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目的:通过观察川芎嗪对顺铂耳毒性作用的拮抗作用,及对Bax、Bcl-2 mRNA及蛋白表达水平的影响,探讨川芎嗪拮抗顺铂耳毒性作用的部分机制。方法:40只豚鼠随机分为空白组(B)和模型组(M),每组20只,对M组豚鼠进行耳聋模型复制,模型复制成功后,将B组及M组进行第2次随机分组,各自再次随机分为两组,分别为:空白对照组(B1)、空白+川芎嗪组(B2)、模型对照组(M1)、模型+川芎嗪(M2),B2及M2组豚鼠均经腹腔注射川芎嗪注射液,剂量为140 mg/kg,每日1次,连续2周。B1及M1组豚鼠经腹腔注射等量生理盐水,每日1次,连续2周。末次给药后1 h,处死豚鼠,采用荧光定量PCR法对各组豚鼠耳蜗组织中Bax、Bcl-2 mRNA表达水平进行检测;采用western-blot法对各组豚鼠耳蜗组织中Bax、Bcl-2蛋白表达水平进行检测。结果:模型评价,空白组和模型组豚鼠ABR阈值具有显著性差异(P<0.0001);模型组豚鼠耳蜗毛细胞变形明显,排列不均,溶解破坏的毛细胞较多。空白对照组和空白+川芎嗪组豚鼠耳蜗毛细胞偶见凋亡阳性细胞,而模型对照组毛细胞凋亡数量最多,可见大量阳性细胞出现,与空白对照组和空白+川芎嗪组比较,有显著性差异(P<0.001);与模型对照组比较,模型+川芎嗪组豚鼠耳蜗毛细胞凋亡数量明显减少,两者比较,有统计学差异(P<0.05)。与空白对照组和空白+川芎嗪组比较,模型对照组豚鼠耳蜗组织中Bax mRNA及蛋白表达水平显著升高(P<0.001),而Bcl-2 mRNA及蛋白表达水平显著降低(P<0.001);与模型对照组比较,模型+川芎嗪组豚鼠耳蜗组织中Bax mRNA及蛋白表达水平显著降低(P<0.01),而Bcl-2 mRNA及蛋白表达水平显著升高(P<0.05)。结论:顺铂具有诱导豚鼠耳蜗毛细胞凋亡的耳毒性作用,其作用机制与顺铂能够影响Bax、Bcl-2 mRNA及蛋白表达相关,而川芎嗪能够很好的拮抗顺铂这一作用。
OBJECTIVE: To investigate the antagonistic effect of tetramethylpyrazine on cisplatin ototoxicity and its effect on mRNA and protein expression of Bax and Bcl-2, and explore the mechanism of tetramethylpyrazine antagonizing cisplatin ototoxicity. Methods: Forty guinea pigs were randomly divided into blank group (B) and model group (M), with 20 mice in each group. The deafness model was replicated in M guinea pigs. After the model was successfully replicated, They were randomly divided into two groups: blank control group (B1), blank + ligustrazine group (B2), model control group (M1), model group + ligustrazine group Ligustrazine injection by intraperitoneal injection, the dose of 140 mg / kg, once daily for 2 weeks. The B1 and M1 guinea pigs were injected intraperitoneally with normal saline once a day for 2 weeks. At 1 h after the last administration, guinea pigs were sacrificed and the expression of Bax and Bcl-2 mRNA in cochleas of each group were detected by real-time PCR. The expressions of Bax and Bcl-2 Protein expression levels were tested. Results: In the model evaluation, the ABR threshold of guinea pigs in the blank group and the model group was significantly different (P <0.0001). The cochlear hair cells of the model group were obviously deformed and arranged unevenly with more damaged hair cells. Compared with blank control group and blank + ligustrazine group, apoptotic cells were found occasionally in control group and blank + ligustrazine group, while the number of apoptotic cells in model control group was the highest, showing a large number of positive cells. Compared with blank control group and blank + (P <0.001). Compared with the model control group, the number of apoptotic hair cells in guinea pig cochlea significantly decreased in model group and ligustrazine group, and the difference was statistically significant (P <0.05). Compared with the blank control group and the blank + ligustrazine group, Bax mRNA and protein expression in the cochlear of model control group were significantly increased (P <0.001), while the expression of Bcl-2 mRNA and protein was significantly decreased (P <0.001) Compared with the model control group, Bax mRNA and protein expression in cochlear of model + ligustrazine group was significantly decreased (P <0.01), while the expression of Bcl-2 mRNA and protein was significantly increased (P <0.05). CONCLUSION: Cisplatin can induce the ototoxicity of cochlear hair cells in guinea pigs. Its mechanism may be related to the effect of cisplatin on Bax and Bcl-2 mRNA and protein expression, while tetramethylpyrazine can antagonize cisplatin.