Effect of electroacupuncture at Wnt/β-catenin signaling pathway on inhibiting cartilage degeneration in rats with knee osteoarthritis / 中国针灸

OBJECTIVE@#To explore the action mechanism of electroacupuncture (EA) for knee osteoarthritis (KOA) based on Wnt/beta-catenin (Wnt/β-catenin) signaling pathway.@*METHODS@#Ten rats were randomly selected into a sham-operation group among 50 male 2-month-old SD rats, and the KOA model was established in the remaining 40 rats by modified Hulth method. Four weeks after the model establishment, the rats were randomly divided into a model group, an experimental A group, an experimental B group and an experimental C group, 10 rats in each group. The rats in the sham-operation group and model group did not receive any intervention. The rats in the experimental A group were treated with EA at "Neixiyan" (EX-LE 4) and "Dubi"(ST 35) for 15 min. The rats in the experimental B group were treated with EA at "Neixiyan" (EX-LE 4) and "Dubi"(ST 35) for 30 min. The rats in the experimental C group were treated with EA at non-acupoint for 15 min. EA intervention was given once a day, five times a week, and totally 12-week treatment was given. After 12 weeks, the knee cartilage tissues were stained and the morphological changes were observed under light microscopy; the severity of cartilage degeneration was evaluated by modified Mankin's score; the content of interleukin-1β (IL-1β) in synovium tissues was detected by ELISA method; the content of Wnt-4, β-catenin and matrix metalloprotein-13 (MMP-13) in cartilage tissues was detected by Western blot method.@*RESULTS@#Compared with the sham-operation group, in the model group the morphology and structure of cartilage were disordered, the number of cells was significantly reduced, the matrix was decontaminated and tidal line was incomplete; the Mankin's score was significantly increased (0.05).@*CONCLUSION@#EA at "Neixiyan" (EX-LE 4) and "Dubi"(ST 35) may reduce the expression of MMP-13 and the production of inflammatory factor IL-1β through Wnt/β-catenin signaling pathway, thus inhibit the degradation of cartilage matrix and the apoptosis of chondrocyte, and improve the morphology and structure of cartilage.

Effect of water extracts from duhuo jisheng decoction on expression of chondrocyte G1 phase regulator mRNA / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effect of water extracts from Duhuo Jisheng decoction on chondrocyte G1 phase.</p><p><b>METHODS</b>Chondrocytes were collected from four-week-old SD rats to establish the chondrocyte in vitro culture system. The third generation of chondrocytes was intervened. MTT method was used to measure the effect of water extracts from different concentrations of Duhuo Jisheng decoction on chondrocyte activity. The expressions of Chondrocyte Cyclin D1, CDK4, CDK6 and P21 mRNA in the blank group and low, middle and high-dose groups (100, 200, 400 mg x L(-1)) were detected by RT-PCR method.</p><p><b>RESULT</b>The MTT assay showed that the chondrocyte activity significantly increased within specific drug concentrations (50-800 mg x L(-1)) (P < 0.01); After the intervention for 24 h, the expressions of CyclinD1, CDK4 and CDK6 mRNA in all dose groups notably increased (P < 0.05), with the maximum expressions at the concentration of 200 mg x L(-1); The expression of P21 mRNA decreased, particularly at the concentration of 200 mg x L(-1) (P < 0.01).</p><p><b>CONCLUSION</b>Water extracts from Duhuo Jisheng decoction can promote chondrocyte proliferation by effecting the expression of chondrocyte G1 phase regulator mRNA.</p>

