| Objective:To research the effect of chinese medicine for ephedrine addiction animal，and supply theoretical basis to develop no side effects and withdrawal effects chinese herba preparation for addiction effect, the weight of brain and body, histological structure, expression of Bax protein, c-Fos and TGF-β1of frontal lobe of ephedrine addiction mice, activities of Superoxide dismutase(SOD), Catalase(CAT) and content of Maleic Dialdehyde(MDA) in brian of ephedrine addiction mice were measured.Methods:Age of5days Kunming filial mice were chosen，randomly divided into control group and another4model groups(model group, chinese medicine A group, chinese medicine B group and complex prescription group). The model groups were intraperitoneal injected escalation doses (2.0,3.0,4.0g/L) of ephedrine in l~15d with0.2ml, two times per day(9:00am，16:00pm), the control group was injected same amount of saline to establish ephedrine addiction animal model，th en were given a gavage of chinese medicine for withdrawal respectively for15days with0.2ml, two times per day(9:00am，16:00pm), the control group was given a gavage of saline similarly．The samples were collected at5，10and15d，then the body weight and the brain weight were measured，the change of frontal lobe structure was observed by optical microscopy and the changes of mice frontal lobe thickness were measured，t he activities of SOD, CAT as well as content of MDA were detected by colcrimrtry. In addition, the expression of Bax, c-Fos and TGF-β1were measured by immunohistochemistry in frontal lobe of mice.Results:1. The chinese medicine affected body weight and brain weight of ephedrine withdrawal mice. The body weight and brain weight of model group in5~15d both are lower than that of control group after giving a gavage of chinese medicine for withdrawal, the body weight and brain weight of chinese medicine A group, chinese medicine B group and complex prescription group all are higher than model group, however, lower than control group. 2. The chinese medicine affected the antioxidant enzymes activity in brain of ephedrine withdrawal mice．The activity of SOD and CAT increased in the early time，then decreased with the extension of time, and activity is lower than control group all along, very significant(P<0.01) when compared with control group. Compared with model group, SOD activity of chinese medicine B group and complex prescription group in5d, chinese medicine A group in10d, significant difference (P<0.05) chinese medicine B group and complex prescription group in10d, chinese medicine groups in15d, very significant(P<0.01). Compared with model group, CAT activity of chinese medicine A group and chinese medicine B group in5d, significant difference (P<0.05) chinese medicine A group and chinese medicine B group in10d and15d, complex prescription group in5~15d, very significant(P<0.01).The content of MDA in brain of ephedrine withdrawal mice increased all along, very significant(P<0.01) when compared with control group. Compared with model group, the content of MDA of chinese medicine A group, chinese medicine B group and complex prescription group in10d, chinese medicine A group in15d, significant difference (P<0.05) chinese medicine B group and complex prescription group in15d, very significant(P<0.01).3. The chinese medicine remited the frontal lobe shrinking of ephedrine addiction mice. After ephedrine addiction, the frontal lobe thickness of mice was lower than control group all along, very significant(P<0.01). Compared with model group, chinese medicine A group, chinese medicine B group in5d and10d, complex prescription group in5d, the thickness of frontal lobe increased, no significant difference(P>0.05) chinese medicine A group, chinese medicine B group in15d, complex prescription group in10d, significant difference(P<0.05) complex prescription group in15d, very significant(P<0.01).4. The chinese medicine relieved damage of fontal lobe histological structure in ephedrine addiction mice. Fontal lobe cortex histological structure of model group was unclear, boundary line was fuzzy. Cell scattered, shape changed obviously, cell number decreased seriously, cell arranged irregularly, and the damage of external pyramidal layer, internal granular layer, internal pyramidal layer and polymorphic layer were most obviously. After chinese medicine withdrawal, the shape of part cell recoverd, every layer was more clear than before.5. The chinese medicine affected expression intensity of Bax protein, c-Fos and TGF-β1in fontal lobe cortex of ephedrine addiction mice. The expression intensity of Bax protein, c-Fos and TGF-β1of model group were higher than control group, very significant (P<0.01). After chinese medicine withdrawal, the expression intensity of specific protein weakened varying degrees in chinese medicine groups: compared with model group, the expression intensity of Bax protein and c-Fos of chinese medicine A group in5d, significant difference(P<0.05) other groups were very significant(P<0.01) the expression intensity of TGF-β1of chinese medicine A group in5d, significant difference(P<0.05) other groups were very significant(P<0.01).6. The chinese medicine affected positive expression cell number of Bax protein, c-Fos and TGF-β1in fontal lobe cortex of ephedrine addiction mice. The positive expression cell number of Bax protein, c-Fos and TGF-β1in fontal lobe cortex in model group was more than control group, very significant(P<0.01). After chinese medicine withdrawal, the positive expression cell number of each group decreased: compared with model group, the positive expression cell number of Bax protein of chinese medicine B group and complex prescription group in10d, chinese medicine A group and chinese medicine B group in15d, significant difference(P<0.05) complex prescription group in15d, very significant(P<0.01) the positive expression cell number of c-Fos of chinese medicine B group and complex prescription group in5d, chinese medicine B group in10d，chinese medicine A group in15d， significant difference(P<0.05), chinese medicine B group in15d，complex prescription group in10d and15d, very significant(P<0.01) the positive expression cell number of TGF-β1in complex prescription group in5d, chinese medicine A and B group in10d， significant difference(P<0.05), complex prescription group in10d and each chinese medicine group in15d, very significant(P<0.01).Conclusion:The body weight and brain weight of ephedrine addiction mice both were lower than control group. The antioxidant system of mice organism was destroyed, lead to the activity of antioxidant enzyme decline, free radicals and lipid peroxide exceeded body clearance ability. In addition, as a kind of harmful factor from outside, ephedrine infected fontal lobe histological structure of mice fontal lobe cortex, disturbed and retarded cortical development, enhanced the expression intensity of Bax protein, c-Fos and TGF-β1, strengthened cell apoptosis, enhanced damage of brain tissue as well. Chinese medicine A and B contain vary kinds of microelement and active material, which could improve body immunity, regulate body metabolism, perfect microcirculation and cerebral blood flow, protect nerve cell. Astragalus polysaccharide, tetramethylpyrazine, ferulic acid and flavone so on are able to clear radical, increase SOD activity, promote producing the enzyme which can clear radical, in addition, they could clear superoxide radical, peroxide subunits, hydrogen peroxide and hydroxyl radical, at the same time, reduce lipid peroxidation damage effectively because of good antioxidant function. Chinese medicine A and B could restrain NO level in damaged tissue, withstand Ca2+, reduce calcium overload, inhibit inflammatory reaction and cell apoptosis, they could be antianxiety and antidepression, central sedation and analgesia as well. By the way above, chinese medicine could reduce the brain damage of ephedrine addiction mice, increase activity of antioxidant enzyme, reduce the level of lipid peroxidation, decrease the expression intensity of Bax protein, c-Fos and TGF-β1in fontal lobe cortex, repair brain tissue damage and relieve withdrawal symptom caused by ephedrine. |
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