| The incidence of spinal cord injury is increasing with the development of society; the incidence of SCI in Beijing was 60 per million people in 2002, while the incidence was 6.8 per million people in the 1980s. It has already become a medical problem urgently need to be solved since it mainly attacks the young and middle-aged population; it costs a huge amount of money for its treatment and daily care; and the lethality and disability rate stay at high level, more than 50% of the patients was suffering paraplegia. So far, however, there is no really effective treatment for SCI all around the world. High dose of methylprednisolone during the early stage of SCI hast definite curative effect, but with a series of side effects, inspite of the fact that it is still questionable in some literatures. The reasons behind this phenomenon lie in the extremely complicated mechanism of SCI. Except for the"primary mechanical injuries", there is still a series of complicated biochemical reactions taking place in the injury region, that is,"secondary injury". The"secondary injury"involves microcirculation dysfunction, electrolyte disorder, lipid peroxidation, inflammation, apoptosis and glial scar and so on. The current symptomatic treatments aim at one or two of the injury mechanisms, and have little effects.The traditional Chinese medicine compound prescriptions have complex ingredients, aiming at the multifaceted injury mechanisms with relatively lower price and fewer side effects. And the effects of some traditional Chinese medicine or the active agents of them against SCI have already been confirmed by some literatures. Besed on that, the development of new traditional Chinese medicine compound prescription against SCI seems rather promising in this field.Objective: To find the pharmacological theoretical evidence of the constitution of the compound prescription based on the Molecular Composition Theory of Chinese Medicinal Materials (MCTCMM).Methods: The related literatures and basic traditional Chinese medicine theory were reviewed to select the promising crude drugs. The selected crude drugs were grouped based on the Molecular Composition Theory of Chinese Medicinal Materials, all the groups were extracted by water decoction method and 70% ethanol reflux method, and the contents of the active agents were tested by the TLC and HPLC methods.Results:①Salvia Miltiorrhiza, Panax Notoginseng and Rhizome Chuanxiong were select as the promising crude drugs.②The contents of the active agents were generally higher in the groups extracted by 70% ethanol reflux method.③ Co-extracted with Panax Notoginseng, the extraction rate of salvianolic acid B stayed almost the same level in the water decoction group, but increased by 28.35% in the ethanol reflux group; Co-extracted with Rhizome Chuanxiong, the extraction rate of salvianolic acid B stayed almost the same level in both water decoction group and ethanol reflux group; Co-extracted with Panax Notoginseng and Rhizome Chuanxiong, the extraction rate of salvianolic acid B stayed almost the same level in the water decoction group, but increased by 23.69% in the ethanol reflux group.④TanshinoneⅡA can hardly be detected in the water decoction group; Co-extracted with Panax Notoginseng, the extraction rate of tanshinoneⅡA increased by 16.66% in the ethanol reflux group; Co-extracted with Rhizome Chuanxiong, the extraction rate of salvianolic acid B decreased by 57.34% in the ethanol reflux group; Co-extracted with Rhizome Chuanxiong, when added Panax Notoginseng, the extraction rate increased by 54.97%.⑤Co-extracted with Panax Notoginseng, the extraction rate of ferulic acid increased by 116.91% in the water decoction group and 64.73% in the ethanol reflux group; Co-extracted with Salvia Miltiorrhiza the extraction rate of ferulic acid increased by 381.94% in the water decoction group and 171.84% in the ethanol reflux group; Co-extracted with Panax Notoginseng and Salvia Miltiorrhiza together, the extraction rate of ferulic acid increased by 255.25% in the water decoction group and 82.56% in the ethanol reflux group.⑥Co-extracted with Salvia Miltiorrhiza, the extraction rate of Panax Notoginseng saponins R1 decreased by 38.52% in the water decoction group, but stayed almost the same level in the ethanol reflux group; Co-extracted with Rhizome Chuanxiong, the extraction rate of Panax Notoginseng saponins R1 increased by 128.79% in the water decoction group, but stayed almost the same level in the ethanol reflux group; Co-extracted with Salvia Miltiorrhiza and Rhizome Chuanxiong, the extraction rate of Panax Notoginseng saponins R1 stayed almost the same level in the water decoction group, but increased by 95.38% in the ethanol reflux group.⑦Co-extracted with Salvia Miltiorrhiza, the extraction rate of ginsenoside Rg1 decreased by 150.49% in the water decoction group, but increased by 20.57% in the ethanol reflux group; Co-extracted with Rhizome Chuanxiong, the extraction rate of ginsenoside Rg1 increased by 45.38% in the water decoction group, but decreased by 50.52% in the ethanol reflux group; Co-extracted with Salvia Miltiorrhiza and Rhizome Chuanxiong, the extraction rate of ginsenoside Rg1 decreased by 40.33% in the water decoction group and 17.69% in the ethanol reflux group.⑧Co-extracted with Salvia Miltiorrhiza, the extraction rate of ginsenoside Rb1 decreased by 53.78% in the water decoction group, but increased by 59.13% in the ethanol reflux group; Co-extracted with Rhizome Chuanxiong, the extraction rate of ginsenoside Rb1 increased by 44.31% in the water decoction group, but decreased by 48.25% in the ethanol reflux group; Co-extracted with Salvia Miltiorrhiza and Rhizome Chuanxiong, the extraction rate of ginsenoside Rb1 decreased by 50.75% in the water decoction group, but increased by 17.39% in the ethanol reflux group.Conclusion: Salvia Miltiorrhiza, Panax Notoginseng, Rhizome Chuanxiong, Salvia Miltiorrhiza + Panax Notoginseng + Rhizome Chuanxiong extracted with ethanol reflux method were selected as promising groups.②The active agents were extracted the most in the Salvia Miltiorrhiza+Panax Notoginseng +Rhizome Chuanxiong extracted by ethanol reflux method group Objective: To study the treatment effect of the compound prescriptions chosen in part one on the acute spinal cord injury models. Methods: 132 female SD rats were divided into 6 groups: Salvia Miltiorrhiza group, Panax Notoginseng group, Rhizome Chuanxiong group, Salvia Miltiorrhiza + Panax Notoginseng + Rhizome Chuanxiong group, control group, and sham-operation group, with 22 rats in each group. Intragastric administration was given to all the rats in the compound prescription groups from ten days before Allen's contusion spinal cord injury model was preformed on the rats once a day until the rats were sacrificed. BBB score were used to assess the recovery of the locomotive function of the rats at 1d, 3d, 5d, 1w, 2w, 4w, 6w, 8w after injury. Histopathological changes were observed by the HE staining method, and expression of GFAP was observed by the immunofluorescence methods at 1w, 2w, 4w, 8w after injury (4 rats in each group). The contents of caspase-3, IL-1β, and IL-10 in the injured spinal cord tissue were detected by the ELISA method, and the contents of SOD, MDA and iNOS in serum and injured spinal cord tissue were detected at 3d after injury (6 rats in each group).Results: The content of SOD in serum was higher in the compound prescription group compared with the control group (P<0.05), while the contents of MDA and iNOS in serum were lower in the compound prescription group compared with the control group (P<0.05). There was no statistical difference of other indicators between the compound prescription groups and the control group. Conclusion:Under the present combination and extraction method of the compound prescription, better recovery of the neurological functions was not observed in each medication groups. |
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