Department of Cell and Molecular Biology, Sanjeev Biomedical Research Centre, 1/15, Kumaran Nagar, Keelkattalai, Chennai, 600117, Tamilnadu, India.
State Licensing Authority, Directorate of Indian Medicine (Govt. of Tamilnadu), State Licensing Authority (IM), Chennai, 600106, Tamilnadu, India.
J Ethnopharmacol. 2023 Apr 6;305:116032. doi: 10.1016/j.jep.2022.116032. Epub 2022 Dec 29.
Kabasura Kudineer (KK), the traditional Indian medicine of Siddha, effectively manages common respiratory symptoms such as flu, cold, and fever. However, there is no evidence of the immunomodulatory capacity of KK in the cultured Jurkat T-lymphocytes under the LPS insult studied.
Assess the effect of the traditional Indian medicine of Siddha, Kabasura Kudineer (KK) on immunomodulation by suppressing oxidative damage in cultured Jurkat T cells in vitro. The miRNA activity on anti-inflammatory gene receptors and cellular nitric oxide levels also was studied.
Jurkat T cells were exposed to LPS treatment in the presence or absence of KK. Cell viability and nitric oxide (NO) were measured with MTT and Griess assay. Cellular antioxidant systems (glutathione and SOD) were determined using glutathione and SOD assay. Lipid peroxidation was measured using an MDA assay. MiRNA-15a-5p expression was performed using microRNA qPCR Assays. Both inflammatory and anti-inflammatory genes (IL-6, IL-1, IL-10, IL-13) were performed using a qPCR and ELISA assay.
The data showed that reduced cell proliferation and exaggerated NO production was observed in LPS treated condition compared to the control condition. Further, LPS treatment increased lipid peroxidation and reduced antioxidant enzyme activities (SOD and glutathione) in cultured Jurkat T cells. However, treatment with KK or N-acetyl cysteine (NAC; antioxidant) treatment mitigates the above effect. Mechanistically, LPS-induced oxidative stress upregulated miR- 15-5p expression and suppressed IL-10 Receptor alpha (IL-10Rα) by binding to its 3'-UTR region. The deregulated expression of IL-10Rα expression leads to increased IL-6 and IL-1β expression in LPS-induced Jurkat T cells; however, treatment with KK or NAC reversed the above effects.
Collectively, our study revealed the previously undefined mechanistic role of Kabasura Kudineer (KK) that alleviates the LPS-induced oxidative damage associated with inflammation by inhibiting the miRNA-15-5p/IL-10Rα axis.
Kabasura Kudineer（KK）是 Siddha 的传统印度药物，可有效治疗流感、感冒和发烧等常见呼吸道症状。然而，在 LPS 刺激下培养的 Jurkat T 淋巴细胞中，没有证据表明 KK 具有免疫调节能力。
评估 Siddha 的传统印度药物 Kabasura Kudineer（KK）对培养的 Jurkat T 细胞的免疫调节作用，抑制体外 LPS 诱导的氧化损伤。还研究了抗炎基因受体和细胞内一氧化氮水平的 miRNA 活性。
将 Jurkat T 细胞暴露于 LPS 处理中，有或没有 KK。使用 MTT 和 Griess 测定法测量细胞活力和一氧化氮（NO）。使用谷胱甘肽和 SOD 测定法测定细胞抗氧化系统（谷胱甘肽和 SOD）。使用 MDA 测定法测量脂质过氧化。使用 microRNA qPCR 测定法进行 miRNA-15a-5p 表达。使用 qPCR 和 ELISA 测定法进行炎症和抗炎基因（IL-6、IL-1、IL-10、IL-13）。
数据显示，与对照条件相比，LPS 处理条件下观察到细胞增殖减少和 NO 产生增加。此外，LPS 处理增加了培养的 Jurkat T 细胞中的脂质过氧化和降低了抗氧化酶活性（SOD 和谷胱甘肽）。然而，用 KK 或 N-乙酰半胱氨酸（NAC；抗氧化剂）治疗减轻了上述作用。从机制上讲，LPS 诱导的氧化应激上调了 miR-15-5p 的表达，并通过结合其 3'-UTR 区域抑制了 IL-10 受体 alpha（IL-10Rα）。IL-10Rα 表达的失调导致 LPS 诱导的 Jurkat T 细胞中 IL-6 和 IL-1β 表达增加；然而，用 KK 或 NAC 治疗逆转了上述作用。
总的来说，我们的研究揭示了 Kabasura Kudineer（KK）的先前未定义的机制作用，通过抑制 miRNA-15-5p/IL-10Rα 轴减轻 LPS 诱导的与炎症相关的氧化损伤。