Clinical observation of warm needling therapy for treating knee osteoarthritis of different Chinese medical syndrome types / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the difference in the therapeutic efficacy of warm needling therapy on knee osteoarthritis (KOA) patients of different Chinese medical syndrome types.</p><p><b>METHODS</b>Totally 197 KOA outpatients [including wind-cold-damp retention syndrome (50 cases, 61 knees), yang deficiency cold coagulation syndrome (48 cases, 58 knees), stagnation of blood stasis syndrome (49 cases, 63 knees), and insufficiency of Shen-essence syndrome (50 cases, 66 knees)] were treated with warm needling therapy, 10 days as one therapeutic course, 3 courses in total. The symptom score and changes of clinical efficacy were assessed. The contents of interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha) in the synovial fluid were determined before and after treatment.</p><p><b>RESULTS</b>The symptom scores and the clinical efficacy were improved in all syndrome groups after treatment (P < 0.05), with the best effects shown in the yang deficiency cold coagulation syndrome (P < 0.05) and the worst effects shown in the stagnation of blood stasis syndrome (P < 0.05). The contents of IL-1, IL-6, and TNF-alpha in the synovial fluid decreased after treatment in all syndrome groups (P < 0.05), with the best effects shown in the yang deficiency cold coagulation syndrome (P < 0.05).</p><p><b>CONCLUSION</b>Warm needling therapy had favorable therapeutic effects on KOA patients of wind-cold-damp retention syndrome, yang deficiency cold coagulation syndrome, and insufficiency of Shen-essence syndrome, with the best effects shown on KOA patients of yang deficiency cold coagulation syndrome.</p>

Experimental study on the suppression of sodium nitroprussiate-induced chondrocyte apoptosis by Tougu Xiaotong Capsule (透骨消痛胶囊)-containing serum / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the mechanism of action of Tougu Xiaotong Capsule (透骨消痛胶囊, TGXTC) ex vivo in suppressing chondrocyte (CD) apoptosis induced by sodium nitroprussiate (SNP).</p><p><b>METHODS</b>Thirty New Zealand rabbits, 2 months old, were randomized by lottery into five groups, six in each: the blank group treated with saline, the positive control group treated with Zhuanggu Guanjie Pill (壮骨关节丸, 70 mg/kg), and the three experimental groups, EGA, EGB, and EGC, treated with low dose (35 mg/kg), moderate dose (70 mg/kg), and high dose (140 mg/kg) of TGXTC, respectively. All treatments were administered via gastrogavage twice a day for 3 days. Arterial blood was collected from the abdominal aorta and drug or drug metabolites-containing serum was prepared. CDs obtained from knee joints of 16 four-week-old New Zealand rabbits were cultured to the third passage and confirmed by toluidine blue staining. SNP of various final concentrations (0, 0.5, 1.0, and 2.0 mmol/L) was used to induce CD apoptosis, and the dosage-effect relationship of SNP in inducing CD apoptosis was determined. Serum samples from the blank, control, and three dosages of TGXTC-treated rabbits were tested in the CD culture in the presence of SNP. Cell apoptosis was determined by Hoechst 33342 staining, viability of CDs was quantified by MTT, CD apoptosis rate was determined by annexin V-FITC/PI staining, levels of p53 and Bcl-2 mRNA expression in CDs were determined with RT-PCR, and contents of caspase-3 and caspase-9 proteins were determined by colorimetry.</p><p><b>RESULTS</b>CD apoptosis was induced by SNP at all concentrations tested and in a dose-dependent manner. The SNP concentration of 1 mmol/L and treatment duration of 24 h appeared to be optimal and were selected for the study. Serum samples from the positive control rabbits and from the two higher doses of TGXTC-treated rabbits showed reduction of SNP-induced CD apoptosis, decrease in p53 mRNA expression, inhibition of catalytic activities of caspase-3 and caspase-9, and increase in Bcl-2 mRNA expression when compared with the serum from the blank group (P<0.05).</p><p><b>CONCLUSION</b>TGXTC-containing sera antagonized SNP-induced CD apoptosis and the molecular basis for the action was associated with up-regulation of Bcl-2, down-regulation of p53 expression, and inhibition of caspase-3 and caspase-9 catalytic activities.</p>

