1.河南省洛阳正骨医院,河南省骨科医院(河南 洛阳 471000)
谢艳,女,副研究员,主要从事骨科基础研究与药物研究工作
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谢艳, 张云芳, 宁桃丽, 等. 酸枣仁提取物调控cAMP-PKA通路促进小鼠体长增长及大鼠慢波睡眠的机制研究[J]. 上海中医药杂志, 2021,55(5):77-83.
Yan XIE, Yunfang ZHANG, Taoli NING, et al. Mechanism of Ziziphi Spinosae Semen extract promoting bone growth in mice and slow wave sleep of rats by regulating cAMP-PKA pathway[J]. Shanghai Journal of Traditional Chinese Medicine, 2021,55(5):77-83.
谢艳, 张云芳, 宁桃丽, 等. 酸枣仁提取物调控cAMP-PKA通路促进小鼠体长增长及大鼠慢波睡眠的机制研究[J]. 上海中医药杂志, 2021,55(5):77-83. DOI: 10.16305/j.1007-1334.2021.2004008.
Yan XIE, Yunfang ZHANG, Taoli NING, et al. Mechanism of Ziziphi Spinosae Semen extract promoting bone growth in mice and slow wave sleep of rats by regulating cAMP-PKA pathway[J]. Shanghai Journal of Traditional Chinese Medicine, 2021,55(5):77-83. DOI: 10.16305/j.1007-1334.2021.2004008.
目的,2,探讨酸枣仁提取物调控cAMP-PKA通路影响小鼠体长增长及大鼠慢波睡眠的具体机制。,方法,2,①48只昆明种雄性小鼠随机分为6组,分别为空白对照组(灌胃去离子水,并于末次灌胃后侧脑室注射灭菌的0.9%NaCl溶液)、蓝墨水组(灌胃去离子水,并于末次灌胃后侧脑室注射过滤灭菌的蓝墨水)、用药组(灌胃酸枣仁提取物水溶液,并于末次灌胃后侧脑室注射过滤灭菌的蓝墨水)、阳性对照组(灌胃酸枣仁皂苷A水溶液,并于末次灌胃后侧脑室注射过滤灭菌的蓝墨水)、Gi/o蛋白抑制剂+用药组(灌胃酸枣仁提取物水溶液,并于末次灌胃后侧脑室注射加有过滤灭菌蓝墨水的PT)和5-HT1AR抑制剂+用药组(灌胃酸枣仁提取物水溶液,并于末次灌胃后侧脑室注射加有过滤灭菌蓝墨水的P-MPPF)。各组小鼠灌胃相应药物,连续20 d。测定小鼠体长差异,采用ELISA法测定小鼠血清生长激素(GH)水平、脑组织5-羟色胺1A受体(5-HT1AR)活性、PKA的含量和Gi/o蛋白的含量;Western blot 法测定小鼠脑组织5-HT1AR蛋白的表达。②36只SD大鼠随机分成6组,分组及给药方式同小鼠,连续给药3 d后,采用WXL睡眠自动分析系统,分析大鼠睡眠时相差异。,结果,2,①与空白对照组和蓝墨水组比较,用药组、Gi/o蛋白抑制剂+用药组、5-HT1AR抑制剂+用药组、阳性对照组小鼠体长均明显增长(,P,<,0.01);与空白对照组比较,用药组和阳性对照组小鼠血清GH水平、5-HT1AR蛋白含量、Gi/o蛋白含量均明显升高(,P,<,0.01),5-HT1AR活性升高(,P,<,0.05),PKA含量明显降低(,P,<,0.01);与蓝墨水组比较,用药组和阳性对照组小鼠血清GH水平明显升高(,P,<,0.01),Gi/o蛋白含量、5-HT1AR活性升高(,P,<,0.05),PKA含量明显降低(,P,<,0.01);与Gi/o蛋白抑制剂+用药组和5-HT1AR抑制剂+用药组比较,用药组和阳性对照组小鼠血清GH水平、5-HT1AR蛋白含量均明显升高(,P,<,0.01),5-HT1AR活性升高(,P,<,0.05)。用药组与阳性对照组比较,小鼠体长、GH水平、5-HT1AR活性、PKA的含量、Gi/o蛋白含量和5-HT1AR蛋白含量差异均无统计学意义(,P,>,0.05)。②与空白对照组比较,用药组大鼠总睡眠时间明显增长(,P,<,0.01);阳性对照组大鼠总睡眠时间增长(,P,<,0.05)。与蓝墨水组比较,用药组大鼠总睡眠时间明显增长(,P,<,0.01);阳性对照组大鼠总睡眠时间增长(,P,<,0.05)。与Gi/o蛋白抑制剂+用药组和5-HT1AR抑制剂+用药组比较,用药组和阳性对照组大鼠总睡眠时间增长(,P,<,0.05)。用药组与阳性对照组比较,大鼠总睡眠时间差异无统计学意义(,P,>,0.05)。,结论,2,酸枣仁提取物可以通过提高Gi/o蛋白的表达,使之通过cAMP-PKA途径促进5-HT1AR在脑内的含量和活性提高,还可延长慢波睡眠,进而促进GH的分泌发挥促进骨骼增长的生物学效应。
Objective,2,To explore the mechanism of Ziziphi Spinosae Semen extract affecting the mice growth in length and slow wave sleep of rats by regulating the cAMP-PKA pathway.,Methods,2,①Forty-eight Kunming male mice were randomly divided into 6 groups: blank control group (intragastrically administered with deionized water, and injected with sterilized 0.9% NaCl in the lateral ventricle after the last intragastric administration), blue ink group (intragastrically administered with deionized water, and injected with filtered and sterilized blue ink in the lateral ventricle after the last intragastric administration), medication group (intragastric administration of the Ziziphi Spinosae Semen extract, and injection of filtered and sterilized blue ink in the lateral ventricle after the last intragastric administration), positive control group (intragastric administration of jujube saponin A, and injection of filtered and sterilized blue ink in the lateral ventricle after the last intragastric administration), Gi/o protein inhibitor + medication group (intragastric administration of Ziziphi Spinosae Semen extract, and injection of PT with filtered and sterilized blue ink in the lateral ventricle after the last intragastric administration) and 5-HT1AR inhibitor + medication group (intragastric administration of Ziziphi