Acetyl-11-Keto-Beta-Boswellic Acid Activates the Nrf2/HO-1 Signaling Pathway in Schwann Cells to Reduce Oxidative Stress and Promote Sciatic Nerve Injury Repair.

Boswellia is a traditional medicine for bruises and injuries. Its main active ingredient, acetyl-11-keto-beta-boswellic acid, has antioxidant and antiapoptotic effects. In this experiment, we used Sprague-Dawley rats to make a sciatic nerve injury model to detect the transcription factor NF-E2-related factor 2/heme oxygenase 1 signaling pathway and apoptosis, combined with clinical indicators, for testing whether acetyl-11-keto-beta-boswellic acid can reduce oxidative stress and promote sciatic nerve repair. Our results showed that acetyl-11-keto-beta-boswellic acid administration promoted myelin regeneration and functional recovery in the rat sciatic nerve, reduced lipid peroxidation levels, upregulated the expression of various antioxidant enzymes and enhanced enzyme activity, decreased the expression levels of apoptosis-related proteins, and promoted nuclear translocation of the transcription factor NF-E2-related factor 2 protein. In vitro studies revealed that acetyl-11-keto-beta-boswellic acid reduced H2O2-induced reactive oxygen species production, restored mitochondrial membrane potential, upregulated the expression of various antioxidant enzymes, and downregulated apoptosis-related indicators in Schwann cells, and these therapeutic effects of acetyl-11-keto-beta-boswellic acid were reversed after ML385 treatment in Schwann cells. In summary, acetyl-11-keto-beta-boswellic acid alleviates oxidative stress and apoptosis caused by sciatic nerve injury in rats by activating the transcription factor NF-E2-related factor 2/heme oxygenase 1 signaling pathway, promotes the recovery of sciatic nerve function in rats, and is a promising therapeutic agent to promote sciatic nerve repair by alleviating excessive oxidative stress.

Network pharmacology integrated with experimental validation revealed potential molecular mechanisms of Camellia nitidissima C. W. Chi in the treatment of lung cancer.

ETHNOPHARMACOLOGICAL RELEVANCE: Camellia nitidissima C.W.Chi (CNC), an ethnomedicine mainly distributed in Southern China's Guangxi Zhuang Autonomous Region, is known as "Panda in plants" and "Camellias Queen" due to its golden blossom. CNC has been applied as a traditional folk medicine in cancer therapy. AIM OF THE STUDY: This study utilized network pharmacology analysis combined with experimental validation to identify the substance basis and potential molecular mechanism of CNC against lung cancer. MATERIALS AND METHODS: The active ingredients of CNC were identified based on published literature. The associated potential targets of CNC in lung cancer treatment were predicted using integrated network pharmacology analysis and molecular docking. The underlying molecular mechanism of CNC in lung cancer were validated in human lung cancer cell lines. RESULTS: A total of 30 active ingredients and 53 targets of CNC were screened. An enrichment analysis of Gene Ontology (GO) revealed that the effects of CNC in lung cancer mainly involve protein binding, regulation of cell proliferation and apoptosis, and signal transduction. KEGG pathways analysis suggested that CNC might exert cancer suppression effects mainly through pathways in cancer, PI3K/AKT signaling pathway. Molecular docking revealed that CNC has high affinity for binding of EGFR, SRC, AKT1, and CCND1 to the key active ingredients including luteolin, kaempferol, quercetin, eriodictyol and 3'4-O-dimethylcedrusin. In in vitro experiments, CNC played the inhibitory roles in lung cancer cells by inducing cell apoptosis, causing G0/G1 and S cell cycle arrest, increasing intracellular ROS levels, and promoting the apoptotic proteins Bax and Caspase-3. Meanwhile, CNC also regulated the expression of core proteins EGFR, SRC, and AKT. CONCLUSION: These results comprehensively clarified the associated substance basis and underlying molecular mechanism of CNC against lung cancer, which would be contributed to develop promising anti-cancer pharmaceuticals or therapeutic approaches for lung cancer therapy.

