BACKGROUND:Inflammation is an important factor in cervical spondylotic vertebral arteriopathy, bone marrow mesenchymal stem cells have the potential to treat cervical spondylotic vertebral arteriopathy because of the immunomodulatory effects to inhibit inflammation.
OBJECTIVE:To investigate the injury mechanism of vascular injury in the model of cervical spondylotic vertebral arteriopathy and to study the therapeutic effects of bone marrow mesenchymal stem cells on it.
METHODS:Forty Japanese big ear rabbits were randomly divided into four groups:control group, model group, tanshinone group, and stem cellgroup. After modeling, the control and model groups were not given intervention, while the tanshinone and stem cellgroups were injected with tanshinone II A sodium sulfonate solution (10 mL) and bone marrow mesenchymal stem cellsuspension (10 mL) along the ear vein, respectively. After 2 weeks, the routine pathological examination was done to observe the vascular morphological changes, immunofluorescence staining was done to observe the cathepsin B expression in the vertebral artery, and ELISA was used to detect the expression of tumor necrosis factor-αand interleukin-6 in the vertebral artery.
RESULTS AND CONCLUSION:Compared with the model group, the arterial smooth muscle cellhypertrophy and hyperplasia was obviously restrained in stem cellgroup, and vascular endothelial fold was in symmetry, while no significant difference was found between stem cellgroup and tanshinone group. Compared with the model group, expressions of cathepsin B, interleukin 6 and tumor necrosis factor-αexpression were reduced significantly in the stem cellgroup (P<0.05), while there was no difference between the model and tanshinone groups (P>0.05). Inflammatory reaction may be one of mechanisms for vertebral artery damage, and bone marrow mesenchymal stem cells can effectively inhibit inflammation of the vertebral artery and improve vascular remodeling.

The efficacy of injecting sclerosing agent next to transverse process of cervical vertebra to induce vertebral artery type of cervical syndrome (CSA) was observed. Twenty rabbits were randomly divided into two groups: the model group and the control group. The rabbits in the model group were injected with sclerosing agent next to transverse process of cervical vertebray, on the contrary, the rabbits in the control group were injected with nothing. Transcranial Doppler (TCD) was used to detect the average speed of blood (Vm), pulsatility index (Pi) and the resistant index (Ri) of the vertebral artery, hematoxylin and eosin (HE) staining was used to observe the morphological changes, and immunohistochemistry was used to detect the expression of alpha-smooth muscle actin (alpha-SMA) and matrix metalloproteinase-2(MMP-2). TCD showed increased Pi, Ri and decreased Vm in the model group (P<0.05) compared with the control group. HE staining revealed hyperplasia and hypertrophied smooth muscle cells in the model group (P<0.05). Immunohistochemistry displayed up-regulation of alpha-SMA and MMP-2 in the model group (P<0.05). It was concluded that injecting sclerosing agent next to transverse process of cervical vertebra induces remodeling of vertebral artery in rabbits, suggesting it is a practical method to establish CSA animal model.

