仙灵骨葆胶囊对去势大鼠骨组织H3K36me3、H3K9me3及JMJD2A表达的影响

Effect of Xianlinggubao capsule on H3K36me2,H3K9me2,and JMJD2A expression in bone tissues of ovariectomized rats

目的研究仙灵骨葆胶囊对去势大鼠骨组织中组蛋白H3第36位点赖氨酸三甲基化(histone H3 lysine36 trimethylation,H3K36me3)、组蛋白H3第9位点赖氨酸三甲基化(histone H3 lysine9 trimethylation,H3K9me3)和组蛋白去甲基化酶JMJD2A表达水平的影响。方法参照随机数字表法将24只SD雌性大鼠分为4组:空白对照组、阴性对照组、实验组、阳性对照组,每组6只。手术处理:空白对照组切除卵巢旁部分脂肪,另外3组大鼠经手术切除双侧卵巢。干预措施:术后1周,实验组给予仙灵骨葆胶囊(30.85 mg/100 g),阳性对照组给予福美加(11.7 mg/100 g),其余2组给予同体积蒸馏水。持续给药3个月后,HE染色观察骨微结构;采用逆转录-聚合酶链式反应(reverse transcription-polymerase chain reaction,RT-PCR)测组蛋白H3、JMJD2A mRNA表达,免疫印迹试验(Western blot)检测大鼠股骨髁骨组织的H3K36me3、H3K9me3、JMJD2A蛋白表达。结果空白组、实验组及阳性对照组骨微结构优于阴性对照组;空白组、实验组及阳性对照组骨组织中JMJD2A表达高于阴性对照组(P<0.05),空白组、实验组及阳性对照组各组组蛋白H3mRNA表达低于阴性对照组(P<0.05),空白组、实验组及阳性对照组各组H3K9me3及H3K36me3蛋白表达低于阴性对照组(P<0.05)。结论仙灵骨葆胶囊可改善去势大鼠骨质疏松症状,其机制可能与其提高去势大鼠骨组织中JMJD2A表达,抑制H3K36me3和H3K9me3的表达有关。

Objective To investigate the effect of Xianlinggubao capsule on the expression of H3K36me3(histone H3 lysine 36 trimethylation),H3K9me3(histone H3 lysine 9 trimethylation),and JMJD2A in bone tissues of ovariectomized rats.Methods Twenty-four female SD rats were randomly divided into four groups:blank control group,negative control group,experimental group,and positive control group,with 6 rats in each group.The rats in the blank control group retained bilateral ovaries and only some fat beside the ovaries were excised.Osteoporosis model was established in the other 3 groups of rats by resection of bilateral ovaries.One week after the operation,rats in the experimental group received 30.85 mg/100 g of Xianlinggubao capsule.Rats in the positive control group received 11.7 mg/100 g of Fumeijia.Rats in the blank control and the negative control group received the same volume of distilled water at the same time.After 3 months of continuous administration,HE staining was used to observe the microstructure of bone;RT-PCR was used to detect the mRNA expressions of histone H3 and JMJD2A.Western blotting(WB)was used to detect the protein expressions of H3K36me3,H3K9me3,and JMJD2A.Results The bone microstructure in the blank group,experimental group,and positive control group was superior to that in the negative control group.The expression of JMJD2A in bone tissue of blank group,experimental group,and positive control group was significantly higher than that in the negative control group(P<0.05).The expression of histone H3 mRNA in blank group,experimental group,and positive control group was lower than that in the negative control group(P<0.05).The protein expressions of H3K9me3 and H3K36me3 in the blank group,experimental group and positive control group were lower than that in the negative control group(P<0.05).Conclusion Xianlinggubao capsule relieves osteoporosis symptoms in ovariectomized rats.Its mechanism may be related to the increase of JMJD2A expression in bone tissues of ovariectomized rats and the inhibition of H3K36me3 and H3K9me3 expression.