Dracorhodin Perchlorate Regulates the Expression of Inflammatory Cytokines through the TLR4 Pathway and Improves Skin Wound Healing in Diabetic Rats.

Background: Dragon's blood is a natural medicine with hemostatic and blood-activating effects and is used to promote wound healing. Dracorhodin perchlorate (DP) is a stable form of dracarhod and is used as a substitute for cochinchinenin. DP promotes the proliferation of rat fibroblasts and promotes wound healing in rats. Methods: DP ointment (0.2 mg/mL) was applied to the skin wounds of nondiabetic and diabetic rats, and the skin of the wound was collected. Wound healing rate, H&E staining, Masson staining, TLR4 pathway, related inflammatory factors, nitric oxide synthase, and so forth were detected. Results: DP treatment alleviated the prolonged inflammatory cell infiltration time and the increase in the TLR4 pathway and inflammatory factors caused by diabetes. DP also promoted wound healing by increasing eNOS protein expression and NO content in the later stage of wound healing. Conclusion: DP promotes wound healing in diabetic rats by regulating the TLR4 pathway and related inflammatory factors. Therefore, adjuvant treatment of DP can be developed for diabetic wound healing.

Based on proteomics to explore the mechanism of mecobalamin promoting the repair of injured peripheral nerves.

Mecobalamin is commonly used in the adjuvant intervention of various peripheral nerve injuries. Actin cytoskeleton plays a role in the regeneration of myelin and axon. Therefore, the purpose of this study was to explore the possibility of mecobalamin regulating actin cytoskeleton in repairing nerve injury. In this study, a crush injury on the right sciatic nerve of two groups of rats (12 in each group) was established. The control group was only given normal saline (i.g.), and the intervention group was given mecobalamin 1 mg/kg (i.g.). The rats were sacrificed on 28th day and the injured nerves were collected for proteomics. The result shows that regulation of actin cytoskeleton pathway changed significantly. The expression of protein Vav1 was verified by Western blot and immunofluorescence. In the intervention group, the nerve fiber structure was complete, the axons were dense and symmetrical, and the myelin sheath was compact and uniform in thickness. The positive rate of myelin basic protein and ßⅢ-tubulin was higher than that in the control group. The findings of the study show that mecobalamin regulates the actin cytoskeleton in the repair of nerve damage and upregulates Vav1 in the regulation of actin cytoskeleton pathway.

[Technical elements and clinical application of umbilical refining of heat-sensitive moxibustion].

Under the guidance of the theory and technique of arrival of qi of heat-sensitive moxibustion, the technical elements of umbilical refining of moxibustion proposed in The Guide to Medicine by LI Yan were analyzed. It is believed that the parameters of moxibustion temperature and time are the key points. The standard of "quantitative moxa" is established to achieve the appropriate moxibustion temperature and moxibustion time. The umbilical refining of heat-sensitive moxibustion is established to reappear the magic effect of LI Yan's umbilical refining of moxibustion. The umbilical refining of heat-sensitive moxibustion is recommended for stomachache, diarrhea, constipation, dysmenorrhea, impotence, etc. with significant curative effect.

[Effects of Yiqi Huayu prescription on knee cartilage degeneration in HIF-1alpha gene knockout mice].