BACKGROUND: Previous animal experiments have shown that epimedium total flavonoids can exhibit preventive effects on estrogen-related bone loss, but few data are available in clinical research.OBJECTIVE: To investigate the effects of pimedium total flavonoids on bone mineral density (BMD) inprimary osteoporosis.DESIGN, TIME AND SETTING: A randomized, double-blinded, positively controlled clinical observation was performed at the Department of Traditional Chinese Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology between June 2005 and September 2007.PARTICIPANTS: A total of 64 patients with primary osteoporosis consisting of 11 males and 53 females were included in this study. METHODS: All patients were randomly and evenly assigned into a treatment group and a control group. The treatment group was orally administered epimedium total flavonoids, 0.45 g once, three times daily, for a total of 6 months. Simultaneously, the control group was orally administered Gushukang, 10 g once, twice daily, for a total of 6 months. MAIN OUTCOME MEASURES: BMD in the sites of lumbar vertebrae (L1-4), neck of femur, Wards triangle, greater trochanter, and left hip was measured, and simultaneously serum levels of calcium, phosphorus, and alkaline phosphatase (ALP) were detected pdor to and after treatment in each group.RESULTS: All 64 patients with primary osteoporosis were included In the final analysis, The BMD in the lumbar vertebrae (L1-4) was significantly higher in the treatment group than in the control group (P<0.05) and there was no significant difference In BMD in other sites between the two groups (P>0.05). Compared with pdor to drug application, BMD in the treatment group did not present obvious change after epimedium total flavonoids application, while in the control group, BMD in the sites of lumbar vertebrae (L1-4), neck of femur, Wards triangle, and greater trochanter was significantly decreased (P<0.01-0.05). After drug application, serum level of calcium was significantly increased in each group, compared with prior to drug application (P<0.05), and no significant difference existed between the treatment and control groups (P>0.05). Tn the treatment group, serum levels of phosphorus and ALP did not alter significantly compared with prior to epimedium total flavonoids application (P>0.05).CONCLUSION: Epimedium total flavonoids exhibit effective effects on the maintenance of BMD in primary osteoporosis.

Objective_To investigate the potential effect of bone marrow mesenchymal stem cells ( MSCs) on repai-ring the colon epithelial cell,and on the treatment of rats with ulcerative colitis ( UC) .Methods_Monocytes were purified from bone marrow, amplified and identified as MSCs in vitro.Thirty female Wistar rats were randomly di-vided into 3 groups, the normal control, model and MSCs groups (10 rats/group).The rats in model and MSCs groups were induced colitis with trinitro-benzene-sulfonic acid;The rats in normal control group and model groups were injected with 1mL saline via tail vein, while those in MSCs group with 1 mL MSCs suspension.After two weeks, colon tissue samples were analyzed for histopathology,and the colon tissues were made into serial section for determining the distribution of Y chromosome and CK20 double positive cells,analyzing the mRNA levels of CK20,NF-κB, IL-4 by RT-PCR,and assaying colonic NF-κB protein expression with Westen blot,detecting colon tissues IL-4 content with ELISA.Results_Y chromosome and CK20 double positive cells were found in MSCs transplanted colon tissues.The expression of CK20 increased in the colon tissues of UC rats(P<0.01) and in MSCs group in-creased as compared with model group( P<0.01) .The expression of NF-κB increased in the colon tissues of UC rats (P<0.01), but decreased MSCs group as compared with model group (P<0.01).The expression of IL-4 was decreased in the colon tissues of UC rats ( P<0.01) , while in MSCs group it was increased as compared with model group ( P<0.01) .Conclusions_MSCs may have therapeutic efficacy on colitis in rats through differentiating into colon epithelial cells.

Inflammatory bowel disease is thought to be regulated by the balance between Th1 and Th2 cytokines secreted by T cells, and NF-kappaB p65 also plays a predominant role in the intestinal inflammation. We evaluated the potency of oxymatrine, one of active components of Sophora Root, in inhibiting the immune responses and inflammation in 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis. The inflammation was markedly ameliorated in the oxymatrine-treated rats. The level of IL-2 was increased and that of IL-10 was decreased in colon tissue in the rat model, which was reversed by the treatment of oxymatrine. Moreover, the elevated expression of NF-kappaB p65 in colon tissue in the model was also improved by oxymatrine treatment. Our results suggest that oxymatrine might be beneficial for the abnormal immune responses and inflammation by regulating the unbalance of Th1 and Th2 cytokines secretion and inhibiting the expression of NF-kappaB p65 in colon tissue.

