Hepatoprotection of Jianpi Qingre Lishi prescription (健脾清热利湿法) on non-alcoholic steatohepatitis via miRNA-27/peroxisome proliferator-activated receptor gamma axis

doi:10.19852/j.cnki.jtcm.2026.03.004

Abstract

Abstract:

OBJECTIVE: To elucidate the molecular targets and physiological mechanisms underlying the therapeutic efficacy of Jianpi Qingre Lishi prescription (健脾清热利湿法, JQLP) in the treatment of non-alcoholic steatohepatitis (NASH).

METHODS: This study used 120 Sprague-Dawley rats to establish six groups at random (n = 20): blank control (BC), model, low-dose JQLP (L-JQLP), medium-dose JQLP (M-JQLP), high-dose JQLP (H-JQLP), and positive control (PC) groups. Rats in the BC group received a diet with methionine- and choline-sufficient (MCS), while those in the remaining groups were fed with methionine- and choline-deficient (MCD) diet. Blood samples were collected for biochemical analyses and inflammatory factors determination; while liver tissues were harvested to identify the histological alterations through hematoxylin-eosin staining and oil red O staining. In addition, the levels of miRNA-27 (miR-27) and peroxisome proliferator-activated receptor γ (PPARγ) were determined utilizing quantitative real-time polymerase chain reaction and Western blotting.

RESULTS: Compared to the BC group, the model group showed no significant changes in fasting blood glucose (FBG), which was substantially reduced after both M-JQLP and H-JQLP treatments. MCD diet induced a series of pathological alterations, such as extensive vacuole-like steatosis, disorganized hepatic plates, and significant inflammatory cell infiltration in liver tissues, which were dose-dependently weakened by JQLP intervention. Rats fed with MCD diet demonstrated increase in total cholesterol, triglyceride, low-density lipoprotein cholesterol, alanine aminotransferase and aspartate aminotransferase activities, but decrease in high-density lipoprotein cholesterol. JQLP treatment effectively normalized these parameters in a dose-dependent manner. Furthermore, rats with MCD diet were detected with largely secreted tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6); while JQLP intervention decreased TNF-α and IL-6 secretion, but enhanced IL-4 content. Additionally, miR-27 upregulation and PPARγ downregulation were induced in liver tissues of rats with MCD diet, which were counteracted by JQLP treatment.

CONCLUSIONS: JQLP can ameliorate NASH progression, potentially through the regulation of miR-27/ PPARγ axis. JQLP may be a promising therapeutic candidate worthy of further clinical investigation for NASH treatment.

Key words: non-alcoholic fatty liver disease, PPAR gamma, inflammation, miRNA-27, Jianpi Qingre Lishi prescription

XU Mengjun, YAN Zixing, CHEN Xi, CAI Juanjuan, ZHANG Haiou, LIN Zhenwen. Hepatoprotection of Jianpi Qingre Lishi prescription (健脾清热利湿法) on non-alcoholic steatohepatitis via miRNA-27/peroxisome proliferator-activated receptor gamma axis[J]. Journal of Traditional Chinese Medicine, 2026, 46(3): 561-570.

Figures/Tables 4

Figure 1 Effect of JQLP on pathological changes of liver tissues in MCD diet-induced NASH rats A: pathological changes of liver tissues (hematoxylin-eosin staining, × 200); B: adipogenesis in liver tissues (oil red O staining, × 200); A1, B1: BC group; A2, B2: model group; A3, B3: L-JQLP group; A4, B4: M-JQLP group; A5, B5: H-JQLP group; A6, B6: PC group. BC group: feeding using methionine- and choline-sufficient diet combined with the administration of normal saline; model group: feeding using methionine- and choline-deficient diet combined with the administration of normal saline; L-JQLP group: feeding using methionine- and choline-deficient diet combined with the administration of 6.489 g/kg of JQLP once daily for 8 weeks; M-JQLP group: feeding using methionine- and choline-deficient diet combined with the administration of 12.978 g/kg of JQLP once daily for 8 weeks; H-JQLP group: feeding using methionine- and choline-deficient diet combined with the administration of 19.467 g/kg of JQLP once daily for 8 weeks; PC group: feeding using methionine- and choline-deficient diet combined with the administration of 0.18 g/kg dosage of polyene phosphatidylcholine capsule once daily for 8 weeks. BC: blank control; L-JQLP: low-dose of Jianpi Qingre Lishi prescription; M-JQLP: medium-dose of Jianpi Qingre Lishi prescription; H-JQLP: high-dose of Jianpi Qingre Lishi prescription; PC: positive control.

