Qingjie decoction attenuated E.coli-induced diarrhea by regulating energy metabolism and alleviating inflammation based on network analysis and metabolomics.

ETHNOPHARMACOLOGICAL RELEVANCE: Diarrhea is a frequently encountered gastrointestinal complication in clinical practice, and E. coli is one of the main causative agents. Although Qingjie decoction (QJD) has been shown to be highly effective in treating diarrhea by eliminating heat-toxin, the underlying molecular mechanisms and pathways of QJD remain unclear. AIM OF REVIEW: The aim of this research was to explore the effects and fundamental mechanism of QJD on diarrhea induced by E.coli in rats. MATERIALS AND METHODS: Initially, we used UHPLC-MS/MS analysis to identify the chemical composition of QJD. Then, we constructed a visualization network using network pharmacology. Next, we utilized metabolomics to identify differentially expressed metabolites of QJD that are effective in treating diarrhea. RESULTS: The chemical composition of QJD was analyzed using UHPLC-MS/MS, which identified a total of 292 components. Using a network pharmacology approach, 127 bioactive compounds of QJD were screened, targeting 171 potential diarrhea treatment targets. TNF-α, IL-6, IL-1ß, and CAT were identified as important targets through visualizing the PPI network. Enrichment analysis demonstrated significant enrichment in the TNF signaling pathway, IL-17 signaling pathway, and PI3K-Akt signaling pathway. QJD showed beneficial effects, such as increased body weight, decreased fecal water content, and reduced inflammatory cell infiltration in the duodenum and colon, as well as maintaining the structure of the duodenum and colon. Metabolomic analysis revealed 32 differentially expressed metabolites in the control, model and QJD-H groups, including glucose, valine, and cysteine. Functional analysis indicated that differential metabolites were related to energy metabolism, including glucose metabolism, TCA cycle, and amino acid metabolism. CONCLUSION: QJD significantly increased body weight, decreased water content in feces, relieved inflammatory cell infiltration, maintained the structure of duodenum and colon. Combining network analysis and metabolomics, QJD exerted therapeutic effects by inhibiting inflammation and oxidative stress, regulating glucose metabolism, tricarboxylic acid metabolism, and amino acid metabolism.

Inhibitory effect of Portulaca oleracea L. aqueous extract and juice on NLRP3 inflammasome activation in an ulcerative colitis mouse model.

Portulaca oleracea L. (PO) is an edible and medicinal plant used for treating gastrointestinal diseases. However, the effects of PO on ulcerative colitis (UC) and underlying mechanisms remain unclear. This study investigated the effects of PO aqueous extract (POE) and PO juice (PJ) on dextran sulfate sodium (DSS)-induced UC in a mouse model and attempted to unravel their underlying mechanisms. The results revealed that PJ contains more bioactive compounds and has more overlapping targets with UC than POE. Both POE and PJ effectively reduced Disease Activity Index scores and inflammatory cell infiltration in the UC mouse model, but PJ had a better effect than POE. Furthermore, PJ inhibited pyroptosis by decreasing the expression of the NLRP3 inflammasome, while also repairing the dysfunction of the intestinal barrier by upregulating the expression of tight junction proteins. Therefore, based on the study findings, we concluded that PJ can improve DSS-induced UC and may suppress pyroptosis by interfering with the activation of the NLRP3 inflammasome.

Mori fructus aqueous extracts attenuate carbon tetrachloride-induced renal injury via the Nrf2 pathway and intestinal flora.

Mori fructus aqueous extracts (MFAEs) have been used as a traditional Chinese medicine for thousands of years with the function of strengthening the liver and tonifying the kidney. However, its inner mechanism to alleviative renal injury is unclear. To investigate the attenuation of MFAEs on nephrotoxicity and uncover its potential molecular mechanism, we established a nephrotoxicity model induced by carbon tetrachloride (CCl4). The mice were randomly divided into control group, CCl4 model group (10% CCl4), CCl4 + low and high MFAEs groups (10% CCl4 + 100 mg/kg and 200 mg/kg MFAEs). We found that MFAEs decreased the kidney index of mice, restored the pathological changes of renal structure induced by CCl4, reduced cystatin C, neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule 1 (Kim-1) blood urea nitrogen and creatinine contents in serum, promoted the nuclear transportation of Nrf2 (nuclear factor erythroid derived 2 like 2), elevated the expression of HO-1 (heme oxygenase 1), GPX4 (glutathione peroxidase 4), SLC7A11 (solute carrier family 7 member 11), ZO-1 (zonula occludens-1) and Occludin, suppressed the expression of Keap1 (kelch-like ECH-associated protein 1), HMGB1 (High Mobility Group Protein 1), ACSL4 (acyl-CoA synthetase long chain family member 4) and TXNIP (thioredoxin interacting protein), upregulated the flora of Akkermansia, Anaerotruncus, Clostridium_sensu_stricto, Ihubacter, Alcaligenes, Dysosmobacter, and downregulated the flora of Clostridium_XlVa, Helicobacter, Paramuribaculum. Overlapped with Disbiome database, Clostridium_XlVa, Akkermansia and Anaerotruncus may be the potential genera treated with renal injury. It indicated that MFAEs could ameliorate kidney injury caused by CCl4 via Nrf2 signaling.

