Shikonin restrains TGF-β1/Smad signaling pathway to improve renal fibrosis

doi:10.12092/j.issn.1009-2501.2026.01.005

Abstract

Abstract:

AIM: To explore the effect of shikonin (SK) in the treatment of renal fibrosis in mice. METHODS: C57BL/6 mice were randomly divided into sham group, UUO group, shikonin low-dose group (5 mg·kg^?1·d^?1) and high-dose group (20 mg·kg^?1·d^?1), with 8 mice in each group. Mice in the experimental group were given different concentrations of shikonin, sham group and UUO group were given equal volume of normal saline, continuous intervention for 10 days. Serum urea nitrogen (BUN), creatinine (Cr) and uric acid (UA) levels were detected by automatic differentiation analyzer, serum inflammatory factors IL-6, IL-1β and TNF-α levels were detected by enzyme-linked immunosorbent assay (ELISA), and renal tissue damage and collagen deposition were detected by HE staining, Masson staining and qRT-PCR. Use protein blotting to detect the expression levels of TGF-β1, p-Smad2, Smad2, p-Smad3, Smad3. RESULTS: Compared with the sham group, the levels of inflammatory factors, renal function indexes, fibrosis-related proteins and TGF-β1/Smad signaling pathway proteins in UUO group were significantly increased (P<0.05), and the degree of renal fibrosis was aggravated. Compared with UUO group, the levels of inflammatory factors, renal function indexes, fibrosis-related proteins and TGF-β1/Smad signaling pathway proteins in the SK group were significantly decreased, and the degree of renal fibrosis was improved, and the improvement was more obvious in the SK high-dose group, with statistical significance (P<0.05). CONCLUSION: Shikonin may alleviate renal inflammation and fibrosis by inhibiting the TGF-β1/Smad signaling pathway, improve renal function and kidney injury status, and play a role in alleviating the progression of renal fibrosis.

Key words: shikonin, renal fibrosis, UUO, collagen deposition, TGF-β1/Smad signaling pathway

CLC Number:

R965.2

Tao WANG, Xiaowen CHENG, Zhirui ZHANG, Hao JIAO. Shikonin restrains TGF-β1/Smad signaling pathway to improve renal fibrosis[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2026, 31(1): 48-54.

Figures/Tables 6

Table 1 Comparison of serum interleukin-6 (IL-6), interleukin-1β (IL-1β) and tumor necrosis factor α (TNF-α) levels in each group of mice (n=5, $\overline x $±s)

Group	IL-6 (pg/mL)	IL-1β (ng/L)	TNF-α (ng/L)
Sham	19.89±4.54	23.88±5.13	343.85±33.36
UUO	112.25±11.76^b	69.66±4.61^b	903.84±32.09^b
SK-L	85.01±5.86^e	55.76±3.22^e	712.27±62.80^e
SK-H	59.37±9.51^e	43.71±4.18^e	548.81±76.54^e

Table 2 Comparion of serum blood urea nitrogen (BUN), creatinine (Cr) and Uric Acid (UA) levels in each group of mice (n=5, $\overline x $±s)

Group	BUN (mmol/L)	Cr (μmol/L)	UA (μmol/L)
Sham	3.02±0.27	2.88±0.55	91.00±10.12
UUO	10.70±1.71^b	153.4±11.55^b	215.20±11.63^b
SK-L	7.94±0.62^e	112.20±8.81^e	162.00±11.02^e
SK-H	5.22±0.98^e	82.40±9.53^e	128.60±11.44^e

Fig.1 Observation of renal pathology morphology by hematoxylin and eosin (HE) staining (×200) A: Sham group; B: UUO group; C: 5 mg·kg−1·d−1 shikonin group; D: 20 mg·kg−1·d−1 shikonin group.

Fig.2 Observation of renal fibrosis by Masson staining (×200) A: sham group; B: UUO group; C: 5 mg·kg−1·d−1 shikonin group; D: 20 mg·kg−1·d−1 shikonin group.

Fig.3 Collagen I and α-SMA expression in kidney tissues of each group (×200) A: sham group; B: UUO group; C: 5 mg·kg−1·d−1 shikonin group; D: 20 mg·kg−1·d−1 shikonin group.

Fig.4 Western blot analysis of the effects of SK on the expression of TGF-β1/Smad signaling pathway in mouse kidney ($\bar x $±s, n=3) bP<0.05,cP<0.01, compared with sham group; eP<0.05,fP<0.01, compared with UUO group.