OBJECTIVE: To study the role of hypoxia-inducible factor 1 alpha (HIF1alpha) on knee cartilage degeneration,and to explore the effects and mechanisms of Chinese herbal compound Yiqi Huayu prescription on HIF-1alpha gene knockout mice on knee cartilage degeneration. METHODS: The 4-month and 6-month HIF-1alpha gene knock out mice were obtained by interbreeding, and divided into HIF-1alpha +/+ 4-month mice group,HIF-1alpha -/- 4-month mice group,HIF-1alpha +/+ 6-month mice group and HIF-1alpha -/- 6-month mice group, 3 mice in each group. And then the 2-month-old HIF-1alpha gene knock out mice were randomly divided into Yiqi Huayu prescription group and physiological saline group. There were 6 mice in each group. After 2 months' drug administration, the knee joint of mice was collected, and the Mankin score were evaluated; Safranine-fast green staining, HE Staining, and immunohistochemistry analysis for VEGF, Col X, Col II, MMP-13 and Sox-9 were performed erespectively. RESULTS: (1) Compared to the results in the HIF-1alpha+/+ mice groups, the HIF-1alpha-/- mice developed aging related cartilage loss and bony tissue appearance, cartilage defects increased,and cells reduced. In HIF-1alpha-/-4-month mice and 6-month mice group, the expresion of Col II and Sox9 decreased, and the expression of Col X, MMP-13 and VEGF increased. (2) Compared to the physiological saline group, the ossification and defect of knee joint cartilage reduced of mice in the Yiqi Huayu prescription group, the cartilage cell distribution was more uniform, and the total number of cells increased. The expression of type II collagen and Sox9 protein increased, expression of Col X, MMP-13 and VEGF protein decreased of mice in the Yiqi Huayu prescription group. CONCLUSION: The knee cartilage degenerates in the HIF-1alpha cKO mice, and the degeneration increased with age adding. The Yiqi Huayu prescription can delay the degeneration of knee cartilage of HIF-1alpha cKO mice.

Potential therapeutic efficacy of curcumin in liver cancer.

PURPOSE: Liver cancer, one of the most common cancers in China, is reported to feature relatively high morbidity and mortality. Curcumin (Cum) is considered as a drug possessing anti-angiogenic, anti-inflammation and anti-oxidation effect. Previous research has demonstrated antitumor effects in a series of cancers. MATERIALS AND METHODS: In this study the in vitro cytotoxicity of Cum was measured by MTT assay and pro-apoptotic effects were assessed by DAPI staining and measurement of caspase-3 activity. In vivo anti-hepatoma efficacy of Cum was assessed with HepG2 xenografts. RESULTS: It is found that Cum dose-dependently inhibited cell growth in HepG2 cells with activation of apoptosis. Moreover, Cum delayed the growth of liver cancer in a dose-dependent manner in nude mice. CONCLUSIONS: Cum might be a promising phytomedicine in cancer therapy and further efforts are needed to explore this therapeutic strategy.

[Professor Shi Qi's experience of applying herbal paste for treating chronic musculoskeletal conditions].

Professor Shi Qi is a famous traditional Chinese medicine doctor specializing in orthopaedics and traumatology, who has formatted a set of systematic protocols for the diagnosis and treatment of chronic musculoskeletal conditions. When it is time for using tonics in winter, he advocates applying herbal paste for treating chronic musculoskeletal diseases. This paper introduces Professor Shi Qi's commonly used prescription for treating chronic musculoskeletal conditions and puts forward demands and understandings in concocting herbal paste, experience in herbal paste for treating chronic musculoskeletal diseases such as cervical spondylosis, lumbar disc herniation, lumbar spinal stenosis, lumbar muscle strain, ankylosing spondylitis, osteoporosis, knee osteoarthritis and avascular necrosis of femoral head, and the advantages of herbal paste for treating chronic musculoskeletal conditions as opposed to alternative treatments.

[The relationship between hypoxia-inducible factor-1α and cartilage degeneration in osteoarthritis: a review].

Osteoarthritis is one of the most common diseases seen in clinical practice. Cartilage survives in the hypoxic microenvironment. Hypoxia-inducible factor-1α (HIF-1α) is a key nuclear transcription factor which mediates the hypoxic response of cells. HIF-1α gene is an important regulator for the adaptation of articular cartilage to the hypoxic environment. It is important for formation of articular cartilage, energy metabolism and matrix synthesis. If the HIF-1α gene is knocked out, the cartilage can not maintain their normal morphology and function, which may lead to cartilage degeneration, and result in diseases such as osteoarthritis. Chinese herbal medicines can regulate the expression of HIF-1α gene and supply a therapy method for osteoarthritis. In this paper, the authors review the situation of the correlation between HIF-1α and osteoarthritis cartilage degeneration examined in recent years.

Inhibitory effect of YQHYRJ recipe on osteoblast differentiation induced by BMP-2 in fibroblasts from posterior longitudinal ligament of mice.

