The Second Clinical College of Beijing University of Chinese Medicine, Beijing 100029, China.
Department of Acupuncture-moxibustion, Dongfang Hospital, Beijing University of Chinese Medicine, Beijing 100078.
Zhen Ci Yan Jiu. 2023 Nov 25;48(11):1103-1110. doi: 10.13702/j.1000-0607.20221180.
To observe the effect of acupuncture stimulation of "Yanglingquan"(GB34), "Zusanli"(ST36) and "Xuanzhong" (GB39) on arthritis index (AI), joint synovial membrane pathology, serum-related immunoinflammatory factors, and expressions of tumor suppressor gene mt-p53, nuclear factor kappa B (NF-κB) and peroxisome proliferator activated receptor gamma (PPARγ) in knee joint synovial tissue of rats with type Ⅱ collagen-induced arthritis (CIA), so as to explore its possible mechanisms underlying improvement of rheumatoid arthritis (RA).
Male SD rats were used in the present study. The CIA model was established by subcutaneous injection of collagen emulsion (200 μL/rat) in the tail root region on the first day and repeat (100 μL/rat) once on the 9 day. Eighteen successful CIA rats were randomized into model, medication and acupuncture groups, with 6 rats in each group. Other 6 normal rats were used as the normal control group. For rats of the medication group, leflunomide (1.9 mg/kg) was administrated by gavage, once a day, and for rats of the acupuncture group, manual acupuncture stimulation was applied to bilateral GB34, ST36, GB39 for 30 min, once a day, for 12 weeks. The arthritis index (AI) score (0-4 points) was evaluated once every week. The contents of IL-6, IL-17 and TNF-α in the serum were determined by ELISA. Histopathological changes of the ankle joint were observed by H.E. staining. The protein and mRNA expression levels of mt-p53, NF-κB p65, and PPARγ in the knee joint synovial tissue were determined by Western blot and quantitative real time PCR, separately.
Compared with the normal control group, the AI scores at different time-points after modeling, contents of serum TNF-α, IL-6 and IL-17, expression levels of mt-p53, NF-κB p65, PPARγ proteins and mRNAs were significantly increased in the model group (<0.01, <0.05). In comparison with the model group, the AI scores at the 10 week in the medication group and at the 3, 9 and 10 week in the acupuncture group, contents of serum TNF-α, IL-6 and IL-17, and the expression levels of mt-p53 and NF-κB p65 proteins in both medication and acupuncture groups, as well as mt-p53 and NF-κB p65 mRNAs in the medication group were apparently decreased (<0.01, <0.05), while the expression levels of PPARγ protein in both medication and acupuncture group and PPARγ mRNA in the medication group were significantly up-regulated (<0.05, <0.01). No significant differences were found between the acupuncture and medication groups in down-regulating the AI score and serum TNF-α, IL-6 and IL-17 contents. The effect of acupuncture was weaker than that of medication in down-regulating the expression of mt-p53 and NF-κB p65 proteins and mRNAs and in up-regulating PPARγ mRNA (<0.01). H.E. results showed ankle cartilage hyperplasia, reduced joint cavity, mild fibroproliferation and inflammatory cell infiltration in the surrounding soft tissue of the ankle joint in rats of the model group, which was milder in both medication and acupuncture groups.
Acupuncture stimulation can improve the degree of joint inflammation and swelling in CIA rats, which may be related to its effects in inhibiting the overexpression of immunoinflammatory factors in serum and regulating expression of mt-p53, NF-κB p65, PPARγ mRNAs and proteins in the synovial tissue.
观察针刺“阳陵泉”（GB34）、“足三里”（ST36）和“悬钟”（GB39）对关节炎指数（AI）、关节滑膜病理、血清相关免疫炎症因子、肿瘤抑制基因 mt-p53、核因子 κB（NF-κB）和过氧化物酶体增殖物激活受体 γ（PPARγ）在 2 型胶原诱导关节炎（CIA）大鼠膝关节滑膜组织中的表达，探讨其改善类风湿关节炎（RA）的可能机制。
雄性 SD 大鼠用于本研究。在第一天尾根区域皮下注射胶原乳液（200 μL/只）建立 CIA 模型，第 9 天重复（100 μL/只）。成功建立 18 只 CIA 大鼠随机分为模型组、药物组和针刺组，每组 6 只。另 6 只正常大鼠作为正常对照组。药物组给予来氟米特（1.9 mg/kg）灌胃，每天 1 次，针刺组给予双侧 GB34、ST36、GB39 针刺刺激 30 min，每天 1 次，共 12 周。每周评估一次关节炎指数（AI）评分（0-4 分）。通过 ELISA 测定血清中 IL-6、IL-17 和 TNF-α 的含量。通过 H.E.染色观察踝关节的组织学变化。通过 Western blot 和定量实时 PCR 分别测定膝关节滑膜组织中 mt-p53、NF-κB p65 和 PPARγ 蛋白和 mRNA 的表达水平。
与正常对照组相比，模型组在建模后不同时间点的 AI 评分、血清 TNF-α、IL-6 和 IL-17 含量、mt-p53、NF-κB p65、PPARγ 蛋白和 mRNAs 表达水平均明显升高（<0.01，<0.05）。与模型组相比，药物组在第 10 周、针刺组在第 3、9 和 10 周的 AI 评分、血清 TNF-α、IL-6 和 IL-17 含量以及药物和针刺组的 mt-p53 和 NF-κB p65 蛋白表达水平以及药物组的 mt-p53 和 NF-κB p65 mRNAs 表达水平均明显降低（<0.01，<0.05），而药物和针刺组的 PPARγ 蛋白表达水平和药物组的 PPARγ mRNA 表达水平均明显升高（<0.05，<0.01）。针刺组和药物组在降低 AI 评分和血清 TNF-α、IL-6 和 IL-17 含量方面无明显差异。针刺组下调 mt-p53 和 NF-κB p65 蛋白和 mRNAs 表达及上调 PPARγ mRNA 的作用弱于药物组（<0.01）。H.E.结果显示，模型组大鼠踝关节软骨增生，关节腔减少，踝关节周围软组织轻度纤维增生和炎症细胞浸润，药物组和针刺组均较轻。
针刺刺激可改善 CIA 大鼠关节炎症和肿胀程度，可能与其抑制血清中免疫炎症因子的过度表达以及调节滑膜组织中 mt-p53、NF-κB p65、PPARγ mRNAs 和蛋白表达有关。
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