Clinical study on the treatment of knee osteoarthritis of Shen-Sui insufficiency syndrome type by electroacupuncture / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the clinical effificacy of electroacupuncture (EA) on treating knee osteoarthritis (KOA) of Shen ()-Sui () insuffificiency (SSI) syndrome type.</p><p><b>METHODS</b>A total of 245 patients (279 knees) of KOA-SSI were randomly assigned to two groups by lottery: 141 knees in the treatment group and 138 knees in the control group. The treatment group was managed with EA at the dominant points of Neixiyan (Ex-LE4) and Waixiyan (Ex-LE5) as well as the conjugate points of Xuanzhong (GB39) and Taixi (KI3) for 30 min, once a day, with 15 days as one course; 2 courses were applied with a 5-day interval in between. The control group was treated with intra-articular injection of 2 mL hyaluronic acid into the affected joint every 7 days for 5 times in total. The clinical effects on the patients in different stages were observed, and their symptom scores of knee and contents of cytokines, including interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), prostaglandin E(2alpha) (PGE(2alpha)) and matrix metalloproteinases-3 (MMP-3), in the knee joint fluid were measured before and after treatment.</p><p><b>RESULTS</b>The study was completed in 235 patients (263 knees); four patients (7 knees) in the treatment group and six patients (9 knees) in the control group dropped out. Comparison of therapeutic effects (excellent and effective rates) between the two groups showed insignificant differences (P>0.05). Symptom scores of knee and contents of cytokines in the knee flfluid after treatment were lowered signifificantly in the patients of stage I-III in both groups (P<0.05 or P<0.01). However, the lowering of the total symptom score of knee in the patients of stage III in the treatment group was more signifificant (P<0.05).</p><p><b>CONCLUSIONS</b>EA could effectively alleviate the clinical symptoms in KOA patients of stage III, showing an effect superior to that of hyaluronic acid. EA also shows action in suppressing the secretion of IL-1, IL-6, TNF-alpha, PGE(2alpha) and MMP-3 in the knee flfluid.</p>

Effect of OA kneepad on apoptosis genes Bcl-2 and p53 expression in articular cartilage cells of experimental knee osteoarthritis / 中国骨伤

<p><b>OBJECTIVE</b>To study the effects of kneepad on expression of Bcl-2 and p53 mRNA of chondrocyte in white rabbits with knee osteoarthritis, so as to explore and treatment mechanism of OA kneepad on apoptosis of chondrocytes of rabbits with knee osteoarthritis in molecular degree.</p><p><b>METHODS</b>Forty-four Japanese healthy 6-month-old rabbits (equal male and female,the weight ranging from 2 to 2.2 kg) were used to establish knee osteoarthritis models by modified Hulth method. The rabbits were randomly divided into 6 groups: normal group, model group, control group (microwave), experimental group 1 (electricity), experimental group 2 (thermal), experimental group 3 (kneepad). Ten rabbits in the normal group were breed with conventional method; 9 rabbits in the model group were breed with conventional method after model made; 9 rabbits in the control group were treated with microwave for 30 minutes, one time daily; 9 rabbits in the experimental group 1 were treated with electricity (density wave) for 30 minutes,one time daily;8 rabbits in the experimental group 2 were treated with hot (hot soft membrane) for 30 minutes, one time daily; 9 rabbits in the experiment group 3 were treated with electrothermal (OA knee pad) for 30 minutes, one time daily. All the rabbits were treated for 16 weeks and then sacrificed. The expressions of Bcl-2 and p53 mRNA of chondrocytes in knee joint were detected by using fluorescence quantitative RT-PCR method.</p><p><b>RESULTS</b>At the 16 hthek,th e OD260/OD280 value range of total RNA extracted from rabbit articular cartilage tissue in each group were all at 1.80 to 2.00,wh ich indicates high RNA purity. The p53 relative mRNA in articular cartilage cells of model group,th e control group,th e experimental group 1 ,r oup 2,gr oup 3 were overexpressed,an d Belc2 mRNA expression levels of articular cartilage cells were low expression,an d compared with the normal group there were significant differences (P < 0.01). Belc2, p53 mRNA expression in articular cartilage cells,th ere were significant differences (P < 0.01) between the control group, experimental group 1, group 2, group 3 and model group. The results between the control group, experimental group 1 ,group 2 and group 3 had significant differences (P < 0.01).</p><p><b>CONCLUSION</b>OA-kneepad can up-regulate the mRNA expression of Bcl-2 as well as down-regulate the mRNA expression of p53, thereby to inhibit the apoptosis of cartilage cells and delay the degeneration of articular cartilage changes.</p>

