Effect of Longbie Capsule on p38MAPK pathway and nuclear factor-kappa B p65 in chondrocytes of osteoarthritis

doi:10.3969/j.issn.2095-4344.2016.46.004

Abstract

Abstract:

BACKGROUND: Previous studies found that Longbie Capsule can relieve knee joint swelling of rats with knee osteoarthritis, and decrease the serum levels of interleukin-1β, interleukin-6 and interleukin-10.
OBJECTIVE: To observe the influence of Longbie Capsule on p38MAPK pathway and nuclear factor-κB p65 (NF-κB p65) in chondrocytes of osteoarthritis.
METHODS: Sixty Sprague-Dawley rats were equivalently randomized into five groups, and the rat models of right knee osteoarthritis were established in model, low-dose, middle-dose and high-dose groups. Three days later, rats in the latter three groups were respectively treated with 0.312 5, 0.625, and 0.937 5 g/kg Longbie Capsule via gastric lavage, twice daily for 4 consecutive weeks; while those in the control group received no intervention. At 2 and 4 weeks after treatment, the rat right knee joint was removed and made into paraffin tissue sections, and the expressions of MEK-3/6, p38, activating transcription factor 2 (ATF2), NF-κBp65, and their phosphorylated products of P-MEK-3/6, P-p38, P-ATF2, P-NF-κB p65 in cartilage tissue were detected by immunohistochemistry streptavidin-perosidase method.
RESULTS AND CONCLUSION: After 2 and 4 weeks of treatment, chondrocytes in each group could express MEK-3/6, p38, ATF2, NF-κB p65, P-MEK-3/6, P-p38, P-ATF2 and P-NF-κB p65, and the percentage of positive cells in the model group was significantly higher than that in the control group (P < 0.01), and the treatment groups (P < 0.05). These results indicate that Longbie Capsule cannot only inhibit the over expressions of MEK-3/6, p38, ATF2, NF-κB p65 and their phosphorylated products, but also inhibit the activation of p38MAPKs and NF-κB signaling pathway in chondrocytes of osteoarthritis, which is beneficial for cartilage degeneration and protecting the cartilage.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Drugs, Chinese Herbal, Osteoarthritis, Tissue Engineering

CLC Number:

R318

Pan Jian-ke, Cao Xue-Wei, Liu Jun, Xie Hui, Yang Wei-yi, Zhang Xian. Effect of Longbie Capsule on p38MAPK pathway and nuclear factor-kappa B p65 in chondrocytes of osteoarthritis[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(46): 6868-6877.

Figures/Tables 5

2.2 龙鳖胶囊对软骨组织MEK-3/6表达的影响见图1，2。给药2，4周后，各组细胞均有MEK-3/6阳性表达，其中模型组MEK-3/6阳性细胞百分比高于对照组(P < 0.01)，低、中、高剂量组MEK-3/6阳性细胞百分比均低于模型组(P < 0.05)，见表1。 2.3 龙鳖胶囊对软骨组织P-MEK-3/6表达的影响见图3，4。给药2，4周后，各组细胞均有P-MEK-3/6阳性表达，其中模型组P-MEK-3/6阳性细胞百分比均高于对照组(P < 0.01)，低、中、高剂量组P-MEK-3/6阳性细胞百分比均低于模型组(P < 0.01)；给药4周后，中剂量组P-MEK-3/6阳性细胞百分比与对照组比较差异无显著性意义(P > 0.05)，见表2。 2.4 龙鳖胶囊对软骨组织p38表达的影响见图5，6。给药2，4周后，各组细胞均有p38阳性表达，其中模型组p38阳性细胞百分比均高于对照组(P < 0.01)，低、中、高剂量组p38阳性细胞百分比均低于模型组(P < 0.01)，低、中、高剂量组p38阳性细胞百分比分别与对照组比较差异均无显著性意义(P > 0.05)，见表3。 2.5 龙鳖胶囊对软骨组织P-p38表达的影响见图7，8。给药2，4周后，各组细胞均有P-p38阳性表达，其中模型组P-p38阳性细胞百分比均高于对照组(P < 0.01)，低、中、高剂量组P-p38阳性细胞百分比均低于模型组 (P < 0.01)；给药4周后，中剂量组P-p38阳性细胞百分比与对照组比较差异无显著性意义(P > 0.05)，见表4。 2.6 龙鳖胶囊对软骨组织ATF2表达的影响见图9，10。给药2，4周后，各组细胞均有ATF2弱阳性表达，且模型组ATF2阳性细胞百分比均高于对照组(P < 0.01)；给药2周后，低、中、高剂量组ATF2阳性细胞百分比均低于模型组(P < 0.01)；给药4周后，龙鳖低、中、高剂量组ATF2阳性细胞百分比与造模组比较，无统计学差异(P > 0.05)，见表5。 2.7 龙鳖胶囊对软骨组织P-ATF2表达的影响见图11，12。给药2，4周后，各组细胞均有P-ATF2弱阳性表达，且模型组P-ATF2阳性细胞百分比均高于对照组(P < 0.01)，低、中、高剂量组P-ATF2阳性细胞百分比均低于模型组(P < 0.05)；给药4周后，低、中、高剂量组P-ATF2阳性细胞百分比与对照组比较差异无显著性意义(P > 0.05)，见表6。 2.8 龙鳖胶囊对软骨组织NF-κB p65表达的影响见图13，14。给药2，4周后，各组细胞均有NF-κB p65阳性表达，且模型组NF-κBp 65阳性细胞百分比均高于对照组(P < 0.01)，低、中、高剂量组NF-κBp65阳性细胞百分比均低于模型组(P < 0.01)；给药2周后，中、高剂量组NF-κB p65阳性细胞百分比与对照组比较差异均无显著性意义(P > 0.05)；给药4周后，低、中、高剂量组NF-κB p65阳性细胞百分比与对照组比较差异均无显著性意义(P > 0.05)，见表7。 2.9 龙鳖胶囊对软骨组织P-NF-κB p65表达的影响见图15，16。给药2，4周后，各组细胞均有P-NF-κB p65阳性表达，且模型组P-NF-κB p65阳性细胞百分比均高于对照组(P < 0.01)，低、中、高剂量组P-NF-κB p65阳性细胞百分比均低于模型组(P < 0.01)，见表8。"

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