Protective effect of resveratrol on estrogen deficiency-induced osteoporosis though attenuating NADPH oxidase 4/nuclear factor kappa B pathway by increasing miR-92b-3p expression

Introduction: Resveratrol (RES) exhibits estrogen-like effects and has potential applications to treatment of osteoporosis caused by estrogen deficiency; however, the specific mechanism of action of RES remains unclear. Here, we examined the therapeutic effects of RES on ovariectomized (OVX) rats with osteoporosis and determined the underlying mechanism. Methods: We established an OVX rat model to study osteoporosis caused by estrogen deficiency. The treatment groups were given orally with RES (50, 100, and 200 mg/day), the estrogen group received 0.8 mg/kg E2 daily via oral route, and the sham-operated and control groups received an equivalent dose of sodium carboxymethylcellulose orally. After 12 weeks of treatment, we used real-time quantitative polymerase chain reaction (PCR) and Western blot analysis to measure the gene and protein expression of miR-92b-3p, Nox4, NF-κBp65, IκB, BMP2, Smad7, and RUNX-2 in bone tissues. Right femur structural parameters were evaluated by micro-CT. Dual-energy X-ray 4500 W was used to determine systemic bone mineral density (BMD). Enzyme-linked immunosorbent assay (ELISA) kits were used to determine the serum levels of bone alkaline phosphatase (BALP), osteoprotegerin (OPG), anti-tartrate acid phosphatase-5b (PTRA5b), and carboxylated terminal peptide (CTX-I). The rat femoral bone specimens were stained using hematoxylin and eosin for pathological examination Results: We observed increased levels of serum estrogen in both ovaries, elevated miR-92b-3p levels in bone tissues, reduced levels of Nox4, NF-κBp65, p-IκB-a, and cathepsin K, and elevated gene and protein expression of BMP2, Smad7, and RUNX-2 in the OVX rat model of osteoporosis after treatment with RES. Elevated levels of BALP, OPG, ALP, and BMD along with reduced levels of TRAP-5b and CTX-I were also observed. The structural model index (SMI) and the trabecular space (Tb. Sp) decreased, while the trabecular thickness (Tb. Th), bone volume fraction (BV/TV), trabecular number (Tb.N), and tissue bone density (Conn.D) increased, thereby improving osteoporosis induced by estrogen deficiency in both ovaries. Conclusion: Cathepsin K expression and Nox4/NF-κB signaling pathway were suppressed by the elevated expression of miR-92b-3p. This inhibition was pivotal in the protective effect of RES against osteoporosis induced by estrogen deficiency in both ovaries. Thus, RES efficiently alleviated osteoporosis induced by estrogen deficiency in rats.

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Toll-like receptor 2 induced senescence in intervertebral disc cells of patients with back pain can be attenuated by o-vanillin

Arthritis Research & Therapy ◽

10.1186/s13075-021-02504-z ◽

2021 ◽

Vol 23 (1) ◽

Author(s):

Matthew Mannarino ◽

Hosni Cherif ◽

Li Li ◽

Kai Sheng ◽

Oded Rabau ◽

...

Keyword(s):

Gene Expression ◽

Back Pain ◽

Protein Expression ◽

Disc Degeneration ◽

Cell Senescence ◽

Enzyme Linked Immunosorbent Assay ◽

Matrix Synthesis ◽

Gene And Protein Expression ◽

Pain Patients ◽

In Cells

Abstract Background There is an increased level of senescent cells and toll-like teceptor-1, -2, -4, and -6 (TLR) expression in degenerating intervertebral discs (IVDs) from back pain patients. However, it is currently not known if the increase in expression of TLRs is related to the senescent cells or if it is a more general increase on all cells. It is also not known if TLR activation in IVD cells will induce cell senescence. Methods Cells from non-degenerate human IVD were obtained from spine donors and cells from degenerate IVDs came from patients undergoing surgery for low back pain. Gene expression of TLR-1,2,4,6, senescence and senescence-associated secretory phenotype (SASP) markers was evaluated by RT-qPCR in isolated cells. Matrix synthesis was verified with safranin-O staining and Dimethyl-Methylene Blue Assay (DMMB) confirmed proteoglycan content. Protein expression of p16INK4a, SASP factors, and TLR-2 was evaluated by immunocytochemistry (ICC) and/or by enzyme-linked immunosorbent assay (ELISA). Results An increase in senescent cells was found following 48-h induction with a TLR-2/6 agonist in cells from both non-degenerate and degenerating human IVDs. Higher levels of SASP factors, TLR-2 gene expression, and protein expression were found following 48-h induction with TLR-2/6 agonist. Treatment with o-vanillin reduced the number of senescent cells, and increased matrix synthesis in IVD cells from back pain patients. Treatment with o-vanillin after induction with TLR-2/6 agonist reduced gene and protein expression of SASP factors and TLR-2. Co-localized staining of p16INK4a and TLR-2 demonstrated that senescent cells have a high TLR-2 expression. Conclusions Taken together our data demonstrate that activation of TLR-2/6 induce senescence and increase TLR-2 and SASP expression in cells from non-degenerate IVDs of organ donors without degeneration and back pain and in cells from degenerating human IVD of patients with disc degeneration and back pain. The senescent cells showed high TLR-2 expression suggesting a link between TLR activation and cell senescence in human IVD cells. The reduction in senescence, SASP, and TLR-2 expression suggest o-vanillin as a potential disease-modifying drug for patients with disc degeneration and back pain.

