Research On Mechanism Of Sanzi Yangqin Decoction Controlling Asthmatic Based On CysLTs Path

Objective:Research on mechanism of Sanzi Yangqin Decoction controlling asthmatic based on CysLTs pathMethodology:Take sixty-five 4-8-week-old C57BL mice, all female, divided into the normal group and the experimental group. The normal group is composed of 4 mice and the experimental group, further divided into the high-, middle-and low-dose groups of Sanzi Yangqin Decoction (a kind of traditional Chinese medicine) (SZ-H group, SZ-M group and SZ-L group) (totally 36 mice), the Montelukast group (Western medicine) (5 mice) and the model group (20 mice). The model group falls into the control group and the atomized group (5 mice). The Sanzi Yangqin Decoction groups and the Montelukast group consists of ordinary mice and the rest, C57BL mice. And the Sanzi Yangqin Decoction groups and the Montelukast group are the blood drawing groups. Mice in the model group receive the intraperitoneal injection of OVA NS at concentration 0.1%mixed with aluminium hydroxide gel (ratio:1:4, PH7.4) on the day immediately before the experiment, the 7th and 14th days of the experiment, lml of such solution for each mouse, and lml of NS at concentration 0.9% for the normal group. Next, mice in the model group are put into the glass box, and receive spray nasal attack with 2ml OVA NS at concentration 5% once a day,30min for each day and lasting for 14 days. Mice in SZ-H group, SZ-M group and SZ-L group receive gavage of 1ml Sanzi Yangqin Decoction of 10.8g·kg-1·d-1,5.4g·kg-1·d-1 and 2.7g·kg-1·d-1 respectively (equal to 10,5 and 2.5 times of adult dose). And mice in the Montelukast group receive gavage of 1ml montelukast of 0.004g·kg-1·d-1. Then adopt the HE staining method to observe the lung tissue of mice in the model group to see if the modeling is successful. If so, separate the bronchial epithelial cells and the smooth muscle cells of the mice. Detect the EOT-1 and EOT-3 levels in the lung tissue homogenate and the IP3 and DAG levels in the smooth muscle cells using the ELISA method. And detect the expression of CysLTs 1mRNA and CysLTs2mRNA in the smooth muscle cells using RT-PCR method.Result:1. General reaction:Mice in the normal group are sensitive and have a high spirit, good movement, stable breath, normal feeding, glossy fur, and gradually growing weight. Mice in the model group are in low spirit on the 7th day and seriously low on the 14th day. They are short of breath and slow in movement. They curl up. Their fur becomes sparse and lackluster. They eat and drink less than those in the normal group, but their see basically no growth in weight after the modeling.Mice in each medication administration team are relatively high in sprit and have stable breath, glossy fur and fast movement.Pathologic condition:The normal group:The alveolar structure is normal, with no bronchial epithelium cell falling off or out of shape. No inflammatory cells are seen aggregating to lead to microglandular hyperplasia and no inflammatory exudation observed; the model group:Lots of bronchial epithelium cells fall off or go out of shape, with many inflammatory cells seen aggregating. Eosinophilic granulocytes are obviously increased and lots of goblet cells observed in the pulmonary alveoli. The alveolar wall swells, and goes red and thin. Some individual alveolar spaces are expanded.3. CysLTRlmRNA and CysLTR2mRNA:Compared with the model group, CysLTs 1 genetic expression in cells of mice in the Montelukast group and SZ-M group is reduced, which is of statistical significance (p<0.05). CysLTs 1 genetic expression in cells of mice in the SZ-H group is obviously reduced, which is of prominent statistical significance (p<0.01). It’s worth noting that, medicines and therapies applied to the Montelukast group, SZ-M group and the SZ-H group can lower the CysLTs 1 genetic expression in cells of mice. CysLTs 1 genetic expression in cells of mice in the SZ-L group is free from obvious change, having no statistical significance. Compared with the model group, CysLTs 2 genetic expression in cells of mice in the Montelukast group and SZ-H, SZ-M and SZ-L are not obviously different, which is of no statistical significance.4. EOT-1 and EOT-3:Compared with the model group, the Eotaxinl level in the cell supernatant of mice in the Montelukast group and SZ-H group is obviously reduced, which is of prominent statistical significance (p<0.01). Eotaxinl level in the cell supernatant of mice in the SZ-M group is reduced to some extent, which is of statistical significance (p<0.05). Note that medicines and therapies applied to the Montelukast group, SZ-M group and the SZ-H group can lower the Eotaxinl level in the cell supernatant of mice. Eotaxin1 level in the cell supernatant of mice in the SZ-L group is free from noted change, having no statistical significance. Compared with the model group, the Eotaxin3 level in the cell supernatant of mice in the Montelukast group and SZ-H group is obviously reduced, which is of prominent statistical significance (p<0.01). Eotaxin3 level in the cell supernatant of mice in the SZ-M group is lowered to some extent, which is of statistical significance (p<0.05). Note that, medicines and therapies applied to the Montelukast group, SZ-M group and the SZ-H group can lower the Eotaxin3 level in the cell supernatant of mice. Eotaxin3 level in the cell supernatant of mice in the SZ-L group is free from noted change, having no statistical significance.5. IP3 and DAG:Compared with the model group, the IP3 level in the cell supernatant of mice in the Montelukast group and SZ-H group is obviously reduced, which is of prominent statistical significance (p<0.01). IP3 level in the cell supernatant of mice in the SZ-M group is lowered to some extent, which is of statistical significance (p<0.05). Note that medicines and therapies applied to the Montelukast group, SZ-M group and the SZ-H group can lower the IP3 level in the cell supernatant of mice. IP3 level in the cell supernatant of mice in the SZ-L group is free from noted change, having no statistical significance. Compared with the model group, the DAG level in the cell supernatant of mice in the Montelukast group, SZ-M group, SZ-H group and the SZ-L group is free from noted change, having no statistical significance.Conclusion:1、Sanzi Yangqin Decoction could reduce the EOT-1and EOT-3 levels in the lung tissue homogenate of mice with asthma.2、Sanzi Yangqin Decoction could reduce the CysLTR1mRNA, CysLTR2mRNA, IP3 and DAG levels in the smooth muscle cell of mice with asthma.3. Low-dose OVA works better for asthma modeling.4. Sanzi Yangqin Decoction controls the asthmatic attack by restraining the CysLTs path.