Research On Effect Of Salvia Cinnamon Twig Five Worm Powder In The Early Stage Of Rats’ Cervical Intervertebral Disc Degeneration

Objective:Cervical Spondylosis is a common and frequently-occurring senile disease and its main pathogenic basis is the cervical intervertebral disc degeneration. TGF-β1、Type Ⅱ collagen、Sox9、BMP-2and aggrecan are observed on their expression and medical intervention in the degenerative intervertebral disc tissue by establishing rats’cervical intervertebral disc degenerative model of unbalanced dynamic and static forces, thus statements are made on the molecular mechanism of Salvia Cinnamon Twig Five Worm Powder mitigating or conversing the intervertebral disc degeneration in the early stage.Method:1. Take1366-month-old SD rats (225-245grams) and establish cervical intervertebral disc degeneration models by destroying spinal dynamic and static stability system from the posterior passage of cervical spine. Grouping:Divide136SD rats into three groups:normal group (32rats), sham-operated group (32rats) and model group (72rats). Make a vertical cut on the nape of each rat in the model group, and dissociate various muscles thoroughly. Cut transversely the deep groups of splenius cervicis and longissimus capitis, cervicis and atlas, cut off costocervicalis and semispinalis capitis. Then cut off the supraspinal ligament and interspinal ligament from C2to C7, in order to establish the intervertebral disc degeneration animal model with unbalanced dynamic and static forces. In the sham-operated group, the rats are sutured after incision of the skin, without excision of the cervical muscles, nor cutting off the supraspinal and interspinal ligaments from C2to C7. Take X-ray photographs of the cervical vertebrae for the three groups. Sacrifice some rats and make pathological sections of the intervertebral disc tissues from C4to C5. Observe their histomorphological changes and degeneration degree with the light microscope and judge success of the animal models.2. Draw randomly5rats from the normal group and sham-operated group respectively, and draw15rats from the model group, and divide randomly3groups:blank group, Salvia Cinnamon Twig Five Worm Powder group and Jingfukang Granules group,5rats in each group. Intragastric administration lasts8weeks for the medicinal administrating groups. Sacrifice the rats after8weeks and take the intervertebral disc tissues from C4to C5. Make five paraffin sections for each intervertebra, and examine expression of TGF-β1and Type Ⅱ collagen by immunocytochemical staining.3. Draw randomly15rats from normal group and sham-operated group respectively. Draw45rats from the model group and divide them into3groups:blank group, Salvia Cinnamon Twig Five Worm Powder group and Jingfukang Granules group,15rats in each group. Sacrifice5rats from each group after establishing models (3months after operation), the fourth and the eighth week after medical intervention respectively. Take the intervertebral disc tissue and examine expression of BMP-2and aggrecan by RT-PCR.Result: 1. There is obvious degeneration on the X-ray photographs of cervical vertebrae among the intervertebral disc degeneration model with unbalanced dynamic and static forces, normal group and the sham-operated group. X-ray photographs of the model group show changes, such as clearly straightness of the cervical curvature, smaller foramen intervertebrale, narrower intervertebral space, osteophytes on the anterior and posterior side of the vertebral body, blurred zygopophysis and joint spaces, etc.2. Lamellar structural derangement can be seen in the cervical intervertebral disc3months after operation in the model group; the chondrocytes move towards the nucleus pulposus area like "Indian file"; collagen fiber arrangement is irregular; part of nucleus pulposus shrinks; collagenous fiber is a little disordered in the nucleus pulposus, but there are no obvious changes of the cervical intervertebral disc in the normal and the sham-operated group.3. The result of immunocytochemical staining indicates that TGF-B1expresses a little in the normal and sham-operated group; TGF-B1expression increases in the model group, II collagen expression decreases, both Salvia Cinnamon Twig Five Worm Powder and Jingfukang Granules can promote expression of TGF-B1and Type II collagen. Chondrocytes increase and the cytoplasm is abundant with secretory granules in it. What’s more, there are protuberances connecting the cells and matrix-like substances accumulating around.4. The result of RT-PCR indicates that Sox9and BMP-2express a little, and aggrecan expresses a lot in the normal and sham-operated groups; expression of Sox9and BMP-2increase continuously the fourth and eighth week after establishing the models, but expression of aggrecan is less; both Salvia Cinnamon Twig Five Worm Powder and Jingfukang Granules can promote expression of Sox9, BMP-2and aggrecan.Conclusion:1. Cervical intervertebral disc degeneration can be induced by unbalanced dynamic and static forces and causing unbalanced cervical vertebrae. The rats’cervical intervertebral disc degenerative models of unbalanced dynamic and static forces are successful.2. TGF-β1, Sox9and BMP-2are closely related with the cervical intervertebral disc degeneration. Salvia Cinnamon Twig Five Worm Powder may promote production of Type II collagen and aggrecan, restrain degradation of extracellular matrix, and relieve damage of the inflammatory factors such as interleukin in the early stage of intervertebral disc degeneration through promoting expression of TGF-β31, Sox9and BMP-2, thus slow down or reverse the intervertebral disc degeneration.