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83 Spatially resolved transcriptomic and proteomic investigation of breast cancer and its immune microenvironment

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2021-sitc2021.083 ◽

2021 ◽

Vol 9 (Suppl 3) ◽

pp. A91-A91

Author(s):

Jennifer Chew ◽

Cedric Uytingco ◽

Rapolas Spalinskas ◽

Yifeng Yin ◽

Joe Shuga ◽

...

Keyword(s):

Breast Cancer ◽

Gene Expression ◽

Tumor Microenvironment ◽

Protein Expression ◽

Protein Detection ◽

Response To Therapy ◽

Pathological Examination ◽

Multi Drug Resistance ◽

Spatially Resolved ◽

Gene And Protein Expression

BackgroundThe tumor microenvironment (TME) is composed of highly heterogeneous extracellular structures and cell types such as endothelial cells, immune cells, and fibroblasts that dynamically influence and communicate with each other. The constant interaction between a tumor and its microenvironment plays a critical role in cancer development and progression and can significantly affect a tumor’s response to therapy and capacity for multi-drug resistance. High resolution analyses of gene and protein expression with spatial context can provide deeper insights into the interactions between tumor cells and surrounding cells within the TME, where a better understanding of the underlying biology can improve treatment efficacy and patient outcomes. Here, we demonstrated the ability to perform streamlined multi-omic tumor analyses by utilizing the 10X Genomics Visium Spatial Gene Expression Solution for FFPE with multiplex protein enablement. This technique simultaneously assesses gene and protein expression to elucidate the immunological profile and microenvironment of different breast cancer samples in conjunction with standard pathological methods.MethodsSerial (5 µm) sections of FFPE human breast cancer samples were placed on Visium Gene Expression (GEX) slides. The Visium GEX slides incorporate ~5,000 molecularly barcoded, spatially encoded capture spots onto which tissue sections are placed, stained, and imaged. Following incubation with a human whole transcriptome, probe-based RNA panel and an immuno-oncology oligo-tagged antibody panel, developed with Abcam conjugated antibodies, the tissues are permeabilized and the representative probes are captured. Paired GEX and protein libraries are generated for each section and then sequenced on an Illumina NovaSeq at a depth of ~50,000 reads per spot. Resulting reads from both libraries are aligned and overlaid with H&E-stained tissue images, enabling analysis of both mRNA and protein expression. Additional analyses and data visualizations were performed on the Loupe Browser v4.1 desktop software.ConclusionsSpatial transcriptomics technology complements pathological examination by combining histological assessment with the throughput and deep biological insight of highly-multiplexed protein detection and RNA-seq. Taken together, our work demonstrated that Visium Spatial technology provides a spatially-resolved, multi-analyte view of the tumor microenvironment, where a greater understanding of cellular behavior in and around tumors can help drive discovery of new biomarkers and therapeutic targets.

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Effects of Haima Duobian Pill in a Rat Model of Kidney Yang Deficiency Syndrome

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2021/6696234 ◽

2021 ◽

Vol 2021 ◽

pp. 1-13

Author(s):

Yijia Zeng ◽

Tingna Li ◽

Xiaorui Zhang ◽

Yuanyuan Ren ◽

Qinwan Huang ◽

...