Shaoyao decoction attenuates DSS-induced ulcerative colitis, macrophage and NLRP3 inflammasome activation through the MKP1/NF-κB pathway.

BACKGROUND: Shaoyao decoction (SYD), a traditional Chinese medicine prescription that originated in the Jin-Yuan Dynasty, has shown effects in treating ulcerative colitis. However, the underlying mechanism is unclear. We combined network pharmacology with molecular biology technology to detect the mechanism underlying the effect of SYD on ulcerative colitis. We combined network pharmacology with molecular biology technology to detected the further mechanism in SYD effect on ulcerative colitis. PURPOSE: In this study, we investigated the mechanism by which SYD exerts a protective effect against ulcerative colitis in vivo and in vitro. STUDY DESIGN AND METHODS: We focused on two aspects of the mechanism by which SYD relieves ulcerative colitis, regulation of the MAPK cascade and the NF-κB signaling pathway, through analysis of the "active ingredient-target-disease" network followed by GO enrichment and KEGG pathway analysis according to network pharmacology. Mice with ulcerative colitis underwent 5% dextran sulfate sodium (DSS), and the RAW 264.7 cell model was used to identify important targets. RESULTS: We found that after 5% DSS treatment, the inflammation indexes and the expression of NLRP3-related proteins were increased concomitant with the loss of mucins and occludin. Treatment with SYD (2.25 g/kg, BW) significantly improved the expression of mucins and occludin after DSS at the protein and transcriptional levels. Furthermore, SYD treatment significantly reduced NF-κB P65 and P38 expression, thus exerting a great antinecrotic effect, as revealed by TUNEL staining and Western blotting. The beneficial effects of SYD were almost canceled by NSC 95397 (an inhibitor of mitogen-activated protein kinase phosphatase-1 (MKP1)) after DSS treatment in vivo or LPS treatment in vitro. In addition, treatment with SYD reduced caspase-1 activity and rescued the release of ASC and GSDMD, thus inhibiting the assembly of NLRP3 and maintaining the integrity of the intestinal barrier. We also conducted in vitro experiments in the LPS-induced RAW 264.7 cell model and found that cells incubated with 1 mg/ml SYD for 24 h possessed the highest cell viability. Next, we incubated 1 mg/ml SYD for 24 h after treatment with 1 µg/ml LPS for 6 h. We showed that 1 mg/ml SYD displayed anti-inflammatory and anti-necrotic effects through the NLRP3, NF-κB P65 and P38 pathways, and the effects of SYD were also inhibited by 10 nM NSC 95397. CONCLUSION: These results demonstrate that SYD has protective effects against ulcerative colitis and alleviates pyroptosis by inhibiting the MKP1/NF-κB/NLRP3 pathway.

[Effects of pomegranate flowers polyphenols on liver PON expression of diabetes combining non-alcoholic fat liver diseases rats].

To investigate the protective effects and possible mechanism of pomegranate flowers polyphenols (PFP) on liver function of rats with diabetes combining non-alcoholic fat liver diseases, diabetes combining nonalcoholic fat liver disease model rats were established with high calorie feeding and small dose intraperitoneal injection of streptozotocin (STZ). Model rats were randomly divided into: model group, metformin group, pomegranate flowers polyphenols low, medium and high dose group (75, 150 and 300 mg x kg(-1)). After four weeks treatment, the levels of FPG, blood fat profiles and serum insulin, ALT, AST levels, SOD and MDA in the liver and serum separately were analyzed with biochemical methods. Paraoxonase (PON1 and PON3) mRNA and protein expression in liver were checked by RT-PCR and immunohistochemical method. Pathological changes of the liver were observed. FPG, IRI, non-HDL-C and transaminase significantly reduced and HDL-C raised in the each PFP dose group; Furthermore, compared with model group, fat drops in liver cells significantly reduced, antioxidant ability enhanced, PON1 mRNA and protein expression level in liver increased significantly. The protective effects of PFP against diabetes combining non-alcoholic fat liver diseases rats might through the increase liver PON1 mRNA and protein expression further enhanced the body antioxidant capacity and reduced IRI so as to ameliorate the rat hepatic steatosis.

