Full
Outline
To investigate the regulatory effect of Modified Chaihu Shugan Decoction（加味柴胡疏肝汤）on epileptic mice through the protein kinases 1（RIP1）/protein kinases 3（RIP3）/mixed lineage kinase domain-like protein（MLKL）pathway.
Sixty KM mice were randomly divided into normal group，model group，low dose group of Modified Chaihu Shugan Decoction［19.52 g/（kg· d）］，medium dose group of Modified Chaihu Shugan Decoction［39.04 g/（kg·d）］，high dose group of Modified Chaihu Shugan Decoction［78.08 g/（kg·d）］and carbamazepine group［30 mg/（kg·d）］. Epilepsy models were prepared using pilocarpine hydrochloride. After continuous gavage for two weeks，the general condition and electroencephalogram（EEG）of the mice were observed. Hematoxylin-eosin（HE）staining was used to observe pathological and morphological changes. TdT-mediated dUTP Nick-End Labeling（TUNEL）method was used to detect the apoptosis rate of neural cells. Quantitative real time polymerase chain reaction（qRT-PCR）and Western Blot（WB）measured the mRNA and protein expression levels of RIP1，RIP3，MLKL，tumor necrosis factor-α（TNF-α），interleukin-1β（IL-1β）and interleukin-6（IL-6）in the hippocampus of the mice.
Compared with the normal group，the body weight of the model group mice was decreased significantly；EEG can clearly observe continuous epileptic waves；HE staining showed that many neurons were pyknotic，with reduced cell volume and irregular morphology. The boundaries between the nucleus and cytoplasm were unclear，and the staining deepened；The positive rate of apoptotic cells significantly increased；The relative expression levels of RIP1，RIP3，MLKL，TNFα，IL-1βand IL-6 were significantly increased. After drug treatment，the weight of mice in each group increased slightly compared to that of the model group. The frequency and intensity of epileptic discharges in EEG waves were significantly reduced. The positive rates of apoptotic cells in the medium dose and high dose groups of Modified Chaihu Shugan Decoction and carbamazepine group were significantly lower than those in the model group，with statistical significance. The relative expression levels of mRNA and protein of RIP1，RIP3，MLKL，TNF-α，IL-1βand IL-6 in the high dose group of Modified Chaihu Shugan Decoction and the carbamazepine group were significantly lower than those of the model group，indicating statistical significance.
Modified Chaihu Shugan Decoction may exert its anti epileptic effect through the RIP1/RIP3/MLKL signaling pathway.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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