桂枝加葛根汤对大鼠颈椎间盘纤维环细胞超微结构的影响

Effect of Guizhi plus Gegen Decoction on Ultrastructural Changes of Intervertebral Disc Annulus Fibrosus Cells

目的观察桂枝加葛根汤对大鼠颈椎间盘纤维环细胞超微结构的影响。方法采用贴壁筛选法分离培养大鼠颈椎间盘纤维环细胞,通过桂枝加葛根汤进行干预,利用光镜和电镜观察未干预和桂枝加葛根汤含药血清不同浓度干预后纤维环细胞的细微和超微结构变化特点。结果光镜观察显示纤维环细胞大部分不规则多角形,少量呈星形,表面突起丰富;细胞核大而完整,为椭圆形;电镜观察结果显示:经不同浓度含药血清干预后,空白组细胞形态各异,大多呈椭圆形;细胞核大,偏位且不规则,异染色质散在,部分线粒体空泡化,粗面内质网扩张;中药低剂量组见线粒体增多、浓缩密度增高,胞质内粗面内质网扩张,偶见脂滴或糖原;中药中剂量组见内质网扩张增多,密度增高,可见较多中等密度的蛋白质沉淀,线粒体增多,密度增高呈不规则囊状,细胞核大小不一;中药高剂量组见粗面内质网增多,扩张明显,镜下见较多高电子密度的蛋白质沉淀,核偏位,可见较多线粒体和分泌颗粒。结论纤维环细胞经过不同浓度桂枝加葛根汤含药血清干预后的超微结构表现为:(1)中药中、高剂量组细胞内粗面内质网较空白组及低剂量组明显增多,出现电子密度较大的蛋白质沉淀,以高剂量组最为明显;(2)桂枝加葛根汤在防止因纤维环细胞退变而引起的超微结构改变中起重要的作用。

Objective To observe the effect of Guizhi plus Gegen Decoction（GGD） on ultrastructural changes of intervertebral disc annulus fibrosus cells.Methods Rats＇ intervertebral disc annulus fibrosus cells were isolated and cultured using adherence wall screening method.After annulus fibrosus cells were intervened by GGD,the microstructure and ultrastructural features of untreated annulus fibrosus cells and annulus fibrosus cells treated by GGD containing serum at different concentrations were observed under optical microscope and electron microscope.Results Under optical microscope,most annulus fibrosus cells showed irregular polygons and few in star shape with rich superficial ecphyma.The nuclei were oval,large and complete.Under electron microscope,most cells in the blank group were oval after intervened by GGD containing serum at different concentrations.The nucleus was large,deviated,and irregular,the heterochromatin scattered diffusely,partial mitochondria vacuolized,and rough endoplasmic reticulum dilated.In the low dose GGD group,increased mitochondria and condensed density could be seen.The rough endoplasmic reticulum were expanded,lipid drops or glycogen could be occasionally seen.In the middle dose GGD group,increased endoplasmic reticulum expansion and condensed density could be seen.More medium density protein sediment could be seen.Increased mitochondria with condensed density could be seen,showing irregular cystic form with various sizes nucleus.In the high dose GGD group,increased rough endoplasmic reticulum with obvious expansion could be seen.More high density protein sediment could be seen.The nuclei were deviated.More mitochondria could be seen with secretory granules in them.Conclusions After intervened by GGD containing serum at different concentrations,the ultrastructure of annulus fibrosus cells were manifested as follows：（1） The endoplasmic reticulum increased more in the middle and high dose GGD groups than in the blank group and the low dose GGD group.Greater density protein sediment occurred,especially in the high dose GGD group.（2） GGD played an important role in preventing ultrastructural changes induced by the degeneration of annulus fibrosus cells.