Spinosae Semen extract, and injection of P-MPPF with filtered and sterilized blue ink in the lateral ventricle after the last intragastric administration). The mice in each group were given corresponding drugs by intragastric administration for 20 days. The body length of the mice was measured. The level of serum growth hormone (GH), the activity of 5-hydroxytryptamine 1A receptor (5-HT1AR) in brain tissue, the contents of PKA and Gi/o protein in brain tissue were measured by ELISA. The expression of 5-HT1AR protein in mouse brain tissue was determined by Western blot. ②Thirty six SD rats were randomly divided into 6 groups (6 rats in each group). The grouping and administration methods were the same as those of mice. The sleep phase differences of rats were analyzed by WXL sleep automatic analysis system after 3 days of continuous administration.,Results,2,①Compared with the blank control group and the blue ink group, the body length of mice increased significantly in the medication group, the Gi/o protein inhibitor + medication group, the 5-HT1AR inhibitor + medication group and the positive control group (,P,<,0.01); Compared with the blank control group, the serum GH level, 5-HT1AR protein content and Gi/o protein content of mice in the medication group and the positive control group increased significantly (,P,<,0.01), and the 5-HT1AR activity was also increased (,P,<,0.05), while the PKA content was decreased significantly (,P,<,0.01) ; Compared with the blue ink group, the serum GH level was significantly increased (,P,<,0.01) and Gi/o protein content and 5-HT1AR activity were increased(,P,<,0.05), while the PKA content was decreased significantly (,P,<,0.01) in the medication group and the positive control group; Compared with the Gi/o protein inhibitor + medication group and the 5-HT1AR inhibitor + medication group, the serum GH level, the protein content of 5-HT1AR were increased significantly(,P,<,0.01) and 5-HT1AR activity was increased (,P,<,0.05) in the medication group and the positive control group. There were no significant differences in body length, GH level, 5-HT1AR activity, PKA content, Gi/oprotein content and 5-HT1AR protein content between the medication group and the positive control group (,P,>,0.05). ②Compared with the blank control group, the total sleep period was increased significantly in the medication group (,P,<,0.01) and the positive control group (,P,<,0.05). Compared with the blue ink group, the total sleep period was increased significantly in the medication group (,P,<,0.01) and the positive control group (,P,<,0.05). Compared with the Gi/o protein inhibitor + medication group and the 5-HT1AR inhibitor + medication group, the total sleep period increased significantly (,P,<,0.05) in the medication group and the positive control group. There was no significant difference in the total sleep period between the medication group and the positive control group (,P,>,0.05).,Conclusion,2,Ziziphi Spinosae Semen extract can increase the expression of Gi/o protein, increase the content and activity of 5-HT1AR in the brain through cAMP-PKA pathway, prolong slow wave sleep and promote the secretion of growth hormone to play the biological effect of promoting bone growth.
酸枣仁提取物酸枣仁皂苷A骨骼增长cAMP-PKA通路Gi/o蛋白5-HT1A受体模型小鼠模型大鼠慢波睡眠
Ziziphi Spinosae Semen extractJujube saponin Abone growthcAMP-PKA pathwayGi/o protein5-HT1A receptormodel micemodel ratsslow wave sleep
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