Ossification of posterior longitudinal ligament (OPLL) is a common disease in Asian countries. Osteoblast differentiation in posterior longitudinal ligamentous fibroblast is a pathologic basis of OPLL. Nowadays, an effective pharmacotherapy for OPLL is still hunted for. YQHYRJ Recipe (YQHYRJ) is designed based on traditional Chinese medicine (TCM) theories, and previous clinic trials reported its effect on relieving syndromes of cervical spondylopathy. To clarify the YQHYRJ effect of OPLL on a cellular level, we induced mice fibroblasts from posterior longitudinal ligaments to differentiate into osteoblasts by human recombinant BMP-2, and treated them with YQHYRJ and its three sub-compounds: YQ, HY and RJ. YQHYRJ and the sub-compounds reduced the increase of fibroblast proliferation, mineralization, type I collagen secretion induced by BMP-2 via MTT, alizarin red staining and immunochemical examination. Moreover, these agents inhibited BMP-2 induced upregulation of ossification-related genes ALP, Col I and OC as well as BMP signal molecules Smad1, Smad 5 and Runx2 mRNA expression. These results suggested YQHYRJ to be effective in inhibiting osteoblast differentiation induced by BMP-2 in fibroblasts from posterior longitudinal ligament. YQHYRJ might be a promising medicine for preventing OPLL disease.

[Effects of Chinese herbal medicine Yiqi Huayu Recipe on apoptosis of dorsal root ganglion neurons and expression of caspase-3 in rats with lumbar nerve root compression].

OBJECTIVE: To observe the effects of Yiqi Huayu Recipe, a Chinese compound herbal medicine, on apoptosis of dorsal root ganglion (DRG) neurons and expression of caspase-3 in rats after lumbar nerve root compression injury. METHODS: A total of 40 male Sprague-Dawley rats were randomly allocated into 4 groups: control group, untreated group, Methylcobal group and Yiqi Huayu Recipe group. Surgery was performed on rats of untreated group, Methylcobal group and Yiqi Huayu Recipe group to place a micro-silica gel on right L4 DRG, while control group received skin and paravertebral muscle incision only. Rats in Methylcobal group and Yiqi Huayu Recipe group were given Methylcobal by intramuscular injection and Yiqi Huayu Recipe intragastrically respectively. Rats in control group and untreated group received saline intragastrically as equal amount as Yiqi Huayu Recipe group. The compressed nerve roots were harvested at the 10th day after treatment. Apoptosis of DRG neurons was detected by terminal deoxynucleotidyl transferase-mediated nick-end labeling. Caspase-3 activity and mRNA expression in compressed nerve roots were detected with spectrophotography and real-time polymerase chain reaction respectively. RESULTS: Apoptosis of DRG neurons was significantly increased in the rat model. The apoptosis index of untreated group was higher than that of control group (P<0.01). Yiqi Huayu Recipe and Methylcobal could reduce the apoptosis of DRG neurons, and both groups showed a lower apoptosis index than untreated group (P<0.01). Caspase-3 activity and its gene expression were significantly increased in untreated group. The levels of caspase-3 activity and its gene expression in untreated group were higher than those in control group (P<0.05 or P<0.01). Yiqi Huayu Recipe and Methylcobal could reduce the overexpression of caspase-3 mRNA, and statistically significant differences were found between the untreated group and Yiqi Huayu Recipe group or Methylcobal group (P<0.01). CONCLUSION: Lumbar nerve root compression results in overexpression of caspase-3 in nerve root tissue and increase of DRG neuron apoptosis. Yiqi Huayu Recipe can inhibit the overexpression of caspase-3 and alleviate the apoptosis of DRG neurons after nerve injury.

[Effects of Yiqi Huayu Decoction on brain-derived neurotrophic factor expression in rats with lumbar nerve root injury].