References 23

1	Zhang YY, Yu C, Li XG, et al. Kidney aging and chronic kidney disease[J]. Int J Mol Sci, 2024, 25 (12): 6585. doi: 10.3390/ijms25126585
2	Zhao XD, Kong YL, Liang B, et al. Mechanosensitive Piezo1 channels mediate renal fibrosis[J]. JCI Insight, 2022, 7 (7): e152330- e152394. doi: 10.1172/jci.insight.152330
3	Huang SD, Lu HQ, Chen J, et al. Advances in drug delivery-based therapeutic strategies for renal fibrosis treatment[J]. J Mater Chem B, 2024, 12 (27): 6532- 6549. doi: 10.1039/D4TB00737A
4	张明昊, 赵绅, 杜婧雯, 等. 灯盏花素通过调控TGF-β1介导的Smad和ERK通路干预肾纤维化大鼠的作用机制研究[J]. 中药新药与临床药理, 2022, 33 (10): 1347- 1356.
5	Yadav S, Sharma A, Nayik GA, et al. Review of shikonin and derivatives: isolation, chemistry, biosynthesis, pharmacology and toxicology[J]. Front Pharmacol, 2022, 13, 905755. doi: 10.3389/fphar.2022.905755
6	Yan CM, Li QX, Sun Q, et al. Promising nanomedicines of shikonin for cancer therapy[J]. Int J Nanomedicine, 2023, 18, 1195- 1218.
7	Guo CJ, He JL, Song XM, et al. Pharmacological properties and derivatives of shikonin-A review in recent years[J]. Pharmacol, 2019, 149, 104463. doi: 10.1016/j.phrs.2019.104463
8	杨晨宇, 尚新悦, 姚国栋. 中药活性成分通过调控PKM2发挥抗肿瘤作用的研究进展[J]. 中草药, 2022, 53 (2): 590- 598. doi: 10.7501/j.issn.0253-2670.2022.02.029
9	Hu XX, Peng XY, Zhang Y, et al. Shikonin reverses cancer-associated fibroblast-induced gemcitabine resistance in pancreatic cancer cells by suppressing monocarboxylate transporter 4-mediated reverse Warburg effect[J]. Phytomedicine, 2023, 11 (23): 155214- 155228. doi: 10.1016/j.phymed.2023.155214
10	周珍, 蔡坤松, 许雅红, 等. 紫草素对博来霉素诱导肺纤维化小鼠的作用及机制[J]. 西部医学, 2020, 32 (1): 33- 37. doi: 10.3969/j.issn.1672-3511.2020.01.008
11	鲁明霞, 王红岗, 胡仁标. 紫草素对四氯化碳诱导的肝纤维化大鼠TGF-β1的影响[J]. 药学实践杂志, 2018, 36 (5): 453- 456.
12	Elena MK, Omar Emiliano AT, Edilia T, et al. Unilateral ureteral obstruction as a model to investigate fibrosis-attenuating treatments[J]. Biomolecules, 2019, 9 (4): 141- 169. doi: 10.3390/biom9040141
13	邱成英, 邓颖云, 卢磊, 等. 紫草素对紫癜性肾炎大鼠肾脏的保护作用[J]. 中国临床药理学杂志, 2020, 36 (20): 3250- 3252.
14	焦浩, 曹静. 紫草素在治疗小鼠草酸钙肾结石中的作用[J]. 中国临床药理学杂志, 2020, 36 (20): 3253- 3256.
15	刘洋, 张瑞红, 只素娟, 等. 肾纤维化手术动物模型构建及相关机制研究进展[J]. 解剖学杂志, 2022, 45 (2): 157- 160. doi: 10.3969/j.issn.1001-1633.2022.02.014
16	Fang X, Tang CY, Zeng D, et al. CircInpp5b ameliorates renal interstitial fibrosis by promoting the lysosomal degradation of DDX1[J]. Biomolecules, 2024, 14 (6): 613. doi: 10.3390/biom14060613
17	Roccatello D, Lan HY, Sciascia S, et al. From inflammation to renal fibrosis: A one-way road in autoimmunity[J]. Autoimmun Rev, 2024, 23 (4): 103466. doi: 10.1016/j.autrev.2023.103466
18	孙晓怡, 姜玉勤, 唐余燕, 等. 细胞焦亡机制及其在多种肾脏病中的作用[J]. 中国医药导报, 2021, 18 (33): 53- 56. doi: 10.20047/j.issn1673-7210.2021.33.012
19	Chen HS, Yang YX, Zhou XJ, et al. Attenuating renal interstitial fibrosis by shenqi pill via reducing inflammation response regulated by nf-κb pathway in vitro and in vivo[J]. Iran J Kidney Dis, 2024, 18 (2): 87- 98.
20	Guo YM, Zhou MM, Mu ZZ, et al. Recent advances in shikonin for the treatment of immune-related diseases: Anti-inflammatory and immunomodulatory mechanisms[J]. Biomed Pharmacother, 2023, 165, 115138. doi: 10.1016/j.biopha.2023.115138
21	Li JC, Zou Y, Kantapan J, et al. TGF-β/Smad signaling in chronic kidney disease: Exploring post-translational regulatory perspectives (Review)[J]. Mol Med Rep, 2024, 30 (2): 143. doi: 10.3892/mmr.2024.13267
22	周林华, 陈晓. 栀子苷通过TGF-β1/Smad信号通路抑制肝纤维化和肝星状细胞活化[J]. 生理学报, 2022, 74 (2): 217- 224. doi: 10.13294/j.aps.2022.0019
23	应勤丽, 黄月碧, 杨秀翠, 等. 圣草酚通过抑制TGF-β1/Smad3信号通路改善糖尿病肾病大鼠肾纤维化[J]. 中国免疫学杂志, 2023, 39 (4): 693- 697. doi: 10.3969/j.issn.1000-484X.2023.04.004

