Chinese Medicine Involving Triple Rehabilitation Therapy for Knee Osteoarthritis in 696 Outpatients: A Multi-Center, Randomized Controlled Trial.

OBJECTIVE: To determine the effects of Chinese medicine (CM) involving triple rehabilitation therapy on the progression of knee osteoarthritis (KOA). METHODS: A total of 722 patients recruited from 38 community health service centers located in China from March 2013 to March 2017 were randomly divided into treatment and control groups equally, using a cluster randomization design. Health education combined with CM involving triple rehabilitation therapy for KOA (electro-acupuncture, Chinese medicinal herb fumigating-washing, and traditional exercises) was administered in the treatment group while conventional rehabilitation therapy (physical factor therapy, joint movement training, and muscle strength training) was administered in the control group. Patients with a visual analog scale (VAS) scores ≽4 were treated with dispersible meloxicam tablets (7.5 mg, once daily). The Lequesne index scores, VAS scores, range of motion (ROM), lower limb muscle strength, knee joint circumference, quantitative scores of KOA symptoms, and the short-form 36 item health survey questionnaire (SF-36) scores were measured for each patient at 5 checkpoints (before treatment, at the 2nd week and the 4th week during the 4-week treatment period, at 1 month and 3 months after end of treatment), and adverse reactions were observed also. RESULTS: A total of 696 patients completed the entire process, with 351 in the treatment group and 345 in the control group. At all treatment checkpoints, the treatment group demonstrated better outcomes than the control group with regard to the total Lequesne index scores, effective rate and improvement rate of the total Lequesne index scores, VAS scores, lower limb muscle strength, knee circumference, quantitative scores of KOA symptoms, and SF-36 scores as well (P<0.05 or P<0.01). No adverse reactions were encountered in this study. CONCLUSIONS: CM involving triple rehabilitation therapy can alleviate KOA-related pain and swelling, improve lower limb muscle strength, promote flexion and activity of the knee and improve the quality of life in patients undergoing KOA. It is suitable for patients with early or mid-stage KOA. (Registration No. ChiCTR-TRC-12002538).

Zhuanggu Jianxi Decoction () limits interleukin-1 ß-induced degeneration chondrocytes via the caveolin-p38 MAPK signal pathway.

OBJECTIVE: To evaluate the effect of Zhuanggu Jianxi Decoction (, ZGJXD) on interleukin-1 ß (IL-1 ß)-induced degeneration of chondrocytes (CDs) as well as the activation of caveolin-p38 mitogen-activated protein kinase (MAPK) signal pathway, investigating the possible molecular mechanism that ZGJXD treats osteoarthritis. METHODS: Serum pharmacology was applied in the present study, where ZGJXD was orally administrated to New Zealand rabbits and then ZGJXD containing serum (ZGJXD-S) was collected for following in vitro experiments. CDs were isolated aseptically from New Zealand rabbits and then cultured in vitro. Upon IL-1 ß stimulation, the degeneration of CDs was verified by inverted microscope, toluidine blue stain and type II collagen immunocytochemistry. After IL-1 ß-stimulated CDs were intervened with blank control serum, ZGJXD-S, together with or without SB203580 (a specific inhibitor of p38 MAPK) for 48 h, caveolin-1 protein expression and the phosphorylation level of p38 were determined by Western blotting, and the mRNA expression of IL-1 ß, tumor necrosis factor α (TNF-α), matrix metalloproteinase 3 (MMP-3) and MMP-13 were examined by real-time polymerase chain reaction. RESULTS: IL-1 ß stimulation induced degeneration of CDs, increased caveolin-1 expression and p38 phosphorylation, up-regulated the mRNA level of IL-1 ß, TNF-α, MMP-3 and MMP-13. However, the IL-1 ß-induced activation of caveolin-p38 signaling and alteration in the expression of p38 downstream target genes were suppressed by ZGJXD-S and/or SB203580 in CDs. CONCLUSION: ZGJXD can prevent CDs degeneration via inhibition of caveolin-p38 MAPK signal pathway, which might be one of the mechanisms that ZGJXD treats osteoarthritis.

[In vitro culture and identification of IL-1beta induced degeneration of cartilage cells in New Zealand white rabbits knee joint].

OBJECTIVE: To explore and identify the method for IL-1beta induced New Zealand rabbit knee chondrocyte degeneration, thus providing experimental bases for Chinese medical research on osteoarthritis from in vitro cultured chondrocytes. METHODS: Under aseptic conditions, bilateral knee joint cartilage was collected from 4-week old New Zealand rabbits. Chondrocytes were separated by type II collagenase digestion and mechanical blowing method. They were randomly divided into two groups when passaged to the 2nd generation, the normal control group (group Z) and the IL-1beta induced model group (group M). No intervention was given to those in group Z. 10% FBS culture media containing 10 ng/mL IL-1beta was added to group M. All cells were passaged to the 3rd generation. They were compared using morphological observation, toluidine blue staining, type II collagen immunohistochemical staining, and flow cytometry. RESULTS: Under inverted microscope, the second and the 3rd generation chondrocytes' phenotype of group Z was stable with good proliferation. Most cells turned into fusiform and slabstone shaped. In group M, most cells turned into long spindle shape or irregular shape. Results of toluidine blue staining and immunohistochemistry showed that the positive expression of chondrocytes after staining in group Z was superior to that in group M. Results of flow cytometry showed that there was statistical difference in the apoptosis rate of the second generation chondrocytes between group M and group Z (P < 0.01). CONCLUSION: It was obviously seen that chondrocytes in IL-1beta induced New Zealand rabbit knee chondrocyte model obviously degenerated, which could be used in related experimental researches on osteoarthritis.