OBJECTIVE: To observe the effects of Yiqi Huayu Decoction, a compound traditional Chinese herbal medicine, on brain-derived neurotrophic factor (BDNF) expression in lumbar nerve root and soleus of rats after lumbar nerve root compression injury. METHODS: A total of 120 male SD rats were randomly allocated into sham-operated group, untreated group, methycobal injection group and Yiqi Huayu Decoction group. Surgery were performed on rats of untreated group, methycobal injection group and Yiqi Huayu Decoction group by placing a micro-silica gel on right L(4) dorsal root ganglia, while rats in sham-operated group received skin and paravertebral muscle incision only. Rats in the methycobal injection group and the Yiqi Huayu Decoction group were given intramuscular injection of methycobal and intragastric administration of Yiqi Huayu Decoction respectively. Rats in the sham-operated group and the untreated group received intragastric administration of normal saline of equal volume as Yiqi Huayu Decoction. The compressed nerve roots and right soleus were harvested after 10-, 30- and 60-day treatment. Microstructure of the compressed nerve roots were revealed under a transmission electron microscope. BDNF protein expression in the compressed nerve roots and mRNA expression in right solues were detected with enzyme-linked immunosorbent assay (ELISA) and real-time quantitative reverse transcription-polymerase chain reaction respectively. RESULTS: The protein expression level of BDNF in compressed nerve root in the untreated group was slightly changed with no significance as compared with sham-operated group. BDNF protein expression in the Yiqi Huayu Decoction group was gradually increased and reached the peak level after 60-day treatment with significant difference as compared with the untreated group (P<.01). The expression of BDNF mRNA in soleus was significantly up-regulated 10 days after nerve root injury (P<0.01), and was slowly decreased to the normal level at day 60 (P>0.05). Compared with the untreated group, BDNF mRNA expression in soleus muscle in the Yiqi Huayu Decoction group was significantly down-regulated after 10-day treatment (P<0.01), while was increased after 60-day treatment (P<0.01). CONCLUSION: Yiqi Huayu Decoction can inhibit the increase of BDNF expression at the early phase after nerve injury, which may have something to do with alleviation of neuropathic pain, and increase BDNF expression at the late phase to promote nerve regeneration.

[Mechanism of Yiqi Huayu Bushen recipe in treating cervical syndrome in rats with qi deficiency, blood stasis and kidney deficiency].

OBJECTIVE: To explore the mechanism of Yiqi Huayu Bushen Recipe (YHBR), a compound traditional Chinese herbal medicine, in treating cervical syndrome (CS) with qi deficiency, blood stasis and kidney deficiency in rats. METHODS: A total of 30 three-month-old female Sprague-Dawley rats were randomly divided into normal control group, untreated group and YHBR group; there were ten rats in each group. The rat model of cervical syndrome with qi deficiency, blood stasis and kidney deficiency was established by combining disease and syndrome models. After one-month YHBR treatment in YHBR group, all rats were sacrificed. Serum, plasma and cervical intervertebral discs were detected and observed by radio-immunology, histopathology, immunohistochemistry, etc. RESULTS: Compared with those in the normal control group, rats in the untreated group showed obvious signs of deficiency in vital energy, such as tiredness, ptosis, few movement, bluish-purple tongue and tail and weight loss; the weight of uterus and appendages, and the ratio of cyclic adenosine monophosphate/cyclic guanosine monophosphate were decreased; hemorheological parameters and the expression of alpha-granular membrane protein (CD62p) were increased; the content of serum estradiol was decreased. YHBR could improve the above indexes except for the weight of uterus and appendages. HE staining showed annulus fibrosus with cracks, diminish of pulposus nucleus, and decrease in the height of intervertebral disc and the thickness of end-plate in the untreated group; and YHBR could improve the changes of cervical intervertebral discs, but no obvious changes in end-plate. Type II collagen protein (Col2a1)was distributed in every part of the annulus fibrosus in normal control group, but was decreased in untreated group; YHBR could increase Col2a1 expression in annulus fibrosus as compared with the untreated group. The expressions of Col2a1 and tissue inhibitor of metalloproteinase-1 (TIMP-1) mRNAs were decreased in untreated group as compared with the normal control group (P<0.01); the expressions of Col2a1 and TIMP-1 mRNAs were higher and matrix metalloproteinase-13 mRNA was lower in the YHBR group than those in the untreated group (P<0.05, P<0.01). CONCLUSION: YHBR may improve the condition of cervical syndrome with qi deficiency, blood stasis and kidney deficiency by regulating immune system, coagulation system and endocrine system, and delay the degeneration of cervical intervertebral disc by regulating extracellular matrix and metalloproteases in intervertebral disc.