The effects of Wumeiwan (WMW) on TNF-alpha, IL-6, IL-8, IL-10 and NF-kappaBp65 in rats with ulcerative colitis (UC) were investigated, the curative effectiveness of WMW vs salicylazosulfapyridine (SASP) was compared, and the action mechanism was analyzed. Fifty-Six Sprague-Dawley (SD) rats were randomly divided into four groups (n=14 in each group, with equal ratio of male and female): normal control group, model group, SASP group, and WMW group. Except normal control group, the rat UC models in the remaining three groups were established using the method of 2.4-dinitrochlorobenzene (DNCB) immunization and acetic acid local enema. The rats in model group, SASP group, and WMW group were treated with distilled water, SASP, and WMW respectively. The changes in the symptoms and signs were observed, and levels of IL-6, IL-8, TNF-alpha, IL-10 and the expression of NF-kappaBp65 in the colonic tissues were statistically analyzed. The results showed that the levels of IL-6, IL-8, and TNF-alpha were significantly increased (P<0.01), while those of IL-10 significantly reduced (P<0.01) after establishment of rat UC models as compared with normal control group. The levels of IL-6, IL-8, and TNF-alpha were obviously lower, but the level of IL-10 was obviously higher in WMW and SASP groups than those in model group (P<0.05). The levels of IL-6, IL-8, and TNF-alpha were lower, while the level of IL-10 was higher in WMW group than in SASP group. NF-kappaBp65 was expressed negatively or weakly in normal colonic tissues. The positive expression rate of NF-kappaBp65 in WMW group and SASP group was obviously lower than in model group (P<0.01), and there was significant difference between WMW group and SASP group (P<0.05). It was concluded that rat UC model was established successfully. WMW could up-regulate the expression of IL-10, down-regulate the expression of TNF-alpha, IL-6, IL-8, and inhibit the NF-kappaBp65 activity to adjust immune function, indicating WMW had better curative effects on UC in rats.

OBJECTIVETo study the effect of NOD2 on colitis pathogenesis in experimental rats, and discuss therapeutical effect and mechanism of kushenin injection (OMT) on colitis in experimental rats.

METHODFourty Sprague-Dawley (SD) rats were randomly divided into four groups: the normal control group, the model group, the SASP group, and the OMT group, with 10 rats in each group. Except the normal control group, models were established in the remaining three groups with TNBS. The OMT group was injected with kushenin injection, the SASP group was orally administered with mesalazine suspension, the model group and the normal group were orally administered with distilled water for 15 days. Colon lesion score and histological score of experimental rats were observed. Expression of NOD2, NF-kappaB p65 protein in rats colonic mucous was detected by immunohistochemistry. Expression of IL-6 in rat colon mucous was detected by ELISA.

RESULTCompared with normal control group, the expression of NOD2, NF-kappaB p65 and IL-6 in colonic mucosa of the model group were significantly increased (P < 0.01). The SASP group and the OMT group showed lower expressions of NOD2, NF-kappaB p65 and IL-6 in colonic mucosa than the model group (P < 0.01, P < 0.05).

CONCLUSIONThe over expression of colonic mucosa proteins NOD2 and NF-kappaB p65 and increasing secretion of IL-6 take part in the appearance and development of ulcerative colitis. OMT can attenuate ulcerative colitis and protect colonic mucosa by inhibiting expression of NOD2, NF-kappaB p65 and decreasing IL-6.

Animals ; Colitis ; metabolism ; pathology ; physiopathology ; prevention & control ; Colon ; drug effects ; metabolism ; pathology ; physiopathology ; Eating ; drug effects ; Gene Expression Regulation ; drug effects ; Injections ; Intestinal Mucosa ; drug effects ; metabolism ; pathology ; physiopathology ; Male ; Nod2 Signaling Adaptor Protein ; metabolism ; Organ Size ; drug effects ; Pterocarpans ; administration & dosage ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Transcription Factor RelA ; metabolism