Table 1 Blood lipid contents and hepatic function of rats in each group ($\bar{x}$ ± s)

Group	n	TC (mmol/L)	TG (mmol/L)	LDL-C (mmol/L)	HDL-C (mmol/L)	ALT (U/L)	AST (U/L)
BC	20	1.02±0.11	0.53±0.05	0.32±0.05	1.23±0.28	15.38±2.70	12.29±2.37
Model	20	3.07±0.54^a	1.68±0.23^a	1.61±0.18^a	0.38±0.11^a	45.58±6.01^a	43.78±10.03^a
L-JQLP	20	1.92±0.30^b	1.18±0.08^b	0.95±0.09^b	0.43±0.09^b	36.98±5.27^b	37.55±13.66^b
M-JQLP	20	1.64±0.25^b	0.96±0.09^b	0.78±0.08^b	0.49±0.13^b	31.08±4.50^b	37.86±16.31^b
H-JQLP	20	1.44±0.26^b	0.77±0.09^b	0.72±0.08^b	0.59±0.12^b	28.58±3.98^b	36.42±19.21^b
PC	20	1.37±0.24^b	0.73±0.05^b	0.63±0.05^b	0.79±0.26^b	23.34±4.70^b	19.34±5.86^b

Table 2 Inflammatory mediators of rats in each group (pg/mL, $\bar{x}$ ± s)

Group	n	TNF-α	IL-6	IL-4
BC	20	78.5±9.1	6.6±1.3	15.3±2.4
Model	20	152.4±14.8^a	14.7±2.2^a	15.8±2.6
L-JQLP	20	135.7±16.3^b	9.1±1.6^b	18.0±2.6
M-JQLP	20	129.4±12.3^b	8.0±1.3^b	19.6±2.3
H-JQLP	20	116.3±13.7^b	7.4±1.4^b	23.6±2.8^b
PC	20	106.4±11.1^b	6.7±1.6^b	25.8±2.8^b

Figure 2 Expression level of PPARγ in liver tissues of rats in each group A: evaluation of PPARγ protein expression by Western blotting; B: quantification of the relative protein expression of PPARγ. BC group: feeding using methionine- and choline-sufficient diet combined with the administration of normal saline; Model group: feeding using methionine- and choline-deficient diet combined with the administration of normal saline; L-JQLP group: feeding using methionine- and choline-deficient diet combined with the administration of 6.489 g/kg of JQLP once daily for 8 weeks; M-JQLP group: feeding using methionine- and choline-deficient diet combined with the administration of 12.978 g/kg of JQLP once daily for 8 weeks; H-JQLP group: feeding using methionine- and choline-deficient diet combined with the administration of 19.467 g/kg of JQLP once daily for 8 weeks; PC group: feeding using methionine- and choline-deficient diet combined with the administration of 0.18 g/kg dosage of polyene phosphatidylcholine capsule once daily for 8 weeks. BC: blank control; L-JQLP: low-dose of Jianpi Qingre Lishi prescription; M-JQLP: medium-dose of Jianpi Qingre Lishi prescription; H-JQLP: high-dose of Jianpi Qingre Lishi prescription; PC: positive control; One-way analysis of variance was performed to determine inter-group differences, and Tukey's post-hoc comparison test to perform multiple comparisons between groups. aP < 0.05 vs the blank control group; bP < 0.05 vs the model group.

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