[Mechanism of Yiqi Huayu Bushen Recipe in treating cervical syndrome with kidney deficiency in rats].

OBJECTIVE: To research the action mechanism of Yiqi Huayu Bushen Recipe (YHBR), a compound of traditional Chinese herbal medicine, in treating cervical syndrome (CS) with kidney deficiency in rats. METHODS: A total of 30 three-month-old female Sprague-Dawley rats were randomly divided into normal control group, CS with kidney deficiency model group (untreated group) and YHBR group, with 10 rats in each group. Rats in the normal control group received no treatment, while rats of the other two groups underwent resection of both cervical muscles and ovaries to establish the model of CS with kidney deficiency. Three months after surgery, rats in the YHBR group were intragastrically administered YHBR for one month. Another one month later, all rats were sacrificed. The content of serum estradiol (E2) was detected by radio-immunoassay; contents of plasma cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) were tested, and the ratio of cAMP/cGMP was also calculated. Hemorheology was detected by Weissenberg's method; expression of alpha-granular membrane protein (CD62p) was detected by flow cytometry; HE staining was used to detect the histopathology of cervical intervertebral disc degeneration; type II collagen protein was detected by immunohistochemistry and aggrecan-1 (Agc1), type II procollagen gene, Col2a1, tissue inhibitor of metalloproteinase-1 (TIMP-1) and matrix metalloproteinase-13 (MMP-13) mRNAs were detected by fluorescent quantization polymerase chain reaction. RESULTS: Compared with the untreated group, rats in the YHBR group showed an obvious increase in serum E2 content (P<0.05), and an increase in plasma cAMP and cGMP content without significant difference; hemorheological parameters and percentage of CD62p expression were significantly decreased in the YHBR group (P<0.05, P<0.01). YHBR could improve the degeneration of cervical intervertebral discs, decrease the Miyamoto scores (P<0.05), and increase the type II collagen. The expressions of Agc1, Col2a1 and TIMP-1 mRNAs were significantly increased and MMP-13 mRNA significantly decreased in YHBR group (P<0.05, P<0.01). CONCLUSION: YHBR may improve CS with kidney deficiency and delay the degeneration of cervical intervertebral disc by regulating the immune-metabolism system, coagulation system and endocrine system.

[Establishment of a rat model of cervical syndrome with qi deficiency, blood stasis and kidney deficiency].

OBJECTIVE: To establish a rat model of cervical syndrome with qi deficiency, blood stasis and kidney deficiency. METHODS: A total of 30 three-month-old female Sprague-Dawley rats were randomly divided into normal group, cervical syndrome group and cervical syndrome with qi deficiency, blood stasis and kidney deficiency group (combined group), with 10 rats in each group. Rats in the normal group received no treatment, rats in cervical syndrome group underwent resection of cervical muscles and ligaments as unbalanced dynamic and static animal model, and rats in combined group underwent resection of both cervical muscles and ovaries, swimming and irregular diet, and injection of adrenal cortex hormone and adrenaline two and a half months after resection as combined model. The qi deficiency, blood stasis and kidney deficiency were determined by observing behaviors and physical signs of the rats, detecting the contents of plasma cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), the hemorrheology, the expression of alpha-granular membrane protein (CD62p) and the serum estradiol (E(2)) content. The aggrecan-1, type II procollagen gene (Col2a1), matrix metalloproteinases-13 (MMP-13) and tissue inhibitor of metalloproteinases-1 (TIMP-1) mRNAs in cervical intervertebral discs were detected by histopathology, immunohistochemistry and real-time polymerase chain reaction. The cataplasia of the intervertebral discs was determined by detecting the histopathology, the expressions of type II collagen and type X collegen proteins, and the expressions of aggrecan-1 (Agc1), type II procollagen gene (Col2a1), matrix metalloproteinase-13 (MMP-13) and tissue inhibitor of metalloproteinase-1 (TIMP-1) mRNAs. RESULTS: Compared with those in the normal group and cervical syndrome group, the rats in the combined group were noted with obvious signs of deficiency of vital energy, such as depression, tiredness, ptosis, obvious weight loss and blue tail. And the ratio of cAMP/cGMP was decreased; the reducing viscosity was significantly up-regulated; the expression of CD62p was increased; the content of serum E(2) was decreased; the intervertebral disc structure was destructed; the cervical intervertebral disc was more seriously deteriorated. There exhibited a decrease in type II collagen protein expression, an increase in type X collagen protein expression, as well as decreases of Agc1, Col2a1 and TIMP-1 mRNA expressions in intervertebral disc, and the expression of MMP-13 mRNA was noted an increase. CONCLUSION: The rat model of cervical syndrome with qi-deficiency, blood stasis and kidney deficiency is established. Qi deficiency, blood stasis and kidney deficiency can aggravate cervical intervertebral disc degeneration.