Keyword(s):

Rat Model ◽

Cyclic Adenosine Monophosphate ◽

Adenosine Monophosphate ◽

Cyclic Guanosine Monophosphate ◽

Deficiency Syndrome ◽

Guanosine Monophosphate ◽

Network Pharmacology ◽

Enzyme Linked Immunosorbent Assay ◽

Therapeutic Effects ◽

Kidney Yang Deficiency

Objective. Modern research shows that Haima Duobian pill (HDP) can relieve the kidney yang deficiency syndrome (KYDS), but the mechanism is still unclear. The aim of this work was to study the effects of HDP in a rat model of KYDS. Materials and Methods. The network pharmacology methods were used to predict the therapeutic effects of Haima Duobian pill. Adenine was used to establish the rat model of kidney yang deficiency syndrome. The general physical signs of rats were observed after different doses of Haima Duobian pill (HDP) were given. Serum cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone (T), estradiol (E2), and gonadotropin-releasing hormone (GnRH) levels were determined using enzyme-linked immunosorbent assay (ELISA) kits. Then, the histopathologic changes and sperm activity were detected. Results. HDP could improve the general signs of kidney yang deficiency syndrome rats. After the rats were treated with HDP, the expression of cGMP and E2 was significantly inhibited and the expression of cAMP and T was significantly increased. The pathological damage of testis, epididymis, and seminal vesicle was alleviated, and the sperm activity was improved. Conclusion. For adenine-induced kidney yang deficiency syndrome in rats, HDP had a significant therapeutic effect.

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Plasma Level and Expression of Visfatin in the Porcine Hypothalamus During the Estrous Cycle and Early Pregnancy

10.21203/rs.3.rs-93399/v1 ◽

2020 ◽

Author(s):

Tadeusz Kaminski ◽

Marta Kiezun ◽

Ewa Zaobidna ◽

Kamil Dobrzyń ◽

Barbara Wasilewska ◽

...

Keyword(s):

Protein Expression ◽

Blood Plasma ◽

Estrous Cycle ◽

Early Pregnancy ◽

Energy Homeostasis ◽

Plasma Concentrations ◽

Enzyme Linked Immunosorbent Assay ◽

Tissue Samples ◽

Hormone Synthesis ◽

Gene And Protein Expression

Abstract BackgroundVisfatin exists in two forms: the intracellular form which is a rate limiting enzyme engaged in nicotinamide adenine dinucleotide biosynthesis and the extracellular form considered as an adipokine, produced mainly by the adipose tissue. Visfatin could be an energy sensor involved in regulating female fertility, which creates a hormonal link integrating the control of energy homeostasis and reproduction. MethodsThe study compares the expression levels of visfatin gene (quantitative real time PCR) and protein (Western blotting and fluorescent immunohistochemistry) in the selected areas of the porcine hypothalamus responsible for gonadotropin releasing hormone synthesis: the mediobasal hypothalamus (MBH) and preoptic area (POA), and visfatin concentrations in the blood plasma (enzyme-linked immunosorbent assay). The tissue samples were harvested from gilts on days 2-3, 10-12, 14-16 and 17-19 of the estrous cycle, and on days 10-11, 12-13, 15-16, 27-28 of pregnancy. Differences between groups were analyzed by one-way ANOVA followed by Tukey’s post hoc test. ResultsDuring the estrous cycle, visfatin mRNA expression in the MBH was higher on days 2-3 and 17-19, while the visfatin protein concentration on days 17-19. During early pregnancy, the most pronounced gene and protein expression in the MBH was found on days 15-16 and 10-11, respectively. In the POA during the estrous cycle, visfatin gene expression was the highest on days 17-19, and the protein level of visfatin on days 14-16. During early pregnancy, the highest expression of visfatin gene in the POA was observed on days 15-16, whereas the protein concentrations on days 27-28. Blood plasma concentrations of visfatin during the estrous cycle were higher on days 2-3 in relation to other studied phases of the cycle. During early pregnancy, the highest visfatin contents in the blood plasma were observed on days 12-13. Visfatin gene and protein expression in MBH and POA, and visfatin plasma concentrations differed during early pregnancy in relation to days 10-12 of the cycle. ConclusionsThis study demonstrated visfatin expression in the porcine hypothalamus and its dependence on hormonal milieu related to the estrous cycle and early pregnancy.

