Objective To study the effect and potential mechanism of Chaihuang Yishen Granules on renal fibrosis in mice with chronic kidney disease.Methods Thirty-six C57BL/6 male mice were randomly divided into sham group,model group,low-,and high-dose Chaihuang Yishen Granules groups(3.835 g·kg-1 and 7.67 g·kg-1),positive control group and Mincle ligand trehalose-6,6-dibehenate(TDB 10 mg·kg-1)group.Unilateral ureteral obstruction(UUO)was used to establish the model of renal fibrosis in mice.One hour after operation,mice in each drug-treated group were given corresponding drugs by gavage,and sham operation group and model group were given saline by gavage,once a day for 7 consecutive days.The pathological changes and fibrosis in the kidneys of mice in all groups were observed by hematoxylin eosin staining and Sirius red staining.The expressions and secretions of inflammatory factor IL-1β,IL-6 and TNF-α in kidneys of all groups were detected by Real-time PCR and ELISA.Immunohistochemistry and Western Blot were used to detect fibrosis indicators,including α-SMA and Fn protein levels.Immunofluorescence and Western Blot were further used to determine protein levels of Mincle and its downstream signal Syk/NF-κB.The proportion of Mincle positive macrophages was analyzed by flow cytometry.Results Compared with the model group,low-and high-dose Chaihuang Yishen Granules can effectively improve UUO-induced renal injury and fibrosis,and inhibit α-SMA and Fn protein levels in the kidney(P＜0.01).Moreover,high-dose Chaihuang Yishen Granules can effectively reduce the expressions and secretions of inflammatory factor IL-1β,IL-6 and TNF-α of kidney in UUO model(P＜0.01),as well as inhibit phosphorylation of NF-κB(P＜0.01).The mechanism study showed that Chaihuang Yishen Granules could significantly reduce the protein level of Mincle and the activity of its downstream signal Syk in the kidney of UUO mice(P＜0.01).After TDB was used to activate Mincle,the effect of Chaihuang Yishen Granules on improving renal injury,inflammatory factor secretion and fibrosis was significantly reduced(P＜0.05,P＜0.01).Conclusion Chaihuang Yishen Granules can improve UUO-induced renal injury,inflammation and fibrosis,and its mechanism may be related to the down-regulation of Mincle/Syk/NF-κB signaling pathway.

Objective To investigate the role of LncRNA-gm33782 in tubular injury in acute kidney injury(AKI)and the mechanism by which isorhamnetin ameliorates inflammatory cell apoptosis of tubular cells in AKI.Methods Forty-eight male C57BL/6J mice were randomly assigned to four groups:control group,AKI model group,electroporation+AKI group,and treatment group(oral administration of 30 mg/kg isorhamnetin).AKI was induced by a one-time intraperitoneal injection of 20 mg/kg cisplatin.Chromatin isolation by RNA purification was used to capture the LncRNA-gm33782 binding protein in AKI-affected kidneys for mass spectrometry,revealing the direct target protein of LncRNA-gm33782.The role of LncRNA-gm33782 in AKI was evaluated by observing the renal function,pathological structure,and expression of renal inflammatory factors(interleukin-1β,interleukin-6,and tumor necrosis factor-α)in mice after electrotransfection and isorhamnetin treatment.Nuclear factor-κB was detected as a critical mediator of inflammation.The expression levels of Bax and Bcl-2 protein were detected and flow cytometry was performed to evaluate the therapeutic effect of isorhamnetin on tubular cell apoptosis in AKI.Results Kidneys with cisplatin-induced AKI showed severe renal tubule injury,macrophage infiltration,and inflammation.Isorhamnetin treatment and LncRNA-gm33782 electrotransfection knockdown alleviated these signs of AKI.LncRNA-gm33782 was mainly expressed in renal tubular cells with AKI.LncRNA-gm33782 binding protein was detected by mass spectrometry,and complement factor H was found to have a direct binding relationship with LncRNA-gm33782.After LncRNA-gm33782 was overexpressed in vitro,the expression of complement factor-H immediately increased,and the therapeutic effect of isorhamnetin on apoptosis of AKI-affected inflammatory cells was inhibited.Conclusions Isorhamnetin alleviates apoptosis of tubular inflammatory cells in AKI by inhibiting the regulatory effect of LncRNA-gm33782 on complement factor H.