[Effect of naringin of Drynaria Rhizome, a Chinese medical component of Zhuanggu Jianxi Recipe containing serum on caveolin-p38MAPK signal pathway in IL-1ß induced rabbit degenerated chondrocytes].

OBJECTIVE: To observe the effect of naringin of Drynaria Rhizome, a Chinese medical component of Zhuanggu Jianxi Recipe (ZJR) containing serum on caveolin-p38MAPK signal factors (such as caveolin-1, p-p38, p-ATF-2, IL-1ß, and TNF-α) in IL-1ß induced rabbit degenerated chondrocytes, and further to explore its mechanism for protecting articular cartilages. METHODS: Naringin of Drynaria Rhizome was obtained and analyzed by HPLC-TOF/MS. Four weeks old New Zealand rabbits were killed and their bilateral knee joints were isolated aseptically. CDs were isolated and then cultured in vitro. The second generation of CDs were used for later experiment. The effect of naringin on CDs proliferation was detected by MTT assay. The effect of naringin on the expression of IL-1ß-induced collagen II in CDs was detected by immunohistochemical method. The effect of naringin on caveolin-1, p-p38, and p-ATF-2 protein in IL-1ß-induced CDs was detected by Western blot. The effect of naringin on mRNA expression of IL-1ß and TNF-α in IL-1ß-induced CDs was detected by RT-PCR. RESULTS: The appearance time of naringin in flow graphs of naringin standard solution and ZJR containing serum was 23.5 min, and the molecular weight ranged between 581.0 and 581.5 m/z. Naringin could promote the proliferation of CDs, and inhibit the effect of IL-1ß on collagen II in CDs. Compared with the model group, naringin could reduce the expression of caveolin-1, p-p38, p-ATF-2, IL-1ß, and TNF-α in IL-1ß induced CDs (P < 0.05), which was approximate to the level of the normal group. CONCLUSIONS: Naringin could not only promote the proliferation of CDs, but also protect IL-1ß-induced CDs. Its mechanism might be associated with decreasing the expression of caveolin-1, p-p38, and p-ATF-2 proteins, inhibiting caveolin-p38MAPK signal pathway, and further reducing mRNA expression of IL-1ß and TNF-α in the downstream of caveolin-p38MAPK signal pathway.

[Experimental study on the effects of pharmaco-serum of diabetic rats fed with Chinese herbs qianggubao decoction ([Chinese characters: see text]) on osteoblast culture in vitro].

OBJECTIVE: To investigate the optimum phase and dose of pharmaco-serum of diabetic rats fed with Qianggubao decoction ([Chinese characters: see text]) on the differentiation and mineralization of osteoblast (OB). METHODS (OB) was isolated from the skull of 10 newly born SD rats aged 1 to 2 days by means of Trypsin-collagenase digestion. After the OB was identified, different kinds of pharmaco-serum of diabetic rats fed with inactive Qianggubao decoction ([Chinese characters: see text]) of different phase (rats were fed with medicine three days or five days after last fed with medicine one hour or three hours) and concentration (5%, 10%, 20%) were added to the OB and incubated. After 7 days and 18 days of culture,the effects of the differentiation and mineralization of osteoblast were detected. RESULTS: The secretion of ALP and formation of mineralized nodules of osteoblast in the different doses of pharmaco-serum groups were almost the same as that of normal control group, but were superior to that in the model control group. And the group with concentration of 20% pharmaco-serum was the best in the secretion of ALP and formation of mineralized nodules of osteoblast. As to the phases of pharmaco-serum, the best one on the differentiation and mineralization of osteoblast was the serums from diabetic rat-model fed with Qianggubao decoction ([Chinese characters: see text]) three days or five days, after one hour of last fed with medicine. CONCLUSION: The pharmaco-serum of diabetic rats fed with Qianggubao decoction ([Chinese characters: see text]) can promote the differentiation and mineralization of osteoblast. Allow for time and the cost of experiment,we presume that pharmaco-serum of diabetic rats fed with Qianggubao decoction ([Chinese characters: see text]) three days, after one hour of last fed, with concentration of 20% and not-inactivation is the optimum on the differentiation and mineralization of osteoblast.

[Experimental study on the effects of inactivated and un-inactivated pharmaco-serum of diabetic rats fed with Chinese herbs Qianggubao decoction on the proliferation of osteoblast cultured in vitro].

OBJECTIVE: To study the effect of inactivated and un-inactivated pharmaco-serum of diabetic rats fed with Chinese herbs Qianggubao decoction on the proliferation of osteoblast cells (OB)cultured in vitro. METHODS: OB was isolated from the skull of newly born SD rats aged 1 to 2 days by means of Trypsin-collagenase digestion and identified by image analysis under inverted microscope, V-G collagen staining, ALP staining, calcification nod staining etc. After the OB was identified, in activated and un-inactivated pharmaco-serum of diabetic rats fed with Qianggubao decoction of ferent phase (rats were fed with medicine 3 days or 5 days after last fed with medicine 1 hour or 3 hours) and concentration (5%, 10%, 20%) were added to the OB and incubated. After determined times, the effects of the proliferation of osteoblasts were detected by MTT analysis. RESULTS: There was significant difference between un-inactivated pharmaco-serum and inactivated pharmaco-serum on the proliferation of osteoblasts, and un-inactivated serum had stronger effects to improve the proliferation of osteoblasts (P < 0.01 or P < 0.05). CONCLUSION: Un-inactivated and inactivation pharmaco-serum of diabetic rats fed with Chinese herbs Qianggubao decoction can influence the proliferation of, and the un-inactivated pharmaco-serum has stronger effects.