[Effects of two different extracts of total saponins from Cicer arietinum on kidney of T2DM rats].

Protective effects of two different extracts of TSCA (total saponins from Cicer arietinum) were studied on kidney of T2DM rats. The diabetic model group was established with high calorie feeding and small dose injection of streptozotocin (STZ, 45 mg x kg(-1)). DM rats were randomly assigned to model group (feed with propylene glycol 1 mL/100 g), TSCA high dose group (300 mg x kg(-1)), TSCA low dose group (100 mg x kg(-1)) and normal control group (feed with propylene glycol 1 mL/100 g). After four weeks treatment with TSCA I and II, the levels of FPG FIns, BUN, Scr, ATII, ET-1, TXB2 and 6-keto-PGF1alpha in blood and the activities of SOD, GSH-PX and MDA in kidney were analyzed by biochemical methods. After four weeks treatment with TSCA II, the levels of FPG FIns, BUN, Scr, ATII and ET-1 were reduced significantly; and the ratios of TXB2 to 6-keto-PGF1alpha and SOD were effectively alleviated in TSCA II group. While there is no significant change on FPG and BUN in comparison to the rats treated with TSCA I, Scr, ATII, ET-I, GSH-PX and SOD were alleviated. The results suggest that TSCA II could be used to reduce FPG and FIns. According to the result of vasoactive substances index, TSCA II is more effective than TSCA I on renal protection of DM rats.

[Specificity effect of acupuncture at "Shuigou" (GV 26) on brain infarction area in MCAO rats and the influence of acupuncture parameter].

OBJECTIVE: To investigate the specificity of the acupoints and related factors on acupuncture effects. METHODS: The rat model of cerebral median artery occlusion (MCAO) was established with thread ligation according to Zea-Longa method. The acupoint group and the non-acupoint group were stimulated by acupuncture with 9 different parameters (marked group No. I - VI) combined by frequency (60 Hz, 120 Hz, 180 Hz) and time (5 s, 60 s, 180 s). The acupoint of "shuigou" (GV 26) was selected for the acupoint group, the fixed point which was at a distance of 10 mm from the iliac crest below the ribs in the affected side. The acupuncture effect was evaluated with the percentage of brain infarct area after TTC dyeing. RESULTS: (1) Acupoint groups could obviously reduce the brain infarction area in MCAO rats. Under 9 intervention parameters, acupoint groups had much more effectiveness tendency than that of the non-acupoint groups. (2) Acupoint group VI was the most effective in reducing the brain infarction area among 9 acupuncture parameter groups. CONCLUSION: (1) Acupoint has specificity effect on reduction of brain infarct area in MCAO rats. (2) Acupoint group VI (180 Hz; 5 s) could significantly reduce the percentage of cerebral infarction area by TTC dyeing.

[Experimental study on Shuigou (GV 26) of inhibiting effect for neuronal necrosis in rats: morphological evidence of the specificity of acupoint].

OBJECTIVE: To investigate the effect of acupoint Shuigou (GV 26) and non-acupoint on inhibiting the neuronal necrosis induced by the middle cerebral artery obstruction (MCAO) in rats through the morphological observation. METHODS: Forty two healthy adult male Wistar rats were randomly divided into a normal group, false-ope ration group, model control group, non-acupuncture group, Shuigou-acupuncture group and non-acupoint acupuncture group, 7 rats in each group. Besides the normal and false-operation groups, the experimental modal of MCAO was established in the other groups by using the Zea-Longa thread method. Acupoint Shuigou (GV 26) and non-acupoint (below the costal region) were selected respectively in the Shuigou-acupuncture group and non-acupoint acupuncture group for puncturing at 180 times/min for 5 seconds. Its effect was estimated by measuring the rate of the neuronal necrosis at the level of the light microscope and the degree of the neuronal necrosis at the level of the electron microscope. RESULTS: (1) At the level of the light microscope, comparing with the model control group (0.66 +/- 0.18), of the neuronal necrosis there were no significant differences in the rate of neuronal necrosis in the non-acupuncture group (0.67 +/- 0.34) and non-acupoint acupuncture group (0.59 +/- 0.11) (both P > 0.05), while it was significantly decreased in the Shuigou-acupuncture group (0.200 +/- 12) (P < 0.05). It indicates that no intervention and wrong (non-acupoint) intervention could light the damage of the neurons, however, only right intervention (Shuigou, GV 26) can obviously decrease the rate of the neuronal necrosis, showing with the specificity of acupoint. (2) At the level of the electron microscope, the ultrastructure of the involved neurons in the non-acupuncture group and non-acupoint acupuncture group is similar to that of the model control group, showing with the serious edema and structure damage. In contrast, the ultrastructure of the involved neurons in the Shuigou-acupuncture group is similar to that of the normal group and false-operation group, showing with light neuronal damage. CONCLUSION: Acupuncturing Shuigou (GV 26) could protect the involved neurons and inhibit the neuronal necrosis induced by the MCAO in rats, but acupuncturing the non-acupoint couldn't, which imply that there is the specificity of acupoint in the acupuncture.