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Crocin ameliorated skin tissue inflammation in atopic dermatitis in mice

International Journal of Research in Dermatology ◽

10.18203/issn.2455-4529.intjresdermatol20200592 ◽

2020 ◽

Vol 6 (2) ◽

pp. 142

Author(s):

Abdullah Alyoussef

Keyword(s):

Atopic Dermatitis ◽

Protein Expression ◽

Skin Disease ◽

Therapeutic Effects ◽

Inflammatory Skin Disease ◽

Mice Model ◽

Subsequent Reduction ◽

Gardenia Jasminoides ◽

Tnf Α ◽

Gene And Protein Expression

Background: Atopic dermatitis (AD) is considered a chronic recurrent inflammatory skin disease. In addition, crocin is the major carotenoid compound found in Gardenia jasminoides. It is previously proved to produce anti-inflammatory actions. Therefore, we conducted this research to investigate the therapeutic effects of crocin on a mice model of AD.Methods: Mice were investigated for the number of scratches and dermatitis score. Skin was isolated and used for measurements of gene and protein expression of β-catenin, NFκB, TNF-α and IL-1β.Results: Authors found that crocin significantly reduced the number of scratches, ear thickness and dermatitis score. In addition, crocin ameliorated AD-induced elevation in the expression of β-catenin, NFκB, TNF-α and IL-1β.Conclusions: Crocin ameliorated DNCB-induced AD in mice via blockage of β-catenin with subsequent reduction in inflammatory pathway.

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Methamphetamine causes neurotoxicity by promoting polarization of macrophages and inflammatory response

Human & Experimental Toxicology ◽

10.1177/0960327117714039 ◽

2017 ◽

Vol 37 (5) ◽

pp. 486-495 ◽

Cited By ~ 5

Author(s):

X Li ◽

F Wu ◽

L Xue ◽

B Wang ◽

J Li ◽

...

Keyword(s):

Inflammatory Response ◽

Protein Expression ◽

Macrophage Polarization ◽

Enzyme Linked Immunosorbent Assay ◽

Neuronal System ◽

Activated Macrophages ◽

Alternatively Activated Macrophages ◽

Raw264.7 Macrophages ◽

Gene And Protein Expression ◽

Anti Inflammatory

Macrophages, especially their activation state, are closely related to the progression of neurotoxicity. Classically activated macrophages (M1) are proinflammatory effectors, while alternatively activated macrophages (M2) exhibit anti-inflammatory properties. As a powerful addictive psychostimulant drug, coupled with its neurotoxicity, methamphetamine (Meth) abuse may lead to long-lasting abnormalities in the neuronal system. The present study investigated the effect of Meth at subtoxic concentration on macrophage activation state and its underlying toxicity to neuronal cells. PC12 and Murine RAW264.7 cells were coincubated with Meth to test its toxicity. 3-(4,5-Dimethylthiazol)-2,5-diphenyltetrazolium-bromide, enzyme-linked immunosorbent assay, real-time polymerase chain reaction, and Western blot assays were performed to evaluate the toxicity, cytokine secretion, gene, and protein expression. Results showed that cytotoxicity was enhanced on PC12 cells after coculturing with RAW264.7 stimulated with Meth. RAW264.7 macrophages tended to switch to the M1 phenotype, releasing more nitric oxide and proinflammatory cytokines, including tumor necrosis factor α (TNFα), interleukin (IL)-12, and IL-1β, while decreasing the release of anti-inflammatory cytokine IL-10 after treatment with Meth. Meth upregulated the gene expression of IL-6, IL-1β, and TNFα and downregulated the expression of Arg-1, IL-10, and KLF4. Meth could also upregulate the protein expression of IL-1β and TNF α and downregulate the expression of Arg-1 and KLF4. However, the abovementioned effects induced by Meth were abolished by the addition of dopamine receptor D3 antagonist. In conclusion, our study demonstrated that Meth promoted macrophage polarization from M0 to M1 and enhanced inflammatory response, which provided the scientific rationale for the neurotoxicity caused by the chronic use of Meth.

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Effects of Dipsacus asperoides Extract on Monosodium Iodoacetate–Induced Osteoarthritis in Rats Based on Gene Expression Profiling

Frontiers in Pharmacology ◽

10.3389/fphar.2021.615157 ◽

2021 ◽

Vol 12 ◽

Author(s):

Jin Mi Chun ◽

A Yeong Lee ◽

Jae Yong Nam ◽

Kyung Seob Lim ◽

Mu Seog Choe ◽

...