The efficacy of injecting sclerosing agent next to transverse process of cervical vertebra to induce vertebral artery type of cervical syndrome(CSA)was observed.Twenty rabbits were randomly divided into two groups: the model group and the control group.The rabbits in the model group were injected with sclerosing agent next to transverse process of cervical vertebray,on the contrary,the rabbits in the control group were injected with nothing.Transcranial Doppler(TCD)was used to detect the average speed of blood(Vm),pulsatility index(Pi)and the resistant index(Ri)of the vertebral artery,hematoxylin and eosin(HE)staining was used to observe the morphological changes,and immunohistochemistry was used to detect the expression of α-smooth muscle actin(α-SMA)and matrix metalloproteinase-2(MMP-2).TCD showed increased Pi,Ri and decreased Vm in the model group(P<0.05) compared with the control group.HE staining revealed hyperplasia and hypertrophied smooth muscle cells in the model group(P<0.05).Immunohistochemistry displayed up-regulation of α-SMA and MMP-2 in the model group(P<0.05).It was concluded that injecting sclerosing agent next to transverse process of cervical vertebra induces remodeling of vertebral artery in rabbits,suggesting it is a practical method to establish CSA animal model.

Inflammatory bowel disease is thought to be regulated by the balance between Th1 and Th2 cytokines secreted by T cells, and NF-κB p65 also plays a predominant role in the intestinal inflammation. We evaluated the potency of oxymatrine, one of active components of Sophora Root, in inhibiting the immune responses and inflammation in 2,4,6-trinitrobenzene sulfonic acid(TNBS)-induced colitis. The inflammation was markedly ameliorated in the oxymatrine-treated rats.The level of IL-2 was increased and that of IL-10 was decreased in colon tissue in the rat model,which was reversed by the treatment of oxymatrine. Moreover, the elevated expression of NF-κB p65in colon tissue in the model was also improved by oxymatrine treatment. Our results suggest that oxymatrine might be beneficial for the abnormal immune responses and inflammation by regulating the unbalance of Th1 and Th2 cytokines secretion and inhibiting the expression of NF-κB p65 in colon tissue.

The effects of Wumeiwan (WMW) on TNF-α, IL-6, IL-8, IL-10 and NF-κBp65 in rats with ulcerative colitis (UC) were investigated, the curative effectiveness of WMW vs salicylazosul-fapyridine (SASP) was compared, and the action mechanism was analyzed. Fifty-Six Spra-gue-Dawley (SD) rats were randomly divided into four groups (n=14 in each group, with equal ratio of male and female): normal control group, model group, SASP group, and WMW group. Except normal control group, the rat UC models in the remaining three groups were established using the method of 2.4-dinitrochlorobenzene (DNCB) immunization and acetic acid local enema. The rats in model group, SASP group, and WMW group were treated with distilled water, SASP, and WMW re-spectively. The changes in the symptoms and signs were observed, and levels of IL-6, IL-8, TNF-α,IL-10 and the expression of NF-κBp65 in the colonic tissues were statistically analyzed. The results showed that the levels of IL-6, IL-8, and TNF-α were significantly increased (P<0.01), while those of IL-10 significantly reduced (P<0.01) after establishment of rat UC models as compared with normal control group. The levels of IL-6, IL-8, and TNF-α were obviously lower, bat the level of IL-10 was obviously higher in WMW and SASP groups than those in model group (P<0.05). The levels of IL-6,IL-8, and TNF-α were lower, while the level of IL-10 was higher in WMW group than in SASP group.NF-κBp65 was expressed negatively or weakly in normal colonic tissues. The positive expression rate of NF-κBp65 in WMW group and SASP group was obviously lower than in model group (P<0.01), and there was significant difference between WMW group and SASP group (P<0.05). It was concluded that rat UC model was established successfully. WMW could up-regulate the expres-sion oflL-10, down-regulate the expression ofTNF-α, IL-6, IL-8, and inhibit the NF-κBp65 activity to adjust immune function, indicating WMW had better curative effects on UC in rats.