[Effects of acupuncture at different acupoints on cerebral blood flow in cerebral ischemia model rats].

OBJECTIVE: To probe into effective methods for treatment of ischemic cerebral infarction and specificity of acupoints in the acupuncture treatment. METHODS: The rat model of middle cerebral artery occlusion (MCAO) was established with thread ligation according to Zea-Longa's method. They were divided into normal group, non-operation group, model control group, non-acupuncture group and acupuncture group, and the acupuncture group was divided into sham-acupuncture group and four Xingnao Kaiqiao groups (contain Shuigou group, Neiguan group, Chize group, Sanyinjiao group and Weizhong group). Then they were treated by acupuncture at "Shuigou" (GV 26), "Neiguan" (PC 6), "Chize" (LU 5), "Sanyinjiao" (SP 6), "Weizhong" (BL 40) and non-acupoints, 3 times/second, for 5 seconds. Cerebral blood blow (CBF) was used for assessment of the effect. RESULTS: As compared with the model control group, in the non-acupuncture group CBF did not significantly change (P>0.05); compared with the non-acupuncture group, after acupuncture CBF was significantly increased in the Shuigou group and the Neiguan groups (all P<0.05), but did not significantly increased in the Chize group, Sanyinjiao group and Weizhong group (all P>0.05). CONCLUSION: CBF has a tendency of spontaneous cure within 72 h after cerebral infarction in the MCAO rat; acupuncture can significantly improve CBF in the MCAO rat, so it is an effective method for treatment of ischemic stroke; among the acupoints in the "Xingnao Kaiqiao" needling method, "Shuigou" (GV 26) and "Neiguan" (PC 6) have obvious effect in improvement of CBF, indicating acupoint specificity of "Shuigou" (GV 26) and "Neiguan" (PC 6) in treatment of ischemic stroke.

[Status and progress of stimulating parameters in acupuncture treatment of ischemic cerebrovascular disease].

Acute ischemic cerebrovascular disease is one of the critical diseases seriously endangering human health. Acupuncture therapy, an effective treatment method for many types of disorders has been generally acknowledged. In recent years, many scientific researchers have studied the relationship between the effects of acupuncture in relieving cerebral ischemia-induced sequelae and the stimulating parameters. The acupuncture stimulating parameter includes the frequency of electroacupuncture (EA), the frequency of acupuncture treatment, and the acquired quantity of stimulation, etc for clinical patients and experimental animals. It was found that different stimulating parameters may have different efficacies. Current research results provide a good basis not only for analysis of the factors of acupuncture-produced effects, but also for determination of the optimal combination of stimulating parameters. However, acupuncture therapeutic effect involves multiple factors and multiple levels, and current quantitative acupuncture parameter researches have been mainly restricted to animal experiments. Hence, more researches in which statistics specialists take part are definitely needed.

[Study on optimum stimulation parameter for Shuigou (GV 26) in Xingnao Kaiqiao needling method].

OBJECTIVE: to investigate the influence of acupuncture parameter on acupuncture effect and to probe the optimum stimulation parameter for Shuigou (GV 26) in Xingnao Kaiqiao needling method. METHODS: The rat model of middle cerebral artery obstruction (MCAO) was replicated and "Shuigou" (GV 26) was stimulated by 9 acupuncture parameters combined by two factors at 3 levels, i. e. frequency 1, 2, 3 times/second, and needling duration 5 s, 60 s, 180 s. The effects of acupuncture at "Shuigou" (GV 26) with the 9 different acupuncture parameters on cerebral blood flow (CBF) in the MCAO rat were investigated by orthogonal test, factor analysis and one-way ANOVA. RESULTS: Acupuncture effect was influenced by acupuncture frequency, acupuncture duration and their combination. Among them, the acupuncture duration was the most important factor. The best acupuncture effect could be attained by using as long acupuncture duration as possibly with the fastest acupuncture frequency. CONCLUSION: In the Xingnao Kaiqiao needling method, acupuncture at "Shuigou" (GV 26) with fastest frequency and enough long duration can get the best acupuncture effect, which are the optimum stimulation parameters for "Shuigou" (GV 26).