Keyword(s):

Gene Expression ◽

Protective Effect ◽

Gene Expression Profiling ◽

Rat Model ◽

Expression Profiling ◽

Bone Fractures ◽

Cartilage Tissue ◽

Histopathological Analysis ◽

Therapeutic Effects ◽

Monosodium Iodoacetate

The root of Dipsacus asperoides C. Y. Cheng et T. M. Ai is traditionally used as an analgesic and anti-inflammatory agent to treat pain, rheumatoid arthritis, and bone fractures. However, neither its effects on osteoarthritis (OA) nor its effects on the arthritic cartilage tissue transcriptome have not been fully investigated. In this study, we used a rat model of monosodium iodoacetate- (MIA-) induced OA to investigate the therapeutic effects of a Dipsacus asperoides ethanolic extract (DAE, 200 mg/kg for 21 days). The study first assessed joint diameter, micro-CT scans, and histopathological analysis and then conducted gene expression profiling using RNA sequencing in articular cartilage tissue. We found that DAE treatment ameliorates OA disease phenotypes; it reduced the knee joint diameter and prevented changes in the structural and histological features of the joint, thereby showing that DAE has a protective effect against OA. Based on the results of gene expression profiling and subsequent pathway analysis, we found that several canonical pathways were linked to DAE treatment, including WNT/β-catenin signaling. Taken together, the present results suggest molecular mechanism, involving gene expression changes, by which DAE has a protective effect in a rat model of MIA-induced OA.

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ShenLing BaiZhu San alleviates Ulcerative Colitis in Rats by Regulating Gut Microbiota

10.21203/rs.3.rs-90760/v1 ◽

2020 ◽

Author(s):

Daxing Gu ◽

Shanshan Zhou ◽

Lili Yao ◽

Ying Tan ◽

Xingzi Chi ◽

...

Keyword(s):

Ulcerative Colitis ◽

Gut Microbiota ◽

Relative Abundance ◽

Rat Model ◽

High Throughput Sequencing ◽

Intestinal Barrier Function ◽

Enzyme Linked Immunosorbent Assay ◽

Trinitrobenzene Sulfonic Acid ◽

Rrna Gene ◽

Therapeutic Effects

Abstract Background: Recent studies have suggested that Shenling Baizhu San (SLBZS), an complementary and alternative medical therapy for ulcerative colitis (UC), alleviate clinic symptoms by the improvement of biochemical criteria and restoration of the intestinal barrier function. SLBZS as a famous Chinese herbal formula has been reportedly used to treat UC, of which mechanism is unknown. This study investigated the therapeutic effects of SLBZS on restoring the gut microbiota in a UC rat model. Methods: We proposed a 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced UC rat model to monitor the structural modulation of the gut microbiota. The test period was 10 days (observation for two days after modeling, treatment for 8 days by SLBZS). In this study, the level of inflammatory cytokines and activity of antioxidant enzymes in serum were ascertained by enzyme-linked immunosorbent assay (ELISA) and histological changes of colon were observed. Feces were collected for high-throughput sequencing of 16S rRNA gene. Results: SLBZS partly reduced the diversity of the gut microbiota, while the abundance of that is increased. Furthermore, at the genus level, the relative abundance of short chain fatty acids (SCFA) producing bacteria including Prevotella and Oscillospira increased, while the relative abundance of harmful bacteria including Desulfovibrio, and Bilophila decreased. Additionally, SLBZS could improve the lesions of colon and significantly reduce the expression of Interleukin-6 (IL-6) and Myeloperoxidase (MPO) and increase the activities of Superoxide dismutase (SOD) and Catalase (CAT) in rats serum. Conclusions: These results demonstrate that SLBZS may treat UC effectively by inhibiting inflammation, enhancing antioxidant capacity and regulating gut microbiota.

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Cannabidiol Attenuates Methamphetamine-Induced Cardiac Inflammatory Response and Necrosis through The PKA/CREB Signaling Pathway

10.21203/rs.3.rs-1159875/v1 ◽

2021 ◽

Author(s):

Qianyun Nie ◽

Wenjuan Dong ◽

Baoyu Shen ◽

Genmeng Yang ◽

Hao Yu ◽

...

