Susilo Raden Joko Kuncoroningrat, Winarni Dwi, Hayaza Suhailah, Doong Ruey-An, Wahyuningsih Sri Puji Astuti, Darmanto Win
Department of Biology, Faculty of Science and Technology, Universitas Airlangga, Surabaya 60115, Indonesia.
Institute of Analytical and Environmental Sciences, National Tsing Hua University, Sec. 2 Kuang Fu Road, Hsinchu 30013, Taiwan.
Vet World. 2022 Apr;15(4):1022-1030. doi: 10.14202/vetworld.2022.1022-1030. Epub 2022 Apr 23.
Interstitial fibrosis is the final stage of chronic kidney injury, which begins with an inflammatory process. Crude polysaccharides are known to have anti-inflammatory properties. The potential role of crude polysaccharides in renal fibrosis through pro-inflammatory cytokines needs further investigation. This study aimed to determine the renoprotective effect of crude polysaccharide extract in mice with carbon tetrachloride (CCL)-induced early kidney fibrosis.
This study was conducted for 4 weeks using 24 male BALB/c mice selected for their metabolic stability. The mice were randomly divided into six groups, including control (CG), model (MG), silymarin group and crude polysaccharide extract groups comprising doses of 25, 50, and 100 mg/kg body weight. After sacrificing the mice, whole blood was analyzed for urea and creatine levels, and kidney tissue was prepared to assess tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), hyaluronic acid (HA), and laminin levels, both using enzyme-linked immunosorbent assay. Kidney histology was determined using hematoxylin and eosin staining, while the extracellular matrix (ECM) components were stained using Masson's trichome staining. The α-smooth muscle actin (α-SMA) concentration was determined using immunohistochemistry. These parameters were measured to determine the effectiveness of the crude polysaccharide extract in preventing interstitial fibrosis.
Administration of crude polysaccharides effectively prevented increases in kidney weight and physiological enzymes, pro-inflammatory cytokines, and ECM production compared with those in the MG, as evidenced by the low levels of urea, creatinine, TNF-α, IL-6, HA, and laminin. Histopathological results also showed that crude polysaccharides prevented the occurrence of inflammatory infiltration, desquamated nuclei, cytoplasm debris, rupture at the brush border, dilatation of the glomeruli space and lumen of the proximal tubule, and necrotic cells compared with the MG. Masson's trichrome staining revealed lower collagen levels in the interstitial tubules of kidney tissue than those in the MG. Immunohistochemical analysis revealed low α-SMA expression in the crude polysaccharides treatment groups than that in the MG.
The crude polysaccharide extract of has a protective effect that prevents the progression of kidney fibrosis in mice.
间质纤维化是慢性肾损伤的终末期,始于炎症过程。已知粗多糖具有抗炎特性。粗多糖通过促炎细胞因子在肾纤维化中的潜在作用尚需进一步研究。本研究旨在确定粗多糖提取物对四氯化碳(CCL)诱导的小鼠早期肾纤维化的肾脏保护作用。
本研究使用24只因其代谢稳定性而选择的雄性BALB/c小鼠,持续进行4周。小鼠被随机分为六组,包括对照组(CG)、模型组(MG)、水飞蓟宾组以及粗多糖提取物组,粗多糖提取物组的剂量分别为25、50和100mg/kg体重。处死小鼠后,分析全血中的尿素和肌酐水平,并制备肾脏组织以评估肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、透明质酸(HA)和层粘连蛋白水平,均采用酶联免疫吸附测定法。使用苏木精和伊红染色确定肾脏组织学,同时使用马松三色染色对细胞外基质(ECM)成分进行染色。使用免疫组织化学法测定α-平滑肌肌动蛋白(α-SMA)浓度。测量这些参数以确定粗多糖提取物在预防间质纤维化方面的有效性。
与模型组相比,给予粗多糖有效地防止了肾脏重量增加以及生理酶、促炎细胞因子和ECM产生的增加,尿素、肌酐、TNF-α、IL-6、HA和层粘连蛋白水平较低证明了这一点。组织病理学结果还显示,与模型组相比,粗多糖可防止炎症浸润、细胞核脱落、细胞质碎片、刷状缘破裂、肾小球间隙和近端小管管腔扩张以及坏死细胞的发生。马松三色染色显示,肾脏组织间质小管中的胶原水平低于模型组。免疫组织化学分析显示,粗多糖治疗组中的α-SMA表达低于模型组。
粗多糖提取物具有保护作用,可防止小鼠肾纤维化进展。