Prescription on Sensitized Mast Cell Degranulation / 中国实验方剂学杂志
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 48-54, 2020.
Article in Chinese | WPRIM | ID: wpr-872758
ABSTRACT

Objective:

To observe the influence of Chang'an Ⅰ prescription drug-containing serum on IgE-mediated RBL-2H3 cell degranulation model, and explore the mechanism of Chang'an Ⅰ prescription in inhibiting RBL-2H3 activation degranulation and releasing inflammatory mediators with v-yes-1 Yanaguchi sarcoma viral related oncogene homolog (Lyn)/spleen tyrosine protein kinase (Syk)/mitogen-activated protein kinase (MAPK) signal pathway.

Method:

Preparation for Chang'an Ⅰ prescription serum. Animal group, SD male rats were randomly divided into Chang'an Ⅰ prescription serum high, medium, low dose, and blank control groups with 10 rats in each group. Dosage: 10 mL·kg-1 distilled water was given to blank control group, while Chang'an Ⅰ prescription serum high, medium and low dose groups were respectively given to the Chang'an Ⅰ prescription concentrated crude drug with concentration of 1.15,2.30,4.60 g·kg-1, respectively once a day for 7 days continuously and then blood was taken from aorta ventralis and centrifuged. Ketotifen as the positive control drug. Mast cells are counted with toluidine blue staining. Cellular release of β-aminohexose was detected by colorimetric method. Contents of MCT, TNF-α, MCP-1 and histamine were measured by enzyme-linked immunosorbent assayELISA) kits, Lyn/Syk/MAPK protein levels were detected by immunoblotting.

Result:

For cell activation and degranulation, compared with the blank control group, the model group had more cell degranulation (P<0.05), compared with model group, the cell degranulation rate of each dose group of Chang'an Ⅰ prescription decreased (P<0.05). The release rate of β-hexosamine in each dose group of Chang'an Ⅰ prescription decreased significantly (P<0.01). For the release of active mediators, compared with the blank control group, the contents of histamine, MCT, TNF-α and MCP-1 all increased in the model group (P<0.01), compared with the model group, the contents in each dose group of Chang'an Ⅰ prescription all decreased significantly (P<0.01). Compared with the normal group, the phosphorylation levels of Lyn and Syk, extracellular regulatory protein kinase 1/2(ERK1/2), c-Jun N-terminal kinase (JNK), and mitogen-activated protein kinase p38 increased in the model group (P<0.05). Compared with the model group, the Lyn, Syk and ERK1/2, JNK and p38 protein phosphorylation levels reduced in Chang'an Ⅰ prescription group (P<0.05).

Conclusion:

Chang'an Ⅰ prescription drug-containing serum down-regulates the phosphorylation levels of proteins Lyn, Syk, and ERK1/2, JNK, and p38, inhibits RBL-2H3 cell activation and degranulation, reduces the release of cytokines and chemokines, such as histamine, MCT, TNF-α and MCP-1, it may be one of its mechanisms for treating IBS-D visceral hypersensitivity.

Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Experimental Traditional Medical Formulae Year: 2020 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Experimental Traditional Medical Formulae Year: 2020 Type: Article