AIM: To observe the effects and mechanisms of hederagenin (HDG) in improving psoriasis skin lesions and inflammation in mice. METHODS: A mouse model of psoriasis was established by depilation of the back and continuous application of imiquimod for 7 days in C57 mice. After modeling, HDG was administered orally (low dose: 25 mg · kg

OBJECTVE To study the improvement effects of obacunone on renal interstitial fibrosis （RIF） in unilateral ureteral obstruction （UUO） model mice， and to investigate its mechanism based on ferroptosis mediated by nuclear factor erythroid 2- related factor 2（Nrf2）/glutathione peroxidase 4 （GPx4） signaling pathway. METHODS Thirty mice were randomly divided into sham operation group， model group， irbesartan group （positive control， 20 mg/kg）， obacunone low-dose and high-dose groups （10， 40 mg/kg）， with 6 mice in each group. Except for sham operation group， UUO model was established by ligation of unilateral ureter in other groups. After operation， administration groups were given intraperitoneal injection of relevant medicine， and sham operation group and model group were given intraperitoneal injection of constant volume of normal saline， once a day， for 7 consecutive days. The levels of creatinine （Cr） and blood urea nitrogen （BUN）， the content of malondialdehyde （MDA） and the activities of total superoxide dismutase （T-SOD） in serum and the concentration of Fe2+ in renal tissue were all detected. HE staining and Masson staining were performed to observe the morphology and the fibrosis of renal tissues. Immunohisto- chemical staining was used to determine expressions of the fibronectin （Fn）， α-smooth muscle actin （α-SMA）， GPx4 964083717@qq.com and Nrf2 in renal tissue. Western blot and real-time quantitative polymerase chain reaction （RT-qPCR） were used E-mail：834300205@qq.com to detect the protein and mRNA levels of Fn， α-SMA， Nrf2， GPx4 and SLC7A11 in the renal tissues. RESULTS Compared with sham operation group， serum levels of Cr and BUN， the concentration of Fe2+ in renal tissue， the protein and mRNA levels of Fn and α-SMA in model group were increased significantly （P＜0.05）， while the activity of T-SOD in serum， protein and mRNA expressions of Nrf2， GPx4， SLC7A11 in kidney tissue were significantly decreased （P＜0.05）； in the kidney tissue， the renal tubules were dilated， the collagen deposition was obvious， the fibrous bands were thicker and darker， and the renal interstitial inflammatory cells infiltrated significantly. After intervened with obacunone， the levels of above indexes （except for mRNA expression of SLC7A11 in obacunone low-dose group） in serum and renal tissue were reversed significantly （P＜0.05）， and pathological damage and collagen deposition of kidney tissue were alleviated. CONCLUSIONS Obacunone can improve renal interstitial fibrosis of UUO model mice， the mechanism of which may be associated with activating the Nrf2/GPx4 pathway and then inhibiting ferroptosis to relieve RIF in UUO model mice.

OBJECTVE To study the effects of Zhilong huoxue tongyu capsules on intracerebral hemorrhage ，long non-coding RNA（LncRNA）and its target genes in mice. METHODS Twenty-four male C 57BL/6 mice were randomly divided into sham operation group 1，model group 1 and Zhilong huoxue tongyu capsule low-dose and high-dose groups （0.35，1.40 g/kg）. Collagenase was injected into the caudate nucleus to construct the model of intracerebral hemorrhage. One hour after the operation ， the mice in each treatment group were given the corresponding medicinal solution ，and the mice in the sham operation group 1 and the model group 1 were given normal saline intragastrically ，once a day ，for 3 consecutive days. The morphological changes of the brain tissue of the mice in each group were observed by hematoxylin-eosin staining and Nissl staining. The protein and mRNA expression of interleukin- 1β（IL-1β）and tumor necrosis factor-α（TNF-α）in the brain tissue were detected by immunohistoche- mistry，Western blot and real-time quantitative polymerase chain reaction （PCR）. In addition ，9 of male C 57BL/6 mice were randomly divided into sham operation group 2，model group 2 and intervention group （Zhilong huoxue tongyu capsule 1.40 g/kg）. The mice were modeled and administered according to the above method ，and then the whole brain tissue of mice in E-mail：tanruizhi627@swmu.edu.cn each group was isolated ， total RNA was extracted and sequenced，followed by analyzing the different LncRNA. Gene ontology（GO）enrichment was performed to predict effective LncRNA and target genes ，and verified by real-time quantitative PCR. RESULTS Compared with model group 1，the brain tissue pathological damages were significantly improved in Zhilong huoxue tongyu capsule low-dose a nd high-dose groups ，and the IL -1β， TNF-α protein and mRNA expression in brain tissue were significantly decreased （P＜0.05）. Two effective LncRNAs were screened out. The results of in vivo verification test （LncRNA-Dlst-211 was highly expressed in model group 2，and significantly down-regulated after the intervention of Zhilong huoxue tongyu capsules ； LncRNA-Dlst-211 target genes Rps 6kl1 and LncRNA-MSTRG.8169.4 were expressed weakly in model group 2，and strongly up-regulated after intervention ）were consistent with the sequencing results. CONCLUSIONS Zhilong huoxue tongyu capsules can improve the brain injury and inflammatory response in intracerebral hemorrhage model mice ，and its mechanism may be related to down-regulating the expression of LncRNA-Dlst-211 and up-regulating the expression of LncRNA-MSTRG. 8169.4 and Rps 6kl1.