[1]	LIN Jingyi, HAN Rangyue, XU Linghui, TAN Ruizhi, SU Hongwei, WANG Li. Huangqi sanqi mixture inhibits lncRNA Gm51500/Adam12 axis to improve renal fibrosis in CKD [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2025, 30(6): 750-762.
[2]	WU Xiaoxue, YE Yiping, LEI Zhendong, ZHANG Zunjing, CHEN Wenling. Icariin improves hypertensive renal fibrosis and injury through Cx32-Nox4 signaling pathway [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2024, 29(8): 870-878.
[3]	LI Shixu, LI Linyun, WANG Xin, LI Ke, BIAN Hua. Effects of Ginkgo biloba extract on renal injury in rats with experimental renal failure through miR-145/FOXO1 axis [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2023, 28(7): 728-735.
[4]	NIU Pilian, FAN Yongxin, LU Furui, ZHOU Xuezhang, BAI Mingsheng. Effects of liquorice extract on cardiac fibroblasts fibrosis induced by TGF-β1 [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2022, 27(2): 129-135.
[5]	CHEN Xinxin, CHEN Yu, ZHENG Chenfei, ZHOU Ying, CHEN Chaosheng. Expression trend of autophagy in mice with renal interstitial fibrosis [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(3): 260-265.
[6]	CHENG Xiaolong, HU Kunmei, HU Haoran，XU Hanghang, YANG Jieren, HAN Jun. Effects of Chinese medicine Yuxia capsule on renal fibrosis in spontaneously hypertensive rats [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2018, 23(6): 632-639.
[7]	HU Kunmei, HU Haoran, SHEN Yuanyuan, LU Yining, YANG Jieren, HAN Jun. Effect of traditional Chinese medicine Yuxia capsule on renal fibrosis in spontaneously hypertensive rats and its relationship with TGF-β1/Smads signaling pathway [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2017, 22(4): 387-393.
[8]	LIU Yan, YANG Jie-ren, HAO Wei, ZHANG Jun-xiu, TANG Li-juan, CHEN Guo-xiang. Mechanisms and protective effect of compound Shanju on aortal injury in spontaneously hypertensive rats [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2013, 18(7): 732-737.
[9]	SU Jian, YIN Li-ping, ZHANG Xin, LI Bi-bo, LIU Li, LI Hui. Influence of rhein intervention on the expression of HGF and BMP7 in renal tissue of rats with chronical allograft nephropathy [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2011, 16(10): 1114-1120.
[10]	YUAN Ding-fen, DENG Hui, YAN Chun-lin, LIAO Kang-huang. Effect of shikonin on angiogenic ability of endothelial cells [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2006, 11(11): 1253-1255.
[11]	WANG Jian-yun, YIN Xiao-xing. Relationship between transforming growth factor-β₁ signal transduction and renal fibrosis of diabetes [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2005, 10(4): 371-376.
[12]	WU Ji-Ning, LIU Bi-Cheng. Current researches of angiotensin receptor blockers in treatment of patients with chronic progressive renal disease [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2004, 9(11): 1201-1204.