[Establishment of a rat model of cervical syndrome with kidney deficiency].

OBJECTIVE: To establish a rat model of cervical syndrome (CS) with kidney deficiency. METHODS: A group of 30 three-month-old female Sprague-Dawley rats were randomly divided into normal control group, CS group and CS with kidney deficiency group (combined group), with 10 rats in each group. Rats in the normal control group received no treatment, rats in the CS group underwent only resection of cervical muscles and ligaments as unbalanced dynamic and static animal model, and rats in combined group underwent resection of both cervical muscles and ovaries as kidney deficiency model. Serum and cervical intervertebral discs were collected. Kidney deficiency was determined by observing the morphologic changes of uterus and appendages, detecting the weight of uterus and appendages and the content of serum estradiol (E(2)). The degeneration of intervertebral discs was determined by detecting the histopathology, the expressions of type II collagen and type X collagen proteins, and the expressions of aggrecan-1 (Agc1), type II procollagen gene (Col2a1), matrix metalloproteinase-13 (MMP-13) and tissue inhibitor of metalloproteinases-1 (TIMP-1) mRNAs. RESULTS: Compared with those in the normal control group and CS group, the rats in the combined group were noted with the uterus atrophied, the caliber of oviduct thinned, the weight of uterus and appendages diminished obviously, the content of serum E(2) decreased, cervical intervertebral disc degenerated more seriously, type II collagen protein expression decreased, type X collagen protein expression increased, Agc1 and Col2a1 mRNA expressions in intervertebral disc decreased, and the MMP-13 mRNA expression increased. CONCLUSION: The rat model of CS with kidney deficiency is established. Kidney deficiency can aggravate cervical intervertebral disc degeneration.

[A rabbit model of cervical spondylosis established by stimulation of wind, cold and dampness].

OBJECTIVE: To make an animal model of cervical spondylosis (arthralgia syndrome type) with stimulation of wind, cold, and dampness. METHODS: Twenty-four 8 months old male New Zealand white rabbits were randomly allocated into four groups: normal control group, light stimulation group, moderate stimulation group and severe stimulation group. The wind speed was 10.8-13.8 m/s, the temperature was (5+/-0.5)degrees centigrade, and the humidity was 100%. The rabbits of light, moderate, and severe stimulation groups were kept in the above-mentioned environments for 4 hours everyday, and for a total of 32, 64, and 128 hours, respectively. The intervertebral discs were stained with HE method, and observed with a light microscope. Prostaglandin E(2) (PGE(2)), 6-ketone-prostaglandin F1alpha (6-K-PGF(1alpha)) and thromboxane B(2) (TXB(2)) contents were measured by ELISA. Fas and Bcl-2 expressions were examined by immunohistochemical avidin-biotin peroxidose complex technique. Interleukin-1beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), and transforming growth factor beta (TGF-beta) mRNA expressions were examined by reverse transcription-polymerase chain reaction. RESULTS: The nucleus pulposus of rabbits in the light and moderate stimulation groups shrunken, and in the severe stimulation group, the anulus fibrosus loosed or ruptured, and the cartilage end-plate became proliferated. Compared with rabbits in the normal control group, the PGE(2) content rose in the light stimulation group, the contents of PGE(2), 6-K-PGF(1alpha), and TXB(2) increased, the expressions of IL-1beta and TNF-alpha mRNAs and Fas were up-regulated, and the expressions of TGF-beta mRNA and Bcl-2 were down-regulated in the moderate and severe stimulation groups. The expression of Fas was up-regulated mostly and Bcl-2 was down-regulated mostly in the severe group. CONCLUSION: Moderate and severe stimulations of wind, cold and dampness can lead to degeneration of cervical intervertebral discs of rabbits. The model corresponds to the theory of traditional Chinese medicine about arthralgia syndrome caused by wind, cold and dampness.

