The Interference Of Cervical Disc Degeneration On Account Of"Liver Controlling Tendon" Decoction By Pass Through The TNF-α/NF-κB Path

The disc disease is caused by cervical disc degeneration.Intervertebral disc is composed of two major structural components nucleus pulposus and annulus. An important signal of disc degeneration is the loss of proteoglycans in the nucleus pulposus. The disc water lost weakened the pressure, causing disc disease, and led to a series of clinical signs and symptoms, including disc height and T2-weighted MRI signal intensity decreased. It also changes the mechanics of the spine.Currently, there is no good way to block or retard disc degeneration. However, using of Chinese medicine means to prevent disc degenerantion and further study the mechanism is quite significative. However, it is still short of cervical disc degeneration model of large animals. This experiment aim to create a cervical disc degeneration model of the goat cervical intervertebral disc by full endoscopic.Through observing the imaging, histology and molecular biology to compare the changes between the TCM group with the other packet produce pathological in the cervical disc injury.Objective(1). Using the minimally invasive endoscopic techniques to establish an animal model of cervical disc degeneration and observing the preoperative,postoperative and herbs interfere goat cervical disc injury morphological changes, pathological changes and imaging features.(2).Explore the TNF-α and NF-κB signaling transduction pathways in the development of cervical disc degeneration interaction to elucidate the pathogenesis of discogenic pain and provide new therapeutic targets for the disease.(3).Explore the relationship between "Liver Controlling Tendon" theory and NF-κB signaling pathway to promote the modernization of TCM etiology and pathogenesis theory.MethodsSelect eighteen6-month-old Boer-crossbred goats,divided them into three groups,normal sham-operated group (group A),the model blank group (group B) and the model of Huo-Xue-Shu-Jin Decoction treatment group (group C). Group of B and C injury the annulus fibrosus in order to cause the cervical disc degeneration model.Under the general anesthesia, take the neck of right anterolateral,exposure C3/4and C4/5intervertebral disc, endoscope operation system in near the top of the vertebral body under the cartilage endplate left anterolateral annulus fibrosus1/3with a hammer knock2.5mm Kirschner wire lcm. Group A exposure method the same with the group B and C, but do not injury the fibrous ring.One week after the surgery, the animals of group C were fed with the extract of Huo-Xue-Shu-Jin Decoction (according to body surface area of goat), the animals of group A and group B were administered, fed with normal saline.Two months after the administration of anesthesia,cervical disc magnetic resonance T1and T2-weighted imaging were done. The different echo T2-weighted imaging and diffusion (tensor) imaging were seen before the animals were killed. To compare groups injured segments of disc degeneration adjacent normal disc difference by magnetic resonance imaging.The animals were killed immediately then take the interception of cervical specimens rapidly in liquid nitrogen freezing, to save the specimen in-80℃. To examine the groups goats of C3/4and C4/5disc tissue histological and immunochemical examination and observe the process of disc degeneration the Huo-Xue-Shu-Jin Decoction extract infect the cartilage and nucleus pulposus of intervertebral disc endplate morphology, expression of TNF-a, NF-κB, metalloproteinases, apoptosis-related proteins (Bcl-2).Results(1).After two months,MRI examination found the surgical site of cervical disc degeneration of the goats.(2).Naked eye found the intervertebral disc height of blank model group (group B)and the model Huo-Xue-Shu-Jin Decoction treatment group (group C) lost.(3).Histology and MRI showed normal sham-operated group (group A), the blank model group (group B), model Huo-Xue-Shu-Jin Decoction treatment group (group C) C2/3, C3/4, C4/5and C5/6intervertebral disc rear annulus fibrosus injuries was not found during the experiment(4).After surgery for two months, the analysis of MRI found disc of the blank model group (group B)and the model Huo-Xue-Shu-Jin Decoction treatment group (group C) C3/4, C4/5degenerated. The degeneration degree of C3/4and C4/5of blank model group (group B) is more serious than model Huo-Xue-Shu-Jin Decoction treatment group (group C) and the signal of T2is decreased.(5).Pathology observed normal sham-operated group (group A), the model blank group (group B), the model Huo-Xue-Shu-Jin Decoction treatment group (group C).Huo-Xue-Shu-Jin Decoction treatment group compared with the blank model group, the nucleus pulposus full cell degeneration lighter, the Huo-Xue-Shu-Jin Decoction treatment group can delay the degeneration of the nucleus pulposus cells.(6).Tunel staining revealed nucleus pulposus number of apoptotic cells of model blank group (group B) and the Huo-Xue-Shu-Jin Decoction treatment group (group C) were higher than normal sham-operated group (group A) significantly (P<0.05). Herbs interfere the number of apoptotic cells was decreased in the cervical disc nucleus pulposus of Huo-Xue-Shu-Jin Decoction treatment group (group C) significantly, less than the blank model group (group B)(P <0.05).(7).After two months,the sham group (group A), the blank model group (group B), the model Huo-Xue-Shu-Jin Decoction treatment group (group C) were carried out to the polymerase chain reaction (PCR) experiments, there are some gene expression differences of the TNF-α, HIF-1, NF-κB, Bcl-2, MMP-1in the nucleus pulposus(P<0.05).Conclusions(1).After the Kirschner wire injury the lateral of goat cervical disc annulus fibrosus,the early cervical disc degeneration occured.(2).The full endoscope system injury the goats lateral cervical disc annulus fibrosus can make animal model of cervical disc degeneration.(3).TNF-α and NF-κB signaling transduction pathways may play an important role in the development of cervical disc degeneration.(4).TNF-α can activate NF-κB pathway,"Liver Controlling Tendon" Decoction Huo-Xue-Shu-Jin Decoction though the TNF-α and NF-κB signaling transduction pathways relative improve the intervertebral disc nutrition environment and effectively relieve the intervertebral cervical disc degeneration.(5). Huo-Xue-Shu-Jin Decoction can delay apoptosis of cervical disc nucleus pulposus cells relatively.