Research on mechanism of Tougu Xiaotong Granule in preventing and treating knee osteoarthritis / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the mechanism of Tougu Xiaotong Granula (TXG) on prevention and treatment of knee osteoarthritis.</p><p><b>METHODS</b>Fifty New Zealand rabbits were randomly divided into 6 groups. Except those in the normal control group, all the rabbits were replicated into knee osteoarthritis model using modified Hulth method. They were administered by gastrogavage once every day respectively with 100 ml of normal saline to the rabbits in the normal group and those in the model group, with 10 g of Zhuanggu Guanjie Pill to those in the control group, and 5 g, 10 g and 20 g of TXG to those in the three TXG tested groups (tested group 1, 2 and 3). The levels of interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) and metalloproteinase 3 (MMP-3) in joint fluid, the blood content of malondialdehyde (MDA) and nitric oxide (NO) absorbance as well as the SOD activity in synovia were observed.</p><p><b>RESULTS</b>Overexpressions of IL-1, IL-6, TNF-alpha and MMP-3 in joint fluid, increased blood content of NO and MDA were shown in the 8th and 16th week, and decreased SOD activity in synovia was shown in the 16th week of the experiment in all the model rabbits, as compared to those in the normal group, the difference was significant respectively (P < 0.05 or P<0.01). After 8 weeks of treatment, the levels of IL-1, TNF-alpha, MMP-3, NO and MDA in the control group, tested group 2 and 3 were significantly different to those in the model group respectively (P < 0.05, P < 0.01), and significant difference was also shown in the comparisons of those indexes between the control group and the tested group 1 vs the tested group 3 (P < 0.05). As for the level of IL-6, significant difference was shown in comparisons of the model group with the control group, tested group 2 and 3 in the 8th and 16th week of the treatment (P < 0.05 or P < 0.01), also in comparison of the tested group 3 with the tested group 1 in the 8th week, and in that of the tested group 2 with the control group and the tested group 1 in the 16th week (P < 0.05).</p><p><b>CONCLUSION</b>TXG could effectively postpone the degeneration of cartilage through effectively inhibiting the biological effects of cytokines, MMP-3 and oxygen free radical.</p>

Experimental study on replicating knee osteoarthritis by modified Hulth's modeling method / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the method of replicating experimental animal model of knee osteoarthritis.</p><p><b>METHODS</b>Knee osteoarthritis was replicated by modified Hulth's modeling method. X-ray photographic and transmission electron microscopic examination, test of the joint synovial fluid of the modeled joint were performed, and serum contents of malondialdehyde (MDA) and nitric oxide (NO) in blood were measured.</p><p><b>RESULTS</b>(1) In the normal control group, the articular surface is smooth and glossy, with intact cells and cellular membrane. In the model group, the medial space of the knee joint became obvious narrowed with rough and deformed articular surface and osteophytes, as well as the atrophic chondrocytes with pyknotic cell nucleus and broken cellular membrane. (2) Eight weeks and 16 weeks after modeling, in the model group, the contents of interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) and matrix metalloproteinase-3 (MMP-3) in synovial fluid and the levels of serum MDA and NO were obviously raised, and the activity of superoxide dismutase (SOD) in synovial membrane was obviously lowered 16 weeks after modeling, showing significant difference when compared with those in the normal control group (P < 0.01).</p><p><b>CONCLUSION</b>Modified Hulth's modeling method in replicating knee osteoarthritis is simple in manipulation with less wound, and the condition of modeled knee joint could be maintained stable to certain degree, which is advantageous to the success of animal model replicating.</p>