[Study of Yiqi Huayu Bushen recipe and its decomposed formulas in regulating gene expressions in degenerated cervical intervertebral discs of rats].

OBJECTIVE: To investigate the gene expression changes in the degenerated cervical intervertebral discs of rats, and to study the function of Yiqi Huayu Bushen Recipe, a compound Chinese herbal medicine, and its discomposed formulas in regulating gene expressions in the degenerated cervical intervertebral discs. METHODS: The rat model with degenerated cervical intervertebral discs caused by imbalance between the dynamic and static forces was established. The mRNA was extracted from the cervical intervertebral discs of rats in the normal control and experiment groups, and the cDNA probes were obtained by inverse transcript. The cDNA probes were hybridized with the gene chips. The gene expression pattern was gained with a laser scanner. Image analysis, standardized ratio value and cluster analysis were used to investigate the differential of gene expressions between the control and experiment groups. RESULTS: Cluster analysis showed that the gene chips of No.1 (Yiqi Huayu group), No.2 (Yiqi Bushen group) and No.3 (Huayu Bushen group) were in one class, while the gene chips of No.4 (untreated group) and No.5 (Yiqi Huayu Bushen group) were in the other. The gene expression of No.4 was different from the others mostly, and the gene expressions of No.2 and No.3 were similar. There were 96 genes expressed differently in three cases and among them 77 genes were already known and the expression of 48 genes were up-regulated (ratio>1.0), and 29 down-regulated (ratio<0.5). There were 25 genes expressed differently between the untreated group and the herb-treated groups. CONCLUSIONS: The gene expressions of the degenerated rat intervertebral discs are changed. Yiqi Huayu Bushen Recipe and its discomposed formulas have the effect of regulating the expressions of related genes, such as PI(3)K, PTK, ERK3, and PH1B1.

[An in vitro natural degeneration model of chondrocytes derived from endplate of intervertebral discs of rats].

OBJECTIVE: To set up a natural degeneration model of chondrocytes derived from endplate of intervertebral discs of rats in order to offer an appropriate carrier for the study on mechanism of intervertebral disc degeneration. METHODS: The method of enzyme digestion combined with natural subculture was used to set up the in vitro natural degeneration model of chondrocytes derived from the endplate of intervertebral disc of rats. The morphological appearances and microstructures of the chondrocytes of different generations were observed. The expression of collagen II in chondrocytes was detected by immunocytochemical method. RESULTS: The chondrocytes derived from the endplate of intervertebral disc expressed collagen II. After 13 days of culture, the chondrocytes of generation III showed that the ability of cell division descended, the nucleoli became unclear, the cells deformed obviously, fusiform shape with weak optical activity appeared, and the intercellular space was enlarged. There were vacuoles and lipid droplets in cytoplasm. The synthesis of collagen II, as well as the cell proliferation rate, descended notably. All results showed the natural degeneration process of the chondrocytes. CONCLUSION: The in vitro natural degeneration model of chondrocytes derived from endplate of intervertebral discs of rats was successfully established. This can offer the cytological basis for study on the mechanism of intervertebral disc degeneration.