Keyword(s):

Inflammatory Response ◽

Protein Expression ◽

Signaling Pathway ◽

Troponin I ◽

Weight Ratio ◽

Public Health Problem ◽

Enzyme Linked Immunosorbent Assay ◽

Therapeutic Effects ◽

Dependent Manner ◽

Expression Levels

Abstract Methamphetamine (MA) abuse is a major global public health problem, with cardiovascular issues becoming an increasingly recognized complication. Cannabidiol (CBD) has gained recent attention, due to its various pharmacological properties. However, whether CBD has therapeutic effects on MA-induced cardiotoxicity remains unknown. In the present study, we investigated whether CBD has a protective or therapeutic effect on MA-induced cardiac damage in rats via the protein kinase A (PKA)/cyclic adenosine monophosphate response element-binding (CREB) signaling pathway. Thirty rats were randomly divided into five groups. The rats were administered MA by intraperitoneal injection (IP) once a day for 4 weeks, with CBD (40 or 80 mg/kg, IP) treatment 1 h prior to the MA injections. Body and heart weights were measured, and morphological changes were determined using hematoxylin & eosin and Masson’s trichrome staining. The serum levels of interleukin-6 (IL-6) and IL-10 were detected using enzyme linked immunosorbent assay (ELISA) kits. The protein expression levels of PKA, phospho-PKA (p-PKA), CREB, phospho-CREB (p-CREB) and cardiac troponin I (cTnI) in the myocardium were detected by western blot analysis. Results showed that the heart-to-body weight ratio increased significantly following MA administration but decreased with CBD treatment. Chronic administration of MA resulted in a cardiac inflammatory response and progressive development of fibrosis, while CBD treatment attenuated these lesions in a dose-dependent manner. MA administration increased IL-6 but decreased IL-10 levels, which were reversed by CBD pretreatment. Moreover, MA significantly increased the cTnI level, but this was decreased by CBD treatment at 80 mg/kg. The protein expression levels of PKA, p-PKA, CREB, and p-CREB increased following MA administration, but significantly decreased with CBD treatment. Overall, these results indicate that chronic MA administration leads to cardiotoxicity, including cardiac inflammatory response, fibrosis, and myocardial necrosis, but these effects can be attenuated by CBD pretreatment. Our research suggests a potential application of CBD for MA-induced cardiotoxicity, which may attenuate inflammatory response and necrosis through the PKA/CREB signaling pathway.

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Oral Administration of Lactobacillus Rhamnosus Ameliorates the Progression of Osteoarthritis by Inhibiting Joint Pain and Inflammation

10.21203/rs.3.rs-220741/v1 ◽

2021 ◽

Author(s):

JooYeon Jhun ◽

Keun-Hyung Cho ◽

Dong hwan Lee ◽

Ji Ye Kwon ◽

Jin Seok Woo ◽

...

Keyword(s):

Oral Administration ◽

Protein Expression ◽

Rat Model ◽

Inflammatory Cytokine ◽

Lactobacillus Rhamnosus ◽

Cartilage Destruction ◽

Pain Severity ◽

Enzyme Linked Immunosorbent Assay ◽

Intestinal Damage ◽

And Cartilage

Abstract Osteoarthritis (OA) is the most common form of arthritis and age-related degenerative joint disorder, which adversely affects quality of life and causes disability. However, the pathogenesis of OA remains unclear. This study was performed to examine the effects of Lactobacillus rhamnosus in OA progression. OA was induced in 6-week-old male Wistar rats by monosodium iodoacetate (MIA) injection, and the effects of oral administration of L. rhamnosus were examined in this OA rat model. Pain severity, cartilage destruction, and inflammation were measured in MIA-induced OA rats. The intestines were isolated from OA rats, and the intestinal structure and inflammation were measured. Protein expression in the dorsal root ganglion was analyzed by immunohistochemistry. The effects of L. rhamnosus on mRNA and protein expression in chondrocytes stimulated with interleukin (IL)-1β and lipopolysaccharide (LPS) were analyzed by real-time polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). Pain severity was decreased in L. rhamnosus-treated MIA-induced OA rats. The levels of expression of MCP-1, a potential inflammatory cytokine, and its receptor, CCR2, were decreased, and GABA and PPAR-γ expression were increased in L. rhamnosus-treated OA rats. The inflammation, as determined by IL-1β, and cartilage destruction, as determined by MMP3, were also significantly decreased by L. rhamnosus in OA rats. Intestinal damage and inflammation were also improved by L. rhamnosus. In human OA chondrocytes, TIMP1, TIMP3, SOX9 and COL2A1 which are tissue inhibitors of MMP, and IL-10, an anti-inflammatory cytokine, were increased by L. rhamnosus.L. rhamnosus treatment led to decreased pain severity and cartilage destruction in a rat model of OA. Intestinal destruction and inflammation were also decreased by L. rhamnosus treatment. Our findings suggested the therapeutic potential